Carbohydrate binding properties of the envelope glycoproteins of human immunodeficiency virus type 1. Here, we confirm and extend our previous findings on human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein N-acetylglucosaminyl binding properties. We show the occurrence of saturable, temperature, pH, and calcium dependent carbohydrate-specific interactions between recombinant precursor gp160 (rgp160) and two affinity matrices: D-mannose-divinylsulfone-agarose, and natural glycoprotein, fetuin, also coupled to agarose. Binding of rgp160 to the matrices was inhibited by soluble mannosyl derivatives, alpha-D-Man17-BSA and mannan, by beta-D-GlcNAc47-BSA and by glycopeptides from Pronase-treated porcine thyroglobulin, which produces oligomannose and complex N-linked glycans. Glycopeptides from Endoglycosidase H-treated thyroglobulin partially inhibited rgp160 binding, as did the asialo-agalacto-tetraantennary precursor oligosaccharide of human alpha 1-acid glycoprotein for binding to fetuin-agarose. beta-D-Glucan and beta-D-Gal17-BSA had no or only limited effect. Also, surface unit rgp120 specifically interacted with fetuin-agarose and soluble fetuin, but in the latter case with a twofold reduced affinity relative to rgp160. After affinity chromatography, rgp160 was specifically retained by the two matrices and eluted by mannan in both cases, while rgp120 was not retained by fetuin-agarose but only eluted as a significantly retarded peak, which confirms its specific but weak interaction. Thus, rgp160 interacts with both oligomannose type, and the mannosyl core of complex type N-linked glycans, and its gp120 region plays a role in this interaction.(ABSTRACT TRUNCATED AT 250 WORDS)


[The treatment of advanced gastric cancer: total gastrectomy "extended" as needed]. Extended resections do play a definite role in the surgical approach to advanced gastric malignancy. Local extension of distant spread of the tumor is no longer to be considered as a contraindication to aggressive surgery based on the evidence that even for palliation, extensive operations, when feasible, have proved to be beneficial in preventing complications related to the natural history of the disease. Over a nine-year period 105 patients with gastric cancer were surgically treated. Out of the 88 patients undergoing tumor excision, 53 were submitted to total gastrectomy (there were 35 males and 18 females, mean age 63 years). This procedure was considered potentially curative in 37 cases and palliative in 16. In 8 of the patients treated with palliation. An extensive procedure, including splenectomy in 7 cases (1 of which with concomitant distal pancreatic resection) and a colonic resection in 1 case, was undertaken. Alimentary continuity following total gastrectomy was restored by jejunal interposition (Mouchet-type reconstruction) in 3 cases, Roux-en-Y esophagojejunostomy in 7 and simple: end-to-side esophagojejunostomy (Sweet and Allen-type) in 6. No postoperative complications have occurred and mean survival time was 8 months with a maximum of 20 months in a patient with a good nutritional status and quality of life (Karnofsky index 78%) who developed no complications related to tumor recurrence.(ABSTRACT TRUNCATED AT 250 WORDS)


[The effect of convaflavin on lipid peroxidation in the heart in experimental thyrotoxicosis]. The effect of convaflavin in doses of 30 and 45 mg/kg on lipid peroxidation in the heart was studied in white rats with experimental thyrotoxicosis. A noticeable decrease of the concentration of diene conjugates and malonic dialdehyde was revealed as was activation of catalase and peroxidase under the influence of convaflavin administered in a dose of 45 mg/kg.


[The effect of sodium salicylate and phenobarbital on the anti-arrhythmia properties of novocainamide and acetylnovocainamide]. CaCl2 and aconitic models of arrhythmias were reproduced in experiments with rats. Sodium salicylate was found to decrease the preventive effect of ecetylnovocainamidum and increased the effect of novocainamidum on ECG changes. Sodium salicylate diminished the preventive effect of acetyl-novocainamidum by elevating myocardial Na+ concentrations, favoured the decrease in myocardial K+ levels, eliminated the preventive effects of novocainamide and acetylnovocainamide by altering myocardial energy metabolism in CaCl2-induced arrhythmia and improved the preventive effect of acetyl-novocainamide on these parameters in aconitic arrhythmia.


[Surgical procedure in an extensive un-operable decaying breast cancer]. The article indicates necessity of palliative surgical treatment of ulcerating and decaying breast cancers. Ulceration and decay of tumour of the breast make worse the quality of life of the afflicted. The operation like simple mastectomy liquidates the threat of haemorrhage and prevent intoxication from decaying tumour. In the period from 1977 to 1991 year the group of 1075 women with breast cancer were operated. In this number there were 44 women with decaying breast cancer. Sometimes there are difficulties in closing of the operative wound, when women with very extensive tumour ulceration are treated and then reconstructive methods are needed (transposition of a simple skin flap from the opposite breast, a latissimus dorsi myocutaneous flap or omental transposition from the abdomen to the chest wall). All these operated women survived the operation and left surgical ward.


Intraventricular administration of substance P induces unattached Purkinje cell dendritic spines in rats. Substance P was infused in the lateral ventricles of twenty Lewis rats for twenty days. The animals under the influence of the substance P demonstrated grooming of the head, the body and the forepaws. On the twentieth day the animals were sacrificed and the cerebellar cortex was processed for electron microscopy. The ultrastructural analysis revealed that although the granule cells, the parallel fibers and the systems of the afferent fibers were intact, numerous unattached Purkinje cell dendritic spines were seen embedded in the soma of the astrocytes, demonstrating postsynaptic differentiation. Numerous unattached spines of the secondary and tertiary dendritic branches of the Purkinje cells were also seen in the molecular layer surrounded by astrocytic sheath. Free unattached spines were also seen not surrounded by any astrocytic process, which did not demonstrate any postsynaptic specialization. The development of unattached Purkinje cell dendritic spines, in an otherwise intact cerebellar cortex, following the intraventricular administration of substance P, suggests that it may act as local growth factor, enforcing the preprogrammed-capability of the Purkinje cells in developing new synaptic surfaces.


[Value of alpha subunit assay in pituitary pathology]. We evaluated serum alpha-subunit concentrations in 72 patients bearing pituitary adenomas. We conclude that: 1) alpha-subunit hypersecretion is found in all patients with TSH secreting pituitary adenoma (n = 10). Two patients have a predominant secretion of alpha-subunit as compared to TSH secretion. 2) As concerns gonadotropin secreting pituitary adenomas (n = 3), all patient have elevated serum alpha-subunit levels with increased FSH and LT concentrations in 2 cases and 1 case respectively. 3) In prolactinomas (n = 14), alpha-subunit concentrations are not significantly different from that of normal subjects. 4) In 11 acromegalic patients, alpha-subunit hypersecretion is found in 2 patients only with GH-alpha and GH-PRL-alpha secreting pituitary adenomas. 5) Among 34 patients with nonsecreting adenomas, 8 have elevated alpha-subunit concentrations (24%). Positive immunocytochemical staining is found in 70% most often with gonadotropins (55%). Only a few pituitary tumors with positive alpha-subunit immunocytochemical staining have elevated serum alpha-subunit levels. At least, the measurement of basal circulating alpha-subunit levels is very useful in the follow-up of patients with nonsecreting adenomas. In our study, medical treatments including bromocriptine and somatostatin analogues have been found effective in patients with alpha-subunit hypersecretion. Further investigations are necessary to prove if such treatments could control tumoral growth and could prevent recurrences.


Immuno-histological characterization of OVS1 and OVS2 monoclonal antibodies recognizing human ovarian mucinous cystadenocarcinoma. OVS1 and OVS2 monoclonal antibodies (MAbs) were established by fusing murine myeloma cell line NS1/1-Ag4-1 with mouse spleen cells immunized with fresh human ovarian mucinous-cystadenocarcinoma tissue. The selection of the MAbs was assayed by an immuno-histological (streptavidin-biotin) staining of the specific antigen antibody reaction localized on frozen sections of the same tumor. Other paraffin sections and established cell lines were also screened by immuno-histological staining in order to characterize the specificity and sensitivity of these two MAbs. OVS1 MAb showed 96% specificity and 67% sensitivity to mucinous cystadenocarcinoma with no cross reactions to normal tissue, benign tissue, other cancers, or any established cell lines. OVS2 MAb revealed only 8% specificity but 78% sensitivity to mucinous cystadenocarcinoma, however, a cross reaction to some normal and benign tissues or other cancers was shown. The data suggested that OVS1 and OVS2 MAbs could be used in combination to detect ovarian mucinous cystadenocarcinoma.


[Tissue plasminogen activator, its inhibitor and other parameters of fibrinolysis in blood of patients operated for mild hypertrophy of the prostate]. An increased blood fibrinolytic activity manifested by increased tissue plasminogen activator (t-PA) and decreased tissue plasminogen activator inhibitor (PAI-1) and increased FDP levels are seen in 40 patients with mild hypertrophy of prostate. Surgical treatment increased blood fibrinolytic activity manifested in the increase in t-PA, decrease in PAI-1, shortening of ELT, increase in FDP, and decrease in plasminogen and 2-AP activities. Blood fibrinolytic activity was the highest immediately after surgery with tendency to the gradual normalization. Positive ethanol test and decrease in thrombocyte count indicate and activation of blood clotting system induced by the tissue thrombo-elastins released during surgery. Subclinical DCI with the secondary increased fibrinolysis activation is present in patients with mild hypertrophy of the prostate both prior to and after surgery.


Use of endotoxin antigens in enzyme-linked immunosorbent assay for the diagnosis of P. pseudomallei infections (melioidosis). An enzyme-linked immunosorbent assay (ELISA) with endotoxin preparations of P. pseudomallei as antigen was developed for detection of IgG antibodies specific to melioidosis. Forty-seven sera of bacteriologically confirmed melioidosis patients, 55 non-melioidosis sera and 50 sera of healthy blood donors from non-endemic areas were subjected to this assay in comparison with indirect hemagglutination assay (IHA). The data were treated by receiver operating characteristics analysis. The sensitivity, specificity and accuracy in this ELISA were 95.7%, 94.2%, and 94.7%, respectively, with cut-off value of OD = 0.312 at 490 nm. Meanwhile, those in IHA were 81.0%, 91.4%, and 88.1%, respectively, with a cut-off value of > or = 1:160. From these results, the ELISA was judged to be more reliable than IHA as the seroassay for diagnosis of melioidosis.


