Dimorphic male brains and alternative reproductive tactics in a vocalizing fish.

Brains and types of behavior among sexually mature vertebrates are portrayed as having two phenotypic states: male and female. However, mating systems and the behavioral tactics employed by the two sexes are far more diverse than conveyed by this simple intersexual dichotomy. For example, in many species, sexually mature males may practice one of two alternative mating tactics. Recent studies of a sound-producing teleost fish now show that intrasexual dimorphism of vocal motor phenotypes among males accompanies the intrasexual divergence of types of reproductive behavior. Thus, one group of males, like females and juveniles, lacks the behavioral, neurophysiological and morphological traits typical of the sexually differentiated vocal motor pathway of the second group. The results are an explicit demonstration that for any one species: (1) alternative mating tactics can be paralleled by alternative phenotypes for the neurons that determine tactic-specific types of behavior, and (2) reproductive maturation is not obligatorily linked to the expression of neuronal secondary sex characteristics.


Oxygen and acid chemoreception in the carotid body chemoreceptors.

The carotid bodies are arterial chemoreceptors that are sensitive to blood PO2, PCO2 and pH. They are the origin of reflexes that are crucial for maintaining PCO2 and pH in the internal milieu and for adjusting the O2 supply according to the metabolic needs of the organism in situations of increased demand, such as exercise and while breathing at decreased O2 partial pressures during ascent or when living at high altitude. Chemoreceptor cells of the carotid body transduce the blood-borne stimuli into a neurosecretory response that is dependent on external Ca2+. These cells have an O2-sensitive K+ current that is reversibly inhibited by low PO2. It is proposed that the depolarization produced by inhibition of this K+ current activates Ca2+ channels; Ca2+ influx and neurosecretion follow. The cells have also a potent Na(+)-Ca2+ antiporter that could be responsible for the intracellular Ca2+ rise required to trigger the release of neurotransmitters during high PCO2 or low pH stimulation.


Further practical experiences in the recognition and management of carcinoma in situ of the testis.

99 biopsies from the contralateral testis in patients with unilateral germ cell tumor were investigated using the semithin section technique. Four cases (4%) revealed a carcinoma in situ (CIS) pattern. Two patients underwent a local radiation (20 Gy), 2 patients received combination chemotherapy (cisplatin, etoposide and bleomycin; PEB). No tumor cells were found in control biopsies 4-8 months after therapy. Both biopsy specimens taken from radiated patients lacked also germ cells. In contrast, 1 patient who was treated with PEB and also another 1 presenting with a teratocarcinoma of apparently retroperitoneal origin and unilateral CIS revealed germ cells after chemotherapy. The present data suggest radiation therapy to be the first line treatment for CIS-bearing testes. In order to get informations about the distribution of CIS cells in the affected testes, one or two biopsies were additionally taken from macroscopically unsuspicious tissue surrounding various solid germ cell tumors (74 patients). 56 cases (76%) revealed CIS. Even considering the possibility of missing CIS, a screening biopsy is actually the only method for detecting early manifestations of germ cell tumors and should be performed routinely in contralateral testes of patients with germ cell tumors.


Transplantation of human benign hyperplastic prostate tissue into nude mice: first results of systemic therapy.

Xenografts of human benign hyperplastic prostate tissue (BPH) have been established as a model for the investigation of the etiology of BPH. In this paper it is our aim to answer the question of whether this model is useful for the established therapy of the disease. Additionally we try to evaluate the value of a commonly used plant extract for the therapy of BPH. Is there any influence of the extract from Sabal serrulatum on the BPH tissue in our model? Human BPH tissue from 2 patients was transplanted into athymic nude mice and treated with three different regimens. Animals of group I did not receive any treatment and served as control, in groups II and III the tissue was stimulated by application of silicone tubes containing crystalline 5 alpha-dihydrotestosterone and estradiol. Animals of group II additionally were treated orally with the lipophilic extract of S. serrulatum (contained in Prostagutt N), which is commonly used for the treatment of BPH clinically. Significant inhibition of tissue growth was observed in group III when compared to group II. In group I (control) atrophy of the graft was observed as expected. However, histologically no differences were visible between groups II and III. Our experiment shows a significant growth-inhibiting effect of the plant extract for human BPH tissue in our model (p less than 0.05). We conclude that the nude mouse model can be useful for the evaluation of systemic therapy modalities of human BPH and that the plant extract may have a certain value for clinical treatment, which is not only due to subjective criteria.


Prostate-specific antigen test: operating characteristics and assessment criteria in the diagnosis of prostatic cancer.

The sensitivity and specificity of prostate-specific antigen (PSA) in prostatic carcinoma is of considerable interest. In this study, we have assessed PSA and also correlations between positive and negative predictive values of PSA and the prevalence of prostatic cancer. Firstly, cutoff point must be selected as a positivity criterion for prostatic cancer, according to the purpose of the test. Besides sensitivity, all the other operating characteristics are dependent upon the prevalence of the disease. Specificity and positive predictive value increase while negative predictive value decreases when prevalence increases. As the relationship between the age and the prevalence of prostatic carcinoma is well known, the age of the patient becomes a variable of prominent importance when assessing the test.


Serum prostatic acid phosphatase, gamma-seminoprotein and prostatic specific antigen in hemodialysis patients.

Serum concentrations of prostatic acid phosphatase (PAP), gamma-seminoprotein (gamma-Sm) and prostatic specific antigen (PSA) were measured in 31 hemodialysis patients without clinical signs of malignant disease. PAP, gamma-Sm and PSA levels in serum were not significantly different between control and hemodialysis groups. A significant reduction in these tumor markers was not found after dialysis treatment. This indicates that the measurement of PAP, gamma-Sm and PSA in serum is useful for the detection of prostatic cancer in patients undergoing hemodialysis.


Organisms in the prostate and antibiotics in the treatment of postoperative infections.

On 24 benign prostatic hyperplasia patients with preoperatively sterile urine, who also had no history of urinary tract infection, the organisms in the prostate obtained through transurethral resection as well as in the anterior urethra were isolated. In 17 patients out of 24, organisms obtained in the prostate were identical to those in the anterior urethra; therefore, it can be concluded that another 7 patients had organisms in the prostate itself before the operation. These results would suggest that sterile urine did not indicate sterile prostate and that organisms in the prostate did not always ascend from the urethra. In the patients who preoperatively received transurethral catheterizations, Streptococcus faecalis was the most predominantly isolated organism in the prostate, which was highly sensitive to ABPC and minocycline and lower to cephems. On the other hand, in the patients with no history of catheterizations, Staphylococcus epidermidis was the most commonly isolated one, which was highly sensitive to ABPC and minocycline as well as cephems. So, in the treatment of the cases with infectious symptoms after transurethral resection of the prostate, ABPC or minocycline should be selected as first-choice drugs rather than cephems.


Alpha-fetoprotein-producing adenocarcinoma of the ureter.

We report a case of primary ureteral tumor producing alpha-fetoprotein (AFP). Computerized tomography, ultrasonography and endoscopy of the bile duct revealed no obvious tumor in the liver, gallbladder, bile duct, pancreas or ovary. Total nephroureterectomy was performed, and histopathological examination revealed adenocarcinoma of the ureter lined with transitional cell carcinoma in its base. The tumor was stained with immunohistological AFP stain, and the high serum AFP level normalized after resection of the tumor.


Biochemical characterization of an antigenic saline extract of Actinobacillus pleuropneumoniae serotype 5 and identification of a serotype-specific antigen for ELISA serodiagnosis.

A saline extract of boiled-formalinized whole cells from a local strain (81-750; Quebec, Canada) of Actinobacillus pleuropneumoniae, serotype 5b was used as an antigen in an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of swine pleuropneumonia. Characterization of this crude extract was done and proteins, neutral sugars, hexosamines, and 2-keto-3-deoxyoctonate (KDO) were evaluated. On phenol extraction of the crude extract a serotype-specific antigen of polysaccharidic nature was recovered from the aqueous phase. This antigen was characterized using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with Coomassie blue, silver and Schiff stainings. Immunoblots were done using sera of experimentally infected pigs that showed serotype specificity and cross-reactivity. Overall, the results indicate that the O-chain of lipopolysaccharides is a specific antigen that could be used in ELISA for the serodiagnosis of serotype 5 of A. pleuropneumoniae.


Biologically active cymbidium ringspot virus satellite RNA in transgenic plants suppresses accumulation of DI RNA.

A full-length DNA copy of cymbidium ringspot virus (CyRSV) satellite RNA was cloned downstream of the bacteriophage T7 RNA polymerase promoter. In vitro transcripts were biologically active in plants when coinoculated with the helper virus or its RNA. Although the transcripts contained 7 or 29 extra nucleotides at the 3' end, the proper 3' terminus was restored in the satRNA progeny. Full-length cDNA clones of CyRSV satRNA under the control of the cauliflower mosaic virus 35S promoter and terminator were used to transform Nicotiana benthamiana plants. Integration of CyRSV satRNA sequence in the plant genome was tested by PCR amplification of DNA extracts from transformed plants and by detection of satRNA-related transcripts in total RNA extracts. Inoculation of transgenic plants with the helper virus induced replication of satRNA of the same size as the native molecule. Sequence analysis of the satRNA progeny showed that it was identical to natural CyRSV satRNA. Infected transgenic plants were not protected from apical necrosis and death by the presence of satRNA sequences. Rather, replication of satRNA was found to suppress accumulation of defective interfering RNA, which acts in the absence of satRNA as an attenuator of virus replication and disease.


Epitopic mapping of linear and conformation-dependent antigenic determinants on GP5 of five U.S. bluetongue viruses.

Two distinct antigenic determinants of the major outer capsid protein, GP5, of five U.S. bluetongue viruses have been identified and mapped using monoclonal and oligoclonal antibodies. One antigenic site, identified by oligoclonal antibody AK-15, was found to be common and conserved in all five U.S. BTV serotypes. This linear epitope was located between amino acid residues 175 and 189 (ALQREAAERSEDEIK). The second determinant identified by monoclonal antibody 34.7 was present in BTV-2, -10, -11, and -17 but absent in BTV-13. The binding of this monoclonal antibody to GP5 could be blocked specifically by one of three short synthetic peptides located among amino acid residues 33-42 (KAAERFAESE), 159-168 (EKILKEEDSK), and 206-215 (EIERDGMQEE), indicating that this antigenic determinant is conformation-dependent. Oligoclonal antibody (AK-15) reacted with denatured GP5 immobilized on nitrocellulose membrane after Western transfer as well as with native GP5 present on the surface of purified BTV virions. Monoclonal antibody (34.7) reacted only with denatured GP5 but not native GP5 using an ELISA assay. However, these two antigenic epitopes alone did not elicit detectable neutralizing antibodies as determined by plaque reduction assay.


