Inhibition of muscarinic receptor- and G-protein-dependent phosphoinositide metabolism in cerebrocortical membranes from neonatal rats by ethanol. Phosphoinositide (PtdIns) metabolism activated by cholinergic muscarinic receptors appears to play a role in brain development and has been recently suggested as a possible biochemical target for the developmental neurotoxicity of ethanol (EtOH). Recent experimental evidence indicates that, in rat brain, muscarinic receptor stimulation is coupled to PtdIns hydrolysis through regulatory GTP-binding proteins (G-proteins). We investigated the effect of various alcohol concentrations (10-500 mM) on guanine nucleotide-, fluoride-, and muscarinic-dependent PtdIns hydrolysis in [3H]inositol-labelled cerebral cortical membranes from neonatal (7-day-old) and adult rats. At both ages, EtOH exerted slight inhibitory effects on GTP(S) (100 microM)- and NaF (5 mM)-induced [3H]inositol phosphates accumulation. The presence of GTP(S) was necessary to unmask the stimulatory effect of the muscarinic agonist carbachol. Under these experimental conditions EtOH markedly inhibited carbachol (100 microM)-induced PtdIns hydrolysis. This effect was concentration-dependent and was more pronounced in the cortex from immature animals, where a statistically significant inhibition was observed at EtOH concentrations as low as 50 mM, comparable to the hematic concentrations reached following in vivo administration of doses of ETOH able to induce developmental neurotoxicity. These results confirm that EtOH exerts an age-specific inhibition of muscarinic-dependent PtdIns metabolism and suggest that this action might be exerted through an interaction with receptor-G-protein coupling.


Functional neuroteratology of drugs acting on adrenergic receptors. Noradrenaline neurotransmission becomes functional early in the development of the brain. Therefore, interference with this transmission by receptor agonists or antagonists, which are used clinically in various conditions in pregnancy, may alter brain development. Long-term biochemical, morphological, behavioural and electrophysiological effects have been reported after pre- or postnatal exposure of rats to adrenergic drugs, the alpha 2-agonist clonidine and the beta-antagonist propranolol in particular, and are summarized here.


Facilitation of agonist motoneurons upon initiation of rapid and slow voluntary movements in man. The time course of facilitation of the agonist motoneurons upon initiation of voluntary ankle dorsiflexion was investigated in eight healthy subjects. The H-reflex and visually guided tracking methods were used for testing the excitability of the motoneuron pool and for controlling the initiation of movement as well as speed and force. Since the onset of voluntary EMG activity (EMG reaction time: EMGvRT) was delayed and/or obscured by test H-reflexes which were evoked very close to the behavioral responses, the subject was instructed to make response movements bilaterally, and EMGvRT was measured on the side without stimulation. In every subject, the EMGvRT was invariably longer in the ramp movement than in the step movement. The onset of H-reflex facilitation prior to EMG onset, which was regarded as indicating the arrival time of the descending motor command to the motoneuron pool, always started earlier in the ramp movement than in the step movement. The difference in facilitation onset between the two tasks was smaller than that in EMGvRT. Since the amplitude of the H-reflex at the onset of the voluntary EMG was equivalent in both movements, the development of H-reflex towards the behavioral EMG onset was more gradual in the ramp movement than in the step movement. The present results demonstrate that the longer reaction time in the slow ramp task depends on 2 factors: delay in the arrival of descending facilitatory impulses to the agonist motoneuron pool, and its slow recruitment thereafter.


Excitatory amino acid receptors in brains of rats with methylazoxymethanol-induced microencephaly. We used methylazoxymethanol-acetate (MAM), a potent alkylating agent, to produce microencephaly in offspring by injecting it into pregnant rats on day 15 of gestation. Binding activities of central excitatory amino acid receptors were examined in Triton-treated membranes prepared from brains of adult offspring with MAM-induced microencephaly (MAM rats). MAM rats exhibited approximately 40-50% reductions of the wet weights of the cerebral cortex, hippocampus and striatum compared to those in controls. In the cortex and hippocampus of MAM-rats, total bindings of [3H]glutamate (Glu) (which is sensitive to N-methyl-D-aspartate (NMDA) receptor), and strychnine-insensitive [3H]glycine (Gly) and (+)-5-[3H]methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imi ne (MK-801; a noncompetitive antagonist of NMDA receptor), were reduced to approximately 40% of those in controls. Similarly, in both regions of MAM rats, total bindings of [3H]kainate and DL-alpha-amino-3-[3H]hydroxy-5-methylisoxazole-4-propionic acid (an agonist of quisqualate receptors), were reduced to approximately 35-50% of those in controls. However, total bindings of these radioligands in the striatum of MAM rats were more than 65% of those in controls, despite the significant loss of striatum mass. However, specific bindings of radioligands in the striatum of MAM rats were elevated by more than 60% of those in controls, and Scatchard analysis revealed that elevations of [3H]Glu, [3H]Gly and [3H]MK-801 bindings were due to a significant increase in the densities of binding sites, with their affinities remaining unaltered. Spatial recognition ability examined by an 8-armed radial maze task was markedly impaired compared to those in controls. These results suggest that the proliferation of neurons bearing excitatory amino acid receptors (EAA) in the striatum is less affected by MAM treatment on day 15 of gestation than that in the cortex and hippocampus in spite of drastic weight loss in these brain regions. The significant reduction of EAA receptors in the cortex and hippocampus may be involved in the impairment of spatial memory observed in MAM-treated rats.


Protein kinases and phosphatase inhibitors mediating long-term desensitization of glutamate receptors in cerebellar Purkinje cells. Long-term desensitization of the AMPA-selective glutamate receptors in Purkinje cells was examined in rat cerebellar slices by means of the wedge recording method. It was not induced by application of AMPA alone, but occurred regularly when slices were conditioned by perfusion with 0.5 mM 8-bromo-cGMP (but not cAMP derivatives) or the protein phosphatase inhibitors, okadaic acid and calyculin A. Phorbol esters also showed a similar effect. The 8-bromo-cGMP desensitization was antagonized by KT5823, an inhibitor of protein kinase G, while the effect of calyculin A was inhibited by polymyxin B, H-7, or K252a. These results suggest that AMPA receptors are persistently desensitized due to concerted action of both an agonist and an enzymatic system involving protein kinases G and C and a protein phosphatase inhibitor.


Axons from the olfactory bulb transplanted into the cerebellum form synapses with dendrites in the granular layer, as demonstrated by mouse allelic form of Thy-1 and electron microscopy. Transplant-to-host axon projection and synapse formation from the olfactory bulb (OB) transplant to the host cerebellum were studied using the mouse allelic form of Thy-1 (AKR strain of Thy-1.1 was used as host and BALB/c strain of Thy-1.2 as graft). Thy-1.2 immunohistochemical and ultrastructural examination showed that numerous OB axons elongate into the cerebellar granular layer and form asymmetrical synapses there with dendrites of host origin (perhaps of the host granule neuron). Factors which support this mismatched synapse formation are discussed.


Industrial blood lead levels in the South Island during 1988 and 1989: trends and follow up patterns. to assess trends in industrial lead exposure and the monitoring programmes in the South Island of New Zealand. during the period 1 January 1988 to 31 December 1989, industrial lead exposure was analysed in 1425 workers in at risk occupations and the efficiency of retesting programmes was determined. forty-four percent of these workers had red cell lead levels above 1.9 mumol/L, the top of the reference range for an unexposed population, and 71 individuals had levels exceeding the recommended action limits (males greater than 5.0 mumol/L, and females greater than 3.8 mumol/L). Although most occupational groups showed a small decline in mean red cell lead levels, the pattern of exposure was similar to previous reports. On average, only 43% of exposed workers were retested within the recommended period and 32% of these workers were not retested within 2 years of having a raised blood lead level. retesting was inefficient but was most reliable when industrial health nurses were employed for monitoring. Not all lead poisoning comes from the traditional lead based industries and significant decreases were found in workers whose primary exposure is to lead from petrol.


The cutting-grinding technique for histologic preparation of undecalcified bone and bone-anchored implants. Improvements in instrumentation and procedures. An improved technique for histologic sectioning of hard tissues is described. Thin sections (5 to 15 microns) of undecalcified tissues such as bones with ceramic and metallic implants as well as teeth with enamel and fillings can be produced. Implant-tissue and hard tissue-soft tissue interfaces are well preserved. Because of the development of equipment and materials designed specifically for this technique, consistently good results are obtained. Most stains used in the paraffin technique may also be used in these sections.


Orofacial complications of chemotherapy for breast cancer. The National Institutes of Health recently recommended research initiatives to investigate oral complications of cancer chemotherapy. This prospective cohort study investigated orofacial complications of combination chemotherapy (cyclophosphamide, methotrexate, fluorouracil, vincristine, and prednisone) in women with breast cancer. Thirty-four patients were given baseline interviews and examinations. Each patient was given weekly orofacial examinations and biweekly interviews for the first seven cycles of cytotoxic treatment. The orofacial complications included neurotoxicity caused by vincristine, mucositis, and candidiasis. Neurotoxicity affected 22 of 34 (65%) patients, was significantly associated with age less than 50 years (p less than 0.05), and manifested as pain in 19 of 34 (56%) patients. Mucositis affected 7 of 34 (21%) patients and was significantly associated with the occurrence of lesions of the oral mucosa at baseline examination; and smaller body surface area--indicating a dose-related toxicity (p less than 0.05). In four of the patients with mucositis (57%) granulocytopenia developed during the 7 days after the onset of mucositis. Intraoral candidiasis affected 4 of 34 (12%) patients.