Immunolocalization of smooth muscle-like cells in the quail ovary. We localized alpha-smooth muscle actin (alpha-SMA) in the quail ovary, using the peroxidase-anti-peroxidase technique. Special attention was paid to the influence of fixation on the immunoreactivity of the antigen. The immunostaining of alpha-SMA largely depended on the nature of the fixative. The antigen could most successfully be localized in ovaries fixed in Carnoy's fluid. We also localized alpha-SMA and desmin in semi-thin glycol methacrylate sections of the pre-ovulatory follicle, using the immunogold-silver staining method. The sections were pretreated with Lugol's iodine or sodium metaperiodate to enhance the immunoreactivity. alpha-SMA was demonstrated in the cells of the chordae, the tunica albuginea, and the theca externa of each follicle. These structures were inter-connected, forming an ovarian suspensory apparatus. The thecal cells of prelampbrush follicles also expressed alpha-SMA. In the wall of the pre-ovulatory follicle, desmin was found in the cells of the chordae and the tunica albuginea, and in a few cells of the theca externa. In the theca interna, desmin, and sometimes alpha-SMA, was observed in cells adjacent to the endothelium of sinusoids, which are probably pericytes. Our results support the hypothesis that in birds the ovarian follicles possess a thecal contractile system, that is presumably involved in the ovulatory process.


Does microwave irradiation have other than thermal effects on histological staining of the mammalian CNS? A light microscopical study of microwave stimulated staining under isothermal conditions in man and rat. The question whether or not microwave irradiation exerts other than thermal effects on histological staining is still a matter of controversy. The present study was undertaken to reveal or reject such a so far hypothetical non-thermal irradiation effect. A device was developed, which enables exposure of histological sections or tissue pieces to microwave irradiation under isothermal conditions, i.e. with synchronous removal of the internal heat produced. Three classical neuroanatomical staining methods were tested on human and rat CNS. As control, identical procedures were performed without simultaneous microwave irradiation. The experiments were performed at three different temperature levels ranging from 5 to 50 degrees C. In none of the cases studied was a light microscopically appreciable difference observed between the microwave and non-microwave versions of a stain at the same temperature. The hypothesis of a separate non-thermal effect of microwave irradiation on histological staining is therefore rejected.


FK506 reverses adriamycin resistance in a multidrug-resistant human leukemia cell line. We have examined the effect of FK506 on the Adriamycin sensitivity of the multidrug resistant human chronic myelocytic leukemia cell line (K562/ADM). In K562/ADM cells, 1.0 microgram/ml FK506 reversed the resistance of Adriamycin, and increased the IC50 value for Adriamycin up to 17 fold. However, IC50 value for the parent cells (K562) increased only 1.5 fold. By cell cycle analysis, the accumulation in late S-G2M phase was confirmed on K562/ADM cells, treated with 1.0 microgram/ml FK506 and low-dose of Adriamycin. Cyclosporin A (CsA) could also restored the Adriamycin sensitivity in the K562/ADM cells, as previously reported. 1.0 microgram/ml FK506 as well as CsA significantly increased radioactive Adriamycin accumulation in K562/ADM cells and blocked [3H]azidopien photoaffinity labeling of P-glycoprotein. These results suggest that 1.0 microgram/ml FK506 could reverse the Adriamycin resistance in a MDR human leukemia cells through the interaction with P-glycoprotein.


Francis Burian--founder of the treatment of facial clefts in Czechoslovakia. Prof. F. Burian, the founder of Czechoslovak Plastic Surgery, belonged to the world-wide scale among the most prominent pioneers of this field. His main interest and professional concern was devoted to the problems of facial clefts. The present communication reviews the gradual development of surgical repair of cleft lip and palate from the early twenties to the late fifties at the Department for Plastic Surgery in Prague, headed by Prof. F. Burian.


Configuration of facial profile in adults with cleft lip with or without cleft palate. X-ray cephalometric studies were carried out in 114 adult males with cleft lip and with or without cleft palate. According to the type and extent of the cleft they were subdivided into 4 groups and were compared with a control group of 50 normal males matched in age. Investigated were the parameters of the skeletal and soft facial profile. The results showed that cleft lip alone is associated with deviations of local character concerning only soft tissues within the oronasal region. The ascertained deviations included a flattening of the nose, reduction of the height, concavity and prominence of the upper lip, increased height of the upper lip vermilion and a more horizontal slope of the columella leading to a reduction of the nasolabial angle. Cleft lip and palate was associated with deviations of global character related predominantly to the extent of retrusion of the upper and lower jaw. The skeletal profile was altered and its deviations were reflected by changes of the soft profile. Of the deviations of soft tissues per se were most important the flattening of the nose and reduction of the height and thickness of the upper lip, which underlined the presence of retrocheilia. Maxillary retrusion was more marked in complete than in incomplete unilateral clefts, while mandibular retrusion and maxillary dentoalveolar retroinclination were more marked in bilateral than in unilateral clefts. Occlusion was always impaired. In unilateral involvement, especially in complete clefts the more horizontal slope of the columella resulted in a marked reduction of the nasolabial angle. Incomplete clefts, similarly as cleft lip alone were not associated with a reduction of thickness of the upper lip and showed an increase of the vermilion height. Because of the persisting protrusion of the premaxilla bilateral clefts were accompanied by only a slight flattening of the skeletal profile and by an excessive nasal depth after the prolongation of the columella. The nasolabial angle was unchanged. The concavity of the upper lip was reduced in complete unilateral and bilateral clefts. Certain characteristics of the oronasal region disclosed a similarity between incomplete cleft lip and palate (in unilateral involvement) and cleft lip alone, however global deviations (due to skeletal changes) were identical with those recorded in complete clefts.


Configuration of facial profile in adults with isolated cleft palate. X-ray cephalometry was used for the assessment of the configuration of the skeletal and soft tissue profile in 81 adult males with isolated cleft palate. They were assessed in three subgroups, i. e. with complete and incomplete clefts and with clefts of the soft palate alone and were compared with a matched group of controls consisting of 50 normal males, as well as with the situation in the earlier examined series with unilateral cleft lip and palate. In isolated cleft palate both the upper and lower jaws were retruded. The maxilla was most markedly retruded in incomplete clefts and most slightly in clefts of the soft palate alone, while the mandible showed the smallest degree of retrusion in complete clefts. The configuration of the skeletal profile was similar as in unilateral cleft lip and palate, yet positive overjet was on the average restored in all types of isolated cleft palate. A more favourable configuration of the profile was present in patients with cleft soft palate alone, in the absence of an impairment of sagittal jaw relations. Changes in the configuration of the soft profile were due mainly to skeletal deviations. The retrusion of skeletal framework of the middle face was to a large degree masked by the larger thickness of the upper lip, in complete clefts also of the lower lip. Therefore the prominence of the upper lip was only slightly impaired and slight flattening of the face occurred only in complete clefts. The more horizontal slope of the columella resulted in a reduction of the nasolabial angle. Some deviations from the described pattern occurred in clefts of the soft palate alone (steeper slope of the upper lip with an unchanged nasolabial angle, normal thickness of the upper lip a. o.). The findings are suggestive of a differing facial physiognomy in isolated cleft palate.


Late results of the surgical treatment in unilateral complete cleft lip and palate. Occlusal and craniofacial characteristics. Sixty-one UCLP patients at 17-20 years of age treated surgically in childhood by M. Krauss were invited for check-up examination to evaluate their occlusal and craniofacial characteristics. Dental study casts were used to determine the dental arch widths and lengths according to Moorrees points and to classify the occlusion according the descriptive and numerical methods. Conventional distances and angles were measured on 45 radiographs. The maxillary canine arch width was smaller by 4 mm than those of the control group, while the maxillary molar arch width did not differ significantly from the normal value. In contrary, the mandibular molar arch width was considerably greater in the studied group than in the control one. The frequency of anterior crossbite was 7%, the lateral crossbite on the cleft side 30%, bilateral crossbite 26% and the crossbite in all segments 16%. In 21% of cases, no crossbite was observed. These results were confirmed by occlusal scoring. The cephalometric analysis showed a decreased maxillary horizontal length and maxillary protrusion angle when compared with the control group, while the anterior upper facial height and incisor inclination angles did not differ significantly. The midface constituted in men 42.5% and in women 42.6% of the total face height.


Skin necrosis, a rare complication of coumarin therapy. Coumarin congeners are frequently being prescribed in vascular surgery. The complication most often seen in haemorrhage. A less known complication is necrosis of skin and soft tissues. This rare complication is potentially lethal. The etiology is unclear, a relation with protein C deficiency seems likely. Early treatment with vitamin K and heparin may prevent the skin necrosis. After necrosis has occurred, surgical intervention is usually necessary.


Multiple uses of the free radial forearm flap. Presenting clinical cases, the authors show the variety of possible uses of the free radial forearm flap. They employ this sensory flap at sites of increased load such as the area of the planta pedis and in thumb reconstruction. Other potential uses include oesophageal and penile reconstruction.


The technique of osteosynthesis in replantations of the upper limb. The authors present their experience in osteosynthesis in replantations in the area of the wrist in 19 patients. For osteosynthesis they use external fixation device, type Poldi 7, which guarantees the stability not only during the microsurgical procedure but also over the whole period of healing. They point out the importance of the resection of both bones ends, i.e. the resection of the comminuted zone, as well as the importance of an extensive fasciotomy. In all patients but one healing was achieved without infection. The authors consider external fixation for the method of choice in the replantation in the wrist area.


Use of Becker's prosthesis in breast reconstruction. The method of breast reconstruction using a submuscular permanent expander is rapid and simple, imposing minimal stress on the patient. Using it, it is possible to obtain good shape and size of the reconstructed breast; the ptotic position remains to pose a problem. This modality is useful especially in bilateral involvement and in all patients with a concomitant disease or another risk factor.


Staphylococcal L-asparaginase: catabolic repression of synthesis. The regulation mechanism of production of staphylococcal L-asparaginase was investigated. The role of cAMP in regulation of its synthesis was confirmed. Production of L-asparaginase from S. aureus NCTC 4135 was inhibited by all carbon sources, mono- and disaccharides added to the growth medium. The strongest inhibition was caused by saccharose and maltose, whereas weaker by galactose, lactose, mannitol and mannose. It was found that exogenous cAMP in the presence of carbon sources stimulated synthesis of the investigated enzyme.


Proteinases of Streptomyces fradiae. II. Specificity. It has been shown that extracellular proteinases synthesized by a keratinolytic strain of S. fradiae are characterized by diversified activity in the decomposition of both proteins and synthetic substrates. Among the six proteinases isolated, apart from the ones dominating and having relatively low specificity, there are two enzymes characterized by narrow catalytic abilities--extremely similar to those of trypsin. These proteinases intensively degraded all the trypsin substrates studied, but they were inactive or showed slight activity toward others. They were also highly sensitive to such specific inhibitors of trypsin as TLCK, SBTI and TIO.