Activated, HTLV-1-specific cytotoxic T-lymphocytes are found in healthy seropositives as well as in patients with tropical spastic paraparesis.

In human T cell lymphoma/leukemia virus (HTLV-1)-infected people with tropical spastic paraparesis (TSP), there are activated HTLV-1-specific cytotoxic T lymphocytes (CTL) in the circulation and lymphocytic infiltrates in spinal cord lesions that are rich in CD8+ T cells. These observations suggest a role for virus-specific CTL in the pathogenesis of TSP. We have examined the anti-HTLV-1 cytotoxic activity of freshly isolated CD8+ T cells from peripheral blood lymphocytes of eight subjects seropositive for HTLV-1. Four of five subjects with TSP had circulating activated anti-Tax CTL. However, two of three seropositive subjects without TSP also had activated anti-Tax CTL. These observations show that such activated CTL are not confined to patients with TSP and raise some uncertainty about their significance in the pathogenesis of the disease. In cultures of CD8+ T cells from two TSP subjects, we detected CTL with other HTLV-1 specificities, without exogenous antigenic stimulation. A CTL epitope in the middle region of Tax and one in the C terminus of Pol have been mapped at the peptide level and the HLA Class 1 molecules restricting their recognition have been defined.


Isolation and characterization of a retrovirus from the fish genus Xiphophorus.

A cell line (BsT) established from neoplastic embryonal tissues of the platyfish (Xiphophorus maculatus) released spontaneously retrovirus-like particles. The particles have a buoyant density of 1.16 g/ml, a mean diameter of 100 nm and the morphology of immature retroviruses. The particle-associated proteins p70, p65, and p28 react with an antiserum directed against the major internal feline leukemia virus structural protein p27. The particles are associated with a reverse transcriptase. The purified enzyme has a molecular weight of about 70 kDa and prefers the template primers poly(rA):oligo(dT), poly(dC):oligo(dG), and poly(rC):oligo(dG) in the presence of Mn2+. The enzyme activity is inhibited by antibodies directed against the reverse transcriptase of feline leukemia virus and simian sarcoma virus. The particles contain a ribonucleic acid of about 70 S. In an endogenous reverse transcriptase reaction nucleic acids in the range of 0.2 to 0.4 kb were synthesized. In Northern blots with these nucleic acids as probe, three transcripts of about 8.5, 4.2, and 1.5 kb were detected in BsT cells. Southern blot analysis with the same probe demonstrates related sequences in the DNA of BsT cells and the platyfish and swordtail (Xiphophorus helleri). Hybridization experiments with the LTR-gag region of the feline leukemia virus show homologous sequences in the Xiphophorus genome.


Influenza viruses differ in recognition of 4-O-acetyl substitution of sialic acid receptor determinant.

Equine alpha 2-macroglobulin (EM), known to contain both Neu5Ac and Neu4,5Ac2 sialic acid residues, was treated with Vibrio cholerae sialidase for the selective removal of Neu5Ac and was compared with the untreated EM for its binding by a panel of influenza viruses. Type A H3N2 virus strains having Leu in position 226 of their hemagglutinin (HA) changed the affinity for sialidase-treated EM only slightly, if at all, indicative of their ability to bind the 4-O-Ac-substituted Neu5Ac receptor determinant. At the same time, all B and H1N1 viruses, some H2N2 variants, as well as H3N2 strains with 226 Gln studied were unable to recognize Neu4,5Ac2 moieties of EM. Molecular modeling based on the known 3-D structure of H3 HA complexed with sialyllactose (Weis et al. (1988) Nature 333, 426-431) predicts that the 4-O-Ac substituent of sialic acid would protrude with its carbonyl oxygen inside the receptor-binding site of HA, thus possibly interfering with binding.


Resistance of human immunodeficiency virus type 1 reverse transcriptase to TIBO derivatives induced by site-directed mutagenesis.

The reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) is the target enzyme for the tetrahydro-imidazo[4,5,1-jk][1,4]- benzodiazepin-2(1H)one and thione (TIBO) derivatives, a class of highly potent and selective anti-HIV agents that specifically inhibit HIV-1 but not HIV-2 replication. The amino acid sequence divergence may be held responsible for the differential sensitivity of HIV-1 RT and HIV-2 RT to the TIBO derivatives. Using site-directed mutagenesis, we have introduced several amino acid substitutions in the conserved regions of HIV-1 RT. Where applicable, the amino acids were replaced by the corresponding amino acids present in HIV-2 RT. The amino acid residues Y181 and Y188 appeared to be critical for the anti-HIV-1 RT activity of the TIBO derivatives, since substitution of these residues by the corresponding HIV-2 amino acids I181 and L188 resulted in a virtual loss of TIBO sensitivity without loss of enzymatic activity.


Intake of intense sweeteners in Germany.

The dietary intake of aspartame, cyclamate, and saccharin was evaluated in Germany (FRG) in 1988/89. In the first part of the study the sweetener intake was evaluated in a representative sample of the population. Complete 24-h records of the amount and type of all foods and drinks consumed were obtained from 2,291 individuals. The total daily intake was calculated for each person from the sweetener content of each product and was expressed in mg/kg body weight (bw). 35.9% of the participants ingested one or more sweeteners on the examination day. Cyclamate and saccharin were the prominent sweeteners because aspartame was at that time permitted only under special regulatory exemption, and products containing acesulfame were not yet available. For users of intense sweeteners the mean intakes of aspartame, cyclamate, and saccharin were 0.15, 2.62, and 0.250 mg/kg bw/day, respectively. At the 90th percentile of intake, i.e., for the heavy consumer, the ingestion of cyclamate and saccharin was about 2.5 times higher. Persons who adhered to a diet (diabetes, weight control) did not ingest sweeteners in substantially higher amounts. Tabletop sweeteners and beverages were the most important sources of sweeteners, and they contributed more than 80% of the total intake. Consumption of sweeteners in excess of the Acceptable Daily Intake (ADI) was rarely observed (saccharin: one person, cyclamate: 16 persons). In the second part of the study, the sweetener intake was further evaluated during a 7-day period in those subjects who in the 1-day study ingested any of the sweeteners in excess of 75% of the ADI. Complete 7-day food records were available from 40 out of the 41 subjects who fulfilled this criterium. In this selected subgroup in which 19 subjects were less than 19 years old, the mean daily intakes of aspartame, cyclamate, and saccharin were 0.13, 4.53, and 0.42 mg/kg body weight (bw), respectively. These levels correspond to 0.33, 41 and 17% of the corresponding ADI values. No subject exceeded the ADI of aspartame or saccharin on any day of the study. For cyclamate, the mean daily intake over the 7-day period exceeded the ADI in 4 subjects. The results indicate that at the time of the study the then valid German sweetener regulation protected the consumer adequately, and that the sweetener intake was in 99.8% of all examined persons within recommended limits.


[A child with XYY syndrome as experienced by his mother. A report of daily journal recordings].

A mother's experiencing of the development of her son with "XYY syndrome" is presented based on notes from the mother's diary. According to recent findings in unselected longitudinal studies the boy clearly belonged to a subgroup of the more severely affected children. His developmental language and motor delays, other language problems and problems at school were typical, whereas his slightly reduced IQ, in the lower normal range, and aggressive and autistic symptoms were atypical. In many ways the mother's experience is comparable to that of parents of mentally retarded children. Complicating factors were the inconspicuous phenotype, late diagnosis and lack of professional help. It is shown that parental adaptation to a "handicapped" child can occur in phases. These can be understood through models of mourning or coping. It would be of great help to these families if self-help groups would be established.


Embedding of dithranol in lipid crystals.

A new cream base, based on lipid crystals, was used to formulate an antipsoriatic cream containing Dithranol. The cream consists of solid lipid crystals of C12 and C14 monoglycerides. The lipids are solid at room temperature but melt at skin temperature. The cream is easy to wash off, provides good stability of dithranol, and is practically nonstaining.


Argyrophilic proteins of the nucleolar organizer region in acute leukemias and its relation to the S-phase fraction of leukemic cells.

Nucleolar organizer regions (NORs) are loops of DNA which transcribes to ribosomal RNA. The NOR-related protein becomes visible in nucleus by a silver-staining technique under a light microscope, and it has been named argyrophilic protein of NOR (Ag-NOR). In various malignancies, the correlation between the proliferation potential of tumor cells or histological grade and the number of Ag-NORs has been reported. In this study, we investigated the Ag-NOR of acute leukemic cells and its relation to the in vivo proportion of bone marrow leukemic cells in DNA synthetic phase. The number of Ag-NORs in bone marrow leukemic cells was more than that in peripheral blood (means values 2.78 and 2.48, respectively, p less than 0.01). This result shows that the number of Ag-NORs reflects the vigorous proliferative potential of bone marrow leukemic cells. However, no significant correlation was obtained between the number of Ag-NORs and the bromodeoxyuridine-labeling indices (r = 0.2064). These results suggest that Ag-NOR might be one of the markers for cellular proliferation in leukemia, while DNA synthesis of leukemic cells do not seem to be directly related to Ag-NOR. In order to clarify the role of Ag-NOR in leukemia, further studies are needed.


Interleukin-6 reduces the optimal growth in vitro of leukemic blast progenitors from acute myeloblastic leukemia patients.

The combined effects of five cytokines; recombinant human (rHu) granulocyte colony-stimulating factor (G-CSF), rHu granulocyte-macrophage colony-stimulating factor (GM-CSF), rHu interleukin-1 beta (IL-1 beta), rHu interleukin-3 (IL-3), and rHu interleukin-6 (IL-6) on blast colony formation in methylcellulose by leukemic blast progenitors from 10 patients with acute myeloblastic leukemia (AML) were studied. Combination of G-CSF, GM-CSF, IL-1 beta, and IL-3 stimulated maximum blast colony formation in 9 patients. Further addition of IL-6 reduced the combined effect of the four cytokines on blast colony formation. IL-6 regulates the proliferation of leukemic blast progenitors and may play an important role in the regulation of hematopoiesis.


Localization of prostatic glycoconjugates by the lectin-gold method.

