Double dens invaginatus of molarized maxillary central incisors, premolarization of maxillary lateral incisors, multituberculism of the mandibular incisors, canines and first premolar, and sensorineural hearing loss.

Molarization and premolarization of anterior teeth have never been reported before. Double dens invaginatus is an extremely rare condition. We describe an affected female who had developmental delay and congenital progressive sensorineural hearing loss. Double dens invaginatus of molarized maxillary central incisors and premolarized maxillary lateral incisors were present. In addition, multituberculated mandibular incisors, canines, and first premolar were observed. Histologically, tooth structure was unremarkable. Family history of dental abnormalities and hearing loss was denied. Clinical and radiographic examinations were those performed when the patient was 7 and 13 years old. The basic defect is postulated to be of mesenchymal origin.


Autosomal recessive Silver-Russell syndrome.

Six children (5 male, 1 female) of normal first cousin Arab parents were found to have Silver-Russell syndrome. Manifestations included intrauterine and postnatal growth retardation, lateral asymmetry, relatively large head, small triangular face with prominent ears, clinodactyly of the fifth fingers, disproportionate toes, and normal psychomotor development. Intrafamilial variability was minimal. Parental consanguinity and affected siblings of both sexes strongly suggest autosomal recessive inheritance. Similar cases from literature are briefly reviewed.


[Plasma levels of MHPG, HVA and total 5-HIAA in depression. Preliminary study].

This study was aimed at assessing monoamine catabolites plasma levels in depressed patients and healthy volunteers. Plasma levels of 3-methoxy-4-hydroxyphenylglycol (MHPG), homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5-HIAA) of 21 control subjects and 26 depressed patients (according to DSM III-R criteria) were measured at baseline (day 0) and day 4, day 7, day 30 of prescribed antidepressant treatment. The clinical assessment, at baseline as well as during treatment, used the Hamilton depression rating scale and the BPRS. Our data show the interest of these results in predicting response. The respondent patients showed a significant decrease in plasma MHPG level at J7, contrary to non-respondent patients. Moreover, a positive correlation between plasma levels of MHPG and HVA before any prescribed antidepressants was found only with respondent patients. The lack of correlation for non-respondent patients can suggest that the relationships between this monoamine systems should be disrupted in these patients. Significant relationships appear between clinical symptoms and plasma catabolites, allowing us to consider new physiopathological aspects of the depressive picture. The 3 monoamines seemed involved in sleep disorders. Perturbations of norepinephrine and serotonin metabolism could intervene in suicidal ideation and behaviour. Motor activity was associated with a modification in dopamine and serotonin metabolism. Moreover significant correlations were observed between items referring to thought content and monoamine plasma catabolites such as MHPG and blunted affect, 5-HIAA and obsessions, HVA and guilt feelings, devalorization and without hope items.(ABSTRACT TRUNCATED AT 250 WORDS)


In vivo effects of OK-432, inosine pranobex and its salt components on macrophage subpopulations in spleen and peritoneum of mice during the primary humoral immune response.

Subsets of macrophages (BM 8-, la- and esterase-positive subtypes) in the spleen and the peritoneum of mice were affected differently by immunomodulators during the humoral immune response. I.p. application of inosine pranobex (INPX), as well as of its salt moiety (DIPPACBA) and its dimethylaminopropanol (DIP) and acetamidobenzoic acid (PACBA) components, increased the number of nonspecific esterase-bearing cells in the peritoneum on day 3. INPX and DIPPACBA stimulated the la+ macrophage on day 3, DIPPACBA the BM 8+ macrophage only on day 1 and DIP on day 5. In spleen, DIP and PACBA had marked effects on the la+ subset in addition to a marginal effect on the BM 8+ phenotype on day 1. DIPPACBA also influenced the BM 8+ macrophage slightly on day 1. In contrast, the microbial product OK-432 stimulated BM 8+ and la+ macrophages in spleen markedly on day 1, but only marginally on days 3 and 5. However, it exerted a strong effect on both subtypes in peritoneum on days 3 and 5. OK-432 was found to be without any influence on esterase-bearing macrophages. The results show that the heterogeneity of macrophages is not only represented by subset markers, but also by their susceptibility to immunomodulators in different organs and stages of the immune response.


Analysis of gastrinomas by immunohistochemistry and in situ hybridization histochemistry.

Gastrinomas from 25 patients were examined by immunohistochemistry (IHC) and in situ hybridization histochemistry (ISH). Most patients (84%) presented with the Zollinger-Ellison syndrome. Six had multiple endocrine neoplasia type I (MEN-I). Twelve patients (48%) had duodenal primaries and 11 of 12 of these had metastases to regional lymph nodes and/or liver in spite of the small sizes of the primary tumors (mean size of 0.9 cm). Five patients had pancreatic gastrinomas and eight patients had metastatic tumor in regional lymph nodes or liver at surgery but a primary was not found. IHC and ISH analyses showed that all cases were positive for gastrin protein and 24 of 25 (96%) expressed gastrin mRNA that was easily detected in formalin-fixed, paraffin-embedded tissue sections. Both benign and malignant tumors expressed alpha subunit of human chorionic gonadotropin protein (alpha-HCG). However, only malignant gastrinomas (29%) expressed adrenocorticotropic hormone protein or proopiomelanocortin (POMC) mRNA. ISH and Northern hybridization analysis revealed that chromogranin A mRNA was the most common member of the chromogranin/secretogranin (Cg/Sg) family which was expressed in both benign and malignant gastrinomas. These results indicate that duodenal gastrinomas are common in both sporadic and MEN-1-associated cases, and small duodenal primaries may be associated with extensive regional lymph node and liver metastases. Expression of ACTH/POMC protein and mRNA was consistently associated only with malignant gastrinomas while gastrin protein, gastrin mRNA and Cgs/Sgs mRNAs were readily detected in both benign and malignant gastrinomas.


Food allergy as a cause of rhinitis and/or asthma.

We studied the histories of patients who had visited our Department during the last 5 years, presenting food allergy with exclusively respiratory symptoms (18.5%). We studied the correlation between IgE levels and the histamine release test and the type of food implicated. Seventy-seven percent of patients presented sensitization to one food, and 23% presented polysensitization. We found that foods such as snails and eggs have a strong predilection for the bronchial tree as the shock organ. High total IgE was found in 85.2% of patients. The histamine release test was positive in 66% of cases. We conclude that this is a highly sensitive method for the in vitro diagnosis of food allergy.


Localization of Mycobacterium avium-intracellulare within a skin lesion of bacillary angiomatosis in a patient with AIDS.

We report a 39-year-old man who had AIDS and who presented with an unusual cutaneous vascular lesion, which was clinically thought to be Kaposi's sarcoma. Histologically, the lesion was characterized by capillary proliferation and a mixed inflammatory infiltrate that included numerous histiocytes. The lesion was found to contain slender intracellular acid-fast bacilli, as well as plump extracellular Warthin-Starry-positive bacilli. The acid-fast bacilli were confirmed to be Mycobacterium avium-intracellulare by subsequent positive blood cultures for this organism. To further investigate the lesion, polymerase chain reaction DNA amplification and sequencing was performed, and the lesion was found to contain DNA sequences identical to those previously established for the agent of bacillary angiomatosis. The lesion is thought to represent a lesion of bacillary angiomatosis with secondary involvement by M. avium-intracellulare.


[Ventricular extrasystole and coronary disease: to treat or not to treat?].

The prognostic significance of ventricular premature beats (VPB) and their proper management are still a subject of controversy, particularly since during the last few years a number of publications have raised doubts as to the effectiveness and safety of antiarrhythmic drugs. An appropriate assessment of the VPB must consider: 1) electrocardiographic characteristics. 2) Associated symptoms. 3) Presence or absence of underlying disease, and 4) Ventricular function (VF). On this basis VPB may be classified as "Benign" (isolated; asymptomatic; mild or absent cardiopathy, and preserved VF); "Hazardous" (presence of complex VPB; with or without symptoms, and recognized organic heart disease), and "Malignant" (complex VPB and ventricular tachyarrhythmias; symptomatic cardiac disease, and compromised VF). Antiarrhythmic drugs are indicated for "Hazardous" and "Malignant" VPB. However, selection and efficacy must be documented by objective testing (Holter, stress, plasma levels). When utilized in well defined groups of patients, with reduction in the hourly number of VPB (more than 50-70%) and mainly of complex forms, there is clear benefit in terms of long-term survival, including coronary disease and congestive heart failure.


Furosemide inhibits in vitro histamine release induced by antigens and anti-IgE.

We used 56 samples of heparinized blood of patients with asthma and/or allergic rhinitis, which were stimulated with Dermatophagoides pteronyssinus or gramineous pollen and anti-IgE in order to study the effect of furosemide on histamine release in basophils. At the same time, blood samples incubated in furosemide in different concentrations (2.5 microM to 1.49 mM) were also subjected to the same stimuli. Results showed a linear and dose-dependent inhibition of histamine release for antigens (r = 0.96; p < 0.01) and anti-IgE (r = 0.95; p < 0.05), reaching 95% at maximum inhibition. These results support the hypothesis that furosemide may act on mastocytes and basophils, thus inhibiting the release of histamine and other mediators.


Rat stem-cell factor induces splenocytes capable of regenerating the thymus.

Cytokine regulation of prethymic T-lymphoid progenitor-cell proliferation and/or differentiation has not been well-defined, although much is known of cytokine regulation of hemopoietic stem- and progenitor-cell development. Here we use a recently identified hemopoietic growth factor, stem-cell factor (SCF) (a form of the c-kit ligand), and a transplant model of thymocyte regeneration to assess the effect of SCF on the in vivo generation of prethymic, thymocyte progenitor-cell activity. We show that recombinant rat SCF (rrSCF164) administered to weanling rats selectively induces an increase in thymocyte progenitor activity in the spleens of treated rats as compared to rats treated with vehicle, polyethylene glycol (PEG)-conjugated rat albumin, or recombinant human granulocyte colony-stimulating factor (rhG-CSF). These data demonstrate that administration of SCF in vivo affects extrathymic-origin thymocyte regenerating cells and may influence, directly or indirectly, early prethymic stages of T-cell lymphopoiesis in addition to its known effect on early stages of myelopoiesis and erythropoiesis.


Studies on the extraction of DesPro(2)-Val15-Leu17-aprotinin from the culture broth of a recombinant Saccharomyces cerevisiae.

