Perspective for the production of antimalarial drugs in Brazil.

There appears to be no chemical manufacture of antimalarial drugs in Brazil. Technology at the laboratory process level has been developed for chloroquine, mefloquine, pyrimethamine and cycloguanil, but not perfected nor scaled-up, largely for economic reasons and market uncertainty. Development of primaquine has been contracted but it will run into the same difficulty. Manufacturing capacity for sulfadoxine was registered in the SDI by Roche. A project to produce artemisinine and its derivatives is under way at UNICAMP-CPQBA but is hampered by low content in the plant. Proguanil could be produced easily, but apparently no attempt has been made to do so. Quinine is imported on a large scale mostly for soft-drink production. Since malarial treatment falls largely within the responsibility of the Government health authorities, manufacture of drugs in Brazil will depend on an assured medium-term purchase order made to a potential local manufacturer, since competition in the world market is scarcely viable at the present moment.


Efficacy of insecticide impregnated bed-nets to control malaria in a rural forested area in southern Cameroon.

Due to current spreading of chemoresistant strains of Plasmodium falciparum malaria control must incorporate vector control programmes. Due to well known constraints house sprayings cannot be performed as before. Personal protection can be developed and a large scale use of insecticide treated bed-nets appeared to be very useful to reduce man-vector contact in Asia, South America and West and East Africa. No trial has been done in forest Central Africa where transmission is permanent. We performed such a trial in the southern part of Cameroon (using deltamethrin, at 25 mg/m2) and obtained similar data to those observed in The Gambia, Burkina Faso and Tanzania with a noteworthy reduction of both transmission and high parasitaemia of P. falciparum (respectively 78% and 75%) meaning a drop of malaria morbidity.


Various pyrethroids on bednets and curtains.

Verandah trap huts in a Tanzanian village were used to assess the effectiveness of impregnated bednets and curtains in preventing hut entry and feeding by, and in killing of, Anopheles gambiae and An. funestus. Permethrin, deltamethrin, lambdacyhalothrin and pyrethrum were used for impregnation of damaged or undamaged bednets, sisal eaves curtains or bed curtains made of polypropylene fibre. The performance of the three synthetic pyrethroids did not differ statistically significantly, except that on a damaged net permethrin was better at preventing feeding. Sisal eaves curtains deterred mosquitoes from hut entry but did not kill those that had entered. In assessing damaged nets and curtains it must be recognised that anything less than the best vector control may have no appreciable impact on holoendemic malaria.


A malaria merozoite surface protein (MSP1)-structure, processing and function.

Merozoite surface protein-1 (MSP-1, also referred to as P195, PMMSA or MSA 1) is one of the most studied of all malaria proteins. The protein is found in all malaria species investigated and structural studies on the gene indicate that parts of the molecule are well-conserved. Studies on Plasmodium falciparum have shown that the protein is in a processed form on the merozoite surface, a result of proteolytic cleavage of the large precursor molecule. Recent studies have identified some of these cleavage sites. During invasion of the new red cell most of the MSP1 molecule is shed from the parasite surface except for a small C-terminal fragment which can be detected in ring stages. Analysis of the structure of this fragment suggests that it contains two growth factor-like domains that may have a functional role.


Ecology of malaria vectors in the Americas and future direction.

The resurgence of malaria in the Americas has renewed interest in Anopheles biology. Anopheles darlingi, An. albimanus, An. nuneztovai and An. aquasalis are reconfirmed as major malaria vectors and other species are playing important roles in regional malaria transmission. Adult biting activity and larval ecology are discussed in detail. Seasonal abundance and daily biting activity of Anophelines vary considerably among species and geographically for the same species. Anopheles albimanus has the least amount of variation in biting activity over its range and An. darlingi has the greatest. All species studied are more exophilic and exophagic than endophilic and endophagic. Anopheles darlingi is more anthropophilic, endophilic and endophagic than other Anophelines. Larval studies remain more descriptive than comprehensive. Research on Anophelines is becoming more integrated and biologists are using new biochemical techniques and ecological principles to answer critical questions. This "pluralization" will help us understand species complexes, population dynamics and malaria transmission. Integrated control programs will require more regional, in-depth ecological studies.


Transmission factors in malaria epidemiology and control in Africa.

Genetic and environmental components of factors contributing in malaria transmission are reviewed. Particular attention is given to density dependent regulation of vector populations in relation to the survival rate of anophelines. The expectation of vector control activities are different according to the epidemiological characteristics of malaria, mainly its stability. In areas with perennial and high transmission (stable malaria) vector control could reduce malaria related morbidity and mortality, without any effect on the endemicity. However this need further investigations. In areas where the transmission period is very short (unstable malaria), vector control will have an important impact on the disease and on the endemicity. Control projects using indoor spraying with insecticide and impregnated bed nets are discussed.


Vector incrimination and effects of antimalarial drugs on malaria transmission and control in the Amazon basin of Brazil.

World ecosystems differ significantly and a multidisciplinary malaria control approach must be adjusted to meet these requirements. These include a comprehensive understanding of the malaria vectors, their behavior, seasonal distribution and abundance, susceptibility to insecticides (physiological and behavioral), methods to reduce the numbers of human gametocyte carriers through effective health care systems and antimalarial drug treatment, urban malaria transmission versus rural or forest malaria transmission, and the impact of vaccine development. Many malaria vectors are members of species complexes and individual relationships to malaria transmission, seasonal distribution, biting behavior, etc. is poorly understood. Additionally, malaria patients are not examined for circulating gametocytes and both falciparum and vivax malaria patients may be highly infective to mosquitoes after treatment with currently used antimalarial drugs. Studies on the physiological and behavioral effects of DDT and other insecticides are inconclusive and need to be evaluated.


Mechanisms of protective immunity against asexual blood stages of Plasmodium falciparum in the experimental host Saimiri.

In the Saimiri monkey, an experimental host for human malaria, acquired protection against Plasmodium falciparum blood stages depends on the IgG antibody populations developed. In vivo protective anti-falciparum activity of IgG antibodies is correlated with the in vitro opsonizing activity promoting phagocytosis of parasitized red blood cells. In contrast, non protective antibodies inhibit this mechanism by competing at the target level. A similar phenomenon can be observed in human infection. Anti-cytoadherent and anti-rosette antibodies developed by Saimiri and humans prevent the development of physiopathological events like cerebral malaria which can also occur in this experimental host. Furthermore, transfer to protective human anti-falciparum IgG antibodies into infected Saimiri monkeys exerts an anti parasite activity as efficient as that observed when it is transferred into acute falciparum malaria patients, making the Saimiri an even more attractive host. Studies on the role of immunocompetent cells in the protective immune response are still in their infancy, however the existence of a restricted polymorphism of MHC II class molecules in the Saimiri confers additional theoretical and practical importance to this model.


Protection of Aotus monkeys after immunization with recombinant antigens of Plasmodium falciparum.

The genus Aotus spp. (owl monkey) is one of the WHO recommended experimental models for Plasmodium falciparum blood stage infection, especially relevant for vaccination studies with asexual blood stage antigens of this parasite. For several immunization trials with purified recombinant merozoite/schizont antigens, the susceptible Aotus karyotypes II, III, IV and VI were immunized with Escherichia coli derived fusion proteins containing partial sequences of the proteins MSAI (merozoite surface antigen I), SERP (serine-stretch protein) and HRPII (histidine alanine rich protein II) as well as with a group of recombinant antigens obtained by an antiserum raised against a protective 41 kD protein band. The subcutaneous application (3x) of the antigen preparations was carried out in intact animals followed by splenectomy prior to challenge, in order to increase the susceptibility of the experimental hosts to the parasite. A partial sequence of HRPII, the combination of three different fusion proteins of the 41 kD group and a mixture of two sequences of SERP in the presence of a modified Al(OH)3 adjuvant conferred significant protection against a challenge infection with P. falciparum blood stages (2-5 x 10(6)) i. RBC). Monkeys immunized with the MS2-fusion protein carrying the N-terminal part of the 195 kD precursor of the major merozoite surface antigens induced only marginal protection showing some correlation between antibody titer and degree of parasitaemia. Based on the protective capacity of these recombinant antigens we have expressed two hybrid proteins (MS2/SERP/HRPII and SERP/MSAI/HRPII) in E. coli containing selected partial sequences of SERP, HRPII and MSAI. Antibodies raised against both hybrid proteins in rabbits and Aotus monkeys recognize the corresponding schizont polypeptides. In two independent immunization trials using 13 animals (age 7 months to 3 years) we could show that immunization of Aotus monkeys with either of the two hybrid proteins administered in an oil-based well tolerated formulation protected the animals from a severe experimental P. falciparum (strain Palo Alto) infection.


Efficiency of human Plasmodium falciparum malaria vaccine candidates in Aotus lemurinus monkeys.

The protective efficacy of several recombinant and a synthetic Plasmodium falciparum protein was assessed in Aotus monkeys. The rp41 aldolase, the 190L fragment of the MSA-1 protein and fusion 190L-CS. T3 protein containing the CS.T3 helper "universal" epitope were emulsified in Freund's adjuvants and injected 3 times in groups of 4-5 monkeys each one. The synthetic polymer Spf (66)30 also emulsified in Freund's adjuvants was injected 6 times. Control groups for both experiments were immunized with saline solution in the same adjuvant following the same schedules. Serology for malaria specific antibodies showed seroconversion in monkeys immunized with the recombinant proteins but not in those immunized with the polymer nor in the controls. Challenge was performed with the 10(5) parasites from the P. falciparum FVO isolate. Neither rp41 nor Spf(66)30 induced protection, whereas 190L induced significant delay of parasitemia. The fusion of the CS.T3 epitope to 190L significantly increased its protective capacity.


Study of humoral immune response in mammals immunized with Plasmodium falciparum antigenic preparations.

Six Plasmodium falciparum protein fractions, isolated under reducing conditions, were used to immunize mice, rabbits and the squirrel monkey Saimiri sciureus. Five or seven subcutaneous injections of each antigenic preparation, in conjunction with Freund's complete or incomplete adjuvants, were administered. This led to the development of specific antibodies detected by IFAT, ELISA or immunoblotting which inhibited merozoite reinvasion in in vitro P. falciparum cultures. This activity seems to be associated with rhoptry proteins contained in fractions Pf F2 and Pf F4.


Impaired renal function in owl monkeys (Aotus nancymai) infected with Plasmodium falciparum.

Impaired renal function was observed in sixteen Aotus nancymai 25 and 3 months following infection with the Uganda Palo Alto strain of Plasmodium falciparum. Decrease were noted in the clearance of endogenous creatinine, creatinine excretion, and urine volume while increases were observed in serum urea nitrogen, urine protein, urine potassium, fractional excretion of phosphorus and potassium, and activities of urinary enzymes. The results were suggestive of glomerulonephropathy and chronic renal disease.