Proteinases of Streptomyces fradiae. III. Catalytic and some physico- chemical properties of keratinolytic proteinase. It has been shown that keratinase--proteinase PIV, the main enzyme of the proteolytic system of S. fradiae, is characterized by high effectiveness in its action on AcAla3OMe--exceeding elastase in catalytic effectiveness several times. This proteinase also cleaves ester and amide bonds formed by the residues of aromatic and basic amino acids, but with a lower effectiveness than chymotrypsin or trypsin. It has also been shown that proteinase IV is a typical serine enzyme highly sensitive to DFP, acting in strongly alkaline pH (about 11.2), with molecular weight 24 kDa and does not contain cysteine.


Isolation and screening of glucose oxidase producing microorganisms from natural sources. Among 1486 mould strains isolated from natural sources (screened for extracellular glucose oxidase) only 119 (Aspergillus and Penicillium) showed this enzyme activity. As the best glucose oxidase producer, A. niger 0-1 was isolated from decaying tree. The dynamics of glucose oxidase synthesis in A. niger 0-1 during its culture by submerged method show that the intracellular activity of this enzyme is 10-times higher than its extracellular level. Some properties of the crude glucose oxidase preparation, isolated from the postculture liquids by lyophilization, were examined.


Relationship between citric acid production and accumulation of phytate-degrading enzymes in Aspergillus niger mycelia. The ability of eight strains of Aspergillus niger to produce citric acid by the solid surface method were found to correlate with their capabilities to synthesize intracellular enzymes which degrade phytates (phytase and acid phosphatase). Another high correlation was observed between phytase and acid phosphatase activities bound to the cell walls of mycelia.


Comparison of dentrification by Paracoccus denitrificans, Pseudomonas stutzeri and Pseudomonas aeruginosa. The course of denitrification of nitrate, nitrite and both compounds together by static cultures of Paracoccus denitrificans, Pseudomonas stutzeri and Pseudomonas aeruginosa was studied. These strains represent three different types of denitrification: 1. reduction of nitrate to gaseous nitrogen without accumulation of nitrite (P. denitrificans); 2. partial accumulation of nitrite in growing cultures during reduction of nitrate to gaseous nitrogen (P. aeruginosa) and 3. two-phase denitrification that includes reduction of nitrates at the very beginning of the process, and then, after depletion of the former, the reduction of nitrates to gaseous nitrogen (P. stutzeri). These observations differ from the results reported in the literature and possible reasons are discussed.


Cognitive effects of mild head injury in children and adolescents. A comprehensive review of recent neuropsychological studies of mild head injury (MHI) involving children and adolescents is presented. The seminal work of Rutter and his colleagues is reviewed. An alternative conceptualization of MHI as proposed by various researchers is elaborated and further research investigating the cognitive sequelae of MHI is reviewed. MHI is discussed within the context of development and information processing models. Finally, the sequelae of MHI are reviewed with respect to academic functioning. Methodological problems inherent in studies of MHI are identified and discussed. The studies reviewed here support the conclusion that both the cognitive and emotional consequences of MHI should receive serious evaluation.


Myelin palingenesis. 1. Electron microscopical localization of myelin/oligodendrocyte proteins in multilamellar structures by the immunogold method. The issue of the capacity of mature oligodendrocytes to remyelinate naked axons has not been totally resolved. The impression is that for this to happen, oligodendrocytes have to undergo cell division. We are interested in providing an answer to the question: can oligodendrocytes myelinate more than once? To address this question, we are using a model system consisting of pure cultures of postmyelination oligodendrocytes. We have previously shown that these cells have plasticity and are able to regenerate, but for this to occur they need a signal. In vitro, this signal is provided by interaction with a positively charged substratum and a serum factor(s). Over time in culture, without cell division, oligodendrocytes assemble multilamellar structures that ultrastructurally resemble myelin. We have named this process myelin palingenesis--rebirth--to distinguish it from remyelination. In order to ascertain the significance of the regenerative process observed in vitro and its predictive value as to the capabilities of oligodendrocytes to ensheathe axons, we undertook an ultrastructural and immunocytochemical characterization of the multilamellar membranes to assess whether they contain all the myelin-characteristic proteins, i.e., myelin basic protein, proteolipid protein, 2',3'-cyclic nucleotide 3'-phosphodiesterase and myelin-associated glycoprotein. We have used antibodies against these proteins as immunocytochemical probes in conjunction with the immunogold method at the electron microscopic level. Oligodendrocyte cultures were processed for electron microscopy. Blocks were serially sectioned parallel to the culture plate at 0.09-micron spacings. Staining of cells was done prior to embedding. Analysis of these micrographs brought to light the existence of membrane-membrane interaction between multilamellar structures; this is reminiscent of the interactions observed in situ between myelinated axons. We have found that the membranous structures possess all the myelin-characteristic proteins. Furthermore, based on the accessibility of these proteins to the antibodies, e.g., whether or not permeabilization was required, we can surmise that these proteins are incorporated into the membranous structures with the same orientation as in myelin. These results, in conjunction with our earlier work, show that mature oligodendrocytes stripped of their myelin are able to regenerate and reassemble multilamellar membranes that have all the myelin proteins. It remains to be proven that when provided with axons, these oligodendrocytes will generate typical myelin.


Carbohydrate moieties of myelin-associated glycoprotein, major glycoprotein of the peripheral nervous system myelin and other myelin glycoproteins potentially involved in cell adhesion. The myelin-associated glycoprotein (MAG) and the major glycoprotein of the peripheral nervous system myelin (P0) are two members of the family of cell adhesion molecules (CAMs). A role in cell adhesion of the carbohydrate moiety of these molecules has been attributed to the presence of N-glycans bearing the HNK-1 carbohydrate epitope. On the other hand, it has been suggested that these glycoproteins could be ligands of an endogenous mannose-binding lectin present in myelin, the cerebellar soluble lectin (CSL). In order to further document the heterogeneity of the glycans of these two CAMs, we have used several probes: an anti-carbohydrate antibody of the HNK-1 type, called Elec-39, the plant lectin concanavalin A (ConA), and the endogenous lectin CSL involved in myelin compaction. This study shows that CSL binds to a small proportion of the polypeptide chains of MAG found in adult CNS of rats and man and the polypeptide chains of P0 molecules from adult human and rat sciatic nerve. For MAG from adult rat brain, the binding of CSL is restricted to glycans of polypeptide chains which could be separated from the others according to their solubility properties. These MAG molecular entities react also with the Elec-39 antibody and with ConA. These results confirm that P0 and MAG are heterogeneous in their carbohydrate moieties.(ABSTRACT TRUNCATED AT 250 WORDS)


[Alveolar macrophages and granuloma formation]. We measured intracytoplasmic free calcium ion concentration ([Ca3+]i) of alveolar macrophages (AMs) in order to elucidate the mechanism(s) of lipopolysaccharide (LPS)-hyperresponsiveness of AMs in patients with sarcoidosis at the second messenger level. Resting [Ca2+]i was higher in patients with sarcoidosis than in normal subjects. [Ca2+]i increase responses were also elevated in patients with sarcoidosis when AMs were stimulated with either anti-CD14 (a LPS/LPS-binding protein complex receptor) antibody, anti-CD64 (Fc gamma receptor I), antibody or platelet activating factor. After incubation with interferon-gamma, resting [Ca2+]i and increase in [Ca2+]i induced by anti-CD14 antibody stimulation were higher in patients with sarcoidosis as compared with values before incubation. Thus, these data suggest that activation of AMs at the second messenger level induced by IFN gamma, at least in part, accounts for LPS-hyperresponsiveness in sarcoidosis.


[Lipopolysaccharide binding protein enhances intratracheally administrated lipopolysaccharide-induced acute lung inflammation via a CD14 receptor]. We examined the role of lipopolysaccharide binding protein (LBP) in the airspace and the CD14 receptor on alveolar macrophages in TNF alpha production and neutrophil (PMN) sequestration in lungs induced by intratracheal injection of lipopolysaccharide (LPS). LPS alone (Salmonella minnesota wild-type; 20 ng) or LPS + LBP complex [LPS (20 ng) + rabbit LBP (500 ng); preincubated for 30 min at 37 degrees C] was injected intratracheally into isolated rabbit lungs perfused with lactate-Ringer-albumin solution. Human PMN (5 x 10(7)) were added to the perfusate after 2 hr perfusion. Samples of lung perfusate were collected every 30 min for 180 min, after which bronchoalveolar lavage (BAL) was also performed. TNF alpha concentration in the perfusate and BAL fluid were determined using a bioassay with L-929 fibroblasts. PMN accumulation in the lung was determined by myeloperoxidase assay of the lung homogenate. LPS alone did not significantly increase TNF alpha production or PMN accumulation in lungs, whereas LPS/LBP complex increased TNF alpha concentration in the perfusate and PMN accumulation. Intratracheal injection of anti-CD14 antibody (40 micrograms) with LPS/LBP complex prevented TNF alpha production and subsequent PMN sequestration. We conclude that LBP in the airspace enhances the effect of LPS on TNF alpha production via a CD14-dependent pathway, and this subsequently contributes to PMN sequestration in the lungs. Airspace accumulation of LBP secondary to increased vascular and epithelial permeability may play a critical role in the development of septic shock and lung injury by promoting TNF alpha production via a CD14-dependent mechanism.


Heparin-binding (fibroblast) growth factors are potential autocrine regulators of esophageal epithelial cell proliferation. A serum-free culture system supplemented with neural tissue extract for normal human esophagus was applied to the culture of mouse esophageal epithelium. Similar to mouse mesenchyme and skin epithelium, esophageal epithelial lines (MEE) emerged after serial culture. The cells had an apparent unlimited life-span, but retained morphology and other characteristics of normal epithelial cells. A screen for growth factors that stimulated growth of MEE cells in the absence of neural extract revealed that epidermal growth factor (EGF) and heparin-binding (fibroblast) growth factors (HBGF) were most effective. An HBGF-like activity was apparent in extracts of rapidly proliferating, but not quiescent MEE cells at low or confluent densities. A cloned cell line (MEE/C8) was selected from MEE cell cultures in the absence of neural extract. MEE/C8 cells proliferated independent of either EGF or HBGF at rates equal to MEE cells, cell extracts exhibited HBGF-like activity at all stages of proliferation, and the cells formed invasive tumors in syngenic hosts. The HBGF-like activity present in extracts of tumorigenic MEE/C8 cells had properties similar to HBGF-1 (acidic fibroblast growth factor).


Allogeneic bone marrow transplantation for malignant hematologic disorders in children. In the present study we carried out allogeneic bone marrow transplantation (BMT) in 14 leukemia children with high risk prognostic factors. Six patients with acute nonlymphocytic leukemia (ANLL), four with acute lymphocytic leukemia (ALL), two with chronic myelogenous leukemia (CML), and two with myelodysplastic syndrome (MDS). Among these patients, six with ANLL, two with ALL, one with CML and one with MDS were alive in complete remission 8 to 58 months post-BMT. Four patients died of relapse (one with ALL, and one with MDS), and chronic GVHD (one with ALL and one with CML). In six patients recombinant granulocyte colony stimulating factor (rG-CSF) was used to shorten the period of granulocytopenia. The mean time of recovery to granulocyte count of 500/mm3 was 13.2 days in the rG-CSF+ group, being 15.9 days faster than that in the rG-CSF- group. In light of these results, allogeneic BMT is shown to be a choice of treatment for leukemia children with high risk prognostic factors and rG-CSF may be an effective reagent to prevent infectious episodes in BMT.