Analysis of adenoid cystic carcinoma treated by radiotherapy. The records of 41 patients with adenoid cystic carcinoma of the head and neck region who had been treated with radiotherapy were reviewed. Local control was achieved in 72.3% in the cases with primary lesions at 5 years. The prognosis for tumors that arose in the major salivary glands was better than that for tumors that arose in the minor salivary glands; however, the difference was not statistically significant. In the minor salivary glands, early-stage tumors were well controlled with the use of radiation therapy alone. In spite of the high local control rate, the disease-free survival rate of the patients at 10 years was only 20.8%. Lung metastasis determined the prognosis.


Rickets--a complication of ifosfamide chemotherapy for Wilms tumor. Recent reports of metabolic bone disease occurring as a result of drug induced Fanconi syndrome due to ifosfamide have appeared in the pediatric literature. Ifosfamide is a chemotherapeutic agent used for treatment of pediatric solid tumors. We present an additional case of ifosfamide induced rickets, the first in the radiologic literature.


Enhancement of the circulating antibody secreting cell response in human diarrhea by a human Lactobacillus strain. Human Lactobacillus sp strain GG (Lactobacillus GG) administered during acute rotavirus diarrhea has been shown to promote clinical recovery. To elucidate the immune mechanisms behind such a favorable outcome, the ELISPOT (solid phase enzyme-linked immunospot) assay of Ig- and specific antibody-secreting cells among circulating lymphocytes was used, giving indirect evidence of the immunologic events in the gut. After rehydration, 39 children with acute rotavirus diarrhea, mean age 16 (SD 6) mo, randomly received either a Lactobacillus GG fermented milk product (study group) or a pasteurized yogurt (placebo group). The duration of diarrhea was significantly shorter in the study group than in the placebo group [mean 1.1 (SD 0.6) versus 2.5 (SD 1.4)d, p = 0.001]. Lactobacillus GG therapy was associated with a significantly enhanced nonspecific humoral response during the acute phase of the infection, reflected in the IgG, IgA, and IgM Ig-secreting cell numbers. At convalescence, 90% of the study group versus 46% of the placebo group had developed an IgA specific antibody-secreting cell response to rotavirus (p = 0.006). The results indicate that Lactobacillus GG promotes recovery from rotavirus diarrhea via augmentation of the local immune defense. Furthermore, specific IgA response to rotavirus is endorsed, which is possibly relevant in protection against reinfections.


Presence of non-maternal antibodies in newborns of mothers with antibody deficiencies. To explain the mechanism for induction and production of specific antibodies found in the newborn already at birth, without previous known exposure to the antigen, we chose a model that presumably excluded the possibility of specific antibodies being transferred from the mother to the fetus. Specific IgG, IgA, and IgM antibodies against Escherichia coli and poliovirus antigens were determined with ELISA in serum, saliva, and amniotic fluid from hypogammaglobulinemic and IgA-deficient mothers as well as in cord serum, saliva, and meconium from their offspring. All the mothers lacked IgA and some also lacked IgM antibodies, which were found in their healthy newborns. The amniotic fluid from a hypogammaglobulinemic mother lacking IgA contained small amounts of IgA antibodies, which were also found in the neonate, suggesting a fetal origin. There was evidence for the presence of antiidiotypic antibodies to poliovirus in the cord sera. We propose that idiotypic and/or antiidiotypic IgG antibodies transferred via the placenta from the mother to the fetus can initiate specific immune responses seen in the newborn. Thus, it may be that transplacental IgG not only passively protects the newborn, but also actively primes the fetus during fetal life via its content of idiotypic and/or antiidiotypic antibodies.


Activation of human eosinophils in vitro by respiratory syncytial virus. To determine the nature of the interaction between viruses and eosinophils, normodense eosinophils were separated from the blood of healthy volunteers and incubated in vitro with respiratory syncytial virus (RSV). After incubation for 2 h with the virus, 29.5 +/- 15.8% of the eosinophils demonstrated specific binding of the virus to the cell membrane, as detected by fluorescent staining with an anti-RSV MAb. Superoxide production and leukotriene C4 release were measured as determinants of cell activation. Using a cytochrome c reduction assay, superoxide could be detected in the supernatant 30 min after exposure to RSV. Maximal release was reached at 3 h postexposure (5.88 +/- 2.19 nmol cytochrome c reduction/5 x 10(5) cells). The virus-induced superoxide generation varied in magnitude among different subjects and ranged from 0.6 to 11.5 nmol cytochrome c reduction/5 x 10(5) cells. RSV also appeared to prime eosinophils to the effects of other known cell activators, as demonstrated by an increase in superoxide production upon subsequent stimulation of RSV-primed cells with phorbol-12-myristate-13-acetate (21.4 +/- 5.8 versus 9.4 +/- 2.7 nmol cytochrome c reduction/5 x 10(5) cells for primed and unprimed cells, respectively) (p less than 0.04). RSV did not directly induce leukotriene C4 release from the eosinophils but primed the cells to exhibit a more vigorous response on subsequent challenge with the calcium ionophore A23187 (9.16 +/- 1.04 versus 4.2 +/- 1.3 ng leukotriene C4/1 x 10(6) cells) (p less than 0.005). These findings indicate that RSV can activate or prime eosinophils to release various inflammatory mediators.(ABSTRACT TRUNCATED AT 250 WORDS)


[A case with delayed-onset radiation pneumonitis suspected to be induced by oral etoposide]. Radiation pneumonitis usually occurs within 1-3 months after the completion of radiation therapy. A 63-year-old male with primary lung cancer treated by radiation therapy developed radiation pneumonitis 5 months after the completion of radiation therapy. He received 60 Gy to the lung tumor in a conventional fractionation schedule, and then two courses of intravenous chemotherapy using cis-diamine-dichloroplatinum (II) (110-140 mg) and etoposide (140-175 mg). Oral etoposide was initiated for bone metastases on the 104th day after the completion of radiation therapy at a daily dose of 20 mg, to a total dose of 1075 mg. He complained of fever and exertional dyspnea 5 months after the completion of radiation therapy. Chest radiography showed homogeneous infiltrates in the irradiated lung. These clinical signs and symptoms were refractory to antibiotic therapy, but steroid therapy resulted in marked improvement. The development of radiation pneumonitis was suspected to be induced by oral etoposide, which was given before the onset of radiation pneumonitis. These data suggest that etoposide induces a recall phenomenon, as has been demonstrated with such drugs as adriamycin and actinomycin-D.


[Split-course radiation therapy in non-small cell lung cancer]. Split-course radiation therapy (Sp-RT) is based on theoretical differences between the kinetics of normal and malignant cells. A rest interval halfway through the course of treatment permits the normal tissues to recover, while the tumor shows vary little repopulation. Indeed, it shows mostly regression, resulting in shrinkage of the radiation field. From 1976 through 1985, 185 patients with localized but inoperable or unresectable (stage I-III) non-small cell carcinoma of the lung completed high-dose definitive RT delivered by continuous-course or split-course irradiation. Forty-seven patients who had large tumors or atelectases of the lung showing slow radioresponsiveness received Sp-RT over 60 Gy at 2 Gy per fraction. Rest periods were two or three weeks long in the interrupted schedules. The 5-year survival rate was 16% in the Sp-RT group and 13% in the continuous RT group. In the 33 patients that had differentiated epidermoid carcinoma with slow responsiveness to irradiation, the radiation fields could be shrunk by Sp-RT to the same extent as in the continuous group. Sp-RT was considered to be useful in the treatment of well-differentiated epidermoid carcinoma of the lung.


[A clinical trial of whole liver simultaneous dynamic MR imaging and its usefulness]. A clinical trial of whole liver simultaneous dynamic MRI was done. In 23 second whole liver was able to be scanned using parameters of filed echo method as follows: TR = 315 msec, TE = 7 msec, Flip angle = 70 degrees or 90 degrees, MAT = 50%, ECD = 60%, FOV 40 cm and no presaturation. Even 2-3 mm nodules of metastatic tumors and small daughter nodules of hepatocellular carcinoma such as 5 mm were demonstrated in the arterial phase clearly.


[Evaluation of multislice dynamic MR imaging of the whole liver by inversion recovery snap shot FLASH method]. We acquired dynamic images over the whole liver by inversion recovery snap shot FLASH method after a bolus intravenous injection of Gadolinium-DTPA. Each nodule of hepatoma in the liver showed early enhancement and gradually turned to show low intensity. In two cases of hepatoma, small intrahepatic metastases, which were not detected by US, CT and spin-echo image of MRI, were suspected as high intensity nodules on early phase. Also recurrent areas after TAE were enhanced on early phase. This method is practical for improving the detection of lesions and is useful for evaluating the recurrence after TAE.


[Development of a moving target aiming system using an ultrasound imaging unit]. We developed an ultrasound imaging unit for gating the irradiation of proton beams or acquiring CT scan data for treatment planning according to the motion of tumors in the abdomen. In proton therapy, it is essential that the maximum region of dose rate distribution in a body always coincide with the volume of the tumor in motion during irradiation. Gated proton bean irradiation based on tumor motion could solve this problem and minimize undesirable dose distribution to normal tissues in the vicinity of the tumor. This device can generate the TTL level signal of time width corresponding to the period that the tumor would be sited in a region determined in advance, to control the various kinds of machines. Results of our preliminary experiment using X-ray irradiation showed that this aiming device was able to make the width of the gated irradiation area coincide with that of the planned area within a difference of about 0.5 mm.