The application of an aqueous two-phase system is described for the extraction of DesPro(2)-Val15-Leu17-aprotinin from yeast culture supernatant, using native chymotrypsin as affinity ligand. The interaction is driven by hydrophobic forces and leads to the accumulation of the aprotinin-chymotrypsin complex in the salt-rich (bottom) phase of a polyethyleglycol/salt system. The complex may be dissociated at low pH values. The separation of the recombinant aprotinin and the protease required chromatographic processes, which proved difficult to interface with the affinity extraction.


Autoimmunity.

The immune system has evolved to protect an organism from the pathogens that invade it but the effector mechanisms involved in mediating this protection are potentially lethal to the host itself. Consequently it is essential that they are not elicited by the host's own tissues and, because biochemically self and non-self are very similar, the immune system has had to develop an exquisite capability to distinguish relatively minor differences. There has been considerable progress recently in understanding how this discrimination is achieved although many questions remain. The problem is important in that the mechanisms that ensure self tolerance occasionally fail. The consequences of this failure are the autoimmune diseases, many of which afflict Man. This article reviews what is known about the way that the immune system normally avoids self reactivity and how breakdown in self tolerance can occur.


[Urodynamic study of obstructive uropathy due to benign prostatic hyperplasia].

Urodynamic examinations in patients with obstructive uropathy due to prostatic adenoma before and after prostatectomy were done over a 6-year-period (1981-1988) at the Department of Urology, Pula Medical Centre. In a comparative group of 30 healthy examinees and in 125 patients with obstructive uropathy as a result of benign prostatic hypertrophy, a maximal uroflowmetry and flow volume measurement were also performed. Control urodynamic studies were done 12 weeks after prostatectomy in 112 patients, and in 102 patients after 12 months. Preoperative and postoperative mean values of maximal uroflow were comparable. The aim of these urodynamic investigations is to estimate their value for a more complete and precise diagnosis of prostatism, and to determine, by urodynamic flow measurement before and after surgery, an outcome of prostatectomy in removal of prostatism symptoms and for urine flow improvement. Eighty-six patients underwent then open surgery, and 38 were operated transurethrally. The benign hypertrophy diagnosis was confirmed by histological analysis in all patients.


The effect of graded single oral doses of prostacyclin on the nucleic acid content of rat gastric mucosa.

Graded single oral doses of prostacyclin significantly enhance in dose-dependent manner the DNA and RNA content of rat gastric fundic mucosa, - resulting in a decreased RNA/DNA ratio. This latter phenomenon is convincing sign of new cell formation. In the antral region no significant changes could be encountered but the RNA content showed a tendency-like elevation, evoking an elevated RNA/DNA ratio, which corresponds to de novo protein synthesis. The diversity of the noted changes gives further evidence to the tenet the antral and the fundic gastric mucosa of the rat reacts differently toward external stimuli.


[The dynamics of the reparative regeneration of the rat epidermis exposed to lysylproline].

Under study was the spatial distribution of mitotic activity (MA) of the rat epidermis in the process of reparative regeneration of the skin for a distance of 6 mm aside from the wound margin, as well as the action of lysyl-proline (LP) on the process which is a product of natural decay of collagen. The wound was made by cutting a skin flap (0.5 x 3 cm) from the area of the back. A teflon ring was introduced through the incision in the cranial direction at a distance of 3 cm. Control animals were given 1 ml of the isotonic solution of sodium chloride injected in the hole of the ring, for experimental animals the solution additionally contained 20 mkg of LP. Healing of the wound in control and experimental animals had the features generally corresponding to the regularities established earlier. It was also found that mitotic activity was growing irregularly forming waves. During 11 days of observations these waves sometimes came nearer to the center of the wound, sometimes moved away. The influence of LP manifested itself in the waves of MA reaching the wound center 1 day earlier, and the approximation and divergence of the waves were smoother, MA was higher and epithelialization of the wound surface also came 1 day earlier. Since the contraction of the wound in experimental animals was also more rapid (activization from the 3rd day), a conclusion can be made of the stimulating influence of LP on the course of the reparative regeneration of the epidermis observed as long as 1.5 weeks after a single injection of LP.(ABSTRACT TRUNCATED AT 250 WORDS)


[The intermediate nerve system].

A comparative-embryological study (223 fetuses of man, cat, dog, albino rat and mole) has established a similarity in the development and structure of the intermediate nerve in mammalia. The nerve under study represents a complex structural-functional system. Its afferent component includes a geniculate ganglion, a nucleus of the solitary tract, root, peripheral branches. A superior salivary nucleus, a root, pterygopalatine, submandibular, sublingual and auricular (in man) parasympathetic ganglia of the head, peripheral branches. The intermediate nerve fibers are included as additional sources in branches of other cranial nerves, vegetative ganglia and plexuses being spread on a considerable territory of the head. The intermediate nerve is an isolated anatomical structure and deserves being detached into a separate pair of cranial nerves.


[The morphofunctional characteristics of the vascular bed of the lymph nodes].

By means of silver nitrate impregnation and ATPase methods the distribution of all links of the lymph node vascular bed has been investigated. Structure of the blood capillary wall has been studied electron microscopically; they are defined as capillaries of the I type, that is having a continuous endothelial and basal layers and a small diameter. An essential amount of ribosomes is specific for blood capillary endothelium in the lymph nodes. A dendrite-like ramification of the portal veins is described, dichotomic--for the cerebral arteries, as well as the distribution of cortical arteries and arterioles. The postcapillary venules are found to localize around the lymphoid noduli and in the paracortical zone. Morphological bases for better exchange between blood and lymph are demonstrated in the cortex of the lymph node. Morphological signs of littoral cells of the lymphatic sinuses have been investigated and a conclusion is made on similarity in the structure of the lymphatic sinus wall and the wall of the lymphatic capillaries. Tight structural interrelations have been stated to exist between the wall of the lymph node blood vessels, its stroma and the wall of the lymphatic sinuses.


[The topical organization of the nuclear projections of the amygdaloid body, the ventral tegmental field and the substantia nigra into the striopallidum of the cat].

The work has generalized and compared results of studying the topical organization of projections of the amygdaloid body, ventral tegmental area and substantia nigra in the extrapyramidal system of the cat's brain. Methods of retrograde axon transport of horseradish peroxidase and fluochrome were used. In the basal ganglia the limbic and motor areas were found with prevailing projections from structures belonging to the limbic system or from the formations bearing a relation to motor activity. Parallel with this, in considerable areas of the basal ganglia there occur overlapping projection fields of the axons belonging to neurons of the both structures. This fact as well as the widely represented in the striopallidal system divergence of axons of neurons belonging to many structures are discussed in association with the hypothesis of the interaction of the limbic and motor systems in the basal ganglia. General features found in the topical organization of projections from the structures under investigation onto the striopallidum nuclei and specific features for each of the nuclei are discussed in connection with similar and different manifestations of behavioral reactions or clinical disturbances in neurological diseases.


Platelet-derived growth factor gene expression in the kidney of malignant hypertension.

To examine the pathogenetic role of platelet-derived growth factor (PDGF) in hypertensive kidney damage, we studied the gene expression of PDGF A-chain and B-chain in an animal model of malignant hypertension. Experimental malignant hypertension induced by unilateral nephrectomy combined with deoxycorticosterone and salt loading in the spontaneously hypertensive rat resulted in severely elevated blood pressure and renal histological damage, characterized by necrotizing vasculitis. Using reverse transcription-polymerase chain reaction analysis followed by Southern blot analysis, we observed that PDGF B-chain gene expression was increased in the kidney of experimental malignant hypertension and was correlated with the severity of glomerular damage, while PDGF A-chain gene expression was unaffected. Antihypertensive treatment with manidipine reduced glomerular damage and a decreased gene expression of PDGF B-chain. These results suggest that PDGF B-chain may have a role in mediating hypertensive kidney damage.


Microcalorimetric evaluation of the effects of methotrexate and 6-thioguanine on sensitive T-lymphoma cells and on a methotrexate-resistant subline.

Isothermal microcalorimetry was used in order to continuously monitor and quantitatively assess the action of two antineoplastic drugs, methotrexate (MTX) and 6-thioguanine (6-TG), on a human T-lymphoma cell line, CCRF-CEM. The results with MTX were compared with data from experiments with a MTX-resistant subline, CEM/MTX. The slope of the power-time curve after drug injection relative to that obtained during unperturbed growth, was used to construct dose-response curves. The normal cell line was characterized by a D50 value (i.e., the dose producing half the maximal response) of 0.05 microM for MTX and 0.38 microM for 6-TG. For the MTX-resistant subline the D50 value was 8 microM of MTX. Comparisons of the continuous power-time curves showed the inhibitory effect of 6-TG to be faster than that of MTX.


The lateral separation of contacts on erythrocytes agglutinated by polylysine.

The form of contact seam (whether a continuous parallel seam or membranes in spatially periodic contact) has been characterized for normal and for neuraminidase pretreated human erythrocytes following adhesion in solutions of polylysine in the molecular mass range 10-225 kDa at concentrations from 0.5 to 1.0 mg/mL. The adhesion contact seam was spatially periodic for all normal control cells in polylysine. The lateral separation of contacts decreased from 1.6 to 0.8 microns as the concentration of 225 kDa polylysine was increased threefold from the adhesion threshold value. The separation distance did not change further even at high polymer concentrations that increased the electrophoretic velocity to positive values over twice the modulus of the velocity of control cells. The probability of cell adhesion decreased at these high polymer concentrations. The lateral contact separation increased and cell adhesion decreased for cells pretreated with neuraminidase. Cell adhesion did not occur when neuraminidase reduced the cell electrophoretic velocity modulus by 30%. Following neuraminidase pretreatments that allowed a small amount of adhesion, the cell contact seam was continuous rather than spatially periodic. The results show that a procedure that increases (e.g., polymer concentration increase) or decreases (e.g., enzyme removal of polycation crosslinking site) attraction leads to shorter (to a limiting value) or longer lateral contact separation, respectively.


Influence of glutaraldehyde fixation of cells adherent to solid substrata on their detachment during exposure to shear stress.

In order to determine the response of fixed and nonfixed cells adherent to a solid substratum to shear stress, human fibroblasts were allowed to adhere and spread on either hydrophilic glass or hydrophobic Fluoroethylene-propylene (FEP-Teflon) and fixed with glutaraldehyde. Then, the cells were exposed to an incrementally loaded shear stress in a parallel plate flow chamber up to shear stresses of about 500 dynes/cm2, followed by exposure to a liquid-air interface passage. The cellular detachment was compared with the one of nonfixed cells. In case of fixed cells, 50% of the adhering cells detached from FEP-Teflon at a shear stress of 350 dynes/cm2, whereas 50% of the adhering, nonfixed cells detached already at a shear stress of 20 dynes/cm2. No fixed cells detached from glass for shear stresses up to at least 500 dynes/cm2. More than 50% of the nonfixed cells were detached from glass at a shear stress of 350 dynes/cm2. Furthermore, the shape and morphology of fixed cells did not change during the incrementally loaded flow, in contrast to the ones of nonfixed cells, which clearly rounded up prior to detachment.