Plasmodium coatneyi-infected rhesus monkeys: a primate model for human cerebral malaria.

Although several animal models for human cerebral malaria have been proposed in the past, none have shown pathological findings that are similar to those seen in humans. In order to develop an animal model for human cerebral malaria, we studied the pathology of brains of Plasmodium coatneyi (primate malaria parasite)-infected rhesus monkeys. Our study demonstrated parasitized erythrocyte (PRBC) sequestration and cytoadherence of knobs on PRBC to endothelial cells in cerebral microvessels of these monkeys. This is similar to the findings seen in human cerebral malaria. Cerebral microvessels with sequestered PRBC were shown by immunohistochemistry to possess CD36, TSP and ICAM-1. These proteins were not evident in cerebral microvessels of uninfected control monkeys. Our study indicates, for the first time, that rhesus monkeys infected with P. coatneyi can be used as a primate model to study human cerebral malaria.


The role of cytokines in Plasmodium vivax malaria.

The cytokine tumor necrosis factor and other as yet unidentified factor(s) which together mediate the killing of intraerythrocytic malaria parasites are transiently elevated in sera during paroxysms in human Plasmodium vivax infections in non-immunes. These factors which included TNF and parasite killing factor(s) are associated with the clinical disease in malaria to the extent that their transient presence in infection sera coincided with paroxysms, the the most pronounced clinical disturbances of P. vivax malaria and secondly because their levels were markedly lower in paroxysm sera of semi-immune patients who were resident of an endemic area. Further, a close parallel was obtained between serum TNF levels and changes in body temperature that occur during a P. vivax paroxysm in non-immune patients, suggesting a causative role for TNF in the fever in malaria. P. vivax rarely if ever cause complicated clinical syndromes. Nevertheless serum TNF levels reached in acutely ill P. vivax patients were as high as in patients suffering from cerebral complications of P. falciparum malaria as reported in studies from the Gambia. Cytokine profiles and other changes accompanying clinical disease in P. vivax and P. falciparum malaria are compared in this paper with a view to discussing the potential role of cytokines in the causation of disease in malaria.


Molecular approaches to malaria and babesiosis diagnosis.

The development of additional methods for detecting and identifying Babesia and Plasmodium infections may be useful in disease monitoring, management and control efforts. The preliminary evaluate synthetic peptide-based serodiagnosis, a hydrophilic sequence (DDESEFDKEK) was selected from the published BabR gene of B. bovis. Immunization of rabbits and cattle with the hemocyanin-conjugated peptide elicited antibody responses that specifically detected both P. falciparum and B. bovis antigens by immunofluorescence and Western blots. Using a dot-ELISA with this peptide, antisera from immunized and naturally-infected cattle, and immunized rodents, were specifically detected. Reactivity was weak and correlated with peptide immunization or infection. DNA-based detection using repetitive DNA was species-specific in dot-blot formats for B. bovis DNA, and in both dot-blot and in situ formats for P. falciparum; a streamlined enzyme-linked synthetic DNA assay for P. falciparum detected 30 parasites/mm3 from patient blood using either colorimetric (2-15 h color development) or chemiluminescent detection (0.5-6-min exposures). Serodiagnostic and DNA hybridization methods may be complementary in the respective detection of both chronic and acute infections. However, recent improvements in the polymerase chain reaction (PCR) make feasible a more sensitive and uniform approach to the diagnosis of these and other infectious disease complexes, with appropriate primers and processing methods. An analysis of ribosomal DNA genes of Plasmodium and Toxoplasma identified Apicomplexa-conserved sequence regions. Specific and distinctive PCR profiles were obtained for primers spanning the internal transcribed spacer locus for each of several Plasmodium and Babesia species.


Human IgG responses against the N-terminal region of the Merozoite Surface Protein 1 of Plasmodium vivax.

The complete primary structure of the gene encoding the Merozoite Surface Protein 1 of Plasmodium vivax (PvMSP-1) revealed the existence of interspecies conserved regions among the analogous proteins of other Plasmodia species. Here, three DNA recombinant clones expressing 50, 200 and 500 amino acids from the N-terminal region of the PvMSP-1 protein were used on ELISA and protein immunoblotting assays to look at the IgG antibody responses of malaria patients from the Brazilian amazon region of Rondônia. The results showed the existence of P. vivax and P. falciparum IgG antibodies directed against PvMSP-1 antigenic determinants expressed in the clones containing the first 200 and the following 500 amino acids of the molecule, but not within the one expressing the most N-terminal 50 amino acids. Interestingly, there was no correlation between the levels of these IgG antibodies and the previous number of malaria infections.


A chromosome 9 deletion in Plasmodium falciparum results in loss of cytoadherence.

Many lines of Plasmodium falciparum undergo a deletion of the right end of chromosome 9 during in vitro culture accompanied by loss of cytoadherence and gametocytogenesis. Selection of cytoadherent cells from a mixed population co-selects for those with an undeleted chromosome 9 and the selected cells produce gametocytes. The deletion also results in loss of expression of PfEMP1, the putative cytoadherence ligand, suggesting that PfEMP1 or a regulatory gene controlling PfEMP1 expression and gametocytogenesis may be encoded in this region. We have isolated several markers for the deleted region and are currently using a YAC-P. falciparum library to investigate this region of the genome in detail.


Characterization of a Plasmodium falciparium mutant that has deleted the majority of the gametocyte-specific Pf11-1 locus.

We identified a gametocyte-specific protein of Plasmodium falciparum called Pf11-1 and provide experimental evidence that this molecule is involved in the emergence of gametes of the infected erythrocyte (gametogenesis). A mutant parasite clone, which has deleted over 90% of the Pf11-1 gene locus, was an important control to establish the gametocyte-specific expression of the Pf11-1. Molecular analysis of the Pf11-1 deletion indicates that it is presumably due to a chromosome breakage with subsequent 'healing' by the addition of telomeric heptanucleotides. Moreover, similar DNA rearrangements are observed in most of the laboratory isolates during asexual propagation in vitro.


Deletion, insertion and translocation of DNA sequences contribute to chromosome size polymorphism in Plasmodium berghei.

Extensive chromosome size polymorphism arises in Plasmodium berghei during in vivo mitotic multiplication. Size differences between homologous chromosomes mainly involve rearrangements in the subtelomeric regions while internal chromosomal regions are more conserved. Size differences are almost exclusively due to differences in the copy number of a 2.3 kb subtelomeric repeat unit. Not only deletion of 2.3 kb repeats occurs, but addition of new copies of this repeat sometimes results in the formation of enlarged chromosomes. Even chromosomes which originally lack 2.3 kb repeats, can acquire these during mitotic multiplication. In one karyotype mutant, 2.3 kb repeats were inserted within one of the original telomeres of chromosome 4, creating an internal stretch of telomeric repeats. Chromosome translocation can contribute to chromosome size polymorphism as well. We found a karyotype mutant in which chromosome 7 with a size of about 1.4 Mb is translocated to chromosome 13/14 with a size of about 3 Mb, resulting in a rearranged chromosome, which was shown to contain a junction between internal DNA sequences of chromosome 13/14 and subtelomeric 2.3 kb repeats of chromosome 7. In this mutant a new chromosome of 1.4 Mb is present which consists of part of chromosome 13/14.


Effects of thyroidectomy on biochemical and ultrastructural aspects of rat slow and fast muscles.

Thyroidectomy provoked the reduction of the oxygen consumption rate in state 3 respiration in both Extensor Digitorum Longus (EDL, fast twitch) and Soleus (slow twitch) muscles of rat, being the last one the most affected. Morphological alterations after 8 weeks of thyroidectomy were found in mitochondria. These organelles exhibited cristae swelling and formation of autophagic vacuoles in subsarcolemmal and intermyofibrillar spaces. Altered mitochondria were also seen in the axon terminal and the postjunctional region of motor end-plate.


[Bacillary necrosis in larvae of the bivalve Euvola ziczac (Linnaeus, 1758) caused by a Pseudomonas sp].

The incidence of bacteria as etiological agents of diseases in larvae of bivalve mollusks, is well documented in the literature. In this study, a series of test were performed to identify and estimate pathogenic level of marine bacteria isolated during an epizootic in a larval culture system of the tropical scallop Euvola ziczac. Such bacteria was identified as Pseudomonas sp. belonging to the group alcaligenes. The bacterium produced not only lethal effects on the scallop larvae through infection (invasive action), but also a possible pathogenic activity through endotoxins associated to bacterial cell-wall. In addition, it is possible that the isolated bacteria could act as a pathogen with members of the genus Vibrio, which was also present during the epizootic outbreak in the culture system. Susceptibility of Pseudomonas sp. to commercial antibiotics was remarkable.


Hydrolytic activity of an onchocercoma.

The onchocercoma or nodule produced by the nematode Onchocerca volvulus (Filarioidea) in the skin of patients suffering from onchocerciasis has not been examined by histochemical techniques. In this work we have used histochemical techniques to evaluate 5 hydrolytic enzymes, namely phosphatases, esterases and beta-glucuronidase. The results show increased enzymatic activity at the sites of major metabolic activity and within reactive cells including macrophages (mc) and giant cells (gc) of the onchocercoma.


Ultrastructure of an iguana fast twitch denervated muscle.

This ultrastructural study was undertaken to investigate the morphological changes which occur in the fast twitch gastrocnemius muscle of the reptile Iguana iguana after nerve section. It was found that initial degenerative alterations appeared in muscle fibers two weeks after denervation and progressed along the two months of the investigative period. They consisted of disorganization of contractile and sarcotubular elements and the appearance of autophagic vacuoles with mitochondrial debris. However, even two months after nerve section some myofibrils and mitochondria looked normal. Our results suggest that although the general course of denervation atrophy in iguana gastrocnemius is similar to that in other twitch muscles of vertebrates, the chronology of the process shows that iguana fast twitch skeletal muscles exhibit an intermediary position among the vertebrates in relation to their velocity of response to denervation.


[Production and characterization of monoclonal antibodies directed against the hepatitis B surface antigen].

Hepatitis B virus surface antigen (HBsAg) found in a commercial vaccine was use as immunogen and antigen for the production and selection of murine monoclonal antibodies against this viral antigen. Antibody relative avidity was determined based on their capture capacity. Competition studies, and the differential recognition pattern of vaccine preparations showed that high avidity IgG1 antibodies were directed against two distinct antigenic regions. Among them, 6F4 was most suitable for the detection of HBsAg in a sandwich ELISA system, using an IgM antibody, 5E8, as capture. However, the combined use of 6F4 with another which does not recognize the same epitope did not improve the sensitivity of the assay. On the other hand, the combination of 6F4 with a lower avidity IgG1 antibody, 6G10, increased the universe of antigenic diversities recognized by this system. An enzyme immunoassay for detection of HBsAg is proposed, using 3 monoclonal antibodies: one IgM as capture and two IgG1 for detection.