Mechanisms for appearance of no-flow areas in tumor microvascular bed. In order to elucidate the mechanism for appearance of no-flow areas (areas where tumor blood flow temporarily cease), we directly observed the process of tumor-induced neovascularization and measured pressure change in a feeding vessel (starting vessel) which supplies blood to the tumor vascular network. Total length of tumor vascular network from one starting vessel increased exponentially as the tumor increased in size exponentially. The pressure of the starting vessel increased from approximately 40 cmH2O to 120 cmH2O with enlargement of the tumor size. As soon as the pressure of the starting vessel reached a plateau, however, there was a rapid increase in low-flow or no-flow areas in places within the tumor. We considered that no-flow areas were produced by the imbalance between the pressure elevation of a starting vessel and the enlargement of the vascular network from that vessel.


Enhanced learning produced by injection of neurokinin substance P into the region of the nucleus basalis magnocellularis: mediation by the N-terminal sequence. The effect of unilateral injection of the neurokinin substance P (SP) and of certain N- or C-terminal SP-fragments into the region of the nucleus basalis magnocellularis (NBM) on inhibitory avoidance learning was investigated. Rats with chronically implanted cannulae were tested on a one-trial uphill avoidance task. Immediately after the training trial, rats were injected with 0.74 pmol SP or equimolar dosed SP(1-7), DIME-C7, or SP(7-11). Control groups included vehicle-injected rats and a group given an injection of SP(1-7) 5-h after the trial. When tested 24 h later, rats treated with SP or SP(1-7), but not with DIME-C7 or SP(7-11), exhibited longer step-up latencies than vehicle-treated controls. The retention latencies for rats in the SP(1-7) 5-h delay group did not differ from those of vehicle-injected animals, ruling out proactive effects of SP(1-7) on performance. The results show that SP facilitates retention of an inhibitory avoidance response when injected into the NBM. Furthermore, the amino acid sequence that encodes this effect may be located in the N-terminal part of the SP-molecule.


Plasticity of the sympathetic nervous system innervating the cerebral arteries in rats. In order to investigate the neuroplasticity of the peripheral sympathetic nervous system innervating the cerebral blood vessels, we observed and traced the sprouting nerve fibers originating in the contralateral superior cervical ganglion (SCG) into the previously denervated cerebral arteries following unilateral excision of the SCG and/or decentralization of the contralateral SCG in young rats (4 weeks old). These nerve fibers were labeled anterogradely with wheat germ agglutinin-horseradish peroxidase or stained immunohistochemically with anti-tyrosine hydroxylase antibody. Eight weeks after the right SCG excision, reinnervating nerve fibers originating in the contralateral ganglion formed a circular pattern of nerve plexus only on the wall of the main cerebral arteries of the circle of Willis in the ganglionectomized side. However, the decentralization of the contralateral SCG, which was performed simultaneously with a unilateral SCG excision, prevented the nerve sprouting into the denervated regions. Unilateral decentralization of SCG itself failed to affect their distribution pattern or their density of nerve fibers originating in the ganglion. It is concluded that in the young rat the outgrowth of the sympathetic nerve fibers into the denervated cerebral arteries was strongly impeded by the disconnection of ganglion cells from the central nervous system, while the decentralization alone could not affect the innervation pattern of the postganglionic fibers which have been already built-up in the cerebral arterial system.


Beyond three: MCH nurses' assessment leads to early interventions. This study reports the results of a one year trial of a comprehensive developmental assessment, including vision and hearing screening, of three to four year olds in one municipality. Initial results indicated that over 50% of the 790 children assessed had no identifiable problems, this increased a further 15% after retesting. One hundred and eighty-seven children were referred to a variety of agencies, mainly for vision and audiological reassessment. One hundred and thirty-five referrals for further investigation were for children whose problems were confirmed and appropriately managed.


Signals that induce sprouting in the central nervous system: sprouting is delayed in a strain of mouse exhibiting delayed axonal degeneration. This study evaluates whether CNS sprouting is initiated by signals related to the degeneration of presynaptic axons. We evaluate the time course of sprouting of cholinergic septohippocampal fibers after unilateral entorhinal cortex (EC) lesions in a substrain of mice carrying a mutation which leads to a substantial delay in the onset of Wallerian degeneration. We first verified that axonal degeneration resulting from EC lesions was delayed in mutant mice using silver-staining techniques (the Fink-Heimer method). Cholinergic sprouting was then evaluated using a histochemical technique for acetylcholinesterase (AChE) in mutant mice and normal controls. In normal control mice, both axonal degeneration and cholinergic sprouting occurred with a time course that was comparable to that described in rats. Argyrophilic degeneration debris was prominent by 4 days postlesion, and increases in AChE staining in the molecular layer of the dentate gyrus were well developed by 10 days. In mice carrying the "Ola" mutation, however, argyrophilic degeneration debris was not detectable at 4 or 6 days postlesion, began to appear in the dentate gyrus by 8 days postlesion, but did not become prominent until 12 days. Increases in AChE staining in the molecular layer of the dentate gyrus were not detectable even at 12 days postlesion, but developed gradually after 14 days. These results demonstrate that the signals which initiate at least one form of CNS sprouting are related to the degeneration of presynaptic axons.


Monoclonal antibodies against potato virus A--competitive binding tests. Six mouse monoclonal antibodies (MoAbs) against potato virus A (PVA) were tested. One of them (PVA 534) reacted only with complete virions and was apparently specific for epitopes dependent on quaternary structure. MoAb PVA 328 recognized the virus antigen only after its dissociation into subunits. MoAb PVA 328 must have reacted with a cryptotope of the antigen. MoAb PVA 151 and 290 appeared to be conformation independent and reacted with exposed regions on native virus particles as well as on the surface of dissociated coat protein subunits. Two other MoAbs (PVA 187 and 634) did not recognize subunits or the virions adsorbed directly to the microtiter plate. This seems to be an evidence of metatope existence. The results of competitive binding tests combined with the reaction patterns of individual MoAbs to different potyviruses indicate that the MoAbs are specific for 6 distinct epitopes.


Herpes simplex virus type 2 and pseudorabies virus associated growth factors and their role in the latency in vitro. A putative herpes simplex virus type 2 (HSV-2) growth factor (HSGF-2) was detected in a crude extract from virus infected mouse embryo cells. This factor, similar to previously described pseudorabies virus (PRV) associated growth factor (PRGF) was shown to have ability to morphologically transform non-transformed cells and to repress the transformed phenotype of transformed cells. Both activities could be neutralized with two, out of seven monoclonal antibodies directed against glycoprotein B of HSV-2. Both PRGF and HSGF-2 were detected in human embryo lung cells latently infected with PRV or HSV-2 either at 41 degrees C, or in the presence of phosphonoacetic acid. Human alpha-2 interferon, when present in medium of latently infected cells enhanced the production of both HSGF and PRGF. On the contrary, when latently infected cells were treated with 5-azacytidine the synthesis of both PRGF and HSGF-2 was completely blocked and the virus reactivated from latency replicated to higher titers than in non-treated cells. The role of PRGF and HSGF-2 in the establishment, maintenance and reactivation of latency, as well as in cellular transformation is discussed.


Determination of antibodies to non-structural and structural adenovirus antigens by ELISA. We evaluated possibilities to analyze serum antibodies to non-structural (peptides derived from adenovirus E1b protein) and structural (hexon) adenovirus antigens by ELISA. Synthetic dodecapeptides covering a putative A-gliadin cross-reactive antigenic determinant of the E1b protein were used. The aminoterminus of the peptides appeared to be important for antibody binding but the exact sequence of a possible common B-cell epitope within the peptides remained open. Coupling of the peptides to a carrier protein was essential for ELISA analyses of serum antipeptide antibodies. IgA antibodies to both adenovirus derived E1b peptides and hexon antigen could be detected already two weeks after the onset of an acute adenovirus infection, while antipeptide IgG antibodies were seen in a restricted number of patients only.


Preparation and characterization of the monoclonal antibodies against Japanese encephalitis virus. Six mouse monoclonal antibodies (MoAbs) against Japanese encephalitis virus (JEV) were prepared and analyzed with indirect immunofluorescence assay (IFA), enzyme linked immunosorbent assay (ELISA), haemagglutination inhibition test (HI), neutralization test (NT), antibody dependent cell mediated cytotoxicity (ADCC) assay, antigenic site specific analysis and relative affinity measurement. These MoAbs could be divided into three classes by indirect immunofluorescence cross reactivity among four flaviviruses, 2H4, 2F2, and nG2 were type specific; 2D2 and mC3 were subgroup specific; and mG9 was family specific. 2H4 and 2F2 had higher neutralization activity, 2D2 and mC3 had the function of inducing ADCC effect, mG9 had higher titer in HI. The six MoAbs recognized five antigenic sites on JEV envelope glycoprotein, 2H4 and 2F2 recognized the same or very similar antigenic site and their relative affinity was ranked as: nG2 > 2H4 > 2D2 > mG9 > 2F2 > mC3.


Common and different antigenic properties of the rabies virus glycoprotein of strains SAD-Vnukovo and Pitman-Moore. Two fixed rabies virus strains, SAD-Vnukovo and Pitman-Moore (PM) were used as combined immunogens for the generation of hybridomas secreting specific monoclonal antibodies (MoAbs). The obtained hybridomas were primarily screened by an ELISA for production of MoAbs to antigen of SAD-Vnukovo strain. Six positive clones were established. A panel of MoAbs has been characterized according to reactivity in immunofluorescence, immunoblot, ELISA and neutralization tests. All MoAbs were positive in immunofluorescence when cells infected with the SAD-Vnukovo strain were used. By immunoblot, four MoAbs showed specificity for the viral glycoprotein of both SAD-Vnukovo and PM rabies strains. This pattern of reactivity indicated the existence of shared conformation-independent epitopes located on the related antigens. However, in ELISA, the tested MoAbs did not recognize viral glycoproteins of the PM strain. This indicates, that the different strain-specific conformations of the native glycoprotein determine the accessibility of the common linear determinants for respective antibodies. Only one antibody, with conformation-dependent glycoprotein specificity, was capable to neutralize the CVS strain of rabies virus.