Kinetics of interleukin-2 induced changes in rigidity of human natural killer cells.

The success of adoptive immunotherapy is dependent in part on the successful delivery of effector cells to the tumor and the expression of cellular activities, such as adhesion, extravasation, and cytotoxic activity of the effector cells in the tumor. The structural rigidity of the effector cell is an important determinant of these functions. The present study was designed to quantify the changes in cellular rigidity and cytotoxic activity of human natural killer (NK) cells in the presence or absence of interleukin-2 (IL-2). Micropipet aspiration was used to measure the resistance of NK cells to an imposed external deformation. Homogeneous suspensions of NK cells were activated with 1000 U/mL of recombinant IL-2 in vitro and tested for cellular rigidity from 0 to 96 h post stimulation. The IL-2 activated cells increased their rigidity within 24 h and maintained it at this level for 96 h. Prolonged incubation of cells in IL-2 (14 d) resulted in a consistently high rigidity, which was further increased on starvation of the cells from IL-2. The increased rigidity of these cells was maintained throughout 96 h of IL-2 deprivation, although significant relaxation of rigidity was observed between 48 and 96 h. The relaxation of rigidity was associated with an increase in the number of nonviable cells. Reintroduction of IL-2 for 24 h to a culture of NK cells depleted of IL-2 for 48 h did not restore the cells to the pre-depletion level of rigidity. Cytotoxic activity of the activated NK cells following removal of IL-2 decreased to about 60% of the control activity within 24 h and continued through 72 h post-deprivation. These findings suggest that the initial activation of human NK cells by IL-2 will produce a relatively rapid increase in rigidity that may cause entrapment of these cells in small capillaries in vivo and that removal of IL-2 will produce an additional increase in rigidity, which is associated with decreased functional activity.


Inhibitors of HIV-1 reverse transcriptase and fidelity of in vitro DNA replication.

Mechanisms of the effects of the dTTP analogues 3'-azido-3'-deoxythymidine 5'-triphosphate (AZTTP) and 3'-amino-3'-deoxythymidine 5'-triphosphate (NH2 TTP) upon the HIV-1 reverse transcriptase (RT) are discussed. These compounds block the RT in vitro and do so by different kinetic mechanisms. Infidelity of replication is a hallmark of the HIV-1 RT, and replication errors by the enzyme on RNA and DNA templates are discussed. The enzyme's infidelity has ramifications for inhibition: On the one hand, the propensity to produce mutations enhances the ability of the virus to escape inhibitors whereas on the other hand, the infidelity of the reverse transcriptase may allow the development of imaginative inhibitor strategies.


Morphology of cell-substratum adhesion. Influence of receptor heterogeneity and nonspecific forces.

Many cell types modulate growth, differentiation, and motility through changes in cell substrate adhesion, including regulation of focal contact formation. Clustering of cell surface adhesion receptors is an essential early step in the development of focal contacts, and thus may influence cell physiology. In this paper, we present a theoretical framework to examine how cell surface chemistry affects receptor clustering. Our one-dimensional tape-peeling model couples the equations of mechanical equilibrium for a cell membrane with kinetic receptor-ligand binding relations. We considered two distinct model scenarios: Adhesion mediated by multiple receptor-ligand interactions of different length and specific binding of a single receptor type occurs in the presence of van der Waals attraction and nonspecific repulsion. In each case, nonuniform (wave-like) membrane morphologies are observed in certain parameter ranges that support the clustering of adhesion receptors. The formation of these morphologies is described in terms of a balance of membrane stresses; when cell-surface potential as a function of separation distance is symmetric between two potential energy minima, nonuniform morphologies are obtained. Increases in the chemical binding energy between receptor and ligand (e.g., increases in ligand density) or decreases in the membrane rigidity result in smaller wavelengths for nonuniform interfaces. Additionally, we show wave-like geometries appear only when the mechanical compliance of receptor-ligand bonds is within an intermediate range, and examine how the mobility of "repellers"--glycocalyx molecules that exert a nonspecific repulsive force--influences membrane morphology. We find fully mobile repellers always redistribute to prevent nonuniform morphologies.


Inhibitors of HIV-1 protease.

The human immunodeficiency virus (HIV), the etiological agent for the acquired immune deficiency syndrome (AIDS), is a retrovirus which makes use of a virally-encoded aspartic protease to perform specific proteolytic processing of two of its gene products in order to form active enzymes and structural proteins within the mature virion. Accordingly, specific, exogenous inhibition of the HIV-1 protease is thought to be a viable approach for the development of novel therapeutics for the treatment of AIDS. Indeed, this hypothesis has been validated in virally-infected cell culture with synthetic inhibitors of HIV-1 protease. This chapter reviews the current status of the development of inhibitors of this enzyme.


Correlations between the in vitro and in vivo activity of anti-HIV agents: implications for future drug development.

Some 10 years after the first recognition of acquired immunodeficiency syndrome (AIDS) as a new syndrome, we have identified a number of molecular targets to interrupt the replicative cycle of human immunodeficiency virus (HIV), the causative agent. A number of dideoxynucleosides have been identified as having anti-HIV activity in vitro, and several of these have been found to have clinical activity in patients. In contrast, while a number of agents have been found to block viral binding to the target cell in vitro, these agents have generally not shown clear-cut evidence of clinical activity. Agents which act at a variety of steps in the HIV replicative cycle are now under development, and it is likely that we will have an increased armamentarium to fight this disease in the near future.


[Dissection of the ascending aorta (type A): diagnostic aspects, surgical treatment and long-term follow-up].

Aortic replacement is the treatment of choice and improves the natural history of dissections involving the ascending aorta. Forty patients (23 male), aged 49.4 years, have been operated at the hospital Clínico de la Universidad Católica. Twenty six presented with acute dissections. Angiography conformed the dissection in 63.3% and computed axial tomography in 84.6% of patients; lately, transesophageal echocardiography has become the most sensitive diagnostic method. Twenty three patients (57.5%) were subjected to emergency operations and 17 to semielective procedures. In 24 patients (60%) ascending aorta was replaced and in 16 a composite graft was used. Operative mortality was 27.5%. Univariate analysis showed that the period in which the operation was performed and the presence of limb ischemia were the only independent predictors of operative mortality. Long term follow up was achieved in 26 patients (89.6%). Actuarial 5 year survival without considering operative mortality was 87.9%. It is concluded that patients with acute dissections involving the ascending aorta should be operated as soon as the imaging diagnosis is complete and, since this is a palliative procedure, a close follow up is required for early detection of complications.


CD4 subsets in IgA nephropathy.

CD45RA+ (2H4+) and CD45RA-(CD29, 4B4+) antigenic expressions, recognized as a suppressor inducer and a helper inducer, respectively, on CD4+ and T alpha 4+ cells (Fc alpha receptor bearing CD4 T cells) were measured by three color flow cytometry in patients with IgA nephropathy (IgAN). The 4B4+ subsets of CD4 and T alpha 4 T cells were increased significantly in the patient group compared with the control group. The level of the 2H4+ subset did not differ between the two groups. No significant correlation was observed between the increase in the 4B4+ subset and disease severity, as estimated by measuring the proteinuria, levels of serum creatinine and immunoglobulins (Igs) or renal tissue damage. It was concluded that 4B4+ CD4+ and 4B4+ T alpha 4+ subsets might be involved in the mechanism of immunopathogenesis of IgA nephropathy.


Immunohistochemical analysis of GH-producing adenomas--with special emphasis on plurihormonality of individual tumor cells by double staining.

Forty-four human growth hormone (GH)-producing adenomas were investigated immunohistochemically for the concomitant localization of GH, prolactin (PRL), and glycoprotein hormone alpha subunit. Immunoreactivity for GH, PRL, and the alpha subunit was found in 44, 37, and 36 cases, respectively. By double immunohistochemical staining, 24 of 27 cases showed colocalization of GH and alpha subunit, with the numbers of tumor cells showing double staining varying from case to case. The colocalization of GH and alpha subunit was noted in some normal pituitary cells. In adenoma cells, the colocalization of GH and PRL, and PRL and alpha subunit, was observed in 9 and 12 cases, respectively. The normal pituitary gland showed only occasional colocalization of GH and PRL, or PRL and alpha subunit. We found that GH-producing adenomas are plurihormonal at the individual tumor cell level, with coexpression of GH-alpha subunit, GH-PRL, and PRL-alpha subunit. The colocalization of GH and alpha subunit may be an expression of a subpopulation of normal anterior pituitary cells (with GH-alpha subunit coexpression), but the more frequent coexpression of GH-PRL may be pathological, accompanying tumorigenesis of the anterior pituitary cells, in which a pituitary-specific transcriptional factor, pit-1, may play a role.


Calcium permeability and modulation of nicotinic acetylcholine receptor-channels in rat parasympathetic neurons.

Neuronal nicotinic acetylcholine (ACh)-activated currents in rat parasympathetic ganglion cells were examined using whole-cell and single-channel patch clamp recording techniques. The whole-cell current-voltage (I-V) relationship exhibited strong inward rectification and a reversal (zero current) potential of -3.9 mV in nearly symmetrical Na+ solutions (external 140 mM Na+/internal 160 mM Na+). Isosmotic replacement of extracellular Na+ with either Ca2+ or Mg2+ yielded the permeability (Px/PNa) sequence Mg2+ (1.1) > Na+ (1.0) > Ca2+ (0.65). Whole-cell ACh-induced current amplitude decreased as [Ca2+]0 was raised from 2.5 mM to 20 mM, and remained constant at higher [Ca2+]0. Unitary ACh-activated currents recorded in excised outside-out patches had conductances ranging from 15-35 pS with at least three distinct conductance levels (33 pS, 26 pS, 19 pS) observed in most patches. The neuronal nicotinic ACh receptor-channel had a slope conductance of 30 pS in Na+ external solution, which decreased to 20 pS in isotonic Ca2+ and was unchanged by isosmotic replacement of Na+ with Mg2+. ACh-activated single channel currents had an apparent mean open time (tau 0) of 1.15 +/- 0.16 ms and a mean burst length (tau b) of 6.83 +/- 1.76 ms at -60 mV in Na+ external solution. Ca(2+)-free external solutions, or raising [Ca2+]0 to 50-100 mM decreased both the tau 0 and tau b of the nAChR channel. Varying [Ca2+]0 produced a marked decrease in NP0, while substitution of Mg2+ for Na+ increased NP0. These data suggest that activation of the neuronal nAChR channel permits a substantial Ca2+ influx which may modulate Ca(2+)-dependent ion channels and second messenger pathways to affect neuronal excitability in parasympathetic ganglia.