[Biomolecules suppressing myelopoiesis].

Hematopoietic stem cells are capable of self-replication and differentiation to lineage-committed progenitor cells. The progenitors proliferate and differentiate to lineage-specific, morphologically recognizable precursors and, finally, to terminal circulating blood cells. These homeostatic mechanisms are regulated by a complex set of interacting growth stimulatory and inhibitory factors that are produced by, or in collaboration with, the tissue's regulatory microenvironment. A number of well-characterized cytokines have been implicated in the negative regulation of hematopoiesis: ferritin H-subunit (HF), lactoferrin (Lf), prostaglandin E (PGE), tumor necrosis factor (TNF), interferon (IFN), transforming growth factor-beta (TGF beta), acetyl-N-Ser-Asp-Lys-Pro (AcSDKP) or thymosin-beta 4, pyroGlu-Glu-Asp-Cys-Lys (pEEDCK), macrophage inflammatory protein-1 alpha (MIP-1 alpha), inhibin, superoxide dismutase (SOD), glutathione (GSH) and others not well-known yet. The role of inhibitors in restraining stem cells from entering the cell cycle and protecting them from the toxic side effects of chemotherapeutic drugs is opening an alternate strategy for the treatment of cancer patients.


Intramuscular capillary abnormalities in a case of myotonic dystrophy (Steinert's disease).

The structural study of a muscle biopsy from a case of myotonic dystrophy showed endothelial cell and pericyte alterations, and infiltration of lymphocytes, macrophages and mast cells. The histopathological picture was similar to that observed in the muscular compromise of some autoimmune diseases. These findings are interesting because although the aetiology of myotonic dystrophy is still obscure, it has been suggested that in the origin of this disease could be involved either primary muscle fibre damage or that myotonic dystrophy is neurogenic, propositions apparently not directly connected with a relation cause-effect to intramuscular capillary abnormalities.


Trypanosoma cruzi: infectivity and virulence of trypomastigotes incubated with leukocytes and serum from patients with Chagas' disease.

An important issue for the understanding of the host-parasite relationship in Chagas' disease is the extent of the action of the immune system on the parasite. To advance our understanding of this aspect, we evaluated the effect of leukocytes and/or sera from patients with Chagas' disease on trypomastigotes of Trypanosoma cruzi. We incubated, in vitro, leukocytes and sera, from patients with Chagas' disease without evidence of heart disease (INF), patients with Chagasic cardiopathy (CDM) and healthy controls (VOL), with trypomastigotes at 37 degrees C for three hours. Mice were inoculated with parasites at the end of the incubation. We kept a record of the survival time of each animal and every three days we evaluated their Parasitemia. INF (36.4%) and CDM (42.9%) prolonged the prepatent period, but not Vol. Only CDM (57.1%) extended the survival time. The sera from CDM patients that extended the survival of mice prolonged the prepatent period. However serum alone did not extend the survival time, providing evidence that leukocytes are required to decrease the virulence of the inoculum. The capacity of leukocytes and sera, from CDM, to prolong survival time shows that the immune system of patients with Chagasic cardiomyopathy can affect the parasite more intensely than IFN. On the other hand, the higher frequency of positive xenodiagnosis in CDM (11), suggests that, in vivo, occur a down regulation of the immune response.


[In vitro stimulation of Leydig cells with human chorionic gonadotropin in adult rats of different ages].

We studied the functional reserve of Leydig cells in adult rats of different ages in vitro. Suspensions of Leydig cells after digestion by collagenase were stimulated with Human Chorionic Gonadotropin (hCG) during different times (1, 3 and 5 hours). We observed that response of Leydig cells, expressed by the testosterone concentration in the culture medium, showed a similar pattern in all studied group, and the highest response was seen at three hours of incubation. On the other hand, the testosterone response to hCG varied with age of animal. Thus, it was higher in the group of 14 months old (middle aged) than young animals (5 month old) and decreased in old animals (21 months old). These results suggest that a primary testicular lesion occurs with aging.


Vegetable gums modify lectin hemagglutinability.

Arabic gum enhances lectin hemagglutinability. The more glycosylated the lectin, the greater the stimulatory effect of the gum. Evidence presented suggests that the interaction between gum and lectin is of a carbohydrate-carbohydrate nature.


Molecular mimicry between Trypanosoma cruzi and host nervous tissues.

Increasing evidence suggests that in Chagas' disease chronic-phase pathology is autoimmune in nature. There are at least two nonexclusive explanations for the generation of autoimmunity in Chagas disease: a) infection with the parasite perturbs immunoregulation, leading to loss of tolerance for self-antigens; b) immune recognition of T. cruzi antigens is crossreactive with selected mammalian antigens, leading to autoimmunity (molecular mimicry). Through this latter mechanism, T. cruzi antigens that share epitopes with mammalian nervous tissue may drive autoreactive B- or T-cell clones to expand and cause autoimmune lesions in chronic chagasic patients. Several different antigens sharing this characteristic have been studied, as for example the 160-kDa flagellum-associated surface protein (Fl-160), which has a nervous tissue crossreactive epitope composed by twelve amino acids. Additionally, it has been demonstrated that a trypomastigote stage-specific 85kDa surface glycoprotein (Gp85) has terminal galactosyl(alpha 1-3)galactose terminal residues, which are reactive with chronic chagasic sera. Common glycolipid antigens have also been reported, as for example galactocerebroside, sulfogalactocerebroside and sulfoglucuronylcerebroside, all of them specifically present at high concentrations in mammalian nervous system and in T. cruzi trypomastigotes. Chronic chagasic patients produce elevated levels of antibodies against these three glycolipid antigens. They also do against terminal galactosyl(alpha 1-3)galactose residues present on several acid and neutral glycolipids common either to nervous system or parasite. These antibodies are powerful lytic for circulating T. cruzi trypomastigotes. Another common strongly immunogenic residues are galactosyl(alpha 1-2)galactose, galactosyl(alpha 1-6)galactose and galactofuranosyl(beta 1-3)mannose residues present on several glycoinositolphospholipids (GIPL), against which chronic chagasic patients have elevated levels of specific antibodies. In brief, very specific host-parasite relationships existing only in Chagas' disease may explain the particular peripheral nervous tissue damage seen in acute or chronic stages of this disease. This specificity could depend either on invasion of autonomic ganglia by T. cruzi trypomastigotes and modification of nervous cell surface structures by some of the several mechanisms of acquired molecular mimicry.


Characterization of polycyclic aromatic hydrocarbons degradative soil Pseudomonas.

Nine Pseudomonas strains, able to degrade polycycle aromatic hydrocarbons (PAHs), were isolated from enriched cultures with naphthalene, as carbon source, and soil samples from a land farming process applied on oil sludge, as inocula. Degradative tests showed that all the strains were capable to catabolize naphthalene (Nah) and phenanthrene (Phn). U2 strain transferred the selected function (Nah) to P. aeruginosa T1 (Hgr Oct+), however some of the transconjugants lost the Oct character, suggesting that it is of plasmidic nature. T1 derivatives as well the wild strains U28 and U31 transferred Nah function to P. putida AC165. All of the examined transconjugants also catabolized phenanthrene, suggesting that Nah and Phn functions in U2, U28, and U31 strains are linked and probably encoded by transferable plasmids.


[Characterization of Paracoccidioides brasiliensis antigens. Serological immunodiagnosis using western blotting].

The electrophoretic characterization on SDS-PAGE gels of Paracoccidioides brasiliensis antigens, strains 2511 and 6688; and the additional use of immunoblotting has permitted in this study to identify the immunogenic, sensitive and specific antigen fractions for the diagnosis of Paracoccidioidomycosis. The antigenic preparations showed differences depending on the morphologic form of the fungus and the method utilized. The procedure revealed heterogeneity in the humoral immune response of the patients studied and permitted us to establish indices of activity of Paracoccidioidomycosis by the sequential analysis of sera obtained during different stages of the disease. The high sensitivity of this method makes it useful as an additional technique for the serologic immunodiagnosis of Paracoccidioidomycosis.


[Value of combined electro- and vectorcardiography in the estimation of left ventricular mass in the elderly].

Electro- and vectorcardiographic methods describe left ventricle increases only when it has attained a significantly high magnitude, but even in such circumstances the exactitude of such methods is substantially lower than that of echocardiogram (ECHO). On the other hand, in comparison to other age groups, there have been relatively few reports relating electrocardiogram (ECG), vectorcardiogram (VCG) and ECHO with left ventricular mass (LVM) in healthy elderly subjects where increases of the left ventricle mass, if present, would be small or moderate. In this paper LVM as well as LVM index (iLVM) from a group of healthy subjects belonging to a physical training program for elderly, was studied by means of ECHO and computerized ECG and VCG. From ECG, voltage indexes and other LVM associated parameters were extracted; from VCG, planar maximum vectors, areas within VCG loops and maximal spatial magnitude of QRS (SM), were measured. Results of LVM (221 +/- 37.9, g) were higher than figures reported for others groups. Voltage indexes showed normal values, but QRS duration was somewhat prolonged. The best simple linear regression, combining variables from VCG and ECG was maximum horizontal vector (Vmax-Hor) vs Sokolow-Lyon index (SOK) and combining ECHO with ECG or VCG, LVM vs Area inside horizontal loop (AreaHor). A model for estimation of LVM from electrical variables was obtained by multiple linear regression; combining five variables from ECG and VCG. The best model included Sokolow-Lyon index and variables from horizontal and sagittal planes of VCG and spatial magnitude of QRS: LVM = 4.8 SOK-186 VmaxHOR-80 VmaxSAG + 126SM + 340 AreaH + 175.(ABSTRACT TRUNCATED AT 250 WORDS)


[Molecular epidemiology of human rotavirus in Cumaná, Venezuela].

During the period January to December 1987, rotaviruses were obtained from infants hospitalized with acute gastroenteritis in Cumaná, Venezuela, and were compared by analysis of the migration of their double-stranded ribonucleic acid (RNA) genome segments in polyacrylamide gel electrophoresis. Rotavirus RNA was detected in 87 (17.03%) of 511 faecal samples, and at least 12 different electropherotypes were recognized throughout the period of study. Of the rotavirus positive samples, 86 (98.85%) had a "long" electrophoretic pattern characteristic of human rotavirus subgroup 2. The incidence of rotavirus infection was statistically related with the age, sex, and nutritional status of the pediatric patients for epidemiological purposes. This statistical relation was very significant with the age and the nutritional status of the patients but not with their sex.


[Effect of aging on schizophrenia].