Synthesis of 23 K acute-phase protein by HBV genome carrying PLC/PRF/5 human hepatoma cells. Elevated synthesis of 23 K protein by human hepatoma PLC/PRF/5 cells was observed after their treatment with conditioned medium from concanavalin A stimulated peripheral-blood monocytes. Increased amount of this protein was first determined 4 hr after the treatment and its maximal level was reached 48 hr later. The role of the 23 K protein remains so far unknown.


Platelet-derived growth factor is angiogenic in vivo. PDGF receptors have recently been found to be expressed in microvascular endothelium in vivo under circumstances of endothelial cell activation and angiogenesis suggesting that PDGF may have a direct effect on endothelial cells. We have tested the angiogenic activity of PDGF-AA and -BB homodimers in the chick chorioallantoic membrane in vivo. PDGF-BB was found to consistently induce an angiogenic response whereas PDGF-AA was less active. Morphological analyses revealed that there was little inflammation associated with this response but an increase in vessel density suggested a direct effect of PDGF on embryonic chorioallantoic endothelial cells. In vitro, PDGF-BB was found to be more potent than PDGF-AA in stimulating the chemotaxis of rat brain capillary endothelial cells. This is consistent with a direct effect of PDGF on endothelial cells. Thus, this novel angiogenic activity of PDGF has implications for several developmental and pathological events in which PDGF, particularly the B-chain, is expressed.


Insulin-like growth factor-1 stimulates proliferation of myeloid FDC-P1 cells overexpressing the human colony-stimulating factor-1 receptor. Retrovirally expressed human CSF-1 receptor can induce CSF-1-dependent growth of IL-3-dependent hemopoietic cells FDC-P1. Here we show that expression of the human CSF-1 receptor also allowed FDC-P1 cells to grow in response to Insulin-like Growth Factor-1 (IGF-I). The authentic receptor for IGF-I was identified by affinity cross-linking and binding analysis on both control (infected with a neo vector) and CSF-1 receptor expressing FDC-P1 cells. DNA and RNA analysis of these cells and of five clones of IGF-I responsive cells demonstrated that the IGF-I receptor gene was not rearranged nor was it abnormally expressed in IGF-I responsive cells. These results suggest that myeloid cells over-expressing CSF-1R (c-fms protooncogene product) might have a proliferative advantage over normal myeloid cells in a physiological situation, independently of the presence of CSF-1 or the capacity of the cells to respond to CSF-1. This would indicate a possible role for c-fms in human neoplasia.


HLA matching: a comparison of conventional and molecular approaches. 1. The UNOS national sharing program has produced a 5-fold increase in the number of compatible transplants since 1987. 2. Transplants with 0 broad HLA-antigen mismatches had 87% 1-year graft survival and an 18-year half-life, essentially the same result as with conventional matching. 3. Equally impressive results were obtained with public determinant matching: 1-year graft survival ranging from 84 to 88% and a half-life of 12-15 years. 4. Increasing the number of matched determinants improved the results but reduced the number of compatible recipients identified. 5. Less than 10% of the current transplants have 0 public determinants mismatched. This fraction could be increased to 66% with local sharing and to over 90% with national sharing. 6. The results for cadaver transplants would approach those of related kidneys if compatible grafts were transplanted at the 90% rate obtainable with molecular matching.


Results of chemotherapy plus external reirradiation in the treatment of locally advanced recurrences of nasopharyngeal carcinoma. Between 1982 and 1991, 16 patients with recurrent cancer of the nasopharynx were treated with chemotherapy (CT) and radiotherapy (RT). All patients had received prior RT (45-69, 30 Gy). According to rTNM there were three rT2, one rT3 and 12 rT4. 5 patients were N1. Reirradiation (12-46 Gy, mean: 28) started 3-4 weeks after CT (2-6 cycles of different combinations), but 2 cases involved concomitant therapy. Out of 16 patients 7 had complete response (CR) (43.7%), 7 partial response and 2 no response. Statistically significant prognostic factors for obtaining CR were time of relapse and response to initial CT. Median duration of CR was 22+ months (9-64+). Failures at primary site occurred in 3 patients, 2/2 of those receiving CT without platinum compounds and 1/5 of other ones, with statistically significant difference in local recurrence free-survival between the two groups. Two- and 3-year actuarial overall survival were 28% and 10%. Rates of disease-free survival were 17% and 8%, respectively. The acute toxicity was generally mild. No central nervous system damage or radiation-induced myelitis were observed in survivors.


Salivary flow and composition in lymphoma patients before, during and after treatment with cytostatic drugs. To investigate the effect of modern, intensive chemotherapy on salivary flow rate and composition, 79 patients suffering from Hodgkin's disease or non-Hodgkin lymphoma were studied before, during and after administration of cytostatic drugs. 49 patients (mean age 49.9 years, 30 men, 19 women) completed the 1-year study. All patients who received radiotherapy or medication other than cytostatics were excluded. The results showed no marked differences in stimulated salivary flow rates, buffering capacities and amylase and total protein concentrations between the beginning and the end of the 12 month trial. However, significant increases in albumin secretion into saliva and salivary lysozyme concentrations were observed. Total salivary IgG, IgA and IgM concentrations decreased significantly during the cancer therapy but values returned to the baseline levels after termination of treatment. Despite the well-known cytolytic effect of anticancer drugs, chemotherapy need not therefore be permanently detrimental to saliva.


[Substance P and the resistance of biological motivations to ethanol]. Ability of the substance P to enhance the tolerance of feeding and defence motivations of ethanol was studied in rabbits. Ethanol led to disintegration of central mechanisms of both feeding and escape responses elicited by a threshold electrical stimulation of lateral and ventromedial hypothalamic centres. Subsequent i.v. administration of the substance P restored the effects of midbrain RF on the motivational centres. The inhibitory effect of the dorsal hippocampus and the facilitatory effect of the midbrain RF on excitability of the lateral hypothalamus, are also restored. The data obtained suggest that the substance P is able to increase a tolerance of the central mechanisms of biological motivations of ethanol.


[The localization of the neurons in the cat stellate ganglion]. Localisation of HRP-labelled neurons in the stellate ganglion was studied. The neurons in the ganglion seem to be organised according to the topic principle.


[An etiological study on fulminant viral hepatitis]. Viral markers were studied in 79 cases of viral hepatitis with hepatic failure. The results were shown as follows: 8 cases were positive for anti-HAV IgM (10.12%); 76 cases positive for HBsAg or anti-HBc IgM (96.20%) and 41 cases positive for anti-HCV antibodies (51.89%). Among those with anti-HCV positive, 35 cases were co-infected with HBV, 5 cases with HAV and/or HCV, only one was infected with HCV alone 2 cases were HD-Ag positive (2.52%) and one not identified (1.27%). With the reference of clinical findings, patients co-infected with HBV/HCV or anti-HBc IgM positive were more critical and usually entail higher mortality. In cases with HCV co-infections, the positive HBV replication markers seems to be reduced. Hepatic failure without HBV replicative markers had a high rate of hepatic coma as well as poor outcome.


Serosurveillance for Pseudomonas pseudomallei infection in Thailand. A nation-wide survey was conducted to see the prevalence of serosensitivity to Pseudomonas pseudomallei antigens by indirect hemagglutination (IHA) and indirect immunofluorescent assay (IFA) for IgG and IgM. Serum samples were collected from blood donors in eight selected areas and bacteriologically confirmed melioidosis patients in Ubon Ratchathani province. The distribution patterns of antibody titers were compared among the survey areas with cut-off points set at 1:160 for IHA, 1:4 for IFA-IgM and 1:32 for IFA-IgG. These cut-off points were decided by ROC (Receiver Operating Characteristics) analysis. The specificity (% true negative reactions) of each serological test in the general population differed significantly among survey areas, possibly reflecting the extent of inapparent infection in each community. IFA was more successful than IHA in differentiating between negative from positive reactions. The survey classified the areas into endemic (Khon Kaen, Ubon Ratchathani), transported (Bangkok), and non-endemic (other provinces) types.


Serosurveillance for double infection with Pseudomonas pseudomallei in tuberculous patients. The serosensitivity to Pseudomonas pseudomallei antigen in pulmonary tuberculous patients was surveyed in Ubon Ratchathani Province, a melioidosis-endemic and tuberculosis-prevalent area in Thailand. Indirect hemagglutination assay (IHA) and indirect immunofluorescent assay (IFA) for IgM and IgG were employed for the measurement of antibody levels, with cut-off points set at 1:160, 1:8, and 1:32, respectively. Retrospective protocol survey of clinical data was also conducted. From these studies, however, no evidence was obtained to show that tuberculous patients have a disposition to acquire double-infected with P. pseudomallei and to develop melioidosis. The serosensitivity was never higher than that of the general population of the province as represented by healthy blood donors, nor related with the clinical severeness. Tuberculosis and melioidosis appear to be mutually independent diseases without showing interrelated prevalence in the endemic area.


[Drug treatment of chronic ventricular arrhythmia]. The treatment of chronic ventricular arrhythmias depends on the severity and tolerance of the arrhythmia. Extrasystoles, even repetitive, in the healthy heart, are usually respected when asymptomatic or treated with betablockers in first intention when symptomatic. These drugs should also be proposed for patients with ischemic heart disease and non-sustained ventricular tachycardia, a situation in which Class I antiarrhythmics should be avoided. The prevention of sustained ventricular tachycardial may be empirical, with betablockers and/or amiodarone, or guided by the results of pharmacological tests during endocavitary electrophysiological studies.


[Enhancing the reparative potentials of the cellular structures in the hypothalamus by the preliminary adaptation of animals to hypoxia and stress]. Effects of preliminary adaptation of rats to hypoxic hypoxia and short-term stress environments on the dynamics of the repair changes of RNA and proteins in the cellular structures of supraoptic (SON) and paraventricular (PVN) nuclei of hypothalamus are studied. Adaptation to hypoxia and short-term stresses is found to increase the potential of protein-synthetizing system of neuro- and gliocytes of SON and PVN. It prevented a marked decrease in the RNA and proteins contents during a post-stress period and increased an intensity of the repair processes in the nuclei being tested. Experimental findings are indicative of a heterochronic pattern of repair processes in hypothalamic neurosecretory structures which to a great extent can be related with their potentials to rearrange the intracellular metabolism for maximum functional maintenance during stresses.


The immunogenetics of immune senescence. Immune senescence is characterized by a dysregulation of the immune system. With respect to humoral immunity, aging is associated with an increased level of many autoantibodies and a decreased antibody response to most foreign antigens. This observation reflects a decreased capacity to activate antibody production by CD5-negative B cells despite a normal or increased capacity to generate antibodies produced by the CD5-positive B cells. A similar dysregulation of cell-mediated immunity is manifested by an altered balance in cytokine production by T cells from old as compared to young subjects. Thus, the production of interleukin-2 (IL-2), IL-3 and granulocyte-macrophage colony-stimulating factor by T cells from old subjects is decreased although the production of IL-4, IL-5 and IL-6 is undiminished or actually increased.