The glycoconjugates of the lateral prostate were examined ultrastructurally by lectin-gold histochemistry in combination with a low-temperature embedding technique using Lowicryl K4M. The binding patterns of concanavalin A, wheat germ agglutinin, Griffonia simplicifolia, soybean agglutinin, peanut agglutinin, Ricinus communis agglutinin isolectin I, Griffonia simplicifolia isolectin B4, Ulex europaeus isolectin I and Phaseolus vulgaris agglutinin P have been documented in the subcellular compartments of the lateral prostate. The results show that the granular endoplasmic reticulum (GER) is rich in glycoproteins with mannosyl residues while the Golgi cisternae, secretory granules and microvilli are less so. The mannose (Man) and N-acetylglucosamine (GlcNAc) residues present in the GER of the epithelial cells may be associated with the initial assembly of the N-linked oligosaccharides of glycoproteins. The secretory granules exhibited different reactivities to lectins. Most of the lectin-binding sites confined to the limiting membranes may play a role in the transport of plasmalemma glycoconjugates to the apical plasma membrane. The epithelial Golgi stack is rich in GlcNAc, galactose (Gal), N-acetylgalactosamine (GalNAc) and sialic acid residues, and a compartmental organization of the Golgi stack is apparent which might be associated with the sequential addition of sugar residues to the oligosaccharides. The plasma membrane contains abundant Man, GlcNAc, Gal, GalNAc and complex carbohydrates, especially in the microvilli, and a differential lectin labelling was noted between the apical and basolateral plasma membrane. The present study showed that fucose-containing glycoconjugates were detected in the apical plasma membrane of the lateral prostate. The stromal extracellular matrices as well as the epithelial basement membranes demonstrated weak lectin reaction. Man, GlcNAc, Gal residues and complex sugars were also noted in the stromal tissues of the lateral prostate including the extracellular matrix, capillaries and smooth muscle.


Solid cell nests of the thyroid gland. Optic and immunohistochemical study at autopsies.

In order to establish the prevalence of solid cell nests (SCN) in adult thyroids, we studied 100 consecutive glands at necropsy. These were serially sectioned and stained with routine and immunoperoxidase techniques in order to detect calcitonin, carcinoembryonic antigen, thyroglobulin and keratin. SCN may be considered as normal thyroid gland components, and they share with C cells a common origin in the ultimobranchial body.


Social inhibition and motor skill performance in first, third and fifth grade children.

A systematic literature search over a period of 7 years yielded 28 articles about social inhibition with few of them addressing the relationship between social and motor functioning. Two sets of empirical data are reported. Firstly, a replication of the study performed by Zimmer in 1981 on the relationship between social inhibition and motor skill performance has been carried out with first, third and fifth grade children. Contrary to Zimmer's (1981) earlier findings with pre-school children, no relationship was found between motor skills test and social inhibition at any of the three age levels studied. Secondly, a group of children who attended extra physical education classes because of delay in motor performance (called "motoric remedial teaching") was found to score significantly lower on the motor skills test and higher on the social inhibition scale than a matched group of classmates. These findings indicate that although social inhibition appears not to be related to motor skill performance in the normal population, a significant relationship is present in a special sample of motorically delayed children.


Early symptoms in autism from family home movies. Evaluation and comparison between 1st and 2nd year of life using I.B.S.E. scale.

The analysis of 11 home movies taken by parents before the recognition of autistic or atypical disorders of their own child has confirmed the major value of this method for describing early pathological signs. Symptomatology analysis has revealed anomalies of eye contact, a deficiency and variability of emotional expression, a defect of attention and initiation of communication, as well as motor abnormalities. The comparison of the frequency of abnormal behaviour, assessed with a rating scale among three groups of children (autistic, pervasive developmental disorders and normal) revealed behavioural differences as a function of early age and diagnosis, which concern not only social and communicative behaviours, but those of emotion and attention as well. The limits and interest of this methodological approach are discussed and the possibilities of subsequently using these documents in a more complete method, such as blind examination and scoring by uninformed investigators, are suggested.


Galanin receptors and their second messengers in the lumbar dorsal spinal cord.

The ligand binding properties of galanin receptors were examined in crude synaptosomal fraction preparations of lumbar dorsal spinal cord, using chloramin-T mono-iodinated porcine Tyr26 galanin as ligand. The equilibrium binding of [125I]galanin showed the presence of a single population of high-affinity binding sites with KD = 0.6 +/- 0.2 nM in a concentration of 55 +/- 15 fmol mg-1 protein (Bmax). The N-terminal fragments galanin (1-16) and galanin (1-12) fully displaced specific [125I]galanin binding from membranes with IC50 values 6 nM and 4 microM, respectively. The C-terminal fragment galanin (17-29) did not displace [125I]galanin when applied in the concentration range 10(-11)-10(-4) M. GTP inhibited the specific binding of [125I] galanin in a concentration dependent manner, with 54% inhibition at 1 mM, suggesting that the galanin receptor found in lumbar dorsal spinal cord is G-protein coupled. Second messenger systems, through which the galanin receptor in lumbar dorsal spinal cord may exert its effect, were also studied. A galanin (10 microM) produced inhibition (58%) of the depolarization induced cGMP increase was found, whereas galanin (10 microM) did not inhibit the noradrenalin (100 microM) activated cAMP synthesis or phosphoinositide turnover in tissue slices of the spinal cord. Bilateral transection of the sciatic nerve at midthigh level 14 days prior to the binding experiment was performed, a treatment which is known to cause a dramatic increase of galanin-like immunoreactivity in the superficial layers of the dorsal spinal cord, dorsal root ganglion and in galanin mRNA levels, but no significant effect on Bmax or KD of the galanin receptor was found.


Effects of age and streptozotocin-induced diabetes on contents and effects of substance P and vasoactive intestinal polypeptide in the lower urinary tract of the rat.

The urinary bladder and urethral content of substance P and vasoactive intestinal polypeptide and the in vitro effects of the peptides on the bladder were studied at 6 weeks and 6 months of streptozotocin-induced diabetes in the rat. The results were compared with those obtained in age-matched control animals. Both short-term and long-term streptozotocin treatment induced a clearcut increase in bladder weight. Bladder substance P content was increased in both groups of diabetic animals but substance P concentration was similar in control and diabetic animals. Vasoactive intestinal polypeptide content was slightly higher in diabetic animals than in controls but vasoactive intestinal polypeptide concentration was significantly lower in the bladders from both short-term and long-term diabetic animals. The bladder contractile response to substance P was similar in all groups of animals and vasoactive intestinal polypeptide was found to be devoid of contractile or relaxatory effects in the rat bladder. No change in urethral weight was seen with diabetes. There were no clear-cut changes in the urethral contents or concentrations of substance P and vasoactive intestinal polypeptide. The study also enabled comparisons between younger (3 months) and older (9 months) rats. This comparison showed a decrease in the concentrations and contents of substance P and vasoactive intestinal polypeptide between young and older rats. The changes were seen in both the bladder and the urethra and were similar in diabetic and normal animals.


Prevention of carryover contamination in the detection of beta S and beta C genes by polymerase chain reaction.

As the polymerase chain reaction (PCR) process becomes a common tool in genetic diagnostic laboratories, prevention of carryover contamination from previous PCR amplifications has become an urgent topic. A PCR carryover prevention technique, utilizing deoxyuridine triphosphate (dUTP) and uracil DNA glycosylase (UDG), has been described recently. We report on its adaptation to a diagnostic system for detecting hemoglobin SS and SC diseases. Excellent amplification was achieved by increasing the dUTP and MgCl2 concentrations. dU-containing DNA could be analyzed by restriction endonucleases to distinguish the beta S from beta A gene using Ddel, but two other restriction enzymes, Bsu361 (replacement for MstII by the manufacturer) and CvnI, can no longer be used. The dU-containing PCR products retained their hybridization specificity with allele-specific oligonucleotide (ASO) probes for the beta A, beta S, and beta C genes. The PCR carryover prevention technique is easy to use and takes only 20 additional minutes. It should be extremely useful to genetic diagnostic laboratories where PCR is repeated daily and carryover contamination may thus lead to misdiagnoses.


Clinical relevance of immunological dissection in T-ALL: a report on 20 cases with stem cell (CD7+, CD4-, CD8-, CD1-) phenotype.

In a prospective study on 44 cases of T-cell origin acute lymphoblastic leukemia, 20 patients were found to display an immature immunophenotype (CD7+, CD4-, CD8-, CD1-) and were classified as T-stem cell leukemia (T-SCL). Twenty-four patients expressed CD4 and/or CD8 antigens on their blast cells, designated T acute lymphoblastic leukemia (T-ALL). The T-SCL subset showed a significantly higher median age, a more frequent incidence of extramedullary leukemia, a morphology L1 in most cases, and a poor response to treatment in terms of either complete remission rate or median survival duration. In addition, significant differences between the two groups were found in evaluating the number of days of blast disappearance from peripheral blood, of CR achievement, and of neutrophils and platelets recovery. We conclude that T-SCL represents a distinct clinical entity, characterized by a poor response to ALL conventional chemotherapy. Alternative therapeutic approaches should be developed for patients suffering from this form of leukemia, to modify its severe prognosis.


Postmarketing surveillance study of permethrin creme rinse.

An observational, epidemiological study was undertaken to evaluate the safety of permethrin 1% creme rinse (Nix) for treatment of head lice infestations. Thirty-seven local public health departments enrolled a total of 38,160 patients for 47,578 treatments with permethrin or other pediculicides from September 1, 1986, through January 31, 1988. Follow-up safety information was collected between 7 and 14 days following treatment via return visit or telephone contact. One hundred three adverse events were reported among 41,955 evaluable treatments. The rates of reported adverse events were 2.2 per 1000 treatments among permethrin treatments, 3.4 per 1000 treatments among lindane treatments, and 1.5 per 1000 treatments among other over-the-counter treatments. No serious, unexpected adverse events were detected in the 18,950 patients treated with permethrin. This study confirmed the safety profile of permethrin in conditions of general use, as seen in clinical trials. Postmarketing safety monitoring in public health departments of drugs used to treat public health conditions was shown to be feasible.


Evaluation of the allergenicity of infant formulas in a guinea pig model.

In the present study we evaluated the allergenicity of infant formulas using a guinea pig model. Seven groups of ten guinea pigs each maintained on a diet free of cow milk received either water or pasteurized cow milk (PCM) or a conventional cow milk infant formula (CMF) or whey hydrolysate formula (WHF) to drink. On day 37 all the animals were challenged by intravenous injection with 0.5 mL of supernatant of either PCM or WHF. The reactions to the challenge were expressed as none, mild, severe nonfatal, and fatal. A score ranging from 0 to 3 was assigned to each animal and considered in the statistical analysis; P values are expressed as comparisons to the negative water-fed control group. Animals fed WHF and challenged with WHF (group 1), showed 10% fatal reactions and 50% severe but not fatal reactions (P less than .05). When challenged with PCM (group 2), 40% showed a mild reaction. Cow milk formula-fed animals showed 10% fatal reactions and 70% severe reactions when challenged with WHF (group 3). The same challenge caused 40% severe reactions in animals fed PCM (group 4) (P greater than .05). Animals fed and challenged with PCM (group 5) showed 100% fatal reaction (P less than .01). Animals fed water showed no reaction when challenged with either WHF (group 6) or PCM (group 7). The results suggest that WHF has less anaphylactic sensitizing power than PCM and CMF.