[Kurt Goldstein: from aphasia to linguistics].

The authors try to take into account Kurt Goldstein's contribution on the subject of aphasia. They introduce the significance of the phenomenological approach of the language leading them toward the structuralist theme.


Plasmodium falciparum CS protein--prime malaria vaccine candidate: definition of the human CTL domain and analysis of its variation.

Studies in mice have shown that immunity to malaria sporozoites is mediated primarily by cytotoxic T lymphocytes (CTL) specific for epitopes within the circumsporozoite (CS) protein. Humans, however, had never been shown to generate CTL against any malaria or other parasite protein. The design of a sub-unit vaccine for humans relies on the epitopes recognized by CTL being identified and polymorphisms therein being defined. We have developed a novel technique using an entire series of overlapping synthetic peptides to define the epitopes of the Plasmodium falciparum CS protein recognized by human CTL and have analyzed the sequence variation of the protein with respect to the identified CTL epitopic domain. We have demonstrated that some humans can indeed generate CTL against the P. falciparum CS protein. Furthermore, the extent of variation observed for the CTL recognition domain is finite and the combination of peptides necessary for inclusion in a polyvalent vaccine may be small. If ways can be found to increase immune responsiveness, then a vaccine designed to stimulate CS protein-specific CTL activity may prevent malaria.


Molecular mechanisms and biological importance of Plasmodium falciparum erythrocyte rosetting.

Rosetting, i.e. the spontaneous binding of uninfected to malaria infected erythrocytes and endothelial cytoadherence may hinder the blood flow and lead to severe Plasmodium falciparum malaria. Falciparum isolates obtained from unconscious patients all form rosettes and/or express a significantly higher mean rosetting rate than isolates from patients with uncomplicated malaria. Furthermore, sera of patients with cerebral malaria are devoid of anti-rosetting activity while sera from patients with mild disease carry high levels of anti-rosetting antibodies. The presence of anti-rosetting antibodies also seems important for the efficient interaction of rosetting infected rbc and leukocytes. Two parasite derived rosetting ligands of Mr 22K and Mr 28K named "rosettins", have been found on the surface of rosetting infected erythrocytes. CD36 has in at least some strains of parasites been found to function as a rosetting receptor on the uninfected erythrocyte. Heparin disrupts rosettes of P. falciparum in vitro and inhibits the sequestration of rosetting cells ex vivo. In conclusion, rosetting seems a crucial factor in the development of cerebral malaria and treatment of patients with anti-rosetting substances might become an effective adjunct in the treatment of severe malaria.


Epitope recognition in histone H1 by SLE autoantibodies in the presence of a DNA-ligand.

To investigate the specificity of anti H1 antibodies peptides from the N- and C-domain of H1 and the synthetic oligonucleotide (AT)6 were complexed. Circular dichroism (CD) spectroscopy indicated that the free peptides H1(1-16), H1(204-218) and C(121-210) in low salt buffer assume a random structure but become helical when bound to the oligonucleotide. The structured and unstructured H1 fragments were then analyzed by enzyme linked immunosorbent assay (ELISA) with anti-H1 antibodies in sera from patients with systemic lupus erythematosis (SLE) and with the monoclonal anti-H1 antibody MRA-12 derived from MLR lpr/lpr autoimmune mice. Binding of these antibodies to H1(204-218) and C was inhibited to a level of 50% when these H1 peptides were complexed with (AT)6. When the same antibody was tested with H1 fragment GC(34-210), attachment to oligonucleotide (AT)6 did not influence antibody binding. Competition studies with liquid phase GC and C antigen against solid phase GC and C indicated that liquid phase GC was more efficient in displacing antibody binding reactivity than liquid phase C. The displacement effect of both liquid phase antigens was greatest against solid phase C. We conclude that anti-H1 autoantibodies are directed against an epitope located near the junction of the G- and C-domain which is exposed and not masked when H1 is bound to DNA.


Levels of CD5+ B cells are not increased in probands or relatives in a family study of primary Sjögren's syndrome.

Levels of CD5+ B lymphocytes were assayed in a large family study of Primary Sjögren's syndrome. There was no significant difference in CD5 expression by index cases or their relatives when compared to controls. No association between CD5 expression, serological abnormalities or HLA haplotype was found and, furthermore, no evidence of linkage with HLA was observed. There was, however, variation in the expression of CD5+ B cells between the families. Levels in spouses were lower and reached statistical significance. The role for genetic and environmental factors influencing CD5 expression is discussed. Any genetic influence does not appear to involve the HLA region or genes in linkage disequilibrium.


Analysis of hemoglobin gamma chain in patients with Cooley's anemia in Taiwan.

The beta-like globin gene cluster, consisting of five genes and a pseudogene, is located on chromosome 11 and is arranged in the order 5'-epsilon-G gamma-A gamma-phi beta-delta-beta-3'. With the fast and sensitive method of high-performance liquid chromatography (HPLC) in the gradient mobile system, we studied the G gamma values of fetal hemoglobin in 34 patients with Cooley's anemia, 100 normal adults and 100 newborns. There were 84 newborns with a mean G gamma value of 67.41 +/- 2.37%, 15 newborns with a G gamma value of 83.45 +/- 1.92% and one newborn with a G gamma value of 100%. Because the medium value of 67.41% and 100% is near the value of 83.45%, we speculate that there are different g gene genotypes; that is, the group with a G gamma value of 100% has a genotype of G gamma-G gamma/G gamma-G gamma, the group with a G gamma value of 83.45% has a genotype of G gamma-G gamma/G gamma-A gamma, and the group with a G gamma value of 67.41% has a genotype of G gamma-A gamma/G gamma-A gamma. The G gamma value for normal adults was 41.84 +/- 20.91%. While the G gamma values for patients with Cooley's anemia were divided into two groups, 56.23 +/- 6.55% and 80.60 +/- 3.05%. There was no case having a G gamma value of 100%. Our results suggest that the expression of the gamma gene in Cooley's anemia is similar to that of the newborns.


Effect of dexamethasone, dimethylsulfoxide and sodium butyrate on a human hepatoma cell line PLC/PRF/5.

The effects of agents which are known to be differentiation inducers on a human hepatoma cell line PLC/PRF/5 were investigated. Dexamethasone (DEX), sodium butyrate (SB) or dimethylsulfoxide (DMSO) were examined. They all reduced cell proliferation but differ from each other in effect on the secretion of alphafetoprotein (AFP) and hepatitis B surface antigen (HBsAg), changes in morphology and RNA transcription. SB changed the cell from polygonal into a fibroblast-like type and decreased AFP secretion. DMSO decreased the cell size and changed AFP secretion in the same manner as SB. DEX changed the cell into a larger size, as well as increased AFP secretion. HBsAg secretion and also HB virus DNA transcription was enhanced by 3 agents. AFP and myc gene transcriptions were reduced by SB but DMSO reduced only AFP. Albumin gene transcription was enhanced by SB and DEX. These results indicate that the decrease of PLC/PRF/5 proliferation is induced through different mechanisms by these 3 agents.


Expression of a Chlamydia anticarbohydrate single-chain antibody as a maltose binding fusion protein.

A single-chain antibody fragment has been constructed for an antibody that binds to the Chlamydia specific carbohydrate structure of the lipopolysaccharide. Single-chain protein was expressed and secreted into the periplasmic space of E. coli as a fusion protein with the maltose binding protein. The fusion protein was purified in one step by virtue of its ability to bind to maltose. In a sandwich ELISA, the eluted protein bound Chlamydia lipopolysaccharide, which demonstrates that the single-chain protein domain will function as part of a fusion protein. The expression of maltose binding fusion proteins into the periplasmic space could be used for production of other single-chain antibodies or protein fragments requiring appropriate folding and disulfide bond formation.


Human immune response to monoclonal antibody-enzyme conjugates in ADEPT pilot clinical trial.

The human immune response to monoclonal antibody-enzyme conjugates has been studied in patients included in the pilot clinical trial of ADEPT. Each patient received murine monoclonal anti-CEA antibody fragments (A5B7-F(ab')2, conjugated to bacterial enzyme, carboxypeptidase G2 (CPG2) followed by a galactosylated monoclonal anti-CPG2 antibody (SB43), 36-48 h after the conjugate. Some patients were also given a dose of 131I-labeled conjugate (4-8 mg, 7-15 mCi) for blood clearance and gamma camera image studies. All patients studied developed human antimouse antibodies (HAMA) and anti-CPG2 antibodies within 10 d after a single course of treatment with the conjugate. In most cases, IgM response was detected at 7 d after the conjugate followed by the IgG response 14 d later. In one patient, HAMA and anti-CPG2 antibodies of the IgG type could still be detected at 10 mo after treatment. Anti-CPG2 antibodies in serum of one patient were found to inhibit CPG2 activity in vitro. Generation of neutralizing antibodies limits the use of repeat cycles of ADEPT in patients. Use of immunosuppressive agents may allow a useful time window for several ADEPT cycle treatments by delaying the appearance of HAMA and anti-CPG2 antibodies. Patients given cyclosporin A before and during ADEPT are currently being studied for HAMA and anti-CPG2 response.


Rational immunotherapy with ribonuclease chimeras. An approach toward humanizing immunotoxins.

Members of the pancreatic ribonuclease (RNase) family have diverse activities toward RNA that could cause them to function during host defense and physiological cell death pathways. This activity could be harnessed by coupling RNases to cell binding ligands for the purpose of engineering them into cell-type specific cytotoxins. Therefore, the cytotoxic potential of RNase was explored by linking bovine pancreatic ribonuclease A via a disulfide bond to human transferrin or antibodies to the transferrin receptor. The RNase hybrid proteins were cytotoxic to K562 human erythroleukemia cells in vitro with an IC50 around 10(-7) M, whereas > 10(-4) M of native RNase was required to inhibit protein synthesis. Cytotoxicity required both components of the conjugate since excess transferrin or ribonuclease inhibitors added to the medium protected the cells from the transferrin-RNase toxicity. Importantly, the RNase conjugates were found to have potent antitumor effects in vivo. Chimeric RNase fusion proteins were also developed. F(ab')2-like antibody-enzyme fusions were prepared by linking the gene for human RNase to a chimeric antitransferrin receptor heavy chain gene. The antibody enzyme fusion gene was introduced into a transfectoma that secreted the chimeric light chain of the same antibody, and cell lines were cloned that synthesized and secreted the antibody-enzyme fusion protein of the expected size at a concentration of 1-5 ng/mL. Culture supernatants from clones secreting the fusion protein caused inhibition of growth and protein synthesis toward K562 cells that express the human transferrin receptor but not toward a nonhuman derived cell line. Since human ribonucleases coupled to antibodies also exhibited receptor mediated toxicities, a new approach to selective cell killing is provided. This may allow the development of new therapeutics for cancer treatment that exhibit less systemic toxicity and, importantly, less immunogenicity than the currently employed ligand-toxin conjugates.