The problem of the senescence's influence on schizophrenia and the right to detach this variable from those who take care of temporal evolution is complicated. From a population of 99 schizophrenic patients (DSM III criteria's) having most of 60 year old and most of 20 years schizophrenia's disease evolution; we have study the whole or partial imputability to the senescence of residential statute's modification; frequency, reason, duration of hospitalization's change, psychotic's symptom levelling; and therapeutic's instability.


LHRH analogues and PCO disease: validity of the use of enantone depot in the treatment of the disease.

The Authors have been showed the utility of LHRH analogues in the treatment of PCO disease, through hormonal dosages and echographic examination made before, during and after the therapy both with Enantone depot, CPA and EE2, and with only CPA and EE2 of control cases. The association of analogue to traditional therapy has determined a more speedy reduction of haematic levels of LH, T free, DHEA-S, than the ones we have found in control case, an evident reduction (50%) of pilonidal growth and an important improvement of acne and seborrhea.


[Contribution of single photon emission computerized tomography (SPECT-HMPAO) to the study of schizophrenia].

The authors studied a group of thirteen schizophrenic patients with functional brain imaging, using single-photon-emission-computed tomography (S.P.E.C.T.). The radiotracer was the H.M.P.A.O. labelled with 99 m Technetium. All were being given neuroleptic drug. Forty-one lesions demonstrated decreased perfusion. These functional abnormalities are mainly located in the left hemisphere and this asymmetry is more pronounced in positive-symptom schizophrenics. However, at rest, these correlations are limited and finally we demonstrated a wide spectrum of metabolic dysfunctions in the same subtype of schizophrenia.


[Measuring quality of life].

The four broad domains of quality of life are the physical status and functional abilities, the psychological status and well-being, the social interactions and the economic status and factors. Health profiles attempt to measure all important aspects of Quality of Life (QOL). They offer a number of advantages and they also have some limitations. A number of QOL indexes have been recently developed to measure emotional and social functions, well-being as well as overall health status. Some scales used to evaluate QOL are function-specific as social interactions scales or daily living scales; others are disease-specific. For the severely mentally ill patients the most comprehensive and psychometrically best characterized scales have been recently reviewed. The Lehman's Structured Quality of Life interview for example based on comprehensive quality of life models, includes both subjective and objective QOL indicators and should be extensively used in french clinical studies.


[Psychiatric selection of European astronauts: the French experience].

In order to realize the future Hermes and Columbus space-flights, European Space Agency decided to build up a european astronaut corps. Space-flights and training constraints require a strict initial selection among candidates. This paper describes space-flights psychological constraints, the aims of a psychiatric exam and its position in astronaut selection procedures as a whole. This paper also describes means and results of that selection among the candidates seen in Paris (France) in 1991.


Autoimmune vasculitis in MRL/Mp-lpr/lpr mice: orthochromatic basophils participate in the development of delayed-type hypersensitivity angiitis.

The pathogenesis of autoimmune vasculitis is poorly understood. Understanding the immunologic mechanisms governing this disease requires precise identification of the cells which comprise the lesion. In this report, we have evaluated tissue sections from MRL/lpr mice from 16 to 45 weeks of age, representing all stages of clinical vasculitis. We demonstrate that basophil myelocytes participate in the evolution of the delayed-type hypersensitivity (DTH) response which initiates and perpetuates autoimmune vasculitis in these mice. These findings raise questions regarding the immunologic mechanisms by which basophils develop in this lesion and the interaction of basophils. VSMCs and lymphocytes in vasculitic angiodestruction.


Inhibition of experimental autoimmune uveoretinitis by oral administration of S-antigen and synthetic peptides.

S-Antigen, a photoreceptor cell protein, is highly efficient in inducing experimental autoimmune uveoretinitis (EAU), a severe inflammation of the uveal tract and retina of the eye. S-Antigen and six synthetic peptides, all of which correspond to known T-cell or B-cell recognition sites, were tested for their ability to induce oral tolerance to EAU in LEW rats. Feeding three 1-mg doses of native S-Antigen or three doses of one synthetic peptide, designated BSA(343-362) (200 micrograms per dose), reduced the incidence and severity of EAU induced by immunization with 50 micrograms of S-Antigen. Another peptide, BSA(270-289), was able to inhibit EAU only when a low dose (10 micrograms) of the uveitogenic S-Antigen was used to induce EAU. Animals which received 200 micrograms doses of four other immunologically active peptides, BSA(31-51), BSA(143-162), BSA(303-327) and BSA(333-352), were not significantly protected. Furthermore, animals fed BSA(343-362) were significantly less susceptible to EAU induced by adoptive transfer (tEAU) of the uveitogenic R9 T-cell lines. Con A-activated lymphocytes purified from spleens of rats fed peptide BSA(343-362) transferred partial resistance to tEAU induced by adoptive transfer of R9 line cells. The resistance of orally tolerized rats to induction of EAU by adoptive transfer of an activated, pathogenic T-cell line, and the ability of lymphocytes from orally-tolerized animals to transfer resistance to tEAU shows that effector mechanisms can be inhibited by oral feeding as well as the afferent mechanisms reported here and elsewhere. Circulating levels of IgG specific for S-Antigen were not affected by feeding any of the peptides.


Cytokine expression in labial salivary glands from patients with primary Sjögren's syndrome.

The presence and distribution of 7 cytokines was examined immunohistologically in labial salivary gland (LSG) specimens from patients with primary Sjögren's syndrome (SS) and control subjects. The cytokines interleukin (IL)-1 beta IL-6, tumor necrosis factor (TNF) alpha and interferon (IFN) gamma were identified in defined parts of LSG from patients but not in the corresponding parts of control glands: (a) LSG acinar epithelium expressed IL-1 beta, (b) blood vessels located in both normal LSG stroma and within lymphocytic infiltrates expressed IL-1 beta, IL-6 and IFN gamma, and (c) lymphocytic infiltrates expressed IL-1 beta, IL-6 and TNF alpha. All four cytokines were expressed by salivary ducts within both patient and control specimens, but with generally greater intensity in patients. IL-1 alpha, IL-4 and TNF beta (lymphotoxin) could not be detected in any of the specimens from patients or controls. The locations of cytokines in LSG suggests possible mechanisms of immunologically mediated parenchymal damage in primary SS.


Mechanisms of complete Freund's adjuvant protection against diabetes in BB rats: induction of non-specific suppressor cells.

We have previously reported that a single injection of complete Freund's adjuvant (CFA) can prevent diabetes appearance in diabetes-prone (DP) BB rats. In this study, we investigated further the mechanism of CFA-induced protection from diabetes. We found that adoptive transfer of splenic cells from CFA-treated DP rats into young DP rats protected the latter from diabetes development. This suggested that CFA-induced protection from diabetes resulted from activation of regulatory (suppressor) cells. Cell mixing experiments in vitro indicated that CFA activated splenic cells with antigen-nonspecific suppressor activity (suppression of lymphoproliferative responses to lipopolysaccharide and to allogeneic splenic cells). Fractionation of splenic cells on Percoll revealed that the suppressor activity resided in low density cells relatively depleted of T-cells, B-cells, macrophages and NK cells. These results suggest that non-specific (natural) suppressor cells in CFA-treated BB rats may be responsible for suppressing autoimmune responses and preventing insulitis and diabetes development.


Thyroid-associated ophthalmopathy.

Thyroid-associated ophthalmopathy (TAO) is one of the most vexing problems in endocrinology but its relationship to thyroid autoimmunity is becoming more clear with the realisation that almost all patients with Graves' disease have this condition and almost all patients with ophthalmopathy have some form of thyroid involvement. This suggests cross reactivity between thyroid and retrobulbar antigens, of which one potential candidate has recently been cloned. Together with new information on predisposing factors and the important role of the retrobulbar fibroblasts, these developments shed new light on the aetiology and pathogenesis of this enigmatic disorder.


A muscle-specific actin gene from the Mediterranean fruit fly, Ceratitis capitata.

A characterization of an actin gene isolated from the genome of the Mediterranean fruit fly, Ceratitis capitata, including the complete sequencing of the coding, 3' and 5' flanking regions of this gene and a partial cDNA was carried out. The partial cDNA was derived from the 3' untranslated region of the actin gene described here, and has been used to identify this gene uniquely. The DNA sequence data presented here, together with the pattern of expression exhibited by this gene during development, strongly support the interpretation that this is a muscle-specific actin gene. Peaks of expression are seen in tissues and during temporal phases of development where muscle differentiation is occurring. The derived protein sequence of the Medfly acting gene shows the highest degrees of similarity, 98.4 and 96.6% respectively, with the two muscle-specific actin genes 79B and 88F from D. melanogaster. The Medfly actin gene also has a single intervening sequence, and an intron is found at the same position in the 79B and 88F actin genes. In the coding region at the DNA level, 17.2 and 16.4% nucleotide differences, respectively, are observed between the Medfly actin gene and these same two D. melanogaster actin genes. The disparity between the amino acid and nucleotide comparisons can be explained, in part, by a high level of synonymous changes in the DNA sequence. In addition, despite the many similarities, codon usage appears to be very different between the actin genes of these species.


Phylogeny of cytoplasmic incompatibility micro-organisms in the parasitoid wasp genus Nasonia (Hymenoptera: Pteromalidae) based on 16S ribosomal DNA sequences.

Cytoplasmic incompatibility results in embryo mortality in diploids, or all male offspring in haplodiploids, when individuals carrying different cytoplasmic factors are crossed. Cytoplasmic factors have been identified as intracellular micro-organisms. Microbe-induced cytoplasmic incompatibility is found in many insect taxa and may play a role in reproductive isolation between populations. Such micro-organisms cause bidirectional incompatibility between species of the parasitoid wasp genus Nasonia. The phylogenetic relationship of cytoplasmic incompatibility microorganisms (CIM) of different Nasonia species was analysed using their 16S ribosomal DNA (rDNA) sequence. Two 16S rDNA operons were detected in the CIM of each Nasonia species. Sequence analysis indicates that the Nasonia CIM are closely related and belong to the alpha group of the Proteobacteria.


Structure and sequence of the Cu, Zn superoxide dismutase gene of Chymomyza amoena: phylogeny of the genus and codon-use evolution.

We have cloned and sequenced the Cu, Zn superoxide dismutase gene of Chymomyza amoena. The coding sequence has the same length as in Drosophila species and in Ceratitis capitata. There are two introns, located at the same sites as in Ceratitis. The second intron is absent in Drosophila: this places Chymomyza outside the Drosophila lineage, contrary to proposals based on anatomical and other evidence. The nucleotide or amino acid distances support a phylogeny in which Ceratitis first branches off the common stem, then Chymomyza splits before the divergence of the two major Drosophila subgenera. The estimated divergence times are 58 million for Chymomyza-Drosophila; 48 million years for the Drosophila subgenera. During the intervening 10 million years, the Drosophila lineage lost the second intron and evolved distinct codon-preferences: the G + C use in the third coding positions is increased by 69% in Drosophila relative to Chymomyza or Ceratitis.