[Immunological parameters in the differential diagnosis of ascites secondary to peritoneal carcinomatosis, hepatic cirrhosis, and congestive heart failure]. As a contribution to the study of ascites in patients with liver cirrhosis, congestive heart failure and peritoneal carcinomatosis evaluate in serum and ascites the concentrations of alphafetoprotein, carcinoembryonic antigen and fibronectin, they might suggest a diagnosis for the basic pathology. Forty-seven patients were studied, from whom 23 with cirrhosis, 17 peritoneal carcinomatosis and 7 with congestive heart failure. We conclude that: a) none of the tools usually employed in the analysis of ascitic fluid alone can make the base pathological process responsible for producing ascites; b) fibronectins were more useful for differential diagnosis between cirrhosis and carcinomatosis; c) alpha-fetoprotein and carcinoembryonic antigen were not useful for the definition for differential diagnosis.


[Major complications after open prostatectomy: management and prevention]. From Jan. 1976 to Dec. 1989, 829 patients with benign prostatic hyperplasia (BPH) were treated. Open prostatectomy was performed successfully in 571 patients. Secondary hemorrhage was observed in 21 patients (3.7%), bladder neck or posterior urethral stricture in 10 (1.7%), and temporary incontinence in 21 (3.7%). No operative mortality was noted. The pathogenesis, management and prevention of those complications were detailed with review of the literatures. Effective hemostasis, proper enucleation of the enlarged gland and meticulous reconstruction of the bladder neck are claimed to be the key points in reducing complications and increasing cure rate in the treatment of BPH with open prostatectomy.


[The importance of fixation in immunohistochemistry: distribution of vimentin and cytokeratins in samples fixed in alcohol and formol]. The fixation in ethanol or formalin for trypsin digestion in immunohistochemical detection of cytokeratins and vimentin was assessed in a case of ovarian cystadenofibrocarcinoma. Superior reactivity for both markers was achieved in ethanol-fixed sections, even in samples stocked up to 60 days. Cytokeratin reaction in formalin-fixed sections was better when trypsin was used. However, this digestion was deleterious to vimentin detection. These data are presented to alert surgeons and oncologists on the relevance of fixation of specimens suspicious for neoplasia, since different epitopes may require different fixatives and the inadequate choice in the operative room may impart difficulties when immunohistochemistry is necessary.


Comparative study of adenoviruses with monoclonal antibodies. The obtainment of monoclonal antibodies for adenovirus species 4(Ad4) is described. The specificities of selected monoclonal antibodies were determined by means of viral neutralization test in cell culture, immunofluorescence and Enzyme-Linked Immunosorbent Assay (ELISA), in the presence of the following species of human adenovirus: 1, 2, 5 (subgenus C), 4 (subgenus E), 7 and 16 (subgenus B) and 9 (subgenus D). Two monoclonal antibodies species specific to adenovirus 4 (1CIII and 3DIII) and one monoclonal antibody that cross reacted with adenovirus species 4 and 7 (2HIII) were obtained.


Bacteriocin-like antagonism in Yersinia enterocolitica. A total of 121 Yersinia enterocolitica O3 isolates from patients with gastroenteritis and 37 Y. enterocolitica reference strains with different O antigens were tested for bacteriocine production and sensitivity. By using cross-streaking method strains belonging to serogroups of O5; O7,8; O7,13; O11; O11,23; O13,27; O17; O19,8 and O34 produced bacteriocin-like substances. None of the Y. enterocolitica O3 strains produced bacteriocin-like material and most of them were uniformly sensitive against the bacteriocin-like material produced by strains of serogroups O7,8; O7,13; O13,27 and O19,8. By sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) significant differences were demonstrated in the whole cell protein patterns of Y. enterocolitica reference strains belonging to different serogroups in the range of 33-47 kilodalton (kDa). Out of the ten examined bacteriocin-like material producer strains only one strain harboured a plasmid of about 60 megadalton (MDa).


[Thermostability of rabies vaccine, type Fuenzalida & Palacios, for human use]. Ten lots of Fuenzalida & Palacios type antirabies vaccine for human use, produced at the Instituto Butantan (São Paulo, Brazil) were stored at temperatures of 45, 37, 28 and 2-8 degrees C. The potency of each lot was determined in samples taken at varied time intervals using the NIH method and lots presenting antigenic values > or = 0,3 were considered satisfactory for use. After 2 hours at 45 degrees C the antigenic value of one out of 10 lots tested was found to be less than the minimum required value. At 37 degrees C all lots maintained satisfactory antigenic values until the third day of storage, whilst at 28 and 2-8 degrees C the potency was fully maintained, respectively for 10 and 360 days. At the ideal temperature of 2-8 degrees C, 100% of the tested vaccines maintained the minimum required antigenicity for a longer period (16 months) than the expiration time of 6-12 months usually recommended for this type of biological produced in Latin American and Caribbean countries. Thus, the obtained data suggested that in countries still producing Fuenzalida & Palacios type vaccine, the expiration tim could be extended to 16 months, what could prevent the unnecessary discarding of products still in useful condition.


Immunoelectrophoretic study on common antigens of São Lourenço da Mata and Belo Horizonte strains of Schistosoma mansoni adult worms and Biomphalaria snails. Immunoelectrophoretic studies on common antigens were carried out by using rabbits sera immunized against São Lourenço da Mata and Belo Horizonte strains of Schistosoma mansoni adult worms and antigens of Biomphalaria glabrata pigmented (Jaboatão--PE); B. glabrata albino (Belo Horizonte--MG) and B. straminea (São Lourenço da Mata, PE). Furthermore, the reverse approach was proceeded, namely, sera anti Biomphalaria snails produced in rabbits were tested against both strains of Schistosoma adult worm antigens. The analysis of the common antigens between the SLM strains of S. mansoni adult worm and B. glabrata pigmented showed 8 to 9 precipitin bands, 3 bands with B. glabrata albino and only 1 band with B. straminea crude extracts. On the other hand, the BH strain of S. mansoni adult worm antisera produced 6 to 7 bands with B. glabrata pigmented, 5 bands with B. glabrata albino and 1 band with B. straminea antigenic extract. Biomphalaria snails crude extracts were fractionated by Sephadex G-100 column and three fractions were collected from each snail strain. The fractions were tested with anti SLM and BH strains of S. mansoni adult worm sera by immunoelectrophoresis. The common antigens fractionated from Biomphalaria snails crude extracts and those found for both strains of S. mansoni adult worm mostly existed in the first fraction and they were estimated to have molecular weight over 158,000 daltons. In our laboratory, it was found a relationship between the antigenic similarities and experimental infection rates of S. mansoni towards Biomphalaria snails so that more bands were seen with increasing infection rates of S. mansoni.


[The effect of glycosaminoglycans in the pulmonary interstitial fibrosis development]. The pulmonary interstitial fibrosis was caused by injecting Bleomycin into mouse trachea. After the injection, the volume of glycosaminoglycans (GAG) in bronchoalveolar lavage fluid and lung tissues was increased. The observation of histochemical stain and electron microscope showed that the distribution of GAG in lung tissues was varied at different time after the injection, and related to the volume of collagen proteins and the formation of pulmonary interstitial fibrosis.


[The effect of mexiletine on theophylline pharmacokinetics in patients with bronchial asthma]. Reported side effects of mexiletine, a useful drug for managing premature ventricular contractions (PVC) and ventricular tachycardia (VT), are gastrointestinal symptoms including nausea and vomiting. Theophylline, a bronchodilator, also causes similar gastrointestinal symptoms with its frequency proportional to the increases in its serum concentration. The occurrence of gastrointestinal symptoms is known to increase in combination therapy of mexiletine and theophylline. However, the pharmacokinetic interaction between mexiletine and theophylline has not been clarified. We, therefore, investigated the effects of mexiletine on theophylline pharmacokinetics. Three patients with bronchial asthma complicating PVC and/or VT (one male and 2 females, aged 70.0 +/- 13.1 years) were studied. All patients were given prophylactic theophylline anticipatory for asthmatic attacks, which was followed by oral administration of mexiletine, 200-300 mg daily, for PVC and/or VT. The serum theophylline concentrations in 3 cases were increased from 13.8 to 25.3 micrograms/ml, from 14.6 to 27.8 micrograms/ml, and from 10.4 to 15.4 micrograms/ml, respectively, after the administration of mexiletine. However, significant decreases in theophylline clearance were observed after the administration of mexiletine by 46, 47 and 35%, respectively. (p < 0.05) With a decrease in theophylline dosage, the serum theophylline concentrations decreased, and gastrointestinal symptoms resolved. Theophylline is metabolized mainly in the liver through an oxidative reaction of p-450 enzyme, however, its metabolism is affected by many factors, such as medications and complications. The results of this study indicated that mexiletine decreases the theophylline clearance by inhibiting the p-450 oxidative reaction to theophylline.(ABSTRACT TRUNCATED AT 250 WORDS)


[Recent trend in G-CSF therapy and clinical laboratory tests]. Granulocyte colony-stimulating factor (G-CSF) has strong leukopoietic activity and it is used for patients with leukopenia during leukemia chemotherapy. However, some leukemia cells show a high affinity to G-CSF and are driven to proliferative phase. In our laboratory, we developed two testing methods. 1) Flow cytometric method on G-CSF susceptibility of leukemia cells using FITC-labeled G-CSF, and 2) Immunohistochemical method for detecting the ratio of cells driven from dormant phase to proliferative phase by G-CSF with anti-PCNA antibody and Ki-67 antibody. In MDS patients G-CSF administration induced an increase of cells in proliferative phase. The patients treated with cytosine arabinoside following G-CSF showed hematologically good improvement. A new mode of therapy using G-CSF in combination with other cytokines or antileukemic agents will be developed in the near future for treatment of leukemia patients.


[Argyrophilic nucleolar organizer region proteins (Ag-NORs) in human central nervous tumors]. A silver colloid staining technique for identifying nucleolar organizer region-associated proteins (Ag-NORs) was applied to 17 primary gliomas and 16 meningiomas. These comprised 8 glioblastomas, one pleomorphic xanthoastrocytoma, 8 benign astrocytoma, 10 nonrecurrent meningiomas and 6 recurrent meningiomas, in which the mean number of Ag-NORs' per nuclei, were 3.37, 2.34, 2.57, 2.16 and 2.80 respectively, identifying significant differences among these groups. Ag-NORs of four giant cell glioblastomas showed low numbers of Ag-NORs, suggesting better prognosis. In spite of the time consuming and complicated counting method Ag-NORs is reproductive and useful as a tool for estimating the proliferating potential of various brain tumors, and appears to widely applicable in clinical laboratories.