Immunotoxins to human small-cell lung cancer.

Ricin A chain ITs directed against a variety of the common cell-surface antigens associated with SCLC exerted selective toxic effects on SCLC cell lines. The potency of the cytotoxic effects matched or exceeded that previously reported for ricin A chain ITs directed against identical or similar antigens on other types of carcinoma, suggesting that SCLC may be uniquely sensitive to this type of IT.


Optimum combination of targeted 131I therapy and total-body irradiation for treatment of disseminated tumors of differing radiosensitivity.

131I is the radionuclide most commonly used in biologically targeted radiotherapy at the present time. Microdosimetric analysis has shown that microtumors whose diameters are less than the beta-particle maximum range absorb radiation energy inefficiently from targeted radionuclides. Micrometastases of diameters < 1 mm are likely to be spared if targeted 131I is used as a single modality. Because of this, combined modality therapy incorporating targeted 131I, external beam total-body irradiation (TBI), and bone marrow rescue has been proposed. In this study, the minimum necessary TBI component is shown to depend on the radiosensitivity of the tumor cells. The analysis shows that the TBI component, to achieve radiocurability, increases directly with tumor radioresistance. For the most radiosensitive tumors, a whole-body TBI treatment dose 2 x 2 Gy is calculated to be obligatory, whereas practical arguments exist in favor of higher doses. For more radioresistant tumors, the analysis implies that a TBI treatment delivery of 5 x 2 Gy is obligatory. In all situations, external beam TBI appears to be an essential factor in providing reasonable probability of cure of disseminated malignant disease. Reasonable prospects of tumor cure by combination strategies incorporating 131I exist for the more radiosensitive tumor types (e.g., neuroblastoma, lymphoma, leukemia, myeloma, seminoma), but more resistant tumors are unlikely to be curable at present. Superior targeting agents, and the possible use of panels of different radionuclides, may be necessary to achieve high cure probabilities for less radiosensitive tumor types.


The clinical significance of HAMA in patients treated with mouse monoclonal antibodies.

Twenty-four patients were analyzed for the development of HAMA (human antimouse antibodies) after being treated with repeated doses (200-500 mg) of the mouse monoclonal antibody (MAb) 17-1A. All patients developed anti-17-1A IgG antibodies, and most of them also developed IgM antibodies. In only two patients could immune complexes be demonstrated. Allergic reactions were rare (1.9%). In an extended study, a further 19 patient were analyzed for an idiotypic response. Forty-one out of 43 patients developed antiidiotypic antibodies (ab2), and 20 of these also anti-anti-idiotypic antibodies (ab3). Ab3+ patients responded significantly better (p = 0.01) and survived longer (p < 0.001) compared to ab3- patients. In this study, we showed that MAb 17-1A could be repeatedly given on a safe basis. The development of high titers of HAMA did not cause significant clinical problems when further repeated infusions of MAb 17-1A were given. The development of an idiotypic response also indicate that the induction of HAMA might be beneficial and not harmful to the patient.


The p53 tumor suppressor gene. A preliminary clinical study in breast cancer patients.

p53 was originally considered to be a nuclear oncogene, but several convergent lines of research have indicated that the wild-type gene functions as a tumor suppressor gene negatively regulating the cell cycle. Mutations in the p53 gene have been detected in many tumor types and seem to be the most common genetic alterations in human cancer. In this preliminary study, sera of 92 patients (pts) with breast disease were analyzed for the presence of the mutant p53 protein (mp53) with a selective immunoenzyme assay employing a monoclonal antibody (PAb 240) specific for the majority of mammalian m p53 but not for the wild-type protein. Of the 10 patients with benign breast disease, only two (20%) showed detectable m p53 levels in the serum. In the breast cancer group, sera from 7 of the 30 pts (23%) without lymph node involvement were positive for m p53, as were 7 out of the 45 pts (15%) with metastatic lymph nodes and 1 out of the 7 pts (14%) with disseminated disease. The specificity of m p53 assay evaluated in 20 healthy controls was 100%. These preliminary results showed that serum positivity for m p53 is not related to breast disease extension. Further studies to assess the utility of m p53 as a possible prognosis factor in breast cancer are currently in progress.


Use of the multipin peptide synthesis technique for the generation of antipeptide sera.

The multipin peptide synthesis technique has been used to map antigenic sites of proteins (1,2). Antibodies raised to the whole protein are screened on pin-synthesized overlapping octapeptides homologous with the protein of interest, and the peptides that bind antibodies clearly identify the epitopes. What is described in this study is a method using pin-synthesized peptides to generate specific antibodies to many peptides. Cleavable linkers have been developed (3) that, used together with the multipin peptide synthesis technique, allow the synthesis and cleavage of many thousands of peptides into aqueous solutions at physiological pH. This technique is useful for assays requiring peptides in solution, e.g., mapping of T-cell determinants. A technique has been developed for the cleavage of many peptides from pins and simultaneous coupling to immunogenic carriers (4). The conjugates produced are suitable for the generation of antipeptide antibodies. This procedure is illustrated using several 15 amino acid long peptides (15-mers), homologous with the sequence of a model antigen, myohemerythrin (MHr). The resulting antipeptide sera generated were tested by ELISA for titer and specificity on pin-synthesized peptides and beta-amide peptides and the protein antigen coated to microtiter plates.


Regulated secretion and purification of recombinant antibodies in E. coli.

A plasmid for optimized protein expression of recombinant Fv antibodies (pOPE) in E. coli was used to express the variable domains of the murine monoclonal antibody HD39 specific for the human B-cell surface antigen CD22. The production of Fv antibodies by pOPE can be regulated over a wide range by varying the IPTG concentration. Antibodies that can discriminate between secreted and nonsecreted Fv antibody fragments were used to show that secretion is the limiting step for the production of functional Fv antibodies. IPTG concentrations above 20 microM increased the total antibody production, but did not yield larger amounts of secreted Fv antibodies. The addition of five histidines to the C terminus facilitates an easy single-step enrichment procedure based on immobilized metal affinity chromatography.


Tegumental proteins of Schistosoma mansoni: complex biomolecules and potent antigens.

The passive transfer of monoclonal antibodies, direct vaccination and in vitro assays have all shown that antigens associated with the tegumental membranes of Schistosoma mansoni are capable of mediating protective immune responses against the parasite in animal models. Furthermore, the principal antigens are highly antigenic during natural infection in man and stimulate strong humoral and cellular responses although, at present, their role in mediating protective immune responses in man remains equivocal. This presentation will review the current state of knowledge of the structure and expression of the major antigenic tegumental proteins of the schistosome and will attempt to relate the relevance of their structural features to possible function both in terms of protective immunity and the parasite's ability to survive within the definitive host. A focus will be the recent advances that have been made in the identification of means of anchoring of the antigenic proteins to the tegumental membrane. In addition, the implications of the structural complexity of the tegumental proteins in terms of their possible utility in vaccination and diagnosis will be considered.


Recombinant human monoclonal antibodies. Basic principles of the immune system transferred to E. coli.

To produce human monoclonal antibodies in bacteria, a gene repertoire of IgM variable regions was isolated from human peripheral B lymphocytes by the polymerase chain reaction. Alternatively, synthetic antibody genes with random hypervariable regions are being generated that may provide libraries of even higher complexity. For the selection of specific monoclonal antibodies from these libraries, we have developed two E. coli vector systems that facilitate the surface display of an antibody physically linked to its own gene. The phagemid pSEX encodes a fusion protein of an antigen binding domain (Fv-antibody) with the docking protein (pIII) of filamentous phages. Specific antibody genes can therefore be enriched by antigen affinity chromatography. The plasmid pAP1 encodes a fusion protein of an Fv-antibody with a bacterial cell-wall protein. Bacteria carrying this plasmid express functional Fv-antibodies tightly bound to their surface. This should enable the selection of single cells with a fluorescence-assisted cell sorter (FACS) using labeled antigen or by adsorption to immobilized antigen. These vectors permit three major principles of the antibody response to be mimicked in E. coli: 1. Generation of a highly complex antibody repertoire; 2. Clonal selection procedures for library screening; and 3. The possibility of increasing a given affinity by repeated rounds of mutation and selection.


Mechanisms of evasion of Schistosoma mansoni schistosomula to the lethal activity of complement.

Schistosomula of Schistosoma mansoni became resistant to antibody-dependent complement damage in vitro after pre-incubation with normal human erythrocytes (NHuE) whatever the ABO or Rh blood group. Resistant parasites were shown to acquire host decay accelerating factor (DAF), a 70 kDa glycoprotein attached to the membrane of NHuE by a GPI anchor. IgG2a mAb anti-human DAF (IA10) immunoprecipitated a 70 kDa molecule from 125I-labeled schistosomula pre-incubated with NHuE and inhibited their resistance to complement-dependent killing in vitro. Incubation of schistosomula with erythrocytes from patients with paroxysmal nocturnal hemoglobinuria (PNHE) or SRBC, which are DAF-deficient, did not protect the parasites from complement lesion. Supernatant of 100,000 x g collected from NHuE incubated for 24 h in defined medium was shown to contain a soluble form of DAF and to protect schistosomula from complement killing. Schistosomula treated with trypsin before incubation with NHuE ghosts did not become resistant to complement damage. On the other hand, pre-treatment with chymotrypsin did not interfere with the acquisition of resistance by the schistosomula. These results indicate that, in vitro, NHuE DAF can be transferred to schistosomula in a soluble form and that the binding of this molecule to the parasite surface is dependent upon trypsin-sensitive chymotrypsin-insensitive polypeptide(s) present on the surface of the worm.


Radiolabeled octreotide. What lessons for antibody-mediated targeting?