Use of Tranilast [N-(3,4-dimethoxycinnamoyl) anthranilic acid] in secretory otitis media.

Treatment with Tranilast of 45 patients (87 ears) with secretory otitis media was studied. Tranilast was administered orally for at least 1 month. The evaluation of its effectiveness was based on changes in subjective symptoms, tympanic membrane findings, hearing level, and tympanometry. Subjects were divided into three groups: allergy group (group I), nonallergy group (group II) and deformity-disorder group (group III). The respective percentages of Tranilast efficacy for these three groups were 46.2%, 42.1%, and 10.0%, respectively. Equivalent effectiveness was demonstrated whether or not allergies were present. After Tranilast administration, the drug and its metabolites were found in MEE samples from all the patients. Anaphylatoxin (C3a, C5a) activities in the MEE were shown to decrease gradually after Tranilast administration. Tranilast may be effective for treating secretory otitis media because it directly suppresses anaphylatoxin present in middle ear effusion.


Inflammatory cells and mediator release during ragweed challenge: correlation between histamine content in nasal secretions and appearance of inflammatory cells.

The early and late phase responses in the nasal tissues exhibit release of inflammatory mediators and a mixed cellular influx in separate nasal challenges. To explore this phenomenon further, histamine concentration was determined along with characterization of cell influx during dose-dependent ragweed challenges. Ten subjects with allergic rhinitis underwent two unilateral nasal lavages using incremental 3-fold concentrations of short ragweed. Low doses of ragweed (0.016 to 0.114 units Amb a I) rarely induced cell influx (1/18 challenges), whereas moderate doses (0.432 to 1.3 units Amb a I) caused cell influxes in 7/18 and high doses (3.39 to 11.7 units Amb a I) resulted in cell influxes in 8/17. The eluent contained greater than 50% neutrophils in seven challenges; greater than 50% eosinophils in three; and a mixed pattern in six. There was a significant association between the dose of antigen and the level of histamine. Challenges with an eosinophilic influx tended to be associated with higher concentrations of histamine than neutrophilic influxes. Similar to the immediate skin response, the early allergic response in the nose demonstrated a cell influx with release of histamine. Nasal cellular inflammation therefore can occur within minutes of allergen exposure.


Characterization of a feline T-cell-specific monoclonal antibody reactive with a CD5-like molecule.

The 43 monoclonal antibody raised against feline T cells was found to react with a single-chain glycoprotein of Mr 72,000 that is present on most thymocytes, 60% of lymph node cells, 20% of splenocytes, and 45% of blood mononuclear cells. All CD4+ and CD8+ T cells were found to express the 43-reactive determinant, as did a small subpopulation of CD4-/CD8-/IgM- lymphocytes in the periphery. The 43-reactive determinant was not detected on B cells, macrophages, or other types of blood cells. The 43 antigen was phosphorylated in resting and activated T cells. Its expression was upregulated by stimulation with phorbol myristate acetate and with phytohemagglutinin. When added to concanavalin A-stimulated T-cell cultures in low concentrations, the 43 antibody was found to augment mitogenesis. The data indicate that this antibody may identify a CD5 homologue on feline T cells.


The Montgomery T-tube in terminal care.

The use of a Montgomery T-tube as a palliative measure in the treatment of patients with respiratory obstruction due to cancer is described. Six patients with terminal malignant disease presented with airway obstruction caused by direct infiltration of the trachea by tumor. The primary carcinoma originated in the esophagus in five cases, whereas one patient had metastatic carcinoma of the breast. In each case, the airway was initially secured using a rigid ventilating bronchoscope that was advanced past the area of tracheal obstruction. Tumor was removed from the lumen of the bronchoscope with suction and cup forceps. Anesthesia was continued through the bronchoscope while the trachea was exposed through a cervical incision and a window cut in the anterior tracheal wall. A Montgomery T-tube was inserted as the bronchoscope was withdrawn. This technique allowed dramatic relief of airway obstruction in all cases. The tube relieves the obstruction by stenting the airway and permits speech in most patients. The authors stress the need for frank discussion with patients and family when considering the appropriateness for this form of palliation.


Granulocyte colony-stimulating factor does not enhance endotoxin-induced acute lung injury in guinea pigs.

We studied recombinant human granulocyte colony-stimulating factor (G-CSF) in terms of its hematopoietic and neutrophil-activating effects on acute lung injury induced by endotoxin. Guinea pigs were divided into four groups: (1) saline control animals, (2) endotoxin alone, (3) cyclophosphamide (CPA)+endotoxin, and (4) G-CSF+endotoxin. A G-CSF dose of 20 micrograms/kg was given subcutaneously twice a day for 5 days. Animals were observed for 4 h after intravenously administered endotoxin (0.02 and 2.0 mg/kg) with serial measurements of complete blood counts and hemodynamics. Lung extravascular water, [125I]albumin leakage in lung tissue, and histopathologic features were examined at death. The endotoxin-alone group showed peripheral leukopenia, transient hypotension, excess lung water, increased albumin leakage, PMN accumulation in lung tissue, and gross histopathologic edema. G-CSF-treated animals showed attenuated responses in peripheral leukopenia, excess lung water, and albumin leakage in comparison with the endotoxin-alone group. No augmented responses were seen in the G-CSF group. The CPA+endotoxin group also had attenuated lung injury, which was similar to that in the G-CSF group. In conclusion, pretreatment with G-CSF tended to attenuate rather than enhance neutrophil-dependent acute lung responses to endotoxin.


Isolated bronchi from asthmatics are hyperresponsive to adenosine, which apparently acts indirectly by liberation of leukotrienes and histamine.

Bronchial hyperresponsiveness can be demonstrated in asthmatic subjects by inhalation of adenosine, but the action of adenosine at the level of the human airway smooth muscle has received comparatively little attention. We have previously observed that bronchi isolated from one asthmatic patient contracted in response to adenosine. We have therefore, during the course of a 3-yr study, further characterized the effects of adenosine in bronchi prepared from surgical specimens of lung tissue of asthmatics and of nonasthmatics. Contraction responses were always studied in vitro the same day the tissues were obtained. Bronchi from asthmatics (19 strips from six patients) were more sensitive to adenosine than were bronchi from nonasthmatics (21 strips, seven patients). In contrast, there was no difference in sensitivity to histamine or leukotriene C4 between the two groups, nor was the maximal tissue contractility different. The contractile effect of adenosine was inhibited by the adenosine A1-antagonist 2-thio-[(1,3-dipropyl)-8-cyclopentyl]-xanthine as well as by the dual A1 and A2 antagonists 8-(p-sulfo)-phenyltheophylline and theophylline. The combination of leukotriene antagonism (receptor-antagonist ICI 198,615 or biosynthesis inhibitor MK-886) and histamine antagonism (antihistamines mepyramine and metiamide) blocked the contractile effects of adenosine, suggesting that adenosine acts indirectly by liberation of leukotrienes and histamine, possibly from mast cells. The findings of increased sensitivity to adenosine in bronchi from asthmatics to our knowledge represents the first evidence of increased bronchial reactivity in vitro in asthmatics.


CD5-negative cell mediated experimental hypersensitivity pneumonitis.

Experimental hypersensitivity pneumonitis (EHP) can be transferred to Strain 2 guinea pigs by peripheral lymph node (PLN) cells cultured in vitro with antigen. The phenotype of the active cells has not been delineated. In addition, it is not known if cultured lung-associated lymph node (LALN) cells can transfer EHP. PLN and LALN cells from donor animals were cultured with a soluble extract of Micropolyspora faeni (10 micrograms/ml), and blast cells were isolated by centrifugation over Percoll gradients. Cultured PLN cells were passed through nylon wool columns, and the adherent and nonadherent fractions were tested for their ability to transfer EHP. PLN blast cell populations were depleted of CD5+ cells by treatment with monoclonal anti CD5 antibody (8BE6) and complement. Syngeneic recipients received media or LALN or PLN blast cells treated with antibody plus complement, media, or complement intravenously. Recipients were challenged intratracheally with M. faeni 48 h after the cell transfer, and they were killed 4 days later. The nonadherent PLN cell population was enriched for CD5+ (T) cells and depleted of surface immunoglobulin-positive (SIg+) cells. Treatment of the PLN blast cell population with 8BE6 and complement decreased CD5+ cells from 25 to 4% and increased SIg+ cells from 62 to 80%. All animals were maintained in HEPA-filtered air. Randomly selected microscopic fields of the lung (250/animal) were judged to be normal or abnormal without knowledge of treatment. There was a low level of pulmonary response to an intratracheal challenge of M. faeni in media recipients. There was a substantial increase (p less than 0.01) of the extent of pulmonary abnormalities in the animals receiving cultured PLN cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of somatostatin analogue and cholecystokinin receptor antagonist on bile-induced acute canine pancreatitis.

To determine whether a synthetic somatostatin analogue, octreotide, and a cholecystokinin receptor antagonist, L-364,718, may be beneficial in acute pancreatitis, 33 dogs were assigned to four groups. Each dog underwent laparotomy with injection of autologous bile into the dorsal pancreatic duct. Thirty minutes after the induction of pancreatitis, Group 1 received a subcutaneous injection of octreotide (200 micrograms/kg), Group 2 received an equal volume of the octreotide carrier, Group 3 received an hourly intravenous bolus of L-364,718 (60 micrograms/kg), and Group 4 received an equal volume of the L-364,718 carrier. Hemodynamic profiles, arterial blood gases, plasma glucose, and serum amylase were obtained before laparotomy, at bile injection, and at hourly intervals. The pancreas was removed after 8 hours for gross evaluation, measurement of water content, and histologic examination. A significant decrease in cardiac index and a significant increase in serum amylase and pancreatic edema occurred in all four groups 8 hours after the induction of pancreatitis (P less than 0.05), but there was no statistical difference between any group. Likewise, there was no difference in gross or histologic changes in the pancreas of any group. The somatostatin analogue, octreotide, and the cholecystokinin receptor antagonist, L-364,718, did not ameliorate the effects of severe, bile-induced pancreatitis in dogs.


Fabry disease: immunocytochemical characterization of neuronal involvement.