[Dynamic 3DFT FISP MR imaging of hepatocellular carcinoma]. Dynamic 3 dimensional Fourier transformation (3DFT) FISP MR imaging was performed in 5 patients with hepatocellular carcinoma before partial hepatectomy. Immediately after 0.1 mmol/kg of Gd-DTPA was administered intravenously, 3DFT FISP images (TR/TE/flip angle/slice thickness, 20 msec/8 msec/30 degrees/2-4 mm) were obtained every 30 seconds until 150 sec. We correlated dynamic MR images of the 5 patients with gross and microscopic findings. Some regions in the tumor corresponding to viable cells showed high intensity enhancement and other regions corresponding to necrotic regions showed no enhancement on the early phase images. We concluded that dynamic 3DFT FISP MR imaging which had good spatial resolution was useful in evaluating the vascularity of the tumor.


Splice site selection by intron aI3 of the COX1 gene from Saccharomyces cerevisiae. Interactions of the 5' and 3' splice sites with intron internal sequences of the yeast mitochondrial group I intron aI3 were studied using mutation analysis. The results can be fully explained by the splice guide model in which the splice sites are defined by the Internal Guide Sequence. No evidence was found for an alternative interaction between intron nucleotides preceding the 3' splice site and nucleotides in the vicinity of the core region as was found for the Tetrahymena intron. Our results also suggest that binding of the 5' and 3' splice site nucleotides to the IGS can not take place simultaneously. The intron must therefore undergo conformational changes as the reaction proceeds from the first step of self splicing, GTP attack at the 5' splice site, to exon ligation, the second step.


A shuttle vector which facilitates the expression of transfected genes in Trypanosoma cruzi and Leishmania. A Trypanosoma cruzi expression vector has been constructed using sequences derived from the flanking regions of the glyceraldehyde 3-phosphate dehydrogenase (gGAPDH) genes. The neomycin phosphotransferase (neor) gene was incorporated as a selectable marker. Using electroporation we have introduced this vector into both T. cruzi and Leishmania cells and conferred G418 resistance. Transformation is mediated by large extrachromosomal circular elements composed of head-to-tail tandem repeats of the vector. The transformed phenotype is stable for at least 6 months in the absence of G418 and can be maintained during passage through the T. cruzi life-cycle. Foreign genes inserted into an expression site within the vector (pTEX) can be expressed at high levels in transformed cells. To our knowledge this paper describes the first trypanosome shuttle vector and the first vector which functions in both trypanosomes and Leishmania.


In vitro replication of bacteriophage PRD1 DNA. Metal activation of protein-primed initiation and DNA elongation. Bacteriophage PRD1 replicates its DNA by means of a protein-primed replication mechanism. Compared to Mg2+, the use of Mn2+ as the metal activator of the phage DNA polymerase results in a great stimulation of the initiation reaction. The molecular basis of the observed stimulatory effect is an increase in the velocity of the reaction. The phage DNA polymerase is also able to catalyze the formation of the initiation complex in the absence of DNA template. Although the presence of Mn2+ does not affect either the polymerization activity or the processivity of the DNA polymerase, this metal is unable to activate the overall replication of the phage genome. This can be explained by a deleterious effect of Mn2+ on the 3'-5'-exonucleolytic and/or the strand-displacement activity, the latter being an intrinsic function of the viral DNA polymerase required for protein-primed DNA replication.


Ellipticine increases the superhelical density of intracellular SV40 DNA by intercalation. We investigated the in vivo effect of ellipticine, a mammalian topoisomeraseII(topoII) inhibitor, on SV40 DNA topology. In contrast to epipodophyllotoxins, ellipticine did not cause significant double stranded cleavage of intracellular SV40 DNA. Furthermore, ellipticine reduced cleavage induced by epipodophyllotoxins, VP16 and VM26. Unexpectedly, ellipticine dramatically increased the superhelical density of a fraction of intracellular SV40 DNA. Several lines of evidence suggest that the formation of this highly supercoiled DNA species (Ih form DNA) is not due to the inhibition of topoII per se, but is the result of intercalation by ellipticine in a subfraction of the intracellular SV40 chromatin followed by the fixation of DNA linking number by a topoisomerase activity. Based on the linking number change and the known unwinding angle of ellipticine, the intercalation density was calculated as one ellipticine molecule per 10-20 bp in the Ih DNA. This result suggests the existence of different populations of intracellular SV40 chromatin with respect to the accessibility to ellipticine intercalation.


Intron 5 alpha of the COXI gene of yeast mitochondrial DNA is a mobile group I intron. We have found that intron 5 alpha of the COXI gene (al5 alpha) of yeast mtDNA is a mobile group I intron in crosses between strains having or lacking the intron. We have demonstrated the following hallmarks of that process: 1) co-conversion of flanking optional intron markers; 2) mutations that truncate the intron open reading frame block intron mobility; and 3) the intron open reading frame encodes an endonuclease activity that is required for intron movement. The endonuclease activity, termed I-Sce IV, cleaves the COXI allele lacking al5 alpha near the site of intron insertion, making a four-base staggered cut with 3' OH overhangs. Three cloned DNAs derived from different forms of the COXI gene, which differ in primary sequence at up to seven nucleotides around the cleavage site, are all good substrates for in vitro I-Sce IV cleavage activity. Two of the strains from which these substrates were derived were tested in crosses and are comparably efficient as al5 alpha recipients. When compared with omega mobility occurring simultaneously in one cross, al5 alpha is less efficient as a mobile element.


Aberrant splicing of Gs alpha transcript in transformed human astroglial and glioblastoma cell lines. Alpha subunits of G proteins, which play a vital role in signal transduction, display considerable structural and functional diversity. Point mutations in two forms of alpha subunits, Gs alpha and Gi2 alpha, impairing their GTPase activity, have been detected in endocrine disorders. We report here the presence of truncated Gs alpha transcripts in a human glioblastoma cell line, HS683, and in an SV40-transformed human astroglial cell line, SVG. These transcripts were detected by polymerase chain reaction (PCR) amplification of cDNAs from the cell lines. The truncated Gs alpha transcripts, with deletions in the central region of the molecule, seem to have originated due to aberrant splicing within exonic sequences, which did not conform to the consensus GT/AG splice signals. The presence of a smaller size protein of mol.wt. around 25,000 kd in the SVG and HS683 cell lines, detected by antibodies specific for the C-terminal region of the Gs alpha subunit, seems to be consistent with the presence of truncated Gs alpha transcripts in these cell lines. These aberrantly spliced transcripts, if translated, could synthesize potentially oncogenic Gs alpha subunits deficient in GTPase activity. Whether such molecules, with sometimes relatively large deletions, retain some aspects of their function and are biologically significant remains to be seen.


Molecular analysis of DNA junctions produced by illegitimate recombination in human cells. In a human HeLa derived-cell line carrying permanently a single integrated copy of an SV40 shuttle vector, the transient expression of the SV40 T-antigen led to the production of heterogeneous populations of circular DNA molecules which retained both integrated vector and its surrounding cellular sequences. Comparison between the integrated copy and the linear maps of 80 different plasmids rescued in bacteria suggested that the formation of circular DNA was the result of bidirectional replication from the SV40 origin of replication followed by a single intramolecular joining leading to the cyclization of the replicated molecules. Sequence analysis of 45 recombinational junctions demonstrated that the cyclization occurred via illegitimate recombination process which did not require preferential nucleotide sequence at the joining sites. However, extensive characterization of recombination junctions revealed that the sequences involved in the recombination at each side of the SV40 origin of replication were not randomly distributed, suggesting the presence of regions which were more prone to be involved in the illegitimate recombination process in human cells. Search of common features usually implied in illegitimate recombination in mammalian cells revealed some association of these regions with palindromes, A + T-rich DNA segments, alternating purine/pyrimidine sequences and Alu family repeats.


Functional role of the ultraviolet light responsive element (URE; TGACAACA) in the transcription and replication of polyoma DNA. We have previously identified a novel 8 bp sequence (UV-responsive element, URE: TGACAACA) present in the regulatory region of polyoma DNA that interacts with protein factors induced in rat fibroblast cells by exposure to UV light. In the present study, we demonstrate through competitive binding assays that this sequence is distinct from the partially homologous AP1 and CRE target sequences. The proteins that bind to the URE appear to have transcriptional activity in UV-exposed rat fibroblasts. In addition, the URE appears to play a role in promoting the replication of polyoma DNA as determined through two different experimental approaches. Together, these findings suggest that the URE is a novel DNA binding element that interacts with proteins involved in the transcription and replication of polyoma sequences.


Singlet oxygen induced mutation spectrum in mammalian cells. In order to characterize the molecular nature of singlet oxygen (1O2) induced mutations in mammalian cells, a SV40-based shuttle vector (pi SVPC13) was treated with singlet oxygen arising from the thermal decomposition of the water-soluble endoperoxide of 3,3'-(1,4-naphthylidene) dipropionate (NDPO2). After the passage of damaged plasmid through monkey COS7 cells, the vector was shuffled into E. coli cells, allowing the screening of supF mutants. The mutation spectrum analysis shows that single and multiple base substitutions arose in 82.5% of the mutants, the others being rearrangements. The distribution of mutations within the supF gene is not random and some hotspots are evident. Most of the point mutations (98.4%) involve G:C base pairs and G:C to T:A transversion was the most frequent mutation (50.8%), followed by G:C to C:G transversion (32.8%). These results indicate that mutagenesis in mammalian cells, mediated by 1O2-induced DNA damage, is targeted selectively at guanine residues.