A Drosophila metallothionein promoter is inducible in mosquito cells.

Expression from a Drosophila metallothionein promoter (Mtn) was investigated in mosquito cells. Recombinant plasmids carrying a transcription unit comprised of the Escherichia coli beta-galactosidase gene (lacZ) fused to the metallothionein promoter were stably introduced into Aedes albopictus C6/36 cells. A low copy transformant containing approximately 60 copies of plasmid per cell, and a high copy transformant (1-2 x 10(4) copies/cell) were characterized. The expression of beta-galactosidase from the metallothionein promoter could be induced and controlled in this heterologous system, even when the copy number of introduced plasmid was several thousand.


Distribution of split 5.8S ribosomal RNA in Diptera.

Although the 5.8S ribosomal RNA (rRNA) of most eucaryotes consists of 155-170 nucleotides, in two dipterous species the 5.8S rRNA consists of two pieces, 123 and 30 nucleotides in length. The distribution of split 5.8S rRNA was studied in other Diptera and the most closely related order Siphonaptera to learn the origin of split 5.8S rRNA. Four species of mosquitoes, Culex tritaeniorhynchus, Culex pipiens molestus, Aedes albopictus, Anopheles sp. (Culicidae) had a single 5.8S rRNA consisting of approximately 154 nucleotides. A flea Ctenocephalides felis (Siphonaptera: Pulicidae) also had a single RNA of approximately 158 nucleotides. A crane fly (Tipulidae). a midge Orthocladius akamusi (Chironomidae), a robber fly (Asilidae), and a house fly Musca domestica (Muscidae), on the other hand, had divided 5.8S rRNA as did a fruit fly Drosophila melanogaster (Drosophilidae) and a dark-winged fungus gnat Sciara coprophila (Sciaridae). Three hypotheses are proposed on the relationship between the evolution of the 5.8S rRNA and the phylogeny of Diptera.


Expression of the chloramphenicol acetyltransferase gene in Aedes albopictus (C6/36) cells using a non-infectious Sindbis virus expression vector.

Genomic RNA was transcribed in vitro from the non-infectious Sindbis (SIN) virus expression vector (pTRCAT) and introduced into C6/36 (Aedes albopictus) cells by liposome-mediated transfections. The chloramphenicol acetyltransferase (CAT) polypeptide expressed within cells was detected by an indirect immunofluorescent assay directly into 24-well polystyrene tissue culture plates. Approximately 1 in 1000 C6/36 cells showed fluorescence when the transfection was optimized. CAT enzyme activity was also assayed; in C6/36 cells CAT expression was detected as early as 8 h after transfection, peaked at 24 h, and could still be detected at 7 days. At 24 h posttransfection each transfected C6/36 cell was calculated to express 1.3 x 10(7) CAT polypeptides.


Characterization of a cDNA clone encoding a glycine-rich cuticular protein of Tenebrio molitor: developmental expression and effect of a juvenile hormone analogue.

The complete sequence of a cDNA clone, isolated from epidermal mRNA of Tenebrio molitor using a monoclonal antibody raised against an adult-specific cuticular antigen only present in the hard cuticle, was obtained after primer extension at the 5' end. From this cDNA sequence, the deduced protein encompasses 199 amino acids (including a signal peptide) with a total molecular weight of 20.7 kDa. The protein exhibits a bipartite structure: glycine-rich region located in its NH2-terminal part and a carboxy-terminal domain sharing homologies with other cuticular proteins of Orthoptera, Diptera and Lepidoptera. In-situ hybridization analysis shows that the corresponding mRNA is present only in epidermal cells secreting the adult fibrous cuticle destined to become heavily sclerotized. In supernumerary pupae obtained after the application of the juvenile hormone analogue (JHA) to newly ecdysed pupae, the mRNA was undetectable, indicating that JHA can prevent the switch to the adult programme. However, in pupal-adult intermediates, obtained when JHA is applied later, the mRNA is detected.


Gene structure of Bombyx mori larval serum protein (BmLSP).

To understand the molecular mechanisms of the larval-specific transcription of Bombyx mori larval serum protein (BmLSP), we isolated a clone of the BmLSP gene from a genomic library and sequenced a 3.5-kb fragment. An intron was found in the 5' noncoding region of the BmLSP gene. A putative transcription start point was determined by primer extension analysis. Genomic Southern hybridization showed that there is one copy of the BmLSP gene in a haploid genome. A database search revealed that the BmLSP gene has presumptive repetitive sequences found in other B. mori genes, the sequence homologous to ecdysone-responsive elements and a heptamer sequence found in storage protein genes.


Sequence analysis of a mosquito ribosomal protein rpL8 gene and its upstream regulatory region.

The gene encoding Aedes albopictus ribosomal protein L8 was isolated using a cDNA probe. Based on the deduced amino acid sequence, rpL8 has a mass of 28,605 Da, a pI of 11.97, and contains 9.6% Arg and 11.9% Lys. The rpL8 gene spans 1229 nucleotides, and contains three exons measuring 73, 150, and 648 nucleotides. The first intron is 293 nucleotides long and interrupts an 85-nucleotide untranslated leader sequence. The AUG codon is located 12 nucleotides downstream of the 5'-end of the second exon. Separating the second and third exon is a 65-nucleotide intron. The major transcription initiation site, identified by primer extension and polymerase stop reactions, mapped 378 nucleotides upstream from the AUG start codon; minor initiation sites were also detected. The DNA sequence upstream of the rpL8 gene was T-rich, but conventional TATA and CAAT boxes were absent. This is the first molecular analysis of a mosquito ribosomal protein gene.


Transfection of cultured cells of the cotton boll weevil, Anthonomus grandis, with a heat-shock-promoter-chloramphenicol-acetyltransferase construct.

Expression of heat shock proteins (hsp) in the BRL-AG-3C cell line from the cotton boll weevil was examined. It was determined that the maximal expression of endogenous hsp occurred at 41 degrees C. Various transfection methods were then compared using this cell line in conjunction with a transiently expressed bacterial gene marker (chloramphenicol acetyltransferase) which was under the control of the Drosophila hsp 70 gene promoter. The cationic lipid preparation Lipofectin was found to be very efficient at transfecting the boll weevil cells. Polylysine and 20-hydroxyecdysone-conjugated polylysine were moderately effective, whereas polybrene and electroporation, under the conditions reported herein, were ineffective at transfecting this cell line.


Genetic variability in the social bee Lasioglossum marginatum and a cryptic undescribed sibling species, as detected by DNA fingerprinting and allozyme electrophoresis.

DNA fingerprints (DNAfp) were obtained for three widely separated samples of bee related to Lasioglossum marginatum using the M13 sequence as a probe. Bee samples were obtained from France (three localities separated by at most 20 km), Greece and India. All European populations exhibited almost identical profiles with similarity indices (S) of over 98% within a French sample, 94% among Greek bees and 90% between Greek and French bees. The DNAfp profiles of Indian bees showed more polymorphism (intrapopulation S = 77%) and were quite dissimilar to the European samples (S = 55% and 56% to French and Greek samples, respectively). The similarity between populations separated by over 2000 km is higher than among unrelated individuals within a population in two other bee species and the tsetse fly. Data from allozyme electrophoresis shows parallel variation to that obtained with DNAfp and the genetic differences between Indian and European samples are strikingly large (Indian and European populations shared no alleles at 14 out of 47 loci surveyed) such that no more than one species must be involved. Nonetheless, the samples are indistinguishable morphologically. We argue that chronically low effective population size in these species results in low levels of genetic variability and that this, combined with a genetic bottleneck during the speciation event and colonization of Europe, may have resulted in both the extremely low levels of DNAfp variation in European bees and the large number of fixed allelic differences between European and Indian samples.(ABSTRACT TRUNCATED AT 250 WORDS)


Modulation of DNA-binding proteins in Locusta migratoria in relation to juvenile hormone action.

Using a gel mobility shift assay, protein factors have been identified that may participate in the transcriptional regulation of the juvenile hormone (JH) induced vitellogenin A (VgA) gene of the migratory locust. Two DNA-binding proteins were found that were modulated in correlation with VgA gene activity during normal reproductive cycles and experimental stimulation by an active JH analogue (pyriproxyfen). One of these, JH factor (JHF), may be a regulator of VgA and perhaps other JH-responsive genes. The second, mobile factor (MF), appears to have a more general role in cellular responses to external stimuli such as JH and wounding. JHF and MF may represent factors that act together with the JH receptor to regulate VgA gene transcription.


Description of Trichuris travassosi n. sp. (Nematoda: Trichurinae) from a Brazilian rodent, by light and scanning electron microscopy.

A new species of a trichurid nematode Trichuris travassosi n. sp., recovered from a wild rodent in the State of Rio Grande do Sul, Brazil, is described and compared to T. myocastoris (Enigk, 1933) and their differentiation was on the basis of a detailed morphometrical study. Oryzomys nigripes (Olfers, 1818) is a new host record for the genus. The denomination spicular prepuce is proposed to designate the structure previously named spicular sheath and, conversely, spicular sheath to indicate the cuticle that covers the spicule.


[Determination of thermal requirements of Stomoxys calcitrans (L.) (Diptera, Muscidae), under laboratory conditions].

The biology of immature stages of Stomoxys calcitrans (L.) was studied in the laboratory under four constant temperatures. The study was carried out in biological incubators at 20, 25, 30 and 35 degrees C; 65 +/- 10% relative humidity and 14 hours of photophase. The most favorable temperature for developing eggs, larval and pupal was 25 degrees C, while 35 degrees C proved to be harmful for a normal developing of S. calcitrans in larval stage. The incubation periods for egg were 69.90, 42.58, 26.10, 21.78 hours and 2.91, 1.77, 1.08, 0.90 days at 20, 25, 30, 35 degrees C, respectively. The larval stage was 18.40, 11.63, 8.55 days and, the pupal stage, 8.60, 4.54, 3.60 days at 20, 25, 30 degrees C, respectively.. Threshold temperatures for males were a little higher than for females, however, this difference was lesser than 1 decree C. On the other hand, the quantity of energy (GD) for developing females was a little higher than for males.


Ultrastructure of the adults of Bunostomium phlebotomum (Nematoda: Ancylostomatidae).

The morphology of the male and female of Bunostomum phlebotomum are described based on scanning electron microscope (SEM) observations. The attachment of the worms to the small intestinal mucosa and during the copula were observed. Structures of the buccal capsule and genital organs were also studied.


[Sparganosis in some Brazilian vertebrates. Difficulties in the identification of Luheella (Spirometra) species].