Fabry disease is an X-linked glycosphingolipid storage disease caused by deficiency of alpha-galactosidase. Storage of globotriaosylceramide, also known as ceramide trihexoside, is maximal in blood vessels but also occurs in neurons. We performed neuropathological histochemical studies on the brains and spinal cords of 2 patients with confirmed Fabry disease. Luxol fast blue-positive deposits were found in blood vessels throughout the central and peripheral nervous system and within selected neurons in spinal cord and ganglia, brainstem, amygdala, hypothalamus, and entorhinal cortex. Regions adjacent to involved neuronal groups, including nucleus basalis, striatum, globus pallidus, and thalamus, were spared. Electron microscopy showed lamellar cytoplasmic neuronal inclusion bodies. Using a monoclonal antibody reactive with ceramide trihexoside, we found more extensive neuronal deposition than evident by Luxol-fast blue staining and new areas of neuronal storage in the spinal cord and cerebral cortex. Blood vessels throughout the nervous system were strongly immunoreactive. The highly selective pattern of neuronal involvement we found suggests that glycosphingolipid exposure, uptake, or catabolism varies greatly with respect to neuronal morphology and distribution. The degree of toxicity to neurons and the clinical significance of this neuronal storage remains to be defined.


Preferential loss of striato-external pallidal projection neurons in presymptomatic Huntington's disease.

We have reported previously that striatal projection neurons are differentially affected in the course of Huntington's disease, and in a prior patient report we noted that differential loss of striatal projection neurons occurs also in patients with presymptomatic Huntington's disease. Striatal neurons projecting to the external segment of the globus pallidus or the substantia nigra show evident loss, whereas those projecting to the internal segment of the globus pallidus appear relatively spared at presymptomatic and early stages of symptomatic Huntington's disease. We now report similar findings in a second apparently presymptomatic Huntington's disease allele carrier.


Purification and characterization of O-acetylserine (thiol) lyase from spinach chloroplasts.

O-Acetylserine (thiol) lyase, the last enzyme in the cysteine biosynthetic pathway, was purified to homogeneity from spinach leaf chloroplasts. The enzyme has a molecular mass of 68,000 and consists of two identical subunits of Mr 35,000. The absorption spectrum obtained at pH 7.5 exhibited a peak at 407 nm due to pyridoxal phosphate, and addition of O-acetylserine induced a considerable modification of the spectrum. The pyridoxal phosphate content was found to be 1.1 per subunit of 35,000, and the chromophore was displaced from the enzyme by O-acetylserine, leading to a progressive inactivation of the holoenzyme. Upon gel filtration chromatography on Superdex 200, part of the chloroplastic O-acetylserine (thiol) lyase eluted in association with serine acetyltransferase at a position corresponding to a molecular mass of 310,000 (such a complex called cysteine synthase has been characterized in bacteria). The activity of O-acetylserine (thiol) lyase was optimum between pH 7.5 and 8.5. The apparent Km for O-acetylserine was 1.3 mM and for sulfide was 0.25 mM. The calculated activation energy was 12.6 kcal/mol at 10 mM O-acetylserine. The overall amino-acid composition of spinach chloroplast O-acetylserine (thiol) lyase was different than that determined for the same enzyme (cytosolic?) obtained from a crude extract of spinach leaves. A polyclonal antibody prepared against the chloroplastic O-acetylserine (thiol) lyase exhibited a very low cross-reactivity with a preparation of mitochondrial matrix and cytosolic proteins suggesting that the chloroplastic isoform was distinct from the mitochondrial and cytosolic counterparts.


Urinary excretion of diethylphosphorus metabolites in persons poisoned by quinalphos or chlorpyrifos.

The urinary excretion rates of diethyl phosphate and diethyl phosphorothioate and changes in blood cholinesterase activities were studied in fifteen persons self-poisoned either by the organophosphorus pesticide quinalphos (twelve persons) or by chlorpyrifos (three persons). The organophosphate poisoning was always indicated by a significant depression of serum and/or red blood cell cholinesterase activities. The return of serum cholinesterase activity in the range of referent values took more than 30 days and had a different course in different persons. The most rapid increase in red blood cell acetylcholinesterase activity was noted within 24 h after the first treatment with oximes Pralidoxime and/or HI-6. None of the spot urine samples, collected daily after admission of persons to hospital, contained measurable quantities of the parent pesticide. There was no correlation between the maximum concentration of total urinary diethylphosphorus metabolites normalized to creatinine and the initial inhibition of blood cholinesterase activities measured in samples collected on the day of admission to hospital. The excretion of metabolites followed the kinetics of a biphasic reaction. The half-time of urinary metabolites concentration decrease in the fast excretion phase in quinalphos poisoned persons was 5.5-14.2 h (eight persons) and 26.8-53.6 h (four persons) and in chlorpyrifos poisoned persons 3.5-5.5 h. The half-time for the slow excretion phase ranged from 66.5 to 127.9 h in all persons and for both compounds. For a given person, the rates of excretion of diethyl phosphate and diethyl phosphorothioate were about the same. However, in quinalphos poisoned persons the proportions of single metabolites in total diethylphosphorus metabolites varied with the initial maximum concentration of total metabolites.(ABSTRACT TRUNCATED AT 250 WORDS)


Aquatic hazard assessment of the organophosphate insecticide fonofos.

This study determined the acute and chronic toxicity of the organophosphate insecticide fonofos to standard freshwater aquatic organisms under laboratory conditions. Fonofos was acutely toxic to bluegill (Lepomis macrochirus), Daphnia (D. magna), and midge (Chironomous riparius) at 5.3, 2.7, and 39 micrograms/L, respectively. Three fonofos formulations (technical, 94.8% A.I.; 20G, field granular 20% A.I.; and 4E, field liquid 4#/gal A.I.) exhibited similar acute toxicities to bluegill. Exposure to fonofos delayed reproduction and decreased the intrinsic rate of increase of Daphnia during 21-d chronic exposure at the lowest tested concentration (0.08 micrograms/L). The no observable effect concentration (NOEC) for Daphnia survival was 0.42 micrograms/L; 0% survival occurred at the lowest observable effect concentration (LOEC) of 1.45 micrograms/L. The NOEC for midge emergence was 3.42 micrograms/L; only 34% emergence occurred at the LOEC of 8.24 micrograms/L. Chronic 30-d exposure of juvenile bluegills decreased growth and survival at 5.65 micrograms/L (LOEC), but no effects occurred at 2.33 micrograms/L (NOEC). The relative hazard of fonofos to aquatic life is similar to other carbamate and organophosphate corn insecticides.


Sulphydryl reactivity of the HLA-B27 epitope: accessibility of the free cysteine studied by flow cytometry.

HLA-B27 has an unpaired cysteine on or near its serologically defined spondylitis associated epitope, and it has been argued that its sulphydryl side chain may be chemically reactive. In a previous study it was shown that chemical treatment of HLA-B27 cells with the sulphydryl binding agent p-chloromercuriphenylsulphonic acid (pCMPSA) specifically reduced binding of antibodies to HLA-B27 by up to 80%, as measured in a cellular enzyme linked immunosorbent assay (CELISA). The effect of sulphydryl blockade on intact B27 cells was investigated using flow cytometry. Compared with the CELISA, inhibition required higher concentrations of pCMPSA, and the degree of inhibition produced by a greater than or equal to 30 microM solution of pCMPSA as measured by flow cytometry (median 28.9%) was significantly lower than that measured by CELISA (median 73.6%; p = 1.6 x 10(-6)). Analysis of unfixed, cell surface HLA-B27 by flow cytometry suggests that on most B27 molecules the unpaired sulphydryl site is not available. On the basis of this evidence for modification after translation, a new 'altered self' hypothesis is proposed for the part which HLA-B27 plays in inflammatory disease.


Clinical and biologic response to clozapine in patients with schizophrenia. Crossover comparison with fluphenazine.

Twenty-one patients with schizophrenia who met criteria for neuroleptic treatment resistance or intolerance participated in a crossover, placebo-controlled, double-blind comparison of long-term typical neuroleptic and clozapine treatment. Clozapine significantly reduced total as well as positive and negative symptoms in comparison with both fluphenazine and placebo. Of the 21 patients, eight (38%) showed clozapine superiority on the basis of prospective response criteria. High levels of extrapyramidal side effects during fluphenazine treatment and later onset of illness were clinical predictors of clozapine superiority. Clozapine and fluphenazine equally reduced plasma homovanillic acid levels in comparison with placebo, although fluphenazine but not clozapine increased plasma prolactin level. A striking biologic difference between clozapine and fluphenazine was clozapine's enhancement of indexes of noradrenergic activity. Superior clozapine response was predicted by low ratios of cerebrospinal fluid homovanillic acid to 5-hydroxyindoleacetic acid, consistent with the notion that balance between dopaminergic and serotoninergic systems is important for clozapine's mechanism of action.


The effects of sodium lauryl sulphate and its oxidative breakdown products on calcium phosphate precipitation and transformation.

Most commercial dentifrices contain sodium lauryl sulphate, which oxidizes upon storage. The effects of aged sodium lauryl sulphate and its oxidative breakdown products on the conversion of amorphous calcium phosphate to hydroxyapatite were studied in vitro by a pH drop method. Hydroxyapatite was identified from its X-ray diffraction pattern. With storage time, the concentration of dodecanol [CH3(CH2)11OH], a breakdown product of sodium lauryl sulphate, increased. The storage dodecanol-containing sodium lauryl sulphate accelerated the conversion of amorphous calcium phosphate to hydroxyapatite. Dodecanol mixed with sodium lauryl sulphate accelerated the conversion when added both before and after initial formation of amorphous calcium phosphate. A stored commercial dentifrice also accelerated the conversion of amorphous calcium phosphate to hydroxyapatite. It was found that the concentration of dodecanol increased 2-fold over a 2-month period.


Occurrence and distribution of different neurochemical markers in the human dental pulp.

The occurrence and distribution of neuronal markers in human premolar and molar pulps were studied immunohistochemically. In the apical and central parts of the pulp, evenly distributed, thick neurofilament-immunoreactive nerve bundles predominated, which in many instances accompanied blood vessels. In the coronal parts, especially in the pulp horns, such nerve bundles formed a subodontoblastic plexus, while thin neurofilament-immunoreactive fibres projected into the odontoblastic region. In the coronal parts of the pulp, thin, varicose, calcitonin gene-related peptide (CGRP)- and occasionally substance P-immunoreactive fibres were observed in the pulp-dentine zone and also in the vicinity of blood vessels. Vasoactive intestinal polypeptide (VIP) fibres were distributed in several nerve bundles, while single VIP fibres were seen projecting into the odontoblastic region as well as in the vicinity of blood vessels. Peptide histidine isoleucine amide (PHI)-immunoreactive fibres showed a similar distribution as VIP, but were less common. Furthermore, neuropeptide Y-immunoreactive fibres occurred occasionally around blood vessels in the inner parts of the pulp. Tyrosine hydroxylase-immunoreactive nerve fibres with a varicose appearance were observed in some nerve bundles, but were also frequently seen around and in blood vessels. In premolar pulps obtained from teeth with open apices a less dense neurofilament innervation was seen in the coronal pulp. However, no apparent difference in the occurrence and distribution of the other neuronal markers was found compared to mature teeth. The human dental pulp, thus, seems to have a rich occurrence of neuropeptides and tyrosine hydroxylase in thin, varicose fibres. However, the distribution of the fibres expressing immunoreactivity to these neuronal markers seems to be sparse in comparison to neurofilament-immunoreactive fibres.