The role of chemotherapy in the treatment of lung cancer. Lung carcinoma despite treatment continues to be a leading cause of cancer death. The use of chemotherapeutic agents in the treatment of lung cancer does not promise cure but can enhance the quality of life of those patients suffering from this disease. The nursing care plan is an effective tool in managing the patient's side effects of both the disease and its treatment.


Purification and characterization of a mammalian endo-exonuclease. An endo-exonuclease has been purified from cultured monkey (CV-1) cells. The enzyme which was purified to near homogeneity to be a 65 kDa monomeric protein. The single-strand DNase activity is endonucleolytic and nonprocessive, whereas the double-strand DNase activity is exonucleolytic and processive. The enzyme was also found to have RNase activity using poly-rA as substrate. The pH optimum for ss-DNase is 8 and for ds-DNase it is 7.5. Both DNase activities require a divalent metal ion (Mg2+, Mn2+, Ca2+, Zn2+) for activity and exhibit the same kinetics of heat inactivation. The purified protein binds to and cleaves a synthetic Holliday junction substrate. The overall enzymatic characteristics of the mammalian protein are very similar to the putative recombination endo-exonucleases purified from Neurospora crassa, Aspergillus nidulans and Saccharomyces cerevisiae.


Nursing care of patients with ocular manifestations of human immunodeficiency virus infection. With the rising incidence of AIDS in the United States, a larger number of patients with HIV-related infectious and noninfectious ocular conditions will inevitably occur. This necessitates that ophthalmic nurses have knowledge of HIV disease; resultant ophthalmic manifestations; treatment protocols, including unapproved therapies; community resources for persons with vision loss; social issues related to HIV; psychologic responses to the disease; and associated losses for patients and their families. This knowledge base will enable the nurse to recognize disease earlier, to educate patients and their families, to administer treatment, and to provide support to a population in critical need of nursing interventions.


[The use of highly purified glucomannan-based fibers in childhood obesity]. To evaluate the effectiveness of highly purified glucomannan in childhood obesity a study has been carried out in 23 obese children (12 boys and 11 girls, aged 5.2-15.8 years), with excess weight of 51 +/- 16%, treated with 2-3 caps twice a day of glucomannan fibres (DICOMAN 5:2-3 gr/die), and in 30 obese children (aged 5-18 years) with excess weight of 51 +/- 10%, studied as controls. After a three-days food recall, a balanced diet with adequate caloric intake was provided to all obese children. In all patients before and 2-4 months after the auxological data (weight, height, weight excess) and laboratory data (serum levels of cholesterol, HDL, triglycerides, glucose, fructosamine, glycosylated hemoglobin, RBC, WBC, hemoglobin, iron, calcium, Cu and Zn) have been determined. Excess weight and triglycerides levels were significantly decreased in treated obese patients than in obese controls 4 months after the beginning of the study. A decrease of cholesterol levels was also observed in treated obese patients, but not in controls, whereas serum iron, calcium, copper and zinc persisted unchanged. No important side-effects were observed in treated patients. On the basis of our results highly purified glucomannan fibres may be employed with effectiveness in obese and dyslipidemic children together with diet.


[Wilms' tumor, polycythemia, ichthyosis and ear malformations: a new syndrome?]. Wilms' tumor (WT) is frequently associated to congenital malformations, i.e. aniridia (0.8%), hemihypertrophy (2.5%), and genitourinary malformations. Ear malformations have been suggested to be a sign of genitourinary malformations. On the other hand WT cases associated to ichthyosis has never been reported. The authors present a case of Wilms' tumor (WT) associated to ear malformations, ichthyosis and polycythemia, and suggest that such a malformation pattern might represent a new syndrome.


Second derivative of developed tension in classifying cardiotonics with respect to their mechanisms of action: drugs affecting cAMP metabolism. The second derivative of developed tension (T", d2T/dt2) has not come into common use in the analysis of cardiac contractility, although it has been shown to give additional information on the myocardial contraction-relaxation cycle (CRC). In the present study a new way to use T" in the analysis of myocardial mechanics, including the time course of T", is described. Profiles of the T" of the some drugs with established cardiotonic effects are presented. Experiments were carried out in spontaneously beating whole rat atria preparations. The effects of changing contraction frequency on the measured parameters were studied with electrically paced left atria. Qualitative inotropic effects of 1-methyl-3-isobutylxanthine (MIX), theophylline, caffeine, milrinone, isoprenaline and forskolin were studied. Concentrations equieffective with respect to the force of contraction were tested. Inotropic profiles were evaluated at the time of maximal force of contraction. We found that methylxanthines have a mechanical behaviour quite distinct from other inotropic agents acting via cAMP. The effects of MIX were similar to those of theophylline in all respects. A tendency to increase the active relaxation phase of T" was a property common to methylxanthines. Caffeine also prolonged the phases of contraction, whereas MIX and theophylline have opposite effects. Milrinone in turn mimics the effects of isoprenaline and forskolin; abbreviation of the relaxation phase of T" was the feature most typical of them. Caffeine was the only agent which did not shorten the duration of CRC. The method proved valuable in basic research on drug effects on cardiac contractility.


Comparative study of nitroimidazoles on the bioelectric properties of frog skin as a membrane model. The effects of misonidazole (MISO) and two other nitroimidazoles (5-NO2 and 4,5-NO2) on the bioelectric parameters of ion transport (potential difference and short circuit current) across frog skin as a membrane model, were studied in vitro. The nitroimidazoles investigated caused structure dependent effects on the sodium transport function of the membrane. MISO induced a biphasic action following administration on the external side of the membrane: after an initial enhancement, the potential difference and short circuit current signals both decreased. The other imidazole derivatives, 5-NO2 and 4,5-NO2, showed only one phase, whether administered on the external or internal membrane surface. All the nitroimidazoles investigated decreased sodium transport after internal or external surface administration. It was found that the 4,5-NO2 imidazole derivative irreversibly decreased the bioelectric membrane parameters.


Receptor subtypes in opioid and stimulant reward. Studies of the behaviourally-reinforcing actions of opioid and stimulant drugs of abuse are reviewed in an attempt to identify their reward-related brain receptors. We focus on data generated by drug self-administration, brain stimulation reinforcement, and conditioned place preference paradigms. A consistent body of evidence supports a role for mu and delta, but not kappa, receptors in opioid reward. Stimulant reward apparently involves both D1 and D2 receptors; the data favour D2 mediation of stimulant drug reinforcement with a permissive or modulatory role for D1 receptors. The reward-relevant opioid and dopamine receptors, as well as the cannabinoid (marijuana) receptor, share the ability to couple Gi proteins that mediate inhibition of adenylate cyclase and stimulation of K+ conductance. These signal transduction mechanisms thus may be generally implicated in the reinforcing properties of diverse drugs of abuse.


Patterned distributions of chondroitin sulfate isoforms, retinoic acid receptor gamma and cellular retinoic acid binding proteins in the embryonic mouse incisor. Odontogenesis is an example of developmental process whereby pluripotential embryonic ectodermal and ectomesenchymal cells give rise to specific, highly ordered, spatial arrangements of overtly differentiated tissues. This process implies the sequential and coordinated expression of several molecules of defined biological activities. The data presented here demonstrate the occurrence, in the embryonic mouse incisor, of a striking correspondence between gradients of mitoses, chondroitin 6-sulfate distribution, retinoic acid receptor gamma and cellular retinoic acid binding protein I transcripts. Although purely phenomenological, these observations provide an important guide to the design of experiments for understanding the mechanisms of pattern formation during tooth development.


Changes in pulpal nerves with aging.

Changes in pulpal nerves with aging. Aging of cat tooth pulp nerves involves ultrastructural changes, and changes in the expression of some neuropeptides and in the expression of the receptor for nerve growth factor. Electron microscopy of old pulps demonstrates loss and degeneration of unmyelinated and myelinated axons, as well as demyelination. Immunohistochemical findings show a marked age-related decrease in pulpal calcitonin-gene related peptide- and substance P-like immunoreactivity, and a reduction in nerve growth factor receptor-like immunoreactivity. Changes in neuropeptide expression are not entirely due to loss of nerve fibers, since most aging pulps contain nerve growth factor receptor-positive fibers which lack neuropeptide-like immunoreactivity. The changes reported here parallel the observation that there is an age-related reduction in sensitivity to pulpal stimulation, but may also contribute to the development of oral sensory phenomena such as neuropathic pain. Moreover, the senescent transformation of pulpal nerves probably affects hemoregulation of the pulp, and may thus have consequences for pulpal extraneuronal tissue.


Immunocytochemical localization of nerve growth factor receptor (NGFR) in human teeth. The distribution and subcellular localization of nerve growth factor receptor (NGFR) in human teeth has been investigated by immunohistochemistry at the light and electron microscopic levels. Many nerves in the dental pulp were intensely immunoreactive for NGFR. The pattern of distribution was largely similar to that of nerve fibers demonstrated by immunostaining for neurofilament protein (NFP), the most universal marker protein for pulpal nerves. In the predentin and dentin, more nerve fibers were intensely immunoreactive for NGFR than were demonstrable with NFP-antibodies. Immunoelectron microscopy showed that the NGFR-immunoreactivity was localized on the axoplasmic membrane in unmyelinated axons and on the outside of accompanying Schwann cells. No NGFR-immunoreactivity was found on the axons of myelinated nerves nor their supporting cells.