Some species of Amphibia and Reptilia are listed as new hosts of spargana, from material deposited in the Helminthological Collection of Oswaldo Cruz Institute. It is discussed the difficulties in identifying the larvae (Sparganum) and also the identification of adults of Luheella species from South America. The histopathology induced by spargana in the liver of a species of Amphibia is briefly described.


Superoxide dismutase from Trichuris ovis--inhibition by benzimidazoles and pyrimidine derivatives.

Three superoxide dismutase isoenzymes of different cellular location were detected in an homogenate of Trichuris ovis. Each of these molecular forms was purified by differential centrifugation and precipitation with ammonium sulphate, followed by chromatography on DEAE-cellulose and Sephadex G-75 columns. The activity levels of the two molecular forms detected in the mitochondrial (one cyanide sensitive Cu-Zn-SOD and the other cyanide insensitive Mn-SOD) were higher than that of the superoxide dismutase detected in the cytoplasmic fraction (cyanide sensitive Cu-Zn-SOD). All molecular forms present evident differences to the SODs contained in the host liver. Molecular mass and some of the physical and chemical properties of the enzyme was determined for all three molecular forms. An inhibitory effect on the SOD of the parasite an the host was detected with a series of compounds, some of which markedly inhibited parasite enzyme but not host enzyme.


[Geopolitical distribution of the occurrence of Fasciola hepatica in Santa Catarina State, Brazil].

During 12 years feces samples from cows, water buffaloes, sheeps and goats were examined by sequential tamis filtration to show the occurrence of Fasciola hepatica eggs. The material came from 129 municipalities of Santa Catarina State, and 5 g of feces per animal were examined. The occurrence of F. hepatica was confirmed in 64.82% of the municipalities. Considering the host, F. hepatica was confirmed in goats from Florianópolis, São José, São João Batista and Guaramirim municipalities; in sheep from Brusque, Pomerode, Palhoça and São José; in water buffaloes from 9 and in cows from 86 municipalities. For this study, 13,762 feces samples were examined and in 3,814 the presence of eggs of F. hepatica was demonstrated. The percentage of occurrence for host species was 27.86 in cows, 24.72 in water buffaloes, 16.92 in sheep and 15.66 in goats. By the results it was demonstrated that Itajaí Valley at Southeast Hidrographic Basin, in Santa Catarina State is an endemic area of F. hepatica, even though Uruguai Hidrographic Basin was not referred as a geographical record for this parasite.


Natural infection of wild rodents by Schistosoma mansoni. Parasitological aspects.

The evaluation of the role of rodents as natural hosts of Schistosoma mansoni was studied at the Pamparrão Valley, Sumidouro, RJ, with monthly captures and examination of the animals. Twenty-three Nectomys squamipes and 9 Akodon arviculoides with a schistosomal infection rate of 56.5% and 22.2% respectively eliminated a great majority of viable eggs. With a strain isolated from one of the naturally infected N. squamipes, we infected 75% of simpatric Biomphalaria glabrata and in 100% of albino Mus musculus mice. The adult worms, isolated from N. squamipes after perfusion were located mainly in the liver (91.5%) and the mesenteric veins (8.5%). The male/female proportion was of 2:1. The eggs were distributed on small intestine segments (proximal, medial and distal portions) and the large intestine without any significant differences in egg concentration of these segments. In A. arviculoides, the few eggs eliminated by the stools were viable and there was little egg retention on intestinal segments. Considering the ease to complete S. mansoni biological cycle in the Nectomys/Biomphalaria/Nectomys system under laboratory conditions, probably the same is likely to occur in natural conditions. In support to this hypothesis there are also the facts that human mansonic schistosomiasis has a very low prevalence in Sumidouro and endemicity among the rodents has not changed even after repeated treatments of the local patients. Based on our experiments, we conclude that N. squamipes has become a natural host of S. mansoni and possibly may participate in keeping the cycle of schistosomiasis transmission at Pamparrão Valley.


Nectomys squamipes (Rodentia: Cricetidae) as an experimental model for Schistosomiasis mansoni.

Twenty specimens of Nectomys squamipes born in captivity, were infected with 500 cercariae by the transcutaneous route. Coprologic examinations were carried out from the 5th to 23rd week after infection. On the 7th, 8th, 12th, 16th, and 23rd weeks the animals were sacrificed and perfused. The oogram was performed in segments of the small intestine (proximal, medial and distal portions) and the large intestine. The average pre-patent period was of 42 days. The average number of eggs varied from 350 on the 6th week, to 800 on the 13th. From the 14th week on, the average number of eggs eliminated was lower than 50 per gram of feces. The recovery of worms kept steady on the 7th, 8th and 12th week (16.85%; 15.45% and 11.95%), decreasing to 7.70% on the 16th week and 8.45% on the 23rd week. The proportion of male/female worms was about the same on the first two weeks, but from the 12th week on, the proportion was: 1.4/1 on the 12th week; 2.5/1 on the 16th week and 1.8/1 on the 23rd week. These observations suggest that N. squamipes may be used as an experimental model for schistosomiasis mansoni, to which it develops resistance mechanism, useful for immunity studies.


Vaccination in murine schistosomiasis with adult worm derived antigens--II. Protective and immune response in inbred mice.

Previous work in our laboratory, mainly focused the prospects of achieving resistance against Schistosoma mansoni infection with adult worm-derived antigens in the form of a soluble extract (SE). This extract obtained by incubation of living adult schistosomes in saline, contains a large number of distinct molecules and was actually shown to be significantly protective in different outbred animals models such as Swiss mice and rabbits. It thus appeared worthwhile to investigate the potential protective activity of SE in different inbred strains of mice, known to be highly susceptible to the infection. Herein we present data showing that DBA/2 mice, once immunized with SE acquire significant levels of resistance to a S. mansoni cercarial challenge. In addition, preliminary studies on the immune system of immunized animals revealed that, injection of SE caused no general imbalance of B or T cell responses.


Report of Balanorchis anastrophus in Pará State with surface topography by scanning electron microscopy.

Balanorchis anastrophus Fischoeder, 1901, from the reticulum of Bos taurus is reported for the first time in the State of Pará, Brazil. The surface topography as revealed by scanning electron microscopy is presented.


[Natural parasitism of buffaloes in Botucatu, SP, Brazil. III. Dynamics of gastrointestinal parasitism in cows and their calves].

Gastrointestinal parasitism of 24 buffalo cows before parturition, and post-parturition, their infection and that of their respective calves during the following 30 weeks were studied. Willis, Hoffmann and whenever possible, the modified Gordon & Whitlock techniques were used for fecal examinations. Toxocara vitulorum eggs were the earliest forms encountered in calves feces, as follows: during the 1st week after birth, 58.33% of the calves were positive, and in the 4th week, 100% of these animals were positive. Eggs of Strongyloides sp were in the 1st week after birth in two of the calves and in the 5th week, all for them were positive. The next parasites to appear were the Coccidia of which oocysts were detected in the feces of two calves in the 2nd week after birth, and 58.33% of the calves were positive for these in the 3rd week, and in the 6th week, all calves shed oocysts in their feces. On the other hand, eggs of Strongyloides were the last forms to appear in calves feces. However, despite their sporadic appearance in the feces, eggs of these parasites were observed continuously from the 11th week onwards, and at this point, the percentage of positive samples began to increase to reach its peak. Relatively to adult animals, eggs of T. vitulorum were observed in the feces of 11 cows, one or twice at most; eggs of Strongyloides sp were seen only once in the feces of four buffalo cows and eggs of Strongyloides in 21 out of 24 cows. Oocysts of Coccidia were observed in 16 cows. Mechanisms of infestation of calves with these parasites are discussed.


Enhancement of Leishmania amazonensis infection in BCG non-responder mice by BCG-antigen specific vaccine.

Different patterns of cutaneous leishmaniasis can be induced when a challenge of alike dose of Leishmania amazonensis amastigotes in various inbred strains was applied. Two strains of mice, the Balb/c and C57BL/10J, showed exceptional susceptibility, and 10(6) amastigotes infective dose lead, to ulcerative progressive lesions with cutaneous metastasis and loss by necrosis of leg on which the footpad primary lesion occurred. Lesions were also progressive but in a lower degree when C3H/HeN and C57BL/6 were infected. Lesions progress slowly in DBA/2 mice presenting lesions which reach a discreet peak after 12 weeks, do not heal but do not ulcerate. DBA/2 mice is, therefore, a good model for immunomodulation. In attempt to determine the influence of BCG in vaccination schedule using microsomal fraction, DBA/2 became an excellent model, since it is also a non-responder to BCG. Vaccination of DBA/2 mice, receiving the same 10(6) BCG viable dose and 10 micrograms or 50 micrograms of protein content of microsomal fraction, lead to a progressive disease with time course similar to those observed in susceptible non-vaccinated C57BL/10J mice after 6 months of observation. An enhancement of infection in BCG non-responder mice suggests that use of BCG as immunostimulant in humans could be critical for both vaccination and immunoprophylactic strategies.


Cotylophoron Travassosi sp. n. (Trematoda--Paramphistomidae) from cattle.

A new species of the genus Cotylophoron (Trematoda--Paramphistomidae) --Cotylophoron travassosi sp. n.--is described. The measurements of the worm and its structures are compared with the valid known species.


[Bionomic studies of Dipetalogaster maximus (Uhler, 1894) (Hemiptera, Reduviidae, Triatominae). III. Population dynamics].

A population dynamics study of D. maximus was carried out under laboratory conditions (28 degrees C e 65% +/- 5% U.R.), and the methodology was the same that has been used for rearing these insects. In order to evaluate the population growth rate of this species, during a 24 months period, five colonies started with a couple recently emerged were observed. Each couple (a male and a female) was maintained in a glass container measuring 20 cm of diameter and 20 cm in height with filter paper on the bottom. The insects were monthly feeding with normal mice blood, and at this day the number of eggs, nymphal stages and adults was registered. All graphical representations of the populations growth showed the same shape. It was found that the average of nymphal stage represented 64.3% of the holè population whereas the oviposition curve showed to be inverse to this one (28.57%) a small percentage of adults was found: males 3.85% and females 3.12%. In this study observations on the biological cycle, longevity and fertility rates were also carried out.


Schizotrypanids: the occurrence of dermatitis in immunodeficient animals infected with Trypanosoma cruzi.

Congenitally athymic nude Balb/c (nu/nu) and their phenotypically normal adult and neonate littermates (nu/+), the C3H/HeN as well, were intraperitoneally infected with two strains (Y or CL) or Trypanosoma cruzi. The nude mice and the neonates developed a severe parasitaemia, the susceptible C3H/HeN also presented high level and adult Balb/c mice presented parasitaemia similar to that observed in outbred mice. Erythematous skin lesions were observed initially in infected athymic nude and neonates, being characterized by nests of amastigotes in the dermis; in C3H/HeN infected mice no nest of parasite was observed but a low-grade inflammatory reaction was seen. In adult Balb/c or OF1 outbred mice nest was found but discreet inflammatory reaction was observed in severe infections.