[The effects of retinoic acid and recombinant human granulocyte colony-stimulating factor on alkaline phosphatase activity of HL-60 cells]. We examined alkaline phosphatase (ALP) activity in the HL-60 cell induced by retinoic acid (RA) and recombinant human granulocyte colony-stimulating factor (rhG-CSF). rhG-CSF induced a small but significant increase of NBT-reducing ability and ALP activity of the HL-60 cells. Among various inducers of cell differentiation, 1,25(OH)2D3 and dimethylsulfoxide (DMSO) caused the increase of the NBT-reducing ability and the suppression of ALP activity induced by rhG-CSF, while RA enhanced both of them. Protein kinase C inhibitors (H-7 and staurosporine) but not a protein kinase A inhibitor (HA1004) significantly suppressed the ALP activity induced by the simultaneous treatment with RA and rhG-CSF.


[The clinical usefulness of a new (the second generation) antibody to hepatitis C virus]. To clarify the clinical usefulness of a second generation antibody to hepatitis C virus (anti-HCV-2), we tested the anti-HCV-2 by enzyme immunosorbent assay (Imucheck HCV Ab) in comparison to the first generation antibody (anti-HCV-1). Serum samples were obtained from the patients with acute hepatitis, chronic liver diseases, alcoholic liver disease and autoimmune liver diseases. Furthermore, both antibodies were tested in serum samples from individuals in a hyperendemic area of non-A, non-B hepatitis. Anti-HCV-2 was detected earlier than anti-HCV-1 in acute type C hepatitis. The prevalence of anti-HCV-2 was higher than that of anti-HCV-1 in acute and chronic non-A, non-B liver diseases. However, prevalence of anti-HCV-2 was not increased in patients with non-viral liver disease such as pure alcoholic liver disease and autoimmune liver diseases. The HCV-infection rate was increased to about 40% by detection of anti-HCV-2 in individuals in hyperendemic area of non-A, non-B hepatitis. Thus, the assay of anti-HCV-2 is very useful to detect actual HCV infection. The accuracy and sensitivity of anti-HCV-2 were higher than those of anti-HCV-1.


[The significance of the determination of AFP-isoform in the early diagnosis of hepatocellular carcinoma]. To evaluate the significance of the determination of the AFP-isoform in the early diagnosis of hepatocellular carcinoma (HCC), the AFP-isoform was determined in 48 patients with HCC and in 53 patients with liver cirrhosis, using the lectin electrophoresis of AFP by antibody-affinity blotting technique. 1. The HCC-type AFP-isoform was detected in 44 of the 48 patients with HCC, 2. In about 30% of the patients with HCC, the HCC-type AFP-isoform appeared 3-10 months before the graphic diagnosis of HCC, 3. In about 60% of the patients with liver cirrhosis found to have the HCC-type AFP-isoform, HCC developed within one year. Thus, the serial determination of the AFP-isoform in patients with liver cirrhosis was considered very useful for the early diagnosis of HCC.


[Coagulation and fibrinolysis in pregnancy]. We studied on coagulation and fibrinolysis systems during pregnancy by measuring plasma levels of thrombin-antithrombin III complex (TAT), plasmin-alpha 2 plasmin inhibitor complex (PIC), fibrinogen/fibrin degradation products (FDP), plasminogen (PLG) and antithrombin III (AT-III). Ninety seven pregnant, 5 post-delivery and 32 nonpregnant women aged from 20 to 52 years old were included in this study. Plasma concentrations of TAT and PIC in nonpregnant women were 3.38 +/- 1.02 micrograms/l and 0.65 +/- 0.24 micrograms/ml, respectively. TAT gradually increased with the progression of pregnancy and rapidly decreased after the delivery. Whereas PIC and AT-III concentrations did not change significantly during pregnancy. Fibrinogen, PLG and FDP concentrations changed similarly as TAT. Eight pregnants whose plasma PIC concentrations elevated more than 1.0 micrograms/l were further examined. In 6 women out of them (71.5%), FDP concentrations were elevated. In this particular group of subjects, however, they delivered normally without complications such as toxemia. These observations suggest that, at least, a hypercoagulative state progresses with pregnancy, being normalized after the delivery. Although we could not find the relationship between the hypercoagulation and clinical complications such as thrombosis and toxemia of pregnancy, present findings suggest that special caution should be paid on the pregnants whose TAT and FDP levels are elevated.


[Detection of anti-HCV antibody, anti-GOR antibody and autoantibodies in sera of patients with non-A, non-B liver diseases]. Anti-HCV antibody, anti-GOR antibody and autoantibodies were measured in sera of 41 patients with non-A, non-B liver diseases. Anti-HCV antibody (C100-3) was positive in 73.1% of the patients. On the other hand, anti-GOR antibody, rheumatoid factor, anti-ssDNA antibody and anti-dsDNA antibody was positive in 56%, 26.8%, 12.1% and 2.4% of the patients, respectively. In 70.7% of the patients, at least one of these four antibodies was positive. No relationship was observed in both positive rate and antibody titers among these different antibodies. On the other hand, positive rates of anti-HCV and anti-GOR antibody were higher in patients with persistently elevated serum ALT. Anti-GOR antibody appears to be a kind of autoantibody since it recognizes liver cell protein which has amino acid sequence partly similar to an epitope of HCV protein. Therefore, it could be speculated that the frequent appearance of antibodies against self constituents may be related to the chronic inflammation of the liver induced by HCV infection.


[Primary adenocarcinoma of the duodenum: a clinical case and review of the literature]. A case of primary duodenal adenocarcinoma recently observed gives the Authors the chance to review the literature. Diagnosis is often delayed because symptomatology is usually vague and non specific, a fact that is even more in contrast with diagnostic techniques, which, on the contrary, are handy and easily performed (endoscopy, duodenography). Surgical resection is the only proposable treatment while no local and distant diffusion has yet showed, otherwise the operation is limited to a digestive and/or biliary derivation. Anyhow, prognosis remains severe.


[Carcinoma of the gallbladder. Our experience]. A series of 11 patients (3 males and 8 females, mean age 63.4 yrs.) suffering from gallbladder cancer has been reviewed. Authors' purpose in reviewing their experience was to identify factors either in patient characteristics or treatment, that may influence statistic data. Symptoms, signs, laboratory and imaging studies were uniformly unhelpful in determining the diagnosis. Early diagnosis at a stage amenable to surgical excision remains the only therapeutical possibility.


Blockade of potassium or calcium channels provokes modifications in TRH-induced TSH release from rat perifused pituitaries. The aim of the present study was to determine the functional relationship between blockade of potassium or calcium channel activity and the initial burst of TSH secretion in response to TRH. Perifused rat pituitary fragments were stimulated by a 6-min pulse of physiological concentration of TRH (10 nM) in the presence or absence of pharmacological blockers of K+ or Ca2+ channels. Blockade of Ca(2+)-activated K+ channels with TEA (10 mM and 30 mM), apamin (200 nM), or charybdotoxin (50 nM) completely or partially blunted TRH-induced TSH release. By contrast, blockade of voltage-dependent K+ channels with 4-aminopyridine (4-AP) (500 microM) or with dendrotoxin (DTX) (350 nM) significantly increased TSH response. Moreover, blockade of T-type voltage-sensitive Ca2+ channels (VSCC) with NiCl (3 mM) or with diphenylhydantoin (100 microM) significantly (P < 0.01) reduced TSH response to TRH, whereas blockade of L-type Ca2+ channels with verapamil (50 microM) was ineffective. Our results suggest that secretion of TSH in response to nanomolar concentrations of TRH is correlated with stimulation of Ca(2+)-activated K+ channels, and inhibition of 4-AP-and DTX-sensitive voltage-dependent K+ channels; furthermore TSH response seems to depend on the activation of T-type VSCC.


[Use of bacterial toximeters with separate cell cultures for continuous water monitoring]. Two commercially available bacterial toxicity monitors are compared. As test organisms pure cultures of Pseudomonas putida are used in both systems. The bacteria are grown continuously in turbidostatic ("Toxalarm", LAR, Berlin, Germany) or chemostatic ("Stiptox-norm", Siepmann und Teutscher, Gross-Umstadt, Germany) regulated cultures in a selective culture medium in nonsterile devices. Toxic substances can be detected by continuously working bacterial respiration tests. Oxygen consumption is the measuring parameter. The bacterial test suspension is mixed continuously in a fixed proportion with air-saturated test water in a measuring cell. The separate culturing of the bacteria and the carrying out of the tests ensures that neither the bacterial culture is endangered by toxic substances nor the sensitivity can be changed by poison adaptation. Results of "Toxalarm"-tests with several chemicals (e.g. Atrazine, Lindane, 2-Nitro-phenol, Sodiumpentachlorophenolate) are presented. The registration of an alarm event (River Rhine) by "Stiptox-norm" is shown.


[Valve movement of the mussel Dreissena polymorpha as a monitoring system for bodies of water]. A new biological "early warning system" for continuous water control is presented that is based on the valve movements of the freshwater mussel Dreissena polymorpha. For two groups of up to 42 mussels, both the percentage of open mussels and the number of valve movements during a time interval are used to describe the activity behaviour of the zebra mussel. A main prerequisite of such a biomonitor should be the knowledge of the "normal" behaviour in correlation to changing environmental conditions. Preliminary results in Dreissena demonstrate the influence of the temperature, the daily cycle of light and the food concentration on both characteristics of the activity pattern. During toxicity tests, the mussels reacted significantly within 3-30 minutes after the addition of a toxicant. This was normally observed by both the decrease of the percentage of open mussels and the increase of the number of valve movements. Therefore--and against the background of the well-known "normal" behaviour--reliable limits for the detection of a harassing event can be defined. First experiments with Pentachlorophenol, Lindane, Atrazine and 2-Nitrophenol demonstrated to some extent a higher sensitivity compared to other "early warning systems".


The virology and serology of hepatitis: an overview. At least five viruses are recognised as causes of acute hepatitis which, in some instances, can develop into chronic disease. The viruses of hepatitis A (HAV) and hepatitis E (HEV) are spread predominantly by the faecal-oral route, whereas hepatitis B, C and D viruses (HBV, HCV and HDV) are spread by blood and other body fluids. The incubation period ranges from about four weeks for HAV to three months for HBV. The diagnosis mainly depends on the detection of various serological markers. The control of infection in the United Kingdom has been facilitated by the introduction of vaccines for HAV and HBV and screening blood for HBV and HCV. Good hygiene and sanitation, the availability of sterile equipment, and measures to modify the behaviour of high-risk groups such as injecting drug users, are also important in the prevention and control of viral hepatitis.