Octreotide is a synthetic analog of the peptide hormone somatostatin (SMS). A wide variety of tumors express enhanced numbers of SMS receptors, notably neuroendocrine tumors and lymphomas, but also some of the more common adenocarcinomas. Octreotide contains only eight amino acids, some of which are in the (D) configuration in order to enhance the stability of the molecule in vivo. Tyrosine and DTPA-containing analogs of octreotide have been synthesized and labeled with iodine-123 and indium-111, respectively, with the intention of targeting SMS receptor-containing tumors for diagnostic purposes. Both radiopharmaceuticals demonstrate a high sensitivity and specificity for these tumors, indicating a clinical role for these agents in management of these diseases. Lessons can be learned from the success of these agents when designing improved antibody-based molecules. Tumor uptake of radiolabeled octreotide is very rapid, occurring within minutes of administration. Blood clearance is also rapid, such that tumors are soon visible even in areas of high blood background. An interesting finding has been the differences between the pharmacokinetics of the iodinated and indium-labeled species. Although the majority of 123I-Tyr3-octreotide undergoes hepatobiliary excretion, 111In-DTPAPhe1-octreotide is eliminated predominantly by the kidneys. These results suggest that the smallest possible antibody-like tracers are likely to have advantages over native immunoglobulins and conventional Fab-like fragments.


The influence of cell culture and storage conditions on HIV-1 infectivity and fusogenic activity.

We have previously demonstrated that acidic medium inhibits the replication of HIV-1. The present study was designed to examine the effects of other growth conditions and infection of fibroblasts by coculture with HIV infected lymphoid cells. Several lymphoblastoid cell lines normally grown in RPMI-1640 were grown in Eagle's MEM. These cells supported virus replication to higher titres than did RPMI-1640. Peak viral titres were achieved within 24-48 h after newly infected or chronically infected cells were placed in fresh medium. When virus was stored in liquid medium either frozen or at higher temperatures, virus titres were retained for several months while frozen but decreased upon storage at 4 degrees C or higher. If cells were passaged after trypsinization in Ca(++)-depleted medium, then a decreased susceptibility of cells for HIV-1 by 2 log10 at 24 h post infection was observed. Infectivity of cell-free and cell-associated HIV-1 was measured using syncytium formation, reverse transcriptase activity and p24 antigen. No fusion between HIV-1 infected CD4+ lymphoblasts and CD4- fibroblasts was observed but HIV-1 infected lymphoid cells, even in the absence of syncytium formation, exerted a strong toxic effect on fibroblasts. This study extends previous findings that medium acidity was inhibitory to virus replication and survival. Thus, conditions for study of HIV must be well controlled in buffered medium so that misleading results are not obtained regarding virus multiplication and possibly regarding transmission to and pathogenesis in CD4- cells.


Antigenic relationships among the members of adenovirus subgenera determined by monoclonal antibodies.

Epitope composition of human adenovirus (AV) serotype 13 (subgenus A), 19, 26 and 27 (subgenus D), as well as 41 (subgenus F) was studied with 23 selected monoclonal antibodies (MAbs) raised against different hexon types. With the help of three panels of MAbs great differences were shown in the epitope structure of subgenus D hexons. Comparing two AV13 strain hexons (a prototype strain and an intermediate strain), it was shown that the epitopes recognized by the MAbs were present in different combinations even on strains of the same serotype. Therefore the epitope composition of the subgenus D hexons seems to be heterogeneous.


Endocrine events prior to puberty in heifers: role of somatotropin, insulin-like growth factor-I and insulin-like growth factor binding proteins.

We have utilized active immunization against growth hormone releasing factor (GRF) to investigate relationships among somatotropin (ST), insulin-like growth factor-I (IGF-I), IGF binding proteins (IGFBP) and ovarian function in heifers. Active immunization against GRF (GRFi) has been demonstrated to abolish episodic release of ST and decrease serum concentrations of IGF-I. In initial experiments investigating onset of puberty, breeds of heifers differing in growth rate and reproductive traits (Angus, Charolais and Simmental) were immunized against GRF or served as controls (immunized against carrier protein, human serum albumin, HSAi). GRFi decreased rate of muscle and skeletal growth, but increased deposition of adipose tissue. In Angus and Charolais, but not Simmental heifers, GRFi at 6 mo of age significantly delayed onset of puberty beyond 18 mo of age. Retrospective analyses of serum IGF-I revealed that GRFi heifers reaching puberty at a normal age had greater pre-treatment (6 mo of age) IGF-I than GRFi heifers in which puberty was delayed. Collectively, these results strongly indicate that the bovine hypothalamic-hypophyseal-ovarian axis is particularly sensitive to changes in metabolism at or near 6 mo of age. Another series of experiments tested the hypothesis that lowering serum IGF-I via GRFi initially at 3 mo of age would increase the percentage of Angus and Simmental heifers not reaching puberty. Three mo old Angus and Simmental heifers were assigned to GRFi (n = 18), HSAi (n = 14) or received no treatment (controls, n = 16). HSAi and GRFi heifers were unilaterally ovariectomized (ULO) at 6 mo of age. As anticipated, GRFi at a younger age increased percentage of heifers not reaching puberty; over 75% of control and HSAi heifers reached puberty by 14 mo of age compared to 22% of GRFi heifers. Serum and follicular fluid (FFL; follicles < or = 4 mm) concentrations of IGF-I were suppressed by GRFi. Serum, but not FFL concentrations of IGF binding protein-2 (IGFBP-2) were greater in GRFi than in HSAi heifers. GRFi delayed puberty apparently by suppressing follicular growth because number of follicles < or = 7 mm was significantly lower in GRFi than in HSAi heifers. In conclusion, active immunization against GRF at 3 or 6 months of age delays puberty in beef heifers. Delayed puberty was preceded by suppression of follicular growth, and decreased concentrations of IGF-I in serum and follicular fluid.(ABSTRACT TRUNCATED AT 400 WORDS)


Intracellular potentials of visceronociceptive neurons in posterior group of thalamic nuclei and its labeling with HRP intracellularly in cat.

The intracellular potentials (ICP) of 42 visceronociceptive neurons related to stimulating greater splanchnic nerve (SPL) in Posterior group of thalamic nuclei (PO) were investigated. We analyzed the electrophysiological features, such as amplitude of resting potentials and action potentials of the neurons. Following acquisition of electrophysiological data, (HRP) was injected into 3 neurons, whose responses were inducted by stimulating SPL (related neurons) and able to be inhibited by morphine intravenously. Two neurons revealing spontaneous discharges but are not able to evoke response by stimulating SPL (non-related neurons), also were stained by HRP. Two kinds of neurons, related and non-related ones, display different dendrite branching patterns.


Immunodetection of keratin proteins in oral epithelial neoplasms.

Immunoperoxidase technique was used as an adjunct to routine Haematoxylin and Eosin Staining to assess the pattern of distribution and intensity of Keratin in Oral epithelial neoplasms. The specimens of papillomas, verrucous carcinomas and squamous cell carcinomas were studied. The broadly reactive polyclonal antikeratin antibodies were reacted with formalin fixed paraffin embedded sections. The staining of keratin proteins was found to be dependent upon the degree of differentiation of the tumour. It was observed that the pattern of keratin distribution was regular in papillomas while it was disturbed in verrucous carcinomas and squamous cell carcinomas.


Transbronchial biopsy in sarcoidosis: the role of immunohistochemical analysis for granuloma detection.

In this study 27 transbronchial biopsy specimens obtained from patients with clinical and/or laboratory features suggestive for sarcoidosis were analysed with conventional morphology and immunohistochemistry comparing the sensitivity and reproducibility of the two methods. A limited panel of monoclonal antibodies recognizing epitopes resistant to conventional fixation and embedment procedures were used (CD45R0, CD68, anti-cheratin, anti-collagen IV). All specimens were independently observed by two different pathologists. On the basis of the recognition of bona-fide noncaseating granulomas, 9 cases were uniformly judged as positive, 10 cases as negative, but 8 cases were considered doubtful. Immunohistochemical analysis reliably demonstrated the presence of epithelioid cells in 3 of these cases and absence in 5. These data suggest that the use of a limited immunohistochemical analysis can significantly improve histological diagnosis of sarcoidosis on small transbronchial biopsies using conventional routine material.


Intraobserver and interobserver reproducibility of the FAB classification in acute leukaemia.

Intraobserver and interobserver reproducibility of FAB classification for acute leukaemia was assessed using the modified criteria of the FAB classification. Leishman stained peripheral smear and May Grunwald Giemsa stained bone marrow smears from 72 cases of acute leukaemia were used for this purpose. Cytochemical stains used were peroxidase, PAS and Sudan black B. Intraobserver and interobserver concordance/discordance was calculated. Kappa statistic was used to correct the chance expected agreement. Intraobserver and interobserver concordance was 76% which improved to 91% when cytochemistry was included. Lymphocytic/Nonlymphocytic concordance was 87.5% and 90% respectively for intraobserver and interobserver groups.


Tissue plasminogen activator expression in endothelial cells exposed to cyclic strain in vitro.

The effects of cyclic strain on the production of tissue plasminogen activator (tPA) and type 1 plasminogen activator inhibitor (PAI-1) by cultured endothelial cells (EC) were examined. Human saphenous vein EC were seeded in selective areas of culture plates with flexible membrane bottoms (corresponding to specific strain regions) and grown to confluence. Membranes were deformed by vacuum (-20 kPa) at 60 cycles/min (0.5 s strain alternating with 0.5 s relaxation in the neutral position) for 5 days. EC grown in the periphery were subjected to 7-24% strain, while cells grown in the center experienced less than 7% strain. The results show a significant increase in immunoreactive tPA production on days 1, 3 and 5 compared to day 0 in EC subjected to more than 7% cyclic strain. There was no significant elevation of tPA in the medium of EC subjected to less than 7% strain. tPA activity could only be detected in the medium of EC subjected to more than 7% cyclic strain. PAI-1 levels in the medium were not significantly different in either group. In addition, immunocytochemical detection of intracellular tPA and messenger ribonucleic acid (mRNA) expression of tPA (assessed by the reverse transcriptase polymerase chain reaction utilizing tPA specific sense and antisense primers) was significantly increased in EC subjected to more than 7% cyclic strain. We conclude that a 60 cycles/min regimen of strain that is greater than 7% can selectively stimulate tPA production by EC in vitro and may contribute to the relative nonthrombogenicity of the endothelium in vivo.


A porcine model for adipose tissue-derived endothelial cell transplantation.

The transplantation of endothelial cells represents a technology which has been suggested for applications ranging from improvement in function of implanted vascular devices to genetic therapy. The use of microvascular endothelial cell transplantation has seen increased use both in animal studies as well as clinical use. This report describes our techniques for the isolation and establishment of initial cultures of microvascular endothelial cells derived from porcine fat. A variety of anatomic sites within the pig were evaluated to determine the appropriateness of different sources of fat for endothelial cell isolation. The properitoneal fat was determined to be optimal due to the predominance of endothelium in this tissue and the ease of isolation of microvascular endothelium following collagenase digestion. The study of endothelial cell transplantation in the porcine model is now possible using the methods described for adipose tissue-derived microvessel endothelial cell isolation.