Lamella formation and emigration from the water by a laboratory colony of Biomphalaria glabrata (Say) in a flow-through system.

Lamella formation and emigration from the water were investigated in juvenile Biomphalaria glabrata reared at two temperatures in aquaria with a constant water flow. Most snails (97.4%) reared at the lower temperature (21 degrees C) formed lamella at the shell aperture and emigrated from the water, whereas only 10.1% did so at 25 degrees C. Eighty percent of emigrations at 21 degrees C occurred within a period of 15 days, 70-85 days after hatching. A comparison of the studies done so far indicates that the phenomenon may be affected by the ageing of snail colonies kept in the laboratory and their geographic origin, rather than the rearing conditions. This hypothesis, however, requires experimental confirmation.


Description of Raillietia flechtmanni sp. n. (Acari: Gamasida).

Raillietia flechtmanni sp. n. is described from the ear canal of its type host the domestic buffalo. The new species parasitizes cattle as well in Brazil.


Preliminary investigations on transmission and life cycle of the ear mites of the genus Raillietia trouessart (Acari: Gamasida) parasites of cattle.

The life cycle of ear mites of the genus Raillietia Trouessart consists of egg, larva, proto- and deutonymph and adult. The proto- and deutonymph are free living, non feeding instars. The teneral adult is the transfer stage. The minimum period required for completion of the life cycle is approximately eight days.


[The effect of granulocyte elastase inhibitor (urinastatin) vaginal suppository on patients with imminent premature delivery].

Cervical maturation, dilatation and uterine contraction in imminent premature delivery are closely related to chemical mediators from activated granulocytes which infiltrate into the cervix. It is known that urinastatin (urinary trypsin inhibitor, UTI) inhibits many kinds of chemical mediators from granulocytes and macrophages such as granulocyte elastase (elastase) and interleukin 1. We examined the effect of a UTI suppository on uterine contraction and the elastase level in cervical mucus in cases of imminent premature delivery. We treated 43 cases of imminent premature delivery with tocolysis index 3 or 4 with 4 kinds of therapy: Group A (N = 12): ritodorine drop infusion therapy; Group B (N = 9): daily UTI suppository (1,000U) therapy; Group C (N = 14): daily UTI suppository + ritodorine drop infusion therapy; Group D: daily UTI suppository + ritodorine drop infusion + antibiotics (oral cepharosporine) therapy. The elastase level of cervical mucus before treatment was 0.76 +/- 0.40 micrograms/ml in group A, 0.93 +/- 0.43 micrograms/ml in group B, 0.85 +/- 0.40 micrograms/ml in group C and 0.90 +/- 0.41 micrograms/ml in group D. There was no significant difference between these groups. The elastase level in cervical mucus was 0.75 +/- 0.47 micrograms/ml in group A, 0.27 +/- 0.35 micrograms/ml in group B, 0.27 +/- 0.33 micrograms/ml in group C and 0.30 +/- 0.19 micrograms/ml in group D, respectively. The elastase level was decreased significantly in groups B, C and D. The time taken to depress uterine contraction was 65 +/- 66 min in group A, 375 +/- 336 min in group B, 70 +/- 64 min in group C and 58 +/- 53 min in group D, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


IgG subclass deficiency associated with corticosteroids in obstructive lung disease.

IgG subclass levels were measured in three groups of adult patients with obstructive airways disease to discern the relationships among depressed IgG subclass levels, chronic corticosteroid use, and recurrent sinopulmonary infections. Group 1 consisted of patients with corticosteroid-dependent bronchial asthma, group 2 patients had corticosteroid-dependent chronic bronchitis/emphysema, and group 3 was comprised of asthma patients not requiring oral corticosteroids but associated with recurrent sinopulmonary infections. One or more IgG subclass deficiencies were noted in 66.7 percent of group 1, 46.7 percent of group 2, and 6.7 percent of group 3. Significant differences were noted between groups 1 and 3 (p = .0008) and between groups 2 and 3 (p = .018), but not between groups 1 and 2 (p = .5). IgG1 deficiency was the most common subclass deficiency found; 14 (77.8 percent) of 18 patients with detectable subclass deficiency demonstrated IgG1 deficiency. In this study population, IgG subclass level deficiencies appeared to be secondary to long-term low-dose corticosteroid therapy.


The allocation of leprosy patients into paucibacillary and multibacillary groups for multidrug therapy, taking into account the number of body areas affected by skin, or skin and nerve lesions.

In Nepal, the setting up and maintaining of reliable services for slit-skin smears has proven difficult. A clinical classification system for leprosy has therefore been developed to assist in the allocation of patients to either paucibacillary or multibacillary groups for the purpose of multiple drug therapy (MDT), using 9 body areas: head (1), arms (2), legs (2), trunk (4). Patients with more than two areas of the body affected are grouped as multibacillary (MB) and those with only one or two areas affected are paucibacillary (PB). Using a computer simulation model and the data of 53 patients registered at Green Pastures Hospital (GPH) in Pokhara and 703 field patients from the Western Region, different clinical classification systems were evaluated with regard to their sensitivity, specificity, and predictive value for MB or PB classification, as compared with the histological classification for the GPH cases and the bacteriological classification for the field patients. The sensitivity and specificity of the body area system in present use were 93% and 39%, respectively. The low specificity is due to MB overclassification. The sensitivity of the WHO classification system without skin smear facilities is 73% (the difference with the body area system is significant: p < 0.05, McNemar's test). Our histology findings confirm previous publications indicating that, while some borderline-tuberculoid (BT) patients may outwardly have a 'PB appearance' and be skin-smear negative, their nerve biopsy and sometimes skin biopsy may show a 'MB' picture. This is the first publication discussing a 'body area system' for the purpose described, including diagrams of the areas used. In Nepal it has proved easy to use and teach and its use may be justified in other control programmes which implement MDT, particularly if slit-skin smear services are unreliable or nonexistent.


General dental practitioners' evaluation of the need for extraction of asymptomatic mandibular third molars.

Thirty general dental practitioners were asked to evaluate the need for extraction of asymptomatic mandibular third molars. Thirty-six mandibular third molars with equal distribution of angular positions, impaction status, males and females and age groups were selected. To estimate the consistency of the evaluation, the 36 cases were duplicated so that, in all, 72 teeth were evaluated. The number of molars proposed to be extracted by the observers varied from 0 to 26. There was no third molar which all observers agreed should be extracted. The two molars which most observers, 25 and 23 of altogether 30 observers, proposed to be extracted were partially covered by soft tissue. The decision not to extract two molars was unanimous. Both of these were completely covered by bone tissue and positioned vertically. The mean overall intra-observer agreement for the therapeutical decision was 92%, with a range of 69-100%. The length of professional experience of the observer did not influence the evaluation whether or not to extract. We conclude that there is a great variation among general dental practitioners regarding their evaluation on the need for removal of asymptomatic mandibular third molars.


Devic's neuromyelitis optica and Schilder's myelinoclastic diffuse sclerosis.

An adult patient developed both Devic's neuromyelitis optica and Schilder's myelinoclastic diffuse sclerosis, suggesting that these entities represent rare topographical and aggressive variants within the spectrum of multiple sclerosis.


Identification, genetic analysis and characterization of a sugar-non-specific nuclease from the cyanobacterium Anabaena sp. PCC 7120.

A nuclease that could be recovered from the supernatant of cultures, as well as from cell-free extracts, of the cyanobacterium Anabaena sp. PCC 7120 was identified as a 29 kDa polypeptide by its ability to degrade DNA after electrophoresis in DNA-containing SDS-polyacrylamide gels. Some clones of a gene library of strain PCC 7120 established in Escherichia coli were found to produce the 29 kDa nuclease. The nucA gene encoding this nuclease was subcloned and sequenced. The deduced polypeptide, NucA, had a molecular weight of 29,650, presented a presumptive signal peptide in its N-terminal region and showed homology to the products of the nuc gene from Serratia marcescens and the NUC1 gene from Saccharomyces cerevisiae. The NucA protein from Anabaena itself, or from the cloned nucA gene expressed in E. coli, catalysed the degradation of both RNA and DNA, had the potential to act as an endonuclease, and functioned best in the presence of Mn2+ or Mg2+. An Anabaena nucA insertional mutant was generated which failed to produce the 29 kDa nuclease.


Purification and point-mutation analysis of human interleukin-2 (Ser-125).

A new type of IL-2(Ser-125) was purified by semi-preparative RP-HPLC, and its purity was analyzed with microbore HPLC, IEF, SDS-PAGE and capillary electrophoresis (CE). The N-terminal sequencing indicated the microheterogeneity of the N-terminus, i.e., N-Met(2/3) and N-Ala(1/3), which was identified with the results of CE and IEF. The amino acid sequence of the point mutated peptide confirmed the replacement of Cys-125 with Ser.


Cloning and partial sequencing of the porcine growth hormone (pGH) gene from pituitary gland.

High molecular weight DNA was isolated from porcine pituitary gland, and a genomic library was constructed using lambda EMBL3 as cloning vector. Five positive clones were identified by in-situ hybridization with the full-length pGH cDNA as the probe. Dot-blot hybridization, restriction analysis, and Southern hybridization showed that one of the positive clones contained the entire pGH gene. A subclone that contained the upstream sequence from the SmaI site of pGH gene was identified, and part of it was sequenced. We also compared the sequence of our pGH gene with the corresponding sequence of the published pGH gene.


Cloning, mapping, and sequencing of 3' and its flanking region of bovine alpha-s1 casein gene.

A positive clone was isolated from a bovine genomic library in EMBL3 with the probe of alpha-s1 casein gene cDNA. The results from both restriction endonuclease digestion and Southern hybridization probed with the upstream and downstream fragments from the termination codon of bovine alpha-s1 casein gene cDNA suggested that the clone contained 3.5 kb downstream and 10.2 kb upstream fragments from the termination codon of alpha-s1 casein gene. Some divergences in restriction endonucleases sites, a few deletions, and many point mutations were found in introns and exons when compared with those which were published.


Establishment of hybridoma cell line secreting specific monoclonal antibodies against turnip mosaic virus and analysis of properties of the McAb.

Monoclonal antibodies (McAb) of hybridomas derived from the fusing of the mouse splenocytes immunized by TuMV with BALB/c mouse myeloma cells (SP2/0-Ag14). Five kinds of hybridoma cell line were produced by indirect-ELISA screening and cloning three times with limiting dilution. Four kinds of hybridoma produced antibodies respectively reactive to TuMV C1, C3, C4 and C5. One kind was reactive to all five strains of TuMV. In indirect-ELISA and sandwich-ELISA tests, TuMV specific monoclonal antibodies did not react with CaMV, CMV, TMV, PVX, and PVY. Antibody titers of ascitic fluids were about 1:256,000 to 2,048,000 in indirect-ELISA. The biological, physical, and chemical properties of the hybridoma cell lines and McAb were identified. The identification of TuMV strains, the specificity and stability of McAb, the coat proteins, and the antigenic site of TuMV were discussed and analyzed with SDS-PAGE and western-blotting.