Number of mice per cage influences uncoupling protein content of brown adipose tissue. The effect of housing density of mice on the thermogenic state and capacity of their brown adipose tissue was studied. Mice were housed one, two, or six per cage at 28 degrees C for 15 days. Increased housing density suppressed the thermogenic capacity of brown adipose tissue (decreased the total amount of uncoupling protein) and decreased the thermogenic state of brown adipose tissue mitochondria (decreased GDP binding). A density of six mice per cage had a greater effect than a density of two mice per cage. The size of brown adipose tissue (wet weight and protein content), the content of mitochondria in it (cytochrome oxidase content), and the total activity of thyroxine 5'-deiodinase were not altered by housing density. We conclude that even at a temperature close to thermoneutrality (29-33 degrees C for the mouse), the occurrence of social thermoregulation (huddling) reduces the requirement for brown adipose tissue thermogenesis and results in a reduction in its thermogenic capacity. It is clearly of importance that the design of studies of mouse brown adipose tissue take into account not only the temperature at which the mice are housed, but also the number of mice housed per cage.


Reduction of eicosanoid production by essential fatty acid depletion does not attenuate the inflammatory response induced by Pseudomonas aeruginosa pneumonia in rat lung. Sipid mediators of inflammation have been implicated in the pathogenesis of Pseudomonas aeruginosa (PA) related pulmonary damage in patients with cystic fibrosis. We studied the role of these mediators in a rat model of PA endobronchitis using essential fatty acid deficient (EFAD) animals. Whole blood from EFAD animals produced significantly less leukotriene B4 (LTB4) and hydroxyheptadecatrienoic acid when stimulated ex vivo than did whole blood from control animals (p less than 0.005). Similarly, lung lavage fluid from EFAD animals infected with PA contained less LTB4 and thromboxane B2 (TXB2) than that from control animals. Despite these differences, cellular infiltration of airways in response to PA infection was virtually identical in animals from the regular diet and the EFAD groups. Both EFAD and control animals had a significant increase in white blood cells (WBC) in lung lavage fluid at 1, 3 and 6 days following infection with PA when compared to animals receiving sterile beads. Localized areas of consolidation and nodularity were grossly evident in the lungs of all PA infected animals irrespective of their ability to generate the lipid inflammatory mediators. Microscopic examination of lung sections demonstrated similar changes in all infected animals. We conclude that LTB4 and TXB2 production occurs early in the course of PA pulmonary infection in rats. This early rise in lipid mediators is temporally associated with an influx of WBC into the airways. However, attenuation of eicosanoid production by use of an EFAD diet does not lead to a reduction in the inflammatory response to PA infection.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of tumor necrosis factor-alpha on the stimulus-coupled responses of neutrophils and their modulation by various inhibitors. Preincubation of human peripheral blood polymorphonuclear leukocytes (HPPMN) with recombinant human tumor necrosis factor-alpha (rHuTNF-alpha) enhanced the formylmethionyl-leucylphenylalanine (FMLP)-induced superoxide (O2-.) generation in a concentration- and preincubation time-dependent manner. The enhancement was very high for the FMLP- or opsonized zymosan (OZ)-induced O2-. generation, but was low for arachidonic acid (AA)- and phorbol myristate acetate (PMA)-induced O.2- generation. The rHuTNF-alpha has no effect on the steady state of intracellular calcium ion concentration ([Ca2+]i) nor on the membrane potential of neutrophils. The rHuTNF-alpha-primed FMLP-induced O2-. generation was inhibited by nicotineamide (NA), pertussis toxin (PT), and by the tyrosine kinase (TK) inhibitor, genistein, but was enhanced by the protein kinase C (PKC) inhibitor, H-7 (1-(5-isoquinolinesulfonyl)-3-methyl-piperazine). The inhibitory actions of NA and PT were also observed in in vivo primed guinea pig peritoneal neutrophils (GPtPMN). However, FMLP-induced O2-. generation of GPtPMN was enhanced by genistein, but was inhibited by H-7. These data indicate that TNF-alpha does not induce changes in [Ca2+]i nor in membrane potential of HPPMN, and that TNF-alpha-primed FMLP-induced O.2- generation of HPPMN is coupled with ADP-ribosylation and activation of G-proteins, and that protein kinases, especially TK, seem to exert an important role in the priming action of TNF.


Invasive lobular and ductal carcinoma: mammographic findings and stage at diagnosis. The authors reviewed 316 cases of breast carcinoma diagnosed from January 1, 1986, to December 31, 1989. Clinical data and mammograms were available for all patients. Of the 316 carcinomas, 272 (86.1%) were invasive; 37 (13.6%) of these represented pure invasive lobular carcinoma (ILC). Twenty-five (68.5%) of the 37 patients with ILC and 161 (70.3%) of the 229 patients with invasive ductal carcinoma (IDC) presented with clinically palpable masses. Asymmetric opacities and architectural distortion were the predominant mammographic signs in 21 (57%) of the cases of ILC but only 32 (13.6%) of the cases of IDC. Malignant calcifications were not present in any of the patients with ILC but were present in 110 (47%) of those with IDC. Of the ILC lesions, 29 (85%) [corrected] had the same opacity as that of normal fibroglandular tissue, and the mammographic findings were often subtle and seen initially on one view only. There was no substantial difference in the TNM stage at diagnosis between the two study groups.


Fast spin-echo MR imaging of the female pelvis. Part I. Use of a whole-volume coil. In this prospective study, axial and sagittal magnetic resonance (MR) images were obtained with T2-weighted conventional spin-echo (CSE) and fast spin-echo (FSE) sequences in 34 consecutive female patients who underwent clinical pelvic MR examination at 1.5 T. The MR images from each patient were compared side by side, blindly and independently, by two radiologists experienced in MR imaging who used a standardized score sheet for anatomic and pathologic findings. The FSE sequences were rated superior significantly more often than the CSE sequences in most categories of findings (P less than .05), including overall image quality and reduction of motion artifact. The examination time for the FSE sequences was 1 minute 46 seconds versus an examination time of 9 minutes 14 seconds for the CSE sequences. (Both CSE and FSE sequences provided 18 sections.) It is concluded that the FSE sequence provides T2-weighted anatomic and pathologic information superior to that provided by the CSE sequence and requires substantially less imaging time.


Multicoil high-resolution fast spin-echo MR imaging of the female pelvis. A fast spin-echo pulse sequence was combined with multiple surface coils used simultaneously in the form of a "multicoil" in magnetic resonance imaging studies of the female pelvis. This combination allowed maximal resolution with maintenance of the signal-to-noise ratio (S/N) at an acceptable level, and the S/N with the multicoil system was substantially better than that achieved with a body coil. Excellent image quality and demonstration of anatomic detail were afforded by use of this technique.


Focal liver disease: comparison of dynamic contrast-enhanced CT and T2-weighted fat-suppressed, FLASH, and dynamic gadolinium-enhanced MR imaging at 1.5 T. Dynamic contrast medium-enhanced computed tomography (CT), T2-weighted fat-suppressed spin-echo (T2FS) magnetic resonance (MR) imaging, and breath-hold T1-weighted fast low-angle shot (FLASH) MR imaging before and after dynamic gadopentetate dimeglumine injection were compared in 73 patients with clinically suspected liver disease. Observer confidence for presence of focal lesions was determined by using receiver operating characteristic analysis. For all MR images, hepatic lesion-liver signal-to-noise ratios were evaluated qualitatively. and resolution and presence of artifacts were evaluated qualitatively. Lesion detection was greatest with T2FS (n = 272) and enhanced FLASH (n = 244) and was statistically greater with both of these than with CT (n = 220) and FLASH (n = 219) (P less than .03). Correct lesion characterization was greatest with enhanced FLASH (n = 236) (P less than .01), followed by CT (n = 199), FLASH (n = 164), and T2FS (n = 144). Enhanced FLASH was particularly successful in characterization of 5-mm- to 1.5-cm-diameter lesions as cystic or solid.


Factors affecting severity of renal injury and recovery of function in acute renal failure. Severity of renal injury and recovery of function in acute renal failure (ARF) are strongly related not only to the magnitude and nature of ARF insult but also to numerous factors in the host which govern renal susceptibility to the insult and repair of renal lesion. Prior ARF affords resistance to a rechallenge with the same or different ARF insult. The mechanisms for this acquired resistance to ARF have not been well established, but suggested mechanisms include (a) increased resistance of regenerated tubular epithelial cells to a rechallenge, (b) glomerular refractoriness to vasoactive substances, (c) failure of damaged kidney to concentrate the toxic substance, (d) enhanced antioxidant enzyme activity in glomeruli, and (e) increased Na(+)-K(+)-ATPase activity in regenerated tubular epithelial cells. Controversy still exists regarding roles of these factors in the resistance to renal failure. Functional and morphologic recovery of postischemic kidney is enhanced by antecedent unilateral nephrectomy but delayed in the presence of the contralateral kidney. The mechanisms for the effect of uninephrectomy remain unsettled. Recent studies suggest contributions of changes in preglomerular vascular resistance; alterations in the environment which follow ischemia to all functioning excretory renal tissues; and altered production and release of vasoactive substances such as angiotensin, endothelin, thromboxane, and atrial natriuretic peptide.