A biochemical analysis of parotid and submandibular salivary gland function with age after simultaneous stimulation with pilocarpine and isoproterenol in female NIA Fischer 344 rats.

This analysis of physiological, biochemical and molecular changes related to aging was made in 3-, 12- and 24-month-old rats. The salivary gland weight/body weight ratio and the structural membrane proteins did not change with age for either gland, but a significant age-related decline in DNA synthesis for both glands was detected, unrelated to the hormonal responsiveness at the level of the plasma membrane. There was a marked increase in the concentration of soluble proteins in adolescent parotid gland and, for the two older age groups, in submandibular gland. The saliva flow rate was different when expressed as volume per time, as volume per time and g glandular wet weight, and/or kg body weight. The concentration of secreted proteins was not affected by age in either gland. The total amount of proteins secreted over 30 min revealed no age-related perturbation for the parotid gland, but showed a significant age-related increase in submandibular saliva. Sodium dodecyl sulphate-polyacrylamide gel analysis revealed changes in the protein bands between 39 and 50 kDa in the Coomassie blue-stained gels from 12-month-old animals. Amylase showed an initial increase (12 months), followed by a marked decline in its activity in parotid saliva. The glandular supernatant had low residual cellular amylase activity after stimulation. Therefore, secretory impairment with age after pilocarpine-isoproterenol stimulation was excluded. Analysis of total RNA showed a pronounced decrease of amylase mRNA in the parotid gland between 12 and 24 months of age. No amylase mRNA was expressed in any of the submandibular samples. For epidermal growth factor, total saliva showed a decrease with age. It seemed that the submandibular gland followed the same picture with age as the parotid gland, with a specific decline in the biosynthesis of single secretory proteins.


The effects of growth factors on DNA synthesis, proteoglycan synthesis and alkaline phosphatase activity in bovine dental pulp cells.

Platelet-derived growth factor (PDGF), insulin-like growth factor-I and -II (IGF-I and -II), acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) stimulated [125I]-deoxyuridine incorporation about 13, 6.2-, 4.6-, 3.8-, 3.1- and 1.2-fold, respectively, above control values at a concentration of 50 ng/ml. Transforming growth factor-beta (TGF-beta) decreased incorporation about 30% at the same dose. aFGF, IGF-I, IGF-II, bFGF and TGF-beta increased [35S]-sulphate incorporation 231, 71, 64, 42 and 39%, respectively, in proliferating cells, while EGF, IGF-I, TGF-beta and PDGF decreased incorporation about 30%, and aFGF increased incorporation 80% in stationary-stage culture. TGF-beta, PDGF, aFGF and bFGF caused 65-40% inhibition of alkaline phosphatase activity in proliferating and stationary cultures. These findings suggest that the proliferation of pulp cells may be stimulated mainly by PDGF and IGF-I, and the production of extracellular matrix proteoglycan may be enhanced by aFGF, IGF-I and IGF-II. Furthermore, TGF-beta, PDGF, aFGF and bFGF may regulate the differentiation of pulp cells into odontoblasts.


A comparison of bladder neck incision and transurethral prostatic resection.

Between 1978 and 1988, 108 patients underwent bladder neck incision (BNI) for bladder outflow obstruction. These patients were compared to a similar group who underwent transurethral resection of the prostate (TURP), during the same time period. Only patients with minimal prostatic enlargement (less than 10 g) with prominent bladder necks and small lateral lobes were included in the study. In addition, all patients in the resection group had a resection weight of less than 10 g on the histopathology report. Patients were followed up by means of a posted questionnaire to which 59 patients in the BNI group and 86 in the TURP group responded. Pre-operative and peri-operative data were also collected from these respondents by a retrospective case record review. This found both operations to be safe with low morbidity and mortality. BNI was better than TURP in terms of shorter operation length (P less than 0.017) and shorter duration of catheterization (P less than 0.004). No other peri-operative differences were found. Follow-up results from the questionnaire showed no significant differences in symptoms between the two groups. Similarly, there was no difference in the number of re-operations performed over the 10 year period studied. Patient assessment of their operation was initially favourable in both groups (greater than 80% patient approval) however, both treatment groups experienced a gradual drop in patient approval over the 10 year period. There were no differences in the level of approval between the BNI and TURP groups.


Influence of platelet activating factor antagonists on the protamine sulphate stimulated release of histamine from peritoneal mast cells of rats in vitro.

The phospholipase (PL)A2 inhibitor p-bromophenacyl bromide (p-BPB), the antagonists of the platelet activating factor (PAF) 3-(4-(2-chlorophenyl)-9-methyl-6H-thieno(3,2-f)-(1,2,4)triazolo(4, 3-a)(1,4)diazepine-2-yl)-1-(4-morpholinyl)-1-propanone (a thieno-triazolo-diazepine, WEB 2086) and terpene-ginkgolide B and the antihistamine with PAF antagonistic qualities 4,9-dihydro-4-(1-methyl-4-piperidinylidene)-10H-benzo[2,5]cyclo hep ta[1,2-b]thiophen-10-one (ketotifen) inhibit in the order p-BPB greater than terpene-ginkgolide B greater than ketotifen greater than WEB 2086 with decreasing activity the protamine sulphate activated histamine release from peritoneal rat mast cells (pRMC) in vitro. All compounds also inhibit the PAF induced aggregation of human platelets. The order of inhibition of the histamine release by these compounds does not agree with the order of their inhibitory activity on PAF induced aggregation of human platelets, indicating that the inhibition of the mast cell degranulation caused by PAF antagonists does not occur via PAF receptors. A generally membrane stabilizing quality of terpene-ginkgolide B and WEB 2086 may be ruled out as the cause of degranulation inhibition because none of the compounds suppresses the cytotoxic, triton X-100 induced release of histamine from pRMC. The mechanism of mast cell degranulation inhibition by PAF antagonists is unclear. An inhibition of PLA2 and/or inhibition of the Ca(2+)-activation are suggested.


Glutamate-gated ion channels in the brain. Genetic mechanism for generating molecular and functional diversity.

L-glutamate is the major excitatory neurotransmitter in the vertebrate central nervous system. Most cells are responsive to glutamate which activates cation channels with different pharmacological, kinetic, and ion permeability properties. These channels play important roles in neurotransmission, memory acquisition as well as acute and chronic disorders of the brain. The present report summarizes recent knowledge on AMPA (a-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid) receptors which mediate fast synaptic neurotransmission.


Racemic epinephrine use in croup and disposition.

The purpose of this study was to determine the effectiveness of a protocol for the outpatient management of laryngotracheitis (croup) using racemic epinephrine and steroids. The authors retrospectively reviewed fifty consecutive charts of children with croup who were treated under this protocol in the Scottish Rite Children's Medical Center Emergency Department (Atlanta, GA) and discharged to home after 2 hours of observation. Forty-seven of the 50 children had stridor at rest and/or retracting at rest on presentation to the emergency department. Forty-seven of the 50 patients did not require further medical care within 48 hours. One patient required another emergency visit and additional treatment with racemic epinephrine. Two patients were lost to follow-up. This study suggests that selected children presenting with croup and significant distress may be effectively treated with racemic epinephrine and steroids, observed for at least 2 hours, and safely discharged home.


Effects of N-methyl-N-nitrosourea on transcriptionally active and inactive nucleosomes: macromolecular damage and DNA repair.

We previously reported a separation, on an organomercurial column, of transcriptionally inactive nucleosomes (class 1) from those containing active gene sequences (classes 2 and 3). In this paper, we analyzed nucleosomal damage caused by exposure of HeLa S3 cells in suspension culture to the directly alkylating carcinogen N-methyl-N-nitrosourea (MNU). The extent and site of methylation induced by the compound in nucleosomal DNA and RNA were determined by cell incubation in the presence of [3H]MNU. The highest amount of damage was detected in DNA of class 3 nucleosomes, while RNA alkylation was comparable in all nucleosomal classes. Cellular capacity for repair of MNU-induced DNA strand breaks (estimated after a short pulse with [3H]thymidine) was found to be higher in active nucleosomal fractions (classes 2 and 3) than in the inactive fraction (class 1). Our data support the postulate that chromatin primary structure plays a role in modulating carcinogen damage to chromosomal macromolecules and in DNA strand breakage and repair mechanisms. Some of these initial steps are believed to be critical in the process of carcinogenesis.


Testicular adenoma and seminal vesicle engorgement in polyomavirus large-T antigen transgenic mice.

Six lines of transgenic mice harboring the cDNA for polyomavirus large-T antigen (PVLT) linked to the mouse metallothionein-1 promoter were isolated. The transgene was expressed in testes in all lines isolated and in testes and seminal vesicles in two lines. Three lines developed enlarged testes and seminal vesicles. Development of the phenotype was divided into three stages separable by age and pathology. In stage 1, birth to 6 mo, PVLT was expressed in testes but no pathology was noted; in stage 2, 6-10 mo, PVLT was expressed solely in testes and not in seminal vesicles, yet the seminal vesicles were enlarged; and in stage 3, 10 mo and older, both testes and seminal vesicles expressed PVLT and both were enlarged. Testes were up to sevenfold heavier and increased up to fourfold to fivefold in each dimension. Seminal vesicles were enlarged up to 20-fold as the result of an accumulation of seminal vesicle fluid. In addition to the four major proteins of seminal vesicle fluid, extra proteins, initially found in stage 2, were increased in stage 3 seminal vesicle fluid. The Leydig cell was the dominant cell type in affected testes; there were few or no normal Sertoli cells or seminiferous tubules remaining by stage 3. The Leydig cells were physiologically active, as indicated by a 8.5-fold higher testosterone level in sera from stage 3 affected mice compared with sera from age-matched normal males. PVLT was present in the nuclei of the Leydig cells and was able to confer an immortal phenotype in vitro. Formation of the Leydig cell adenoma was dependent on PVLT expression, but since PVLT expression occurred much earlier than did pathology, additional secondary factors must determine the delay in phenotype development.


Correlation of increased levels of Ha-ras T24 protein with extent of loss of gap junction function in rat liver epithelial cells.