[Symptomatic treatment of pain in cervico-facial cancers].

Pain is frequent in patients presenting cervico-facial cancers. It can be acute or persistent, or present at the morbid entity known as chronic pain. The specific anatomical site and the often poor environment explain that the physical disability related to pain is increased by multiple psychosocial problems. Multidisciplinary care management by several actors is required and can be facilitated by a simple decision-making model. The schema presented can be used to prompt discussion and criticism. It needs constant improvement and extensions in order to reach a consensual attitude towards this king of suffering but also towards other situations of pain care management frequently encountered in cancer patients as well as those with other pathologies.


Neovascular glaucoma and intraocular pressure: I. Pathogenesis of increased intraocular pressure and therapy (a review of historical and current therapeutic modalities).

The pathogenesis of neovascular glaucoma is summarized. Elevation of intraocular pressure is mostly associated with mechanical obstruction of the anterior chamber angle by the fibrovascular membrane, as well as with an enhanced ultrafiltration from the newly formed vessels. Drug therapy is insufficient. Standard filtering surgery is unsatisfactory with regard to the control of intraocular pressure and hemorrhagic complications. The drainage implants appear to be promising. Panretinal photocoagulation is unfavorable due to turbidity of the optical media. Favorable therapeutical results were achieved with cryosurgical techniques, namely panretinal cryocoagulation, combined with cyclocryocoagulation. Cyclocryocoagulation alone accounts for a high percentage of complications and therapeutical results are not convincing.


Identification of uncultured microorganisms: expanding the spectrum of characterized microbial pathogens.

The combination of enzymatic nucleic acid amplification techniques with 16S rRNA-based molecular phylogeny has brought about a new approach to the identification of microbial pathogens that can not be cultivated in the laboratory. The applications of this experimental approach to bacillary angiomatosis and to Whipple's disease have revealed the presence of two previously uncharacterized organisms. These results suggest the existence of a far greater microbial diversity among human pathogens than has been so far appreciated with culture-dependent methods. PCR-based studies of aquatic environmental microbial communities have already reached similar conclusions. As a result, new and provocative questions are raised concerning the association of amplified 16S rRNA sequences with diseased tissue. The answers must await the results of further investigations and the expansion of sequence data bases.


Self-expanding mesh stent for endoscopic palliation of rectal obstructing tumors: a preliminary report.

The endoscopic insertion of self-expanding mesh stents in four patients affected by obstructing rectal malignant tumors is reported. The preliminary experience shows that, in the short term, normal defecation was achieved, with no complications. Longer follow-up is necessary to evaluate the duration and the quality of the palliative effect.


Screening Moroccan infants using the Wolański Gross Motor Evaluation: a pilot study.

The purpose of this study was to examine the possibility of using the Wolański Gross Motor Evaluation (WGME), to effectively screen Moroccan infants for motor disabilities. Additionally, the appropriateness of Polish norms for use with Moroccan children was considered. One hundred and twenty subjects, 15 females and 15 males from four age groups (three, six, nine and 12 months), were recruited from well-baby clinics in Marrakech. Subjects were tested for gross motor maturity using the WGME. Differences were found in the sequence of skill acquisition between the Polish norms and the Moroccan sample in sitting, standing, and rolling skills. Differences were also found in the rate of development between the Moroccan and Polish infants. No differences were found between Moroccan females and Moroccan males in the rate of motor development. Results from this study indicate that the grids used to score the WGME are not applicable to Moroccan infants, and would need to be modified for use with a Moroccan population.


Angiofollicular lymph node hyperplasia: a comparison of unicentric, multicentric, hyaline vascular, and plasma cell types of disease by morphometric and clinical analysis.

A morphometric and clinical analysis of 47 cases of angiofollicular lymph node hyperplasia (AFLH) demonstrated morphometric differences between plasma cell (28 cases) and hyaline vascular (19 cases) AFLH. Twenty-one AFLH cases were unicentric, and 26 cases were multicentric. Plasma cell types of AFLH, defined by standard morphologic and objective morphometric assessment, were associated with systemic clinical and laboratory abnormalities. Unicentric or hyaline vascular AFLH was amenable to surgical therapy, and multicentric or plasma cell AFLH without neuropathy was amenable to steroid treatment or chemotherapy with good prognoses. Multicentric or plasma cell AFLH with neuropathy was associated with resistance to steroids and chemotherapy and with a poor prognosis.


Expression of vimentin in proliferating and damaged bile ductules and interlobular bile ducts in nonneoplastic hepatobiliary diseases.

Biliary epithelial cells express characteristically cytokeratin in their cytoplasm in normal and diseased livers. The present study disclosed that vimentin was frequently expressed in the cytoplasm of proliferating and damaged bile ductules and interlobular bile ducts, while their normal counterparts were negative for vimentin. Although this expression itself seemed nonspecific to any of the hepatobiliary diseases examined, bile ductules and interlobular bile ducts were frequently positive in chronic cholestatic and necroinflammatory liver diseases. In biliary epithelial cells, vimentin was localized around the nucleus or in the subnuclear regions, when present. Immunoelectron microscopically, reaction products for vimentin and for cytokeratin were found on bundles of intermediate filaments in the cytoplasm of biliary epithelial cells. The former was found mostly in the paranuclear and subnuclear regions, while the latter detected around the desmosomes, in addition to the paranuclear cytoplasm. Vimentin and cytokeratin were also seen together under immunoelectron microscopy on the same intermediate filaments. It seems likely that aberrant expression of vimentin in bile ductules and interlobular bile ducts and heterogeneous antigenic expression of intermediate filaments in the same biliary epithelial cells may be related to proliferation of, reorganization of, or damage to the ductular and ductal biliary cells in a variety of hepatobiliary diseases.


Paranuclear blue inclusions in metastatic undifferentiated small cell carcinoma in the bone marrow.

Paranuclear blue inclusions (PBIs) are frequently identified within metastatic undifferentiated small cell carcinoma (SCC) cells on air-dried bone marrow aspirates stained with Wright's stain. To determine the sensitivity and specificity of this finding, 116 bone marrow aspirates containing metastatic neoplasms were evaluated for the presence and frequency of PBIs. Bone marrow specimens included 47 cases of metastatic SCC of the lung, 13 cases of large cell lymphoma, 19 cases of neuroblastoma, five cases of small, noncleaved cell lymphoma, seven cases of rhabdomyosarcoma, three cases of Ewing's sarcoma, three cases of other sarcomas, and 19 cases of non-small cell carcinoma (adenocarcinoma). PBIs were identified in 40 of 47 (85%) cases of SCC and their frequency varied from 0 to 24% of tumor cells among different cases. In approximately half the cases of SCC, PBIs were identified in 1 to 4% tumor cells; and in eight cases, PBIs were present in 5% or more of tumor cells. PBIs were also identified in two of seven (29%) cases of rhabdomyosarcoma and one case of malignant peripheral nerve sheath tumor, but they were not seen in Ewing's sarcoma, small non-cleaved cell lymphoma, large cell lymphoma, neuroblastoma, or non-small cell carcinoma. In addition, PBIs were not seen in alcohol-fixed, Papanicolaou-stained cytology specimens containing SCC. Ultrastructurally, PBIs may represent phagocytized nuclear/cellular material. PBIs are a feature of small cell carcinoma on air-dried, cytologic material stained with Romanowsky type stains. Their presence may provide diagnostic information with regard to the differential diagnosis of metastatic SCC in the bone marrow. Future studies evaluating non-bone marrow Wright's stained fine-needle aspiration specimens are needed to determine if PBIs are useful in distinguishing SCC from other poorly differentiated tumors in the cytology laboratory.


Inhibition of tobacco nitrite reductase activity by expression of antisense RNA.

A tobacco nitrite reductase (NiR) cDNA and its corresponding gene were isolated from cDNA and genomic libraries. An NiR antisense mRNA was expressed in transgenic tobacco under the control of a double 35S promoter. Transformants were obtained on a medium containing ammonium as the sole source of nitrogen. One plant growing normally on ammonium but displaying drastically reduced development and chlorotic leaves when grown on nitrate as the sole source of nitrogen was studied further. This plant accumulated nitrite fivefold over wild-type level and showed reduced amounts of ammonium (11% wild-type level), glutamine (19%), and total protein (8%). NiR mRNA and activity were below detectable levels. Under these conditions, nitrate reductase (NR) activity and mRNA were overexpressed, suggesting that N-metabolites resulting from nitrate reduction are responsible for the repression of the expression of the NR gene, independently from the presence or absence of a functional NR protein.


[Occurrence of an epidermoid bronchial carcinoma 8 years after a small cell carcinoma treated with only 3 courses of chemotherapy].

A case of squamous cell bronchial carcinoma developed 8 years after treatment of a small cell carcinoma is reported. Three hypotheses may be put forward to explain this sequence of events: either a squamous cell line was selected from within an initially mixed tumour, or the small cell carcinoma differentiated into a squamous cell carcinoma, or a second primary tumour developed independently. All this is purely speculative, but the long doubling time implied by the first two hypotheses would be in favour of a second primary tumour.


[Pancreatic endocrine tumor with metastases and increase of alpha-fetoprotein. A case report].

The authors report the case of a 48 years old man presenting a pancreatic islet cell carcinoma (gastrinoma) with liver, nodes and peritoneal metastases, associated with an elevated alpha-fetoprotein (AFP) concentration. Incomplete remission was first obtained with a chemotherapy using Streptozotocin combined with 5-Fluorouracil, in association with a Somatostatin analogue (SMS 201-995). But when relapses occur, another chemotherapy was not so effective. Serum gastrin and AFP levels had the same evolution and appear to have the same interest to follow the course of the disease.


Monitoring the efficacy of temephos application and use of fine mesh nylon strainers by examination of drinking water containers in guineaworm endemic villages.

Application of temephos in unsafe water sources for destroying cyclops, the intermediate hosts of guineaworm, and distribution of fine mesh nylon strainers for promoting prophylaxis against guineaworm are accepted methods of guineaworm control in different endemic countries. The existing methods of monitoring the efficacy of these guineaworm control methods are not fully informative. Examination of drinking water stored at household levels for presence of cyclops with or without Dracunculus larvae can provide information on the efficacy of these control/prophylactic methods, besides serving as a means of interpersonal health education to the community. This paper presents observations carried out in two villages in peninsular India in 1991, which revealed that while in one village complete absence of cyclops from stored water containers was attributable to the use of temephos in the village and straining of drinking water, in the other village, with no temephos application, 15.6 per cent of containers contained varying numbers of cyclops in them. Implications of these observations for guineaworm eradication activities are discussed.