Endothelin and cyclosporine nephrotoxicity. Although some humoral and neural factors have been implicated in the persistent vasoconstriction characterizing many forms of acute renal failure, the mechanisms for this abnormal vascular function have remained largely unresolved. Several factors previously postulated to have a role in acute renal failure have been shown to enhance endothelin (Et) production or gene expression. We studied the potential pathophysiologic role for Et in several models of acute renal failure, including postischemia, endotoxin, and cyclosporine (Cy) nephrotoxicity. We have found that, in vivo, Cy (and also endotoxin) elevates circulating Et. We further showed that antagonizing Et's action with Et antibody ameliorates renal vasoconstriction following renal ischemia, Cy, and endotoxin administration. Additionally, our studies showed that even after circulating levels of Et decrease (following Cy), there is upregulation in Et receptors in the kidneys. Overall, endothelin appears to feature prominently in the pathophysiologic processes occurring during several forms of acute renal failure.


Reactive oxygen metabolites in toxic acute renal failure. We have examined the role of reactive oxygen metabolites (ROM) in gentamicin nephrotoxicity and in glycerol-induced acute renal failure, a model for myoglobinuric acute renal failure. Several agents which affect mitochondrial respiration have been shown to enhance the generation of hydrogen peroxide. Based on gentamicin's ability to alter mitochondrial respiration both in vitro and in vivo we postulated that gentamicin may enhance the generation of ROM by renal cortical mitochondria. Gentamicin, in a dose-dependent fashion, enhanced hydrogen peroxide production by rat renal cortical mitochondria as measured by the decrease in scopoletin fluorescence. At the highest concentration of gentamicin tested (4.0 mM), the rate of hydrogen peroxide generation was markedly increased from 0.17 +/- 0.02 to 6.21 +/- 0.67 nmol/mg/min. We next demonstrated that hydroxyl radical scavengers and an iron chelator provide a marked functional and histological protection in gentamicin-induced acute renal failure in rats. Hydroxyl radical scavengers and the iron chelator deferoxamine also protected renal function in glycerol-injected rats, a model for acute renal failure due to muscle injury. Although these data suggest that ROM may be important mediators of toxic renal injury, in vivo generation of ROM by kidney in normal and pathological states has not been previously examined. Aminotriazole (AT) irreversibly inactivates catalase only in the presence of hydrogen peroxide and previous studies have shown that AT-mediated inhibition of catalase in a sensitive measure of in vivo changes in the hydrogen peroxide generation. Using this method, we have demonstrated the in vivo generation of hydrogen peroxide under normal conditions and enhanced generation of hydrogen peroxide in rats treated with gentamicin or glycerol.(ABSTRACT TRUNCATED AT 250 WORDS)


Dense parenchymal accumulation of Lipiodol after transarterial chemoembolization. Dense accumulation of Lipiodol in hepatic segmental parenchyma was studied by computed tomography (CT) after transarterial chemoembolization (TACE) for hepatic tumor. Six patients showed dense accumulation of Lipiodol in hepatic segmental parenchyma on CT two weeks after TACE. Four of the six showed parenchymal accumulation of Lipiodol as dense as that in a tumor at three weeks after TACE. Therefore, it was considered that evaluation with CT should be performed after one month or more in order to differentiate between Lipiodol accumulation in tumors and that in non-neoplastic liver parenchyma.


[Coagulation and bronchopulmonary cancers: from clinical aspects to biology]. Activation of coagulation and of the fibrinolytic system has been identified in small cell and non-small cell cancers respectively. For the clinician this poses the diagnostic problem of a thrombosis, which is most often venous with or without pulmonary emboli, complicating the evolution of an already diagnosed cancer. The inverse is that these features may reveal an underlying neoplasm and amongst the most common of these would be bronchopulmonary cancer. Numerous laboratory studies have shown the existence of a state of hyper-coagulability, with disseminated intra-vascular coagulation, which is more or less compensated and is the more marked, the more advanced the cancer is. One should not fail to recognise that this state of hyper-coagulability may be aggravated by certain cytotoxic drugs. At the level of the tumour itself, there seems to be interactions between the cancer cells and the coagulation and fibrinolytic system: these interactions are very different according to the histological type as to whether they are small cell or non-small cell bronchopulmonary cancers.


[Role of inhalation therapy in the management of childhood asthma]. Inhaled drugs became of great interest in the treatment of childhood asthma. They must be adapted now to age and each form of the disease. The primarily interest of an organ therapy is to lead to a maximal efficacy by bringing locally an optimal quantity of drug without or with very few side effects. The choice of the device depends upon age which determines drug tolerance and quality of the inhalation technique. In infants and young children the use of nebulizers appears to be the most suitable technique; preschool children are capable to use metered-dose inhalers (MDI) with spacers; in older children the use of MDI, without spacers, or dry powder inhalers is allowed. During attacks of asthma, inhaled therapy appears to be effective in most cases using either B2 agonists alone in moderate forms, or B2 agonists associated with oral or parenteral corticoids in more severe forms. For the preventive treatment of asthma, in order to prevent attacks, some inhaled drugs also belong to a first line therapy against either allergy or on specific bronchial hyperreactivity: cromolyn or nedocromil are often used in mild to moderate forms (in association with oral anti-histamine drugs in some cases); in more severe forms we can start with a bronchodilator B2 agonist long-term treatment (associated with sustain-released theophylline in some cases), except in infants before twelve or eighteen months of age; in the most severe forms of chronic asthma an anti-inflammatory long-term treatment with inhaled corticosteroids may be prescribed even in young children.


Parathyroid hormone regulation of matrix degrading enzymes in rat osteoblastic osteosarcoma 17/2.8 cells. The present study was designed to further understand the role of PTH on the secretion of the neutral metalloproteinases, collagenase and gelatinase, from the rat osteosarcoma clonal cell line, ROS 17/2.8. Semiconfluent cells were treated with bovine parathyroid hormone, b-PTH-(1-34) at 100 nM-0.01 nM for 24-96 hours and pooled, concentrated media were analyzed by functional assay for collagenase (3H-methyl collagen) and gelatinase (3H-methyl gelatin). Collagenase activity significantly decreased (P less than 0.01) in the PTH conditioned media in a dose-dependent manner before (98-64%) and after (91-39%) reduction and alkylation. SDS-PAGE and fluorography apparently showed the most degradation to alpha A chains in collagen with controls, whereas this substrate remained intact with PTH (100 nM). PTH (100 nM) media also showed neutral gelatinase activity approximately 2% compared to control before and after reduction and alkylation (P less than 0.01). Significant amounts of an inhibitor to collagenase and gelatinase might have been secreted at 1 nM and 0.01 nM PTH, since collagenase and gelatinase activities were greater after reduction and alkylation. Reduction and alkylation likely destroyed these significant amounts of inhibitor. Polymorphonuclear leukocyte collagenase activity was also inhibited 80% by PTH conditioned media, but not by control. However, upon reduction and alkylation which destroyed inhibitor, the PTH treated media showed only a 14% inhibition against polymorphonuclear leukocyte collagenase (P less than 0.01). PTH appeared to downregulate neutral metalloproteinase activities through its effects on an inhibitor. This downregulation may represent a specific phenotypic response to PTH in ROS 17/2.8 cells.


The effect of dietary selenium deficiency on the antioxidant enzyme system in the gastrointestinal tract of the rat. The effect of selenium deficiency on the glutathione content and antioxidant enzymes in the gastrointestinal mucosa was studied. It was found that under Se-deficiency the level of glutathione increased, whereas the activity of Se-dependent glutathione peroxidase decreased. The activities of catalase and glutathione-S-transferase were unaffected. Malondialdehyde content was markedly increased in the glandular stomach of Se-deficient rats.


[Sensory ataxia in peripheral neuropathies--etiological and pathological analysis of four cases]. Among 85 neuropathy patients admitted and studied in the Department of Neurology, University of Occupational and Environmental Health, Japan, from 1979 to 1990, four patients suffering from sensory ataxia are reported with special reference to their etiological and pathological conditions. All of them were classified as having immune-mediated neuropathy. The first patient, a 56-year-old woman, was diagnosed as having chronic progressive ataxic sensory neuropathy. Her symptoms became progressively worse over a nine-year period after onset, but no evidence of cancer has been revealed. The positive rheumatoid factor was the only other feature noted. The second patient, a 63-year-old woman, after extensive laboratory studies, including the biopsy of the lymph node at the bifurcation of the bronchus in search of the cancer, was diagnosed as having subacute sensory neuropathy with small cell carcinoma of the lung. Chemotherapy was completed without subsequent obvious clinical benefits. The clinical diagnosis was confirmed on autopsy 29 months after the onset. The symptoms of the first patient were indistinguishable from those of the second patient, especially in the early clinical stage. In both patients, the proprioceptive sensations were severely affected and the disturbance of the proprioceptive sensations seemed to be almost parallel with the ataxia signs. The main site of the lesion seemed to be the neuron in the dorsal root ganglion in the first patient, as well as in the second patient who showed a marked loss of neurons in the dorsal root ganglion considered to be the primary lesion on autopsy.(ABSTRACT TRUNCATED AT 250 WORDS)


Using the virus challenge dose in the analysis of virus neutralization assays. We propose a new basis for adjusting the results of virus neutralization assays. These assays consist of two separate experiments performed in parallel: a virus titration experiment and a serum dilution assay. In the virus titration experiment, one estimates the amount of virus used in the assay (the virus challenge dose). In a typical virus neutralization assay, the virus challenge dose may range over an order of magnitude. In the serial dilution assay, one measures the serum neutralizing activity against a particular virus. Most standard methods use the virus titration results only to ensure that the overall experiment is acceptable; the specific results of the virus titration experiment are not used to adjust the estimate of serum neutralizing activity. Although adjustment based on calibration with reference sera could be done, this seldom occurs in practice. We use results from recent studies of the kinetics and stoichiometry of polio virus neutralization with monoclonal antibodies to develop a method to use results from the virus titration experiment to adjust the serum neutralizing activity directly. Our results also indicate that a simple ad hoc procedure can improve the accuracy of the estimated serum neutralizing activity.