Although it is known that cells transformed by ras and other oncogenes show reduced gap junction function, to date there has been no investigation of the quantitative relationship between intracellular levels of ras oncoprotein and loss of cell-cell communication. Using the rat liver epithelial cell line MTR6, which carries a zinc-inducible metallothionein ras T24 (MTrasT24) fusion gene, we showed a direct correlation between the accumulation of ras T24 protein and the loss of dye transfer as measured by interactive laser cytometry. After stimulation with zinc sulfate, changes in both parameters were rapid and measurable by 24 h. Similarly, there was a dose-response relationship between loss of gap junction function and increase in ras T24 protein. Northern analysis of two gap junction proteins (connexins 43 and 32) showed no differences between cells that expressed high levels of ras and control cells. These data demonstrate that the degree of loss of gap junction function is dependent on the amount of increase in ras T24 protein levels, but the mechanism by which these changes are effected remains unclear.


Multidrug resistance: prospects for clinical management.

Clinical success in the treatment of tumors with chemotherapy has significantly improved over the past several years. However, treatment failures due to drug resistance of cancer cells has remained a major problem. The classical form of multiple drug resistance is perhaps also the most common type of drug resistance, and represents the overexpression of a transmembrane glycoprotein pump (P-170) that mediates the efflux of a spectrum of structurally and functionally unrelated drugs. Here, we discuss recent evidence that support the concept that the total phenomenon of multiple drug resistance (MDR) involves several other mechanisms in addition to that underlying "classical" MDR. These include the action of other energy-dependent membrane efflux pumps, elevated levels of GSH for drug conjugation and detoxification to facilitate export, enhanced DNA repair facility, gene amplification and oncogene activation. The combination of mechanisms used by any particular cell line is variable and suggests that many of these mechanisms are independent. Successful reversal of drug resistance appears to require the identification of relevant operative resistance mechanisms. An example is the competitive inhibition of P-170 with verapamil, quinine and tamoxifen. A broadly successful strategy for killing drug-resistant cancer cells, however, could be based on either selective energy depletion of cancer cells or the permeabilization of tumor cells with an effective bypass of efflux pumps, since many mechanisms of drug resistance entail the energy-dependent export of toxins. The latter approach may be achieved via membrane lipid modifications or the introduction of membrane pores by biological or physical (electroporation) means.


Altered expression of ABO (H) carbohydrate antigens is seen in pleomorphic adenomas.

Cell surface carbohydrate antigens show changes in relation to differentiation, maturation and malignant transformation. The expression of type 2 chain ABH carbohydrate structures of the ABO histo-blood group system was investigated in 28 pleomorphic adenomas (PA) and normal parotid glands in order to study possible changes in the glycosylation pattern. The distribution of carbohydrate structures was investigated by immunohistological stainings of formalin-fixed paraffin-embedded material using monoclonal antibodies (MAbs) with well-defined specificity. A strong interindividual variation was found in the normal tissue as well as in the tumors. In normal tissue, acinus and duct cells all expressed elongated carbohydrate structures. The yoepithelial cells did not stain with any of the MAbs investigated. In the PAs, staining was seen in the ductular structures and myoepithelial cells. In contrast to normal tissue, the tumors expressed the short precursor molecule sialylated N-acetyllactosamine. Furthermore, the PAs showed loss of H and A antigens, and a reduced expression of Le(y) compared to normal tissue. The ductular structures as well as the modified myoepithelial cells expressed binary N-acetyllactosamine, which in the normal tissue could only be found in the striated and excretory ducts. Thus our study has shown that aberrant glycosylation is not only a feature of malignant neoplasms but also occurs in pleomorphic adenomas.


Substance P-like immunoreactivity in the brain of the gymnotiform fish Apteronotus leptorhynchus: presence of sex differences.

The distribution of substance P-like immunoreactivity (SPli) was charted in the brain of the gymnotiform fish Apteronotus leptorhynchus, and correlated with the circuitry underlying intraspecific electrocommunication. Cell bodies were found predominantly in the lateral hypothalamus and in certain paraventricular organs: nucleus preopticus periventricularis, anterior subdivision; anterior hypothalamus; nucleus posterioris periventricularis; nucleus recessus lateralis, medial subdivision 2; nucleus recessus posterioris and nucleus recessus lateralis, lateral subdivision. Cell bodies were also found in the rostral olfactory nucleus, ventral telencephalon (ventral and central subdivisions), the habenula, the vagal sensory and motor nuclei and in the subtrigeminal nucleus. The distribution of SPli fibers was similar in some respects to that reported for other vertebrates. SPli was found in the rhombencephalon associated with vagal afferent fibers and in the funicular nucleus (possibly related to nociception). In the diencephalon and midbrain SPli fibers were found in the habenular-interpeduncular tract, in the hypothalamus and pituitary. SPli fibers were also found in preoptic and forebrain areas. The most striking result was the sexually dimorphic SPli innervation of certain hypothalamic and septal nuclei, and of the prepacemaker nucleus (PPn), a diencephalic cell group which controls communication ('chirping') in gymnotiforms. The PPn and septal/hypothalamic nuclei were densely innervated by SPli in males but devoid of SPli in females.


Neural cell adhesion molecules modulate tyrosine phosphorylation of tubulin in nerve growth cone membranes.

Triggering neural cell adhesion molecules of the immunoglobulin superfamily with specific ligands or antibodies inhibited the phosphorylation of tryosyl residues in a subpopulation of alpha- and beta-tubulin associated with membranes from a subcellular fraction of nerve growth cones from fetal rat brain. Preincubation of these membranes with purified extracellular fragments of L1, N-CAM, or myelin-associated glycoprotein, or with antibodies directed against the extracellular domains of L1 or N-CAM, inhibited pp60c-src-dependent phosphorylation of tubulin in an endogenous membrane kinase reaction. Other proteins that affect neurite outgrowth (fibronectin, laminin, antibodies against N-cadherin) had no effect. The results suggest that cell adhesion molecules transduce cell surface events to intracellular signals by modulating the activity of protein tyrosine kinases or phosphatases in axonal membranes to influence cytoskeletal dynamics at the growth cone.


Calcitonin gene-related peptide in the human trigeminal sensory system at developmental and adult life stages: immunohistochemistry, neuronal morphometry and coexistence with substance P.

The distribution of calcitonin gene-related peptide (CGRP) has been examined by the indirect immunofluorescence technique in the Gasserian ganglion and spinal nucleus of the human trigeminal nerve. In the ganglion CGRP is present in almost 50% of primary sensory neurons, in varicose and non-varicose nerve fibres and in pericellular basket-like plexuses around non-immunoreactive ganglionic perikarya. Morphometric analysis reveals that the CGRP-positive neuronal population is heterogeneous in cell size. Observation of specimens from subjects at fetal, perinatal and adult life stages reveals that the percentage of CGRP-immunoreactive cells reaches a maximum at perinatal stages and then remains constant, declining only in old age. Pericellular basket-like nerve fibres are detectable only in fetal and pre-term and full-term newborn tissue. Coexistence between CGRP and substance P (SP) occurs, SP being present in about one quarter of the CGRP-immunoreactive neurons and CGRP being localized in a little more than half of the SP-immunoreactive neurons. However, perikarya, nerve fibres and pericellular fibres containing only one or other peptide are also present. Bundles of immunoreactive fibres and dot-like nerve terminals occur in the spinal tract and superficial and deep regions of the spinal trigeminal nucleus. A particularly dense plexus is present in the peripheral nuclear layers. Double immunostaining shows a similar regional distribution for SP. However, in inner substantia gelatinosa the density of CGRP-immunoreactive fibres is much higher than that of SP-immunoreactive ones. The results obtained add information to our knowledge of the organization of neurochemically identified neurons in the human trigeminal sensory system.


The chicken neural extracellular matrix molecule restrictin: similarity with EGF-, fibronectin type III-, and fibrinogen-like motifs.

Restrictin is a chick neural extracellular matrix protein implicated in neural cell attachment and found to be associated with the cell surface recognition protein F11. Here we show by cDNA cloning that restrictin is a large multidomain protein composed of 4 structural motifs. At the N-terminus restrictin contains a cysteine-rich segment of about 140 aa that might link restrictin monomers into oligomers. This region is followed by 4.5 epidermal growth factor-like repeats and then by 9 consecutive motifs that are similar to fibronectin type III motifs. At the C-terminus restriction is related to the beta and gamma chains of fibrinogen, including similarity to a calcium-binding segment. Restrictin shows substantial sequence similarity with tenascin (cytotactin) throughout the polypeptide, and like tenascin, it forms oligomeric structures, as revealed by electron microscopy of immunoaffinity-purified restriction. The cell attachment site of restrictin is mapped to the C-terminal region by antibody perturbation experiments.


The trkB tyrosine protein kinase is a receptor for neurotrophin-4.

Neurotrophin-4 is a novel member of the nerve growth factor family of neurotrophins recently isolated from Xenopus and viper DNA. We now report that the Xenopus NT-4 protein (XNT-4) can mediate some of its biological properties through gp145trkB, a murine tyrosine protein kinase previously identified as a primary receptor for the related brain-derived neurotrophic factor (BDNF). XNT-4 displaces 125I-labeled BDNF from binding to cells expressing gp145trkB receptors, induces their rapid phosphorylation on tyrosine residues, and causes the morphologic transformation of NIH 3T3 cells when coexpressed with gp145trkB. Moreover, XNT-4 induces the differentiation of PC12 cells into sympathetic-like neurons only if they ectopically express gp145trkB receptors. None of these biochemical or biological effects could be observed when XNT-4 was added to cells expressing the related receptors. Replacement of one of the extracellular cysteines (Cys-345) of gp145trkB by a serine residue prevents its activation by XNT-4 but not by BDNF. Therefore, XNT-4 and BDNF may interact with at least partially distinct domains within the gp145trkB receptor.


The neurotrophins BDNF, NT-3, and NGF display distinct patterns of retrograde axonal transport in peripheral and central neurons.

The pattern of retrograde axonal transport of the target-derived neurotrophic molecule, nerve growth factor (NGF), correlates with its trophic actions in adult neurons. We have determined that the NGF-related neurotrophins, brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3), are also retrogradely transported by distinct populations of peripheral and central nervous system neurons in the adult. All three 125I-labeled neurotrophins are retrogradely transported to sites previously shown to contain neurotrophin-responsive neurons as assessed in vitro, such as dorsal root ganglion and basal forebrain neurons. The patterns of transport also indicate the existence of neuronal populations that selectively transport NT-3 and/or BDNF, but not NGF, such as spinal cord motor neurons, neurons in the entorhinal cortex, thalamus, and neurons within the hippocampus itself. Our observations suggest that neurotrophins are transported by overlapping as well as distinct populations of neurons when injected into a given target field. Retrograde transport may thus be predictive of neuronal types selectively responsive to either BDNF or NT-3 in the adult, as first demonstrated for NGF.