The response of regulatory peptides to moderate hypoglycaemia of short duration in type 1 (insulin-dependent) diabetes mellitus and in normal man.

The changes in plasma gastrin-releasing peptide (GRP), arginine vasopressin (AVP), neuropeptide Y (NPY), corticotropin releasing hormone (CRH), galanin, ACTH, cortisol, delta sleep-inducing peptide (DSIP), adrenaline, noradrenaline and pancreatic polypeptide (PP) were measured after 5 and 15 minutes of acute insulin-induced moderate hypoglycaemia (2.0 mmol/l) in 10 patients with Type 1 diabetes mellitus with no autonomic neuropathy and in 10 healthy subjects. Plasma catecholamine and PP levels rose in both groups in response to hypoglycemia and the secretory response of ACTH was lower in the diabetic subjects (p < 0.01). GRP concentrations increased during hypoglycaemia (p < 0.01) while a reduction in AVP occurred at the start of hypoglycaemia (p < 0.001). The plasma AVP concentrations were higher in the diabetic group compared with those in the normal group (p < 0.05). The NPY concentrations were higher in the normal subjects (p < 0.05) but no change in the mean level occurred in either group during hypoglycaemia. No group differences or changes in mean plasma concentrations were found for galanin, DSIP and CRH. These observations support the view that regulatory peptides, if involved in glucose homeostasis, may rather have a modulatory effect than a direct action in restoring normoglycaemia.


[Biochemical profile of Senegalese women on oral contraceptives].

By its many advantages, pills are one of the most used methods in the world and in Senegal. On the other hand, exist side effects which can limit their utilization. Our study concern 35 women using between 12 and 36 months mini pills, which might produce low side effects. In our users, biochemical parameters are modified but not pathologic. By our results, we think that in the following 5 or 10 years, these alterations might increase; so, we raise the alarm and propose a biological following up of the oral contraceptive users.


Submucosal glands are the predominant site of CFTR expression in the human bronchus.

We have used in situ hybridization and immunocytochemistry to characterize the cellular distribution of cystic fibrosis (CF) gene expression in human bronchus. The cystic fibrosis transmembrane conductance regular (CFTR) was primarily localized to cells of submucosal glands in bronchial tissues from non-CF individuals notably in the serous component of the secretory tubules as well as a subpopulation of cells in ducts. Normal distribution of CFTR mRNA was found in CF tissues while expression of CFTR protein was genotype specific, with delta F508 homozygotes demonstrating no detectable protein and compound heterozygotes expressing decreased levels of normally distributed protein. Our data suggest mechanisms whereby defects in CFTR expression could lead to abnormal production of mucus in human lung.


Histamine release and SOD, allopurinol and ranitidine pretreatment in haemorrhagic shock in the rat.

Histamine release have been demonstrated in haemorrhagic shock. There are some observations that oxygen free radicals can cause histamine release. Oxygen free radicals play a role in the pathogenesis of gastric mucosal lesions. The goal of this study was to determine whether ranitidine or SOD and allopurinol pretreatment modify the histamine release during and after the haemorrhagic shock in the rat. In the anaesthetized rat 0.1 N HCl was instilled into the stomach and the rat was bled to reduce the blood pressure to 30 mmHg for 20 min. The shed blood was reinfused. Twenty min later the stomach was removed. The area of gastric mucosal lesions were measured, histological grading was made. Blood samples taken from the carotid artery were examined by radioimmunoassay (IMMUNOTECH) to determine the plasma histamine level. Plasma histamine level did not change significantly during the preparative surgery, but there was a significant increase of histamine level by the end of shock period. After the reinfusion of the blood the plasma histamine remained essentially at the same level for five min. Oxygen free radicals did not cause an important histamine release. By the end of the experiment the histamine level decreased dramatically. Ranitidine, allopurinol and SOD pretreatment provided significant protection against the gastric mucosal lesions. Allopurinol and SOD did not influence significantly the histamine level. Ranitidine caused significant histamine release immediately after the injection and every histamine value was significantly higher in this group except for the final value which was lower than the control one. The oxygen free radicals were not found as endogenous histamine releasers in this study.


Cholinergic neurons are involved in the effect of substance P on the circular muscle of the guinea pig small intestine.

The mode of action of the excitatory neuropeptide substance P was studied on the circular muscle of the guinea pig ileum in vitro. Atropine or tetrodotoxin strongly inhibited substance P-induced phasic contractions. The atropine-resistant part of the circular response was blocked by tetrodotoxin. A newly-developed method for quantitative evaluation revealed a rightward displacement of the substance P concentration-response curve, as well as a strong depression of the maximum effect, in the presence of atropine. These results indicate that cholinergic (and probably also non-cholinergic) excitatory neurons mediate phasic contractions due to substance P. The tonic component of the substance P-induced contraction was slightly reduced by atropine.


The effect of somatostatin, gabexate mesilate and dextran 40 on the microcirculation in sodium taurocholate-induced pancreatitis.

In vivo microscopy was performed to assess the effect of dextran 40, gabexate mesilate and somatostatin on the microcirculation in sodium taurocholate-induced pancreatitis in rats. Intraductal infusion of 0.4 ml of a 4% solution of sodium taurocholate decreased capillary blood flow, induced capillary stasis and increased vascular permeability in the head of the pancreas. Dextran 40, gabexate mesilate and somatostatin improved capillary blood flow in the initial phase of acute pancreatitis significantly and prevented stasis in 5 of 9, 3 of 8 and 7 of 10 (p < 0.05) cases. Only dextran 40 reduced the increase of vascular permeability. Decrease of capillary blood flow, capillary stasis and vascular permeability changes are important factors contributing to the pathogenesis of sodium taurocholate-induced pancreatitis. Dextran 40, gabexate mesilate and somatostatin exert a beneficial effect on the microcirculatory changes in this model of acute pancreatitis.


Antiinflammatory effect of a protein kinase C inhibitor (K-252a) on the development of the dextran-induced paw edema in the rat (preliminary results).

The effect of a metabolite of Nocardiopsis sp. as a protein kinase C inhibitor from microbial origin was investigated on the onset and development of dextran-induced paw edema in the rat. It was published that this compound (K-252a) interferes with histamine release from mast cells, while dextran-induced paw and nose edema are induced by vasoactive agents, like histamine etc., released from the disrupted mast cells. The antiinflammatory effect of the K-252a is effectuated by the inhibition of protein kinase C. Groups of male Wistar rats with 180-200 g b.w. were used; each group consisted of 10-10 rats. The following groups were consisted: rats given orally DMSO (control), rats given 1 mg/kg, or 3 mg/kg b.w. of K-252a dissolved in DMSO and given p.o. one hour before dextran injection. Dextran (BDH Chem. LTD, molW: 200.000, England) was injected intraperitoneally in 10% solution, in a dose of one ml/100 g b.w. Volume of the hind leg was measured by a mercury plethysmometer. Time-sequence of the edema was followed. Increase in volume of hind leg paw was related to its 0-min volume in %. Results were analyzed by the Kruskal-Wallis-test. Edema of the legs and noses appeared in each of the control rats in one hour. The 1 mg/kg dose of K-252a retarded the appearance of the edema by 1 hour, the 3 mg/kg dose, however, prevented its onset for 4 hours.(ABSTRACT TRUNCATED AT 250 WORDS)


Sensory nerve terminal activity in severe hypertension as reflected by circulating calcitonin gene-related peptide (CGRP) and substance P.

In patients with severe hypertension and in age and sex matched controls the circulating levels of calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) and substance P-LI were measured. Samples were taken before medication, after 2-4 weeks and 2-12 months of pharmacological treatment to normotension. In the control group CGRP-LI levels were significantly higher for females than for males. No such relation was seen for substance P-LI. There were no correlations between CGRP-LI, substance P-LI or blood pressure. In the untreated acute hypertensive group there was a significant correlation between circulating levels of CGRP-LI and both diastolic and systolic blood pressure. No such relationship was seen for substance P-LI. The plasma levels of substance P-LI were significantly elevated (2.8 +/- 4.0) compared to controls (1.3 +/- 1.3, pmol/l, mean +/- S.D., p < 0.01). The levels of CGRP-LI did not differ from the control group. After 2-4 weeks of treatment the blood pressure decreased significantly and the plasma levels of substance P-LI were normalized while the CGRP-LI still did not differ from that of controls. After 2-12 months of treatment the blood pressure was still normalized, and the plasma levels of CGRP-LI and substance P-LI were not different from the control group. In the present study there was a positive correlation in hypertensives between the circulating CGRP-LI levels and diastolic and systolic blood pressure and elevated levels of substance P-LI. This would implicate the existence of a dynamic control through which the sensory system may register and damp the pressure response.


[Upper digestive hemorrhage in acute pancreatitis].

We proposed to investigate the following parameters regarding upper gastrointestinal hemorrhage occurring in the patients with acute pancreatitis: incidence, form of manifestation, severity, correlated to the clinical and morpho-pathological form of acute pancreatitis, cause (direct causality or morbid association). The study was carried out on the inpatients of the internal-medicine and surgery units over a period of one year. For sake of accuracy the study also included the cases submitted to necropsy over a period of 5 years. The practical experience has revealed that though gastrointestinal bleeding is not a frequent complication--9.1% in the patients with acute pancreatitis and 36.3% of the necropsied cases--it is often severe, which imposes the early recognition for its adequate management.


The communicative Effectiveness Index: its use with South African stroke patients.

The functional communication of a group of 28 South African stroke patients was examined using the Communicative Effectiveness Index (CETI). It was translated into Afrikaans, Sotho and Zulu and administered to the significant others of 22 aphasic patients with left hemisphere damage and 6 patients with right hemisphere damage. Results were related to the results of standardized language testing and to case history factors such as cultural factors and time since onset. The CETI was readministered in the case of eight of the aphasic subjects after a mean period of six months in order to assess its sensitivity to recovery. Results showed that the CETI seems applicable across different language groups, that it is sensitive to change across time as well as sensitive to the communication disorders resulting from both right and left hemisphere damage. Further it appears to correlate well with overall level of severity. It does not appear to differentiate patients in terms of time since onset. Its potential use as a relatively culture free assessment tool in the South African context is discussed.


The incompatibility complex of H. plasmids.

This review deals with the general properties of the very large transfer thermosensitivity R. factor belonging to the H. incompatibility complex. This group is of particular interest not only because their temperature sensitivity transfer system but also for the number as well as distinctive resistance determinants being accumulate by them, and their prevalence in Salmonella serotypes and in other Gram-negative non-pathogenic bacteria both in man and animals.