Detection of enteroviruses in faeces by polymerase chain reaction. A polymerase chain reaction (PCR) technique for the detection of human enteroviruses in stool specimens was developed. The test was based on the synthesis of cDNA, followed by PCR and slot blot hybridization. The primers used were selected from a highly conserved sequence in the 5'non-coding region of the enteroviral genome. By this method 27 different enterovirus serotypes (15 echo, 6 coxsackie A, 4 coxsackie B, poliovirus type 2 and enterovirus 71) from 89 patients could be detected. Using positive virus culture as reference, the sensitivity of PCR was 69% after 30 cycles of amplification, 91% using 30 + 10 cycles and 100% following 2 rounds of amplification with ensuing hybridization. None of 23 stool samples from healthy individuals or patients with meningitis of proven non-enteroviral etiology were positive by the PCR. By contrast, 13/26 culture-negative, randomly chosen stool samples from patients with suspected enteroviral disease were positive by the test. These findings demonstrate a high sensitivity and an apparently high specificity of PCR for detection of enteroviruses in stool samples. Therefore, the methodology may be useful in the laboratory diagnosis of enterovirus infections.


Herpes simplex virus--the most frequently isolated pathogen in the lungs of patients with severe respiratory distress. 308 consecutive patients with severe or complicated respiratory tract infections underwent fiber-optic bronchoscopy in the search for a microbiological etiology. Protected brush specimens were used for bacterial cultures and bronchoalveolar lavage (BAL) for virus isolation and cytological examination. Herpes simplex virus (HSV) was the most commonly found pathogen and was isolated in 37 patients. 20 (54%) of them also had serological and/or cytological signs of HSV infection. 132 patients required assisted ventilation (AV) and in this group 34 (92%) of the 37 HSV positive patients were found. Isolation of HSV was significantly (p less than 0.001) associated with AV compared to patients not requiring AV. Of all patients treated with AV 26% had positive HSV isolation in conjunction with suspected acute lower respiratory infection. Coinfection with HSV and bacteria occurred in only 8 (22%) patients. HSV was more common in patients with burns (47%) compared to other patient groups such as patients with AIDS (3%) or other immunodeficiencies (9%).


DNA fingerprinting in the epidemiology of African serogroup A Neisseria meningitidis. The restriction endonuclease (RE) technique was used to compare 172 meningococcal group A strains collected between 1969 and 1990, mainly from countries of the so-called African Meningitis Belt, the Gambia and Ethiopia. The 64 strains from various African countries (Niger, Chad, Burkina Faso, Cameroon, Morocco, Djibouti) were distributed within 3 main restriction enzyme patterns (REPs); the 77 Gambian strains fell into 5 REPs and the 24 Ethiopian strains into 2 such patterns. Several of the main REPs were formed by clusters of closely related clones. Clones, very similar to dominating REPs of the 1960s in Niger, Burkina Faso and Cameroon, were in the 1980s found to be strongly represented in the Gambia to the extreme west of the Meningitis Belt. One of the Gambian clones from 1983-86 was identical to an Indian clone recovered in New Delhi 1986-87. Another clone was detected in 1983 in the Gambia, in 1989 again in the Gambia as well as in Ethiopia, and in 1990 in Tanzania. Our results are largely in line with those of previous studies based on modern techniques of protein and isoenzyme electrophoresis. The RE method is useful mainly for the exact genotypic differentiation of closely related clones, and seems to be a valuable complement to phenotypic tools for epidemiological mapping of Group A meningococcal infection.


Hemiplegia--a rare complication of acute Epstein-Barr virus (EBV) infection. A 32-year-old male presented to hospital with a transient hemiplegia associated with a rash and systemic upset. He was found to have an acute Epstein-Barr virus (EBV) infection. Hemiplegia complicating glandular fever has been described but once previously.


A truncated erythropoietin receptor that fails to prevent programmed cell death of erythroid cells. A form of the human erythropoietin receptor (EPOR) was identified in which the cytoplasmic region is truncated by alternative splicing. The truncated form of the receptor (EPOR-T) is the most prevalent form of EPOR in early-stage erythroid progenitor cells, but the full-length EPOR (EPOR-F) becomes the most prevalent form in late-stage progenitors. EPOR-T can transduce a mitogenic signal. However, cells transfected with EPOR-T are more prone to programmed cell death than those expressing EPOR-F. EPOR-F may transduce a signal to prevent programmed cell death that is independent of the mitogenic signal, and alternative splicing of the EPOR gene may have an important role in erythropoiesis.


[Cytochemical study of human sperm metabolism during in vitro capacitation after cryopreservation]. To study the effects of deep freezing on the energy metabolism of human spermatozoa, we investigated, by cytochemical quantitative methods, cytochrome oxidase and lactate dehydrogenase activities of fresh and frozen human spermatozoa during in vitro capacitation. Fresh and frozen human spermatozoa were incubated in Biggers, Whitten and Wittingham's medium supplemented with 15% heat-inactivated human serum. Both histoenzymological reactions can be quantitated and have been evaluated by microdensitometric method. The results indicate that human spermatozoa depend almost entirely on anaerobic glycolysis during in vitro capacitation and suggest that both aerobic and anaerobic metabolism in spermatozoa are only slightly impaired by freezing-thawing and storage.


[Treatment with rufloxacin of complicated urinary tract infections]. Eighteen patients of either sex (14 M; 4 F), ranging in age from 23 to 79 years, with clinical diagnosis of complicated cystitis due to Rufloxacin sensitive pathogens, were enrolled. Rufloxacin was administered orally at the dosage of 400 mg/die the first day; 200 mg/die the following 6 days or more. The mean duration of treatment was 7.25 +/- 0.78 days. No concomitant antimicrobial therapy was administered during the study. At the end of therapy 5/14 evaluable patients recovered, 9/14 evaluable patients improve; 4 patients were considered by Investigator as "not evaluable". Causative pathogens were isolated in all patients and eradicated in 18 out of 18 bacteriologically evaluable patients (eradication rate = 100%). Neither reinfections nor superinfections occurred. No clinical adverse event related to study medication was reported. The results indicate that Rufloxacin at the oral dose of 200 mg/die is well tolerated and effective in the treatment of complicated cystitis.


The evolving role of systemic therapy in carcinoma of the lung. Locally advanced lung cancer carries a poor prognosis, and its treatment continues to challenge medical, radiation, and surgical oncologists. While systemic chemotherapy has improved the survival of patients with small cell lung cancer (SCLC), the role and timing of thoracic radiotherapy has not been clearly defined. The roles of chemotherapy and radiotherapy appear to be reversed in the treatment of locally advanced non-small cell lung cancer (NSCLC). The routine use of thoracic radiotherapy has been shown to improve local control after surgery without affecting survival, due to a high incidence of distant metastases. This contrasts with the marginal survival benefit seen with chemotherapy in NSCLC. Nevertheless, the results of recent clinical trials in both SCLC and NSCLC are encouraging and support continued investigation. These studies and the results of recent pilot studies suggest that a closer integration of chemotherapy and radiotherapy (concomitant chemoradiotherapy) may be necessary for further improvement in outcome. This review will present the results of recent studies in systemic therapy of lung cancer and the evidence supporting concomitant chemoradiotherapeutic treatment of this disease.


Randomized double-blinded comparison of three intravenous immunoglobulin products in bone marrow transplantation. We have used samples from the in vivo situation to compare antibody levels provided by the infusion of different IVIG products, a measure, albeit indirect, of potential therapeutic efficacy. The further correlation of in vivo antibody titers with functional activities of these antibodies (eg, opsonization and viral neutralization) would provide additive valuable information about the usefulness of this therapy.


Characterization of three mutations causing von Willebrand disease type IIA in five unrelated families. Von Willebrand disease (vWD) type IIA is characterized by decreased ristocetin-induced platelet aggregation, and by the absence from plasma of high molecular weight multimers of von Willebrand factor (vWF). Most mutations causing vWD type IIA are clustered within the A2 domain of the mature vWF subunit that is encoded by exon 28. Using the polymerase chain reaction (PCR), the entire exon 28 from patients with vWD type IIA and normal controls was amplified and sequenced. Three missense mutations were detected that result in the amino acid substitutions were detected that result in the amino acid substitutions Arg(834)----Trp, Gly(742)----Glu, and Ser(743)----Leu. The first mutation occurred independently in three unrelated families; each of the latter mutations was found in one family. By restriction endonuclease analysis and allele-specific oligonucleotide (ASO) hybridization the mutations were confirmed in affected family members and excluded in unaffected members and 50 normal controls. The apparently high frequency of identical independent mutations among patients with vWD type IIA suggests that a precise diagnosis may be possible in a majority of patients using relatively simple recombinant DNA screening assays.