The epidemiology and control of hepatitis A. Hepatitis A virus (HAV) infection is endemic in developing countries and remains a public health problem elsewhere. A total of 7430 cases was notified in England and Wales in 1991. Children and young adults were most frequently affected. Risk factors include travel to countries of high endemicity, contact with an infected person and shellfish consumption. Person-to-person spread is common within families, closely knit communities and schools (especially nurseries). The provision of safe water and sanitary disposal of faeces are prerequisites for control of HAV throughout the world. In developed countries, human normal immunoglobulin (HNIG) is used for post-exposure prophylaxis and is given to those travelling abroad prior to possible exposure. Recommendations for the use of hepatitis A vaccine have have recently been made but it is expensive. Further studies on risk factors for infection, and on the efficacy of hepatitis A vaccine in outbreak control, are needed.


Circannual analysis of pineal gland indoles and vasotocin-like immunoreactivity in male rats kept on constant photoschedules and temperature. Levels of vasotocin-like immunoreactivity (iVT) and the indoles serotonin (5HT), 5-hydroxyindole acetic acid, N-acetyl-serotonin (NAS) and melatonin were measured over 24-hour periods at monthly intervals in the pineal gland of male rats kept under constant laboratory conditions of photoperiod and temperature. The 8- to 10-fold increase in pineal iVT, which occurred during mid-August, was not related to variations over the 24-hour light:dark (L:D) cycle. Tryptophan and 5HT exhibited seasonal variations over the 24 h L:D cycle with a single peak during the winter months. Twenty-four-hour levels of NAS tended to increase during the fall and spring. Seasonal effects on indole metabolism were further suggested by a marked decline during the fall in the correlation between NAS versus melatonin in individual pineal glands. Positive correlations between 5HT versus NAS and 5HT versus melatonin tended to be significant only during the summer months. These data suggest that environmental cues other than photo-period and temperature can regulate pineal neuroendocrine functions.


The diagnosis and management of viral hepatitis. Recent developments make it possible to identify the causative virus in most cases of acute and chronic viral hepatitis. Improvements in supportive care and the availability of hepatic transplantation have considerably reduced mortality for the fulminant forms of acute viral hepatitis, and advances in antiviral therapy have led to the possibility of cure for some patients with chronic viral hepatitis and improvement for many others. Algorithms are presented as a guide to the diagnosis and management of acute and chronic viral hepatitis.


Pertussis toxin stimulates the secretion of [Met5]-enkephalin and the expression of proenkephalin A mRNA in bovine adrenal medullary chromaffin cells. [Met5]-Enkephalin (ME) secretion and the expression of proenkephalin A (proENK) mRNA were studied following long-term exposure of bovine adrenal medullary chromaffin (BAMC) cells to pertussis toxin. Treatment with pertussis toxin for 24 h increased the secretion of ME in a concentration- and time-dependent manner. The magnitude of ME secretion continued to increase with time in the presence of pertussis toxin. The intracellular concentration of ME in the pertussis toxin-treated group was not significantly different from controls, suggesting that elevated levels of ME secretion result from increased biosynthesis of ME rather than from release of stored ME. Prolonged (24 h) stimulation of BAMC cells with pertussis toxin also increased proENK gene expression. Pretreatment with nimodipine (a calcium channel blocker) and calmidazolium (a calmodulin antagonist) inhibited both the secretion of ME and the increase in proENK mRNA levels induced by pertussis toxin, while the intracellular calcium antagonist dantrolene and the protein kinase C inhibitors sphingosine and H7 [1-(5-isoquinolinylsulfonyl)-2-methylpiperazine] were ineffective in blocking pertussis toxin-induced responses. Forskolin (an adenyl cyclase activator) and isobutyl methyl xanthine (a phosphodiesterase inhibitor) increased both ME secretion and proENK mRNA levels; pertussis toxin synergistically increased the secretion of ME with these cyclic AMP-elevating agents but had only an additive effect with these agents on the level of proENK mRNA. Our results suggest that a pertussis toxin-sensitive G protein may tonically regulate the secretion of ME as well as the level of proENK mRNA.(ABSTRACT TRUNCATED AT 250 WORDS)


A pilot study of dried blood spot testing for HIV antibody in neonates. An unlinked anonymous HIV antibody study of neonates, using surplus dried blood spots on Guthrie cards, was introduced as a means of estimating the prevalence of HIV infection in childbearing women. From March 1990 to February 1992, blood spots from 14,520 infants were tested for anti-HIV, using an HIV IgG antibody capture particle-adherence test; only one sample gave a confirmed positive reaction. This type of study involves no interference in routine care and can be operated without difficulty in a district general hospital setting where neonatal metabolic screening is performed.


Performance assessment of neonatal dried blood spot testing for HIV antibody. A study was undertaken to assess the performance of laboratories participating in the UK unlinked anonymised neonatal screening programme for the detection of HIV antibody. A coded panel of 100 contrived dried blood spots prepared form well characterised anti-HIV-1 and anti-HIV-2 positive sera and an anti-HIV negative serum was distributed to eight testing centres. Fifty-one out of 52 anti-HIV-1 positive samples were identified by all eight laboratories: five laboratories identified all 52 specimens. Only one laboratory detected all four anti-HIV-2 positive spots. None of the laboratories reported a seronegative spot as reactive. The sensitivity of anonymised neonatal anti-HIV-1 screening was deemed satisfactory in all participating laboratories but the margin of sensitivity was insufficient to allow pooling of dried blood spot eluates before testing. Technical modifications would be necessary to increase sensitivity for anti-HIV-2 were HIV-2 infection to be reported more frequently in the UK. Performance assessment of the screening programme should be repeated at regular intervals.


Legionnaires' disease surveillance: England and Wales, 1991. One hundred and eleven cases of legionnaires' disease were reported in England and Wales in 1991, a smaller total than in any year since reporting began in the late 1970s. Four cases were hospital acquired and a further nine may have been associated with a hospital stay. Nosocomial legionellosis remains a cause for concern. Fifty-two cases were associated with travel abroad.


The surveillance and control of campylobacter infection. Campylobacter jejuni and C. coli are the most commonly reported bacterial causes of enteritis in man in the United Kingdom. The reported incidence of campylobacter infection varies with place and time. Its epidemiology differs from that of salmonella: campylobacters do not multiply on food, secondary spread is rare, and infections peak at the end of May (salmonellas peak in July). Known sources of infection include poultry, milk, water, pets and other domestic animals. Campylobacters are widespread in the environment, and in domestic and wild birds and mammals. About 10% of infections reported in the UK are acquired abroad. The application of preventive measures can be effective in reducing the incidence of infection and its not inconsiderable financial burden.


Vero cytotoxin-producing Escherichia coli O 157 in the United Kingdom. A total of 226 cultures of Vero cytotoxin-producing Escherichia coli O 157 isolated from humans was received by the PHLS Laboratory of Enteric Pathogens during the six month period January to June 1992. A record monthly total of 122 isolates was received in June. Ten phage types (PT) were identified during this period; PT 2 (45%) and PT 49 (21%) predominated. In addition, 27 human sera with antibodies to E. coli O 157 lipopolysaccharide (LPS) were examined from other cases during this period, making a total of 253 cases of infection associated with E. coli O 157.


Listeriosis surveillance: 1991.

Listeriosis surveillance: 1991. A total of 130 cases of listeriosis was reported in England, Wales and Northern Ireland in 1991. This represents a slight increase on the 1990 total of 118 reports but a marked decline compared with the peak incidence of reporting of 291 in 1988, which was part of an upsurge of cases between 1987 and mid 1989. Two epidemiological features of note in 1991 were the reappearance of a late summer-autumn peak in cases (commonly seen prior to 1987) and an increase in the number of reports among younger patients and children. The decline in listeriosis observed during the second half of 1989, which followed government health warnings about the consumption of pâté, and the continued low level of reporting since 1989 may be due to the disappearance of a common food source. However, in the light of the recent outbreak of listeriosis in France it is important that diagnostic vigilance and warnings about high risk foods are maintained.


The laboratory diagnosis of mycobacterial disease. The diagnosis of mycobacterial disease still relies heavily on conventional microscopy and culture techniques. More modern methods, such as the Bactec 460 radiometric system and the Roche biphasic system, are becoming available but are not yet widely used. Non-culture methods eg, using molecular biology techniques or gas liquid chromatography, are being developed but are currently the province of research and reference laboratories. Disease caused by species other than Mycobacterium tuberculosis poses problems of diagnosis and treatment. These other species can be identified by comparatively simple techniques but, as they are widely distributed in the environment, deciding between casual contamination and true infection can be difficult. Although such infections are not a major problem in numerical terms, the variable response of patients to treatment means that conventional sensitivity testing is of little assistance, and it is necessary to seek evidence of synergy in drug combinations for the effective treatment of these infections.


Food safety: the HACCP approach to hazard control. The Hazard Analysis Critical Control Point (HACCP) approach is becoming increasingly recognised as a valuable means of identifying and controlling hazards in the food production process, and thereby ensuring that food reaching the consumer is safe. The central feature of HACCP analysis is the determination of critical control points (CCPs)--those stages in the process which must be controlled to ensure the safety of the product. Once identified, a monitoring system is set up for each CCP to ensure that correct procedures are maintained and actions taken if CCP criteria are not achieved. The chief advantage of HACCP is that it is proactive; it aims to prevent problems occurring. Although HACCP is most readily applied to manufacturing processes, attempts are being made to adapt the system to other sectors of the food industry, such as catering.


Exposure to hepatitis B virus: guidance on post-exposure prophylaxis. PHLS Hepatitis Subcommittee. This paper summarises the views of the PHLS Hepatitis Subcommittee on prophylaxis after exposure to known and potential sources of hepatitis B virus (HBV) at work and in the community, and expands on the guidance on hepatitis B immunisation and the prevention of occupational exposure to blood given elsewhere. It defines significant exposure and gives guidance on incident recording, risk assessment, testing and storage of incident-related blood specimens and follow-up. It recommends that HBV prophylaxis should be determined by assessment of the likely infectivity of the source and of the HBV status of the person exposed.


Vitamin D3, its receptor and regulation of epidermal keratin gene expression. Among extrinsic modulators of keratinization are certain hormones and vitamins, which makes them potentially important pharmacological tools for treatment of keratinization disorders. Vitamin D3 and vitamin A, and their metabolites, promote and inhibit keratinization, respectively. We have shown that retinoic acid, via its nuclear receptor, directly suppresses the expression of the keratin genes which are markers of keratinocyte differentiation. Here we present evidence that 1,25(OH)2 vitamin D3 and its nuclear receptor do not directly regulate keratin gene expression. Co-transfection of a vector expressing the nuclear receptor for vitamin D3 with responder DNA constructs containing keratin gene promoters had no effect on the level of activity of keratin gene promoters either in the presence or in the absence of vitamin D3. We conclude that vitamin D3, unlike retinoic acid, modifies keratin synthesis indirectly, by changing the differentiation phenotype of the keratinocyte.