Effects of ammonium and lactate on hybridoma cell growth and metabolism.

The effects of ammonium and lactate at different concentrations on the growth of 2F7 hybridoma cells, involving glucose consumption, lactate accumulation and cell viability, are investigated. The results suggest that growth inhibition occurs at 2.5 mM ammonium and 2.5 mg/ml lactate, and that severe inhibition occurs at 5 mM ammonium and 5.0 mg/ml lactate.


Metabolic regulation of pigment formation of Onosma paniculatum cultured cells.

The pigment yields of Onosma paniculatum callus and suspension cultured cells were increased 5.5 and 8.1 fold of that of the control respectively, when 10(-5) M of copper ion was added into the medium. The pigments synthesis was also stimulated greatly when 10(-5) M of L-phenylalanine was added into the medium and the cells were cultured for twenty-one days. The maximum yield of pigments was obtained when 10(-6) M of ascorbic acid was added to the medium.


Partition coefficient of luciferase from photobacteria in PEG/salt two aqueous phase system.

The relationship between the logarithmic partition coefficient (K) of luciferase of photobacteria and PEG MW in the PEG/salt two aqueous phase system was shown to be a linear function. The hydrophilic PEG of lower MW facilitated the partition of luciferase into the top PEG-riching phase and gave a higher K value, while PEG of higher MW, its hydrophobic characteristic, made more enzyme partition into the bottom salt-riching phase and lower K value was obtained. In the PEG/trivalent salt system, such as phosphate and citrate, there was a turning-point on the linear relation between the log K and the PEG MW, but which never appeared if a divalent salt such as sulfate, succinate or tartrate was used in the system. When the system was composed of homogeneous PEG and ammonium sulfate, the K value was increased with the increment of the salt concentration, but after the salt concentration had reached at certain level, the K value was uninfluenced. When two kinds of PEG with different MW were used in this system a minimal K value appeared at certain concentration of ammonium sulfate, and the K value was raised when the salt concentration was either increased or decreased. Neither the proportion of the two kinds of PEG nor their total concentration used in the system showed any effect on the above patterns, although the K value may give some corresponding changes. (ABSTRACT TRUNCATED AT 250 WORDS)


Construction of hybridomas secreting monoclonal antibody against sweet potato feathery mottle virus and use of antibody for detection of SPFMV.

Two hybridomas stably secreting monoclonal antibodies (McAb) against sweet potato feathery mottle virus (SPFMV) were constructed by fusing mouse myeloma (Sp2/0-Ag14) with mouse splenocytes immunized with SPFMV-LF. The McAb secreted by the two hybridomas specifically reacted with both SPFMV-LF (China strain) and SPFMV-RC (American strain) in dot-blot-ELISA. A large amount of McAb was produced with BALB/c mice. The titer of hybridoma cultured supernatant was 1:256, and that of the ascitic fluids was 1:1 x 10(4); and the immunoglobulins of McAb were IgG2a and IgG3 respectively. The detection of SPFMV-infected sweet potato plants with McAb confirmed that the McAb obtained in this test were desirable for large-scale routine surveys of SPFMV. This study on construction of hybridomas and detection of SPFMV using McAb is the first report in the world.


Neuropathology of immunosuppression.

Traditionally, the brain has been considered an "immunologically privileged" organ. Under normal conditions, the blood-brain barrier (BBB) is highly effective in preventing both cellular and humoral constituents of the blood from entering the brain parenchyma. In certain pathological conditions, such as viral infections and demyelinating disorders, the BBB may become altered, activated T cells and monocytes may gain access to the brain parenchyma, and microglia may assume the functions of antigen-presenting cells and macrophages. Naturally-occurring or clinically-induced immunosuppression may dramatically alter various cellular and/or humoral aspects of the immune system. Consequently, the brain may become susceptible to disorders that would otherwise be excluded or may develop more severe manifestations of diseases, such as certain infections. This review considers the neuropathologic aspects of various conditions that may be encountered in the setting of both acquired and inherited immunosuppression. The major categories include infectious, neoplastic, vascular, and metabolic disorders. The review also briefly addresses the neuropathology of complications of chemotherapeutic agents, radiotherapy, and organ transplantation inasmuch as they often occur in the clinical setting of acquired immunosuppression.


Dominant and recessive molecular changes in neuroblastomas.

Of all human tumors, neuroblastomas bear the most prominent genetic changes. Amplifications and deletions of chromosomal DNA can be identified by light microscopy on chromosomal spreads of neuroblastoma cells with remarkable frequency and consistency. Consequently, extensive studies have been undertaken to elucidate the molecular basis of these cytogenetic changes. A rich body of information has accumulated on the role played by dominant oncogenes and recessive tumor suppressor genes in the pathogenesis of this disease. Most notably, it was found that amplification of N-myc is responsible for the presence of double minutes and homogeneously staining regions in neuroblastoma chromosomes. It has also been discovered that N-myc amplification is a prognostic sign of malignancy. More recently, recessive genetic alterations in neuroblastoma, such as deletion of putative tumor-suppressing genes have received increasing attention, and considerable efforts are being made to identify such genetic elements. Finally, the susceptibility of neuroblastoma cells to differentiating stimuli has made them a popular in vitro system for neurobiological and pharmacological research. The need for suitable in vivo systems has spurred the development of several animal models employing tumor viruses and transgenic technologies.


Anatomical features of the developing brain implicated in pathogenesis of hypoxic-ischemic injury.

The developing nervous systems is subject to damage from lack of vital substances necessary for normal maturation and function as well as from trauma or a variety of toxins and infectious agents. By far, the most important of these is inadequate oxygen delivery to the fetus in utero, and/or during the intrapartum and/or early neonatal period. Many types of lesions have been described under the rubric of hypoxic-ischemic encephalopathy, a major proportion of which are found only in the immature nervous system and essentially are never seen later in life. Moreover, a large number are primarily hemorrhagic rather than ischemic in character. The unique character and distribution of these lesions results from a collision of the changing anatomy of the developing nervous system and pathophysiological factors afflicting the immature organism. Whereas the majority of hypoxic-ischemic lesions in the fetus/neonate fall into this group, abnormalities characteristically found in the mature nervous system are also seen. Recognition of the anatomic and physiological features peculiar to the developing nervous system will assist in diagnosis of hypoxic-ischemic damage peculiar to the fetus and neonate.


Effect of cerebral blood flow and cerebrovascular autoregulation on the distribution, type and extent of cerebral injury.

Global cerebral blood flow (GCBF) is low in the human neonate compared to the adult. It is even lower in mechanically ventilated, preterm infants: 10-12 ml/100 g/minute, a level associated with brain infarction in adults. The reactivity, however, of global CBF to changes in cerebral metabolism, PaCO2, and arterial blood pressure is normal, except following severe birth asphyxia, or in mechanically ventilated preterm infants, who subsequently develop major germinal layer hemorrhage. The low level of cerebral blood flow (CBF) matches a low cerebral metabolism of glucose and a relatively small number of cortical synapses in the perinatal period. It has not been possible to define a threshold for GCBF below which electrical dysfunction or brain damage occurs (such as white matter and thalamic-basal ganglia necrosis). Three explanations for the lack of clear relation between GCBF and electrical brain activity of the preterm infant must be examined more closely: 1) low levels of CBF are adequate; 2) GCBF does not adequately reflect critically low perfusion of the white matter, and 3) acute white matter ischemia does not result in electrical silence. Two clinical patterns of brain damage following asphyxia may be explained by changes in the blood flow distribution induced by asphyxia: brainstem sparing and parasagittal cerebral injury. Hours to days after severe asphyxia, a state of marked global hyperperfusion may prevail. It is associated with poor neurological outcome and may be an entry point for trials of interventions aiming sat blocking the translation of asphyctic injury to cellular death and tissue damage.


Cerebral carbohydrate and energy metabolism in perinatal hypoxic-ischemic brain damage.

Cerebral hypoxia-ischemia remains a major cause of acute perinatal brain injury. Research in experimental animals over the past decade has greatly expanded our knowledge of those oxidative events which occur during a hypoxic-ischemic insult to the brain, as well as those metabolic alterations which evolve during the recovery period following resuscitation. The available evidence suggests that hypoxia alone does not lead to brain damage, but rather a combination of hypoxia-ischemia or isolated cerebral ischemia is a necessary prerequisite for tissue injury to occur. Furthermore, hypoxia-ischemia severe enough to produce irreversible tissue injury is always associated with major perturbations in the energy status of the perinatal brain which persists well into the recovery period. The lingering energy depletion sets in motion a cascade of biochemical alterations that are initiated during the course of the insult and proceed well into the recovery period to culminate in either neuronal necrosis or infarction. Unlike the adult, where glucose supplementation prior to or during hypoxia-ischemia accentuates tissue injury, glucose treatment of perinatal animals subjected to a similar insult substantially reduces the extent of tissue injury. The mechanism for the age-specific effect of glucose on hypoxic-ischemic brain damage is discussed in relation to pathogenetic mechanisms responsible for the occurrence of permanent brain damage.


Excitatory amino acids contribute to the pathogenesis of perinatal hypoxic-ischemic brain injury.

A large body of experimental evidence indicates that over-activation of excitatory amino acid (EAA) receptors may mediate irreversible neuronal injury in a variety of pathologic settings including cerebral ischemia, and that the developing brain may be particularly susceptible to the adverse effects of EAA receptor overactivation. In this article, we review current information about EAA receptor pharmacology and EAA neurotoxicity in the immature brain, and summarize recent experimental data indicating that EAA contribute to the pathogenesis of perinatal hypoxic-ischemic brain injury.


Heat shock proteins in hypoxic-ischemic brain injury: a perspective.

There is much to suggest that the induction of heat shock protein synthesis is an important response to injury and stress in the brain. The role of heat shock proteins in neurological disease has been approached from two points-of-view. First, the induction and synthesis of specific proteins after brain cell injury provide a window through which insight on the regulation of gene expression in pathological tissue can be obtained. These studies have broad implications for understanding pathophysiological mechanisms of disease. Second, putative cell protective effects of heat shock proteins in brain tissue provide insight into biochemical mechanisms of selective neuronal vulnerability. These studies have extremely important clinical implications since cell sensitivity to injury can seemingly be modified. The role of heat shock proteins in hypoxic-ischemic brain injury is discussed forthwith.