Localization of acidic fibroblast growth factor within the mouse brain using biochemical and immunocytochemical techniques.

The localization of acidic fibroblast growth factor (aFGF) in the male mouse brain was studied with biochemical and immunocytochemical techniques. Using two peptide-based aFGF antisera directed against independent epitopes, Western gel analysis of dissected brain demonstrated significant levels of aFGF immunoreactivity in the pons-medulla, hypothalamus and cerebellum. The cortex contained much less immunoreactivity. Consistent with the biochemical data, immunocytochemical analysis with the same two antisera demonstrated that aFGF immunoreactivity is localized in neuronal cell bodies in these regions. Numerous immunoreactive neurons were observed in the reticular formation of the pons and medulla, as well as in several other brainstem nuclei and areas. Immunoreactive neurons were also present in the lateral and medial hypothalamus, and some thalamic, subthalamic and epithalamic nuclei. In the basal ganglia, immunoreactive neurons were present in the amygdala and septum. Few intensely stained immunoreactive neurons were observed in the striatum, pallidum and neocortex. Limbic cortices contained more numerous immunoreactive neurons than neocortex. These results support the concept that aFGF is present in the brain, where it is heterogeneously distributed in neuronal cell bodies in regions involved in sensory, extrapyramidal motor, limbic and autonomic functions. The results are consistent with various neurotrophic, mitogenic, and neuromodulatory functions associated with aFGF in the mammalian central nervous system.


Distribution of acidic and basic fibroblast growth factors (FGF) in the foetal rat eye: implications for lens development.

Previously we reported that, in vitro, lens cells proliferate, migrate or differentiate in response to low, medium and high concentrations of FGF respectively. To examine further the role of FGF in lens development we used immunohistochemistry to study the distribution of aFGF and bFGF in the eye of the 20 day rat foetus. Strong aFGF-like reactivity was localised in a band of cells near the lens equator which included the germinative zone where most cell proliferation occurs and the transitional zone where epithelial cells differentiate into fibres. The closely apposed inner epithelial layer of the ciliary and iridial retina also reacted strongly. Reactivity for aFGF was also found in the epidermis and in the corneal and conjunctival epithelia. In the neural retina, reactivity was found in the nerve fibre layer and in isolated cells of the inner plexiform layer. bFGF-like reactivity was found in the retinal ganglion cell layer, extra-ocular muscles and associated with endothelial cells of the hyaloid, lenticular and choroid vasculatures. Pre-digestion of sections with hyaluronidase caused loss of cell-associated reactivity but revealed strong bFGF-like reactivity in ocular basement membranes, in particular, the lens capsule. The sensitivity of this capsular bFGF localisation to heparinase indicates that bFGF in the extracellular matrix is complexed with heparan sulphate proteoglycans. The results of this study are consistent with the hypothesis that FGF plays an important role in lens development via both autocrine and paracrine mechanisms.


Effect of recombinant human granulocyte colony-stimulating factor on survival in lethally irradiated mice.

Repeated injections of recombinant human granulocyte colony-stimulating factor (rhG-CSF) to lethally irradiated mice increased the rate of animal survival. Dose modification factor was 1.20 when 4.5 micrograms/mouse of rhG-CSF was given daily for 14 days after whole body irradiation. Haematological examinations revealed that rhG-CSF increased the number of blood-circulating leukocytes, neutrophils, monocytes and erythrocytes, but not that of lymphocytes and thrombocytes. Spleen weight and number of endogenous spleen colonies were also increased by rhG-CSF treatment compared with the values for mice irradiated only.


Chromium content in human milk, cow's milk, and infant formulas.

Chromium was measured by Electrothermal Atomic Absorption Spectrometry in human milk, cow's milk, and infant formulas. The mean levels found were 1.56, 0.83, and 4.84 (13%) micrograms/L, respectively. According to our data, the daily intake for the Spanish neonates is lower than the Recommended Dietary Allowances (RDAs) (10-40 micrograms/d) for this element.


Dietary chromium intake. Freely chosen diets, institutional diet, and individual foods.

Chromium content of 22 daily diets, designed by nutritionists to be well-balanced, ranged from 8.4 to 23.7 micrograms/1000 cal with a mean +/- SEM chromium content of 13.4 +/- 1.1 micrograms/1000 cal. Most diary products are low in chromium and provide less than 0.6 micrograms/serving. Meats, poultry, and fish are also low in chromium, providing 2 micrograms of chromium or less per serving. Chromium contents of grain products, fruits, and vegetables vary widely, with some foods providing greater than 20 micrograms/serving. In summary, chromium content of individual foods varies, and is dependent upon chromium introduced in the growing, transport, processing, and fortification of the food. Even well-balanced diets may contain suboptimal levels of dietary chromium.


Glucose tolerance factor potentiation of insulin action in adipocytes from rats raised on a torula yeast diet cannot be attributed to a deficiency of chromium or glucose tolerance factor activity in the diet.

The nature of the dietary component responsible for adipocytes having the ability to respond to Glucose Tolerance Factor (GTF) was investigated. Rats were raised on either a control diet or one of three diets differing only in the protein source (torula yeast, brewer's yeast, or casein). Only in adipocytes from rats fed the torula yeast diet did a GTF fraction prepared from brewer's yeast potentiate the action of suboptimal concentrations of insulin in the incorporation of label from D-[1-14C]-glucose and D-[U-14C]-glucose into CO2 and fatty acids. It was concluded that this potentiation was not the result of a deficiency of GTF activity in torula yeast, because a GTF fraction prepared from torula yeast had similar insulin potentiating activity. Differences in response among diets were not owing to differences in levels of amino acids or owing to concentrations of 22 (Al, As, B, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mo, Na, Ni, P, Pb S, Se, Si, Sn, Sr, Zn) of the 23 trace elements investigated. The level of Mn was low in all diets, but particularly low in the torula yeast diet. Mn deficiencies have previously been implicated in perturbations of glucose metabolism, so that it is possible that this deficiency may be responsible for the effects attributed to the torula yeast diet.


A study of chromium in human cataractous lenses and whole blood of diabetics, senile, and normal population.

Chromium (Cr) is of known biological importance, necessary for the maintenance of normal glucose metabolism. There is a lower level of blood Cr concentrations in cases of diabetes. Diabetes carries a risk of cataract development, so the potential effects of Cr on the eye may need to be studied in more depth. The presence of this trace element in both normal and cataractous human lenses has to our knowledge not been investigated so far. The concentration of total Cr in 61 human lenses and 38 blood samples was determined by electrothermal atomic absorption spectrometry with Zeeman effect (EAASZ). Analysis of the levels of Cr in human lenses shows a significant difference between normal and diabetic populations, and an absence of difference between senile and diabetic populations.


Total chromium in the human lens. Determination with Zeeman electrothermal atomic absorption spectrometry following mineralization in a mini-autoclave.

A simple, quick procedure has been developed for preparation of human lenses for total chromium (Cr) analysis by electrothermal atomic absorption spectrometry (EAAS). This procedure involves wet ashing in a mini-autoclave with HNO3 and H2SO4. Recovery was 101.6%. This procedure is simple to carry out, does not generate corrosive fumes, and minimizes contaminations. Measurements obtained by this method give values similar to those found previously by more cumbersome methods. It can be conveniently used to prepare biological samples for ultratrace analysis. The mean Cr concentration in human lenses varied between 0.345 +/- 0.147 micrograms/g dry wt in the normal population and 0.205 +/- 0.160 micrograms/g dry wt in cataractous lenses.


Chromium in the elderly.

Several studies indicate that glucose tolerance improves and lipid levels decline in the elderly after supplementation with Cr-rich brewer's yeast or inorganic Cr. Other studies report equivocal results or no changes. Interpretation of these investigations is hampered by 1. Lack of a marker to identify Cr-deficient people 2. Artifactually high levels of dietary and body Cr, owing to inadequate analytic techniques and 3. The interplay of chronic health problems, medications, institutionalization, and dietary practices. Investigators have studied the effects of aging on Cr in the body. In the rat, tissue retention of 51Cr decreases, and organ distribution changes with age. In humans, plasma Cr levels of healthy elderly subjects are not different from those of young adults. There is no evidence of malabsorption in aged humans or animals. Nevertheless, higher urinary Cr losses are reported in elderly people. These data suggest that Cr retention may decrease with aging and that aging may alter Cr metabolism. Diets of many healthy elderly people contain less than 30 micrograms Cr. Two elderly men living under controlled conditions maintained Cr balance with 37 micrograms/d. However, these levels may be insufficient during the stresses and illnesses associated with aging.


Toxic effects of chromium and its compounds.

Chromium was discovered in 1797 by Vauquelin. Numerous industrial applications raised chromium to a very important economic element. At the same time, with the development of its uses, the adverse effects of chromium compounds in human health were being defined. Trivalent chromium is an essential trace element in humans and in animals. Chromium as pure metal has no adverse effect. Little toxic effect is attributed to trivalent chromium when present in very large quantities. Both acute and chronic toxicity of chromium are mainly caused by hexavalent compounds. The most important toxic effects, after contact, inhalation, or ingestion of hexavalent chromium compounds are the following: dermatitis, allergic and eczematous skin reactions, skin and mucous membrane ulcerations, perforation of the nasal septum, allergic asthmatic reactions, bronchial carcinomas, gastro-enteritis, hepatocellular deficiency, and renal oligo anuric deficiency. Prevention of occupational risks, biological monitoring of workers, and treatment of poisoning are also reported.


Hair, serum, and urine chromium concentrations in former employees of the leather tanning industry.

In this study, we measured concentrations of Cr in hair, serum, and urine of men who were previously employed in the leather tanning industry. Concentrations of Cr in hair and serum were significantly lower than corresponding values obtained during their employment and were comparable to levels obtained for controls in a previous study. These results suggest that Cr III accumulated from employment in the leather tanning industry does not result in long-term elevation of Cr concentrations in the body.


Clinical implications of trace elements in endocrinology.

The implications of essential trace elements in endocrinological processes, mainly thyroid function, growth, gonadal function, adrenal hormones, prolactin, glucose homeostasis, calcium-phosphorus metabolism, and thymulin activity, are reviewed. Most concerned elements in this field include iodine, zinc, selenium, copper, chromium, manganese and vanadium. The minerals are powerful modulators of several physiological functions that can be considerably perturbed in deficiency states. The resulting biochemical and clinical modifications can be prevented and/or corrected by adequate supplementation. Sometimes, however, they act like pharmacological agents when their beneficial effects are not the result of a correction of a nutritional deficiency state. Their potentialities as therapeutic agents are perfectly described in many cases, but some indications deserve further investigations.