Low molecular weight heparin and prevention of postoperative thrombosis in abdominal surgery. In a prospective, double-blind, randomized multicenter trial the efficacy and safety of low molecular weight heparin and unfractionated heparin were compared for the prevention of postoperative deep vein thrombosis in patients undergoing abdominal surgery. Six hundred and seventy-three patients were randomly allocated to the two prophylaxis groups; 20 of these, however, did not undergo surgery and did not receive any prophylaxis. Of the remaining 653 patients 323 received one subcutaneous injection of 3,000 anti-Xa units of low molecular weight heparin and 330 received subcutaneously 5,000 U heparin three times a day. Treatment was initiated 2 h preoperatively and continued for 7 to 10 days. The occurrence of DVT was determined by the 125I-labelled fibrinogen uptake test and phlebography. Venous thrombosis was diagnosed in 24 of 323 patients (7.4%) treated with low molecular weight heparin and in 26 of 330 patients (7.9%) treated with low-dose heparin. DVT of proximal veins was detected in four patients of the low molecular weight heparin group and in three patients of the low-dose heparin group. During the observation period three pulmonary emboli - one fatal and two non-fatal - occurred in patients receiving prophylaxis with low-dose heparin. No pulmonary embolism was found in patients treated with low molecular weight heparin. Both prophylactic schemes were well tolerated. Intra- and postoperative blood loss, incidence of wound hematoma, frequency and volume of intra- and postoperative blood transfusion were similar in both groups with a slight advantage for the low molecular weight heparin group.(ABSTRACT TRUNCATED AT 250 WORDS)


[Ten years of clinical experiences with canine parvovirus infection CPV-2 infection)]. Clinical and laboratory data of 205 dogs with acute canine parvovirus infection were collected. Seventy-six of these dogs underwent 179 clinical reexaminations. Parvovirus infection can be clinically diagnosed if there are typical gastrointestinal symptoms (vomiting, diarrhoea) in combination with a leucopenia of under 4000 cells/microliters and/or a rise in antibody titer of two degrees or more. In typical parvovirus infection a distinct hyperthermia often precedes vomiting and diarrhoea. The characteristic fall in leucocyte concentration is rapid and most marked between the 3rd and 5th day of the disease. In some cases virus-induced encephalitis and moderate coincident involvement of the cardiac muscle were seen. Secondary bacterial infections of subcutaneous tissue and lungs as well as invaginations are important complications. Surviving dogs will not necessarily have digestive problems later. Therapeutical considerations are discussed.


[Parapox in a pigmy chimpanzee].

[Parapox in a pigmy chimpanzee]. Parapoxvirus was found using electron microscopy in skin scrapings from a pygmy chimpanzee with a pustular skin disease. Clinical findings are described and possible ways of transmission are discussed. It seems remarkable that this is the first time parapoxvirus has been diagnosed in a nonhuman primate.


Binding of brevetoxins and ciguatoxin to the voltage-sensitive sodium channel and conformational analysis of brevetoxin B. The marine toxins known generically as brevetoxins, as well as their structural relative ciguatoxin, are known as polyether ladder toxins, and bind uniquely to site 5 of the voltage-sensitive sodium channel. Rat brain synaptosome binding data show similarities in binding affinity for brevetoxins having the same structural (ladder) backbone, but different affinities between brevetoxins having different backbones. Ciguatoxin has a different backbone from the brevetoxins, but binds even more strongly to the same site. Could the flexibility of the backbone be related to their relative toxicities? As part of an effort to identify the common pharmacophore for the toxins, Monte Carlo methods were used to generate conformational models of the polyether ladder toxin brevetoxin B (PbTx-2) which shows significant flexibility at the juncture of the two 7-membered rings.


Systemic antiviral drugs used in ophthalmology. Over the past two decades, the recognition of viral enzymes and proteins that can serve as molecular targets of drugs has revolutionized the treatment of viral infections. Beginning with acyclovir, a number of systemically administered agents which are both relatively safe and effective for the treatment of herpetic infections and human immunodeficiency virus (HIV) infections have become widely available. Because of increased numbers of herpes virus infections, as well as the rising epidemic of HIV infections, the ophthalmologist is, more likely than ever before to be involved in the treatment of severe and frequent ocular infections caused by herpes viruses. In addition, the acute retinal necrosis (ARN) syndrome has been demonstrated to be caused by herpes viruses and a once rare retinal infection caused by cytomegalovirus is common in patients with the acquired immunodeficiency syndrome (AIDS). In this article, four systemic antiviral drugs (Vidarabine, Acyclovir, Ganciclovir, and Foscarnet) that have demonstrated usefulness in the treatment of ophthalmic disease are reviewed in detail with regard to their mechanisms, applications, effectiveness, and side effects.


Effect of beta-endorphin on cAMP accumulation in rat luteal cells. The effect of beta-endorphin on cAMP levels in 4-day-old rat luteal cells was investigated. In both the presence and absence of low doses of human chorionic gonadotropin (hCG, 0.001 IU/ml), beta-endorphin inhibited cAMP accumulation, whereas in the presence of high doses of hCG (0.01 IU/ml) it did not. This inhibitory effect was abolished by pre-treatment with islet-activating protein (IAP). Moreover, treatment with IAP resulted in an overall enhancement of hCG-stimulated cAMP accumulation when compared with untreated controls. These results suggest that beta-endorphin suppresses adenylate cyclase activity via Gi, which may be coupled to the LH receptor.


Effect of ultrasound on the neonatal rat brain. Six-day-old rat neonates were exposed to 2.25 MHz continuous wave (CW) ultrasound for 5 min at an intensity of 2.5 W/cm2 (SATA). The temperatures on the head surface and in the mouth were measured. There was a higher average temperature elevation in the mouth (9 degrees C) than on the head surface (7 degrees C). Survival differs between control and exposed groups at 30 days after exposure. Ninety percent of the control group lived to 30 days, versus 59.7% for the exposed group. At differing times following exposure, the brains of the pups were removed and tested for enzymatic activities. Changes in acetylcholine esterase and in 2',3'-cyclic nucleotide phosphohydrolase activities were not statistically significantly different from controls. There were no significant differences in brain weight and total protein between control and exposed pups.


Human papillomavirus sequences are not detectable by Southern blotting or general primer-mediated polymerase chain reaction in transitional cell tumours of the bladder. A large series of transitional cell tumours has been screened for the presence of human papillomavirus (HPV) sequences using Southern blotting and general primer-mediated polymerase chain reaction (GP-PCR). The latter technique allows the detection of a broad spectrum of both sequenced and unsequenced HPV types using two primer pairs located in the highly conserved L1 and E1 regions of the HPV genome. No evidence for HPV infection was found in 100 transitional cell tumours, 6 cases of carcinoma in situ, 2 adenocarcinomas and a squamous carcinoma of the bladder and 3 cases of cystitis. Similarly, 12 bladder tumour cell lines were HPV-negative in these assays. Cervical carcinoma cell lines containing from 1-3 to 600 copies of the HPV genome were used as positive controls and were scored positive in all assays by both Southern blotting and GP-PCR. It is concluded that despite the close proximity of the urothelium to the genital mucosa and the resemblance of some bladder tumours to known HPV-induced lesions in other tissues, HPV infection is absent or very uncommon in bladder tumours.


Receptor kinetics and concentration-effect relation of cardiac glycosides. Therapeutic and toxic actions of cardiac glycosides are attributed to an inhibition of Na, K-ATPase. The therapeutically relevant range is between 25% and 50% inhibition. There is a good correlation between the average steady state serum concentration of glycosides and their therapeutic action. However, therapeutic and toxic effects set in with a latency and therefore do not follow the daily variations in glycoside concentration. Although the effect follows the average serum concentrations, only the minimal concentration is measured. In principle this is only adequate if the ratio of average/minimal concentration is constant. A model calculation showed that with a constant average steady state concentration an increase in the distribution volume or a decrease in total body clearance with corresponding reduction of the daily dose lead to an increase of the minimal concentrations of 5-7%. This means a corresponding underestimation of the average concentration from the minimum concentration. However, the deviations are too small to be of clinical relevance.


[Possibilities for preventing the epidemic spread of rotavirus infections on neonatal wards]. Rota viruses are the most frequent cause of acute gastroenteritis of infants and toddlers. Small epidemies occur enlarged in institutions for newborns and toddlers. The infection occurs normally via faeces. An aerogenes infection is in discussion. In case of a suspected Rota-Virus-Infection in an infant- or toddler care unit the patients have to be isolated immediately or quarantined. Further essential precautions preventing the spreading are mentioned. With this regime we could only see individual Rota-Virus-Infections in infant care units.


[cAMP-binding protein--a protein kinase of the plague pathogen]. In Y. pestis a cyclic AMP-binding protein was detected, isolated to a homogeneous state, and its physico-chemical properties were studied. The protein is a highly molecular compound with a molecular weight of 180 kD, capable of being released into the environment in the process of cell growth and having protein kinase activity, not depending on the presence of cyclic AMP. Y. pestis neuraminidase is one of the substrates appearing due to the action of protein kinase detected in this study. Y. pestis protein kinase may alter the spectrum of protein phosphorylation in the leukocytes of white mice. The direct participation of this protein in the development of infection is supposed.


[Status of collective immunity to hepatitis A virus in urban residents in Byelorussia]. The large immune stratum and intense collective immunity to virus hepatitis A among the urban population of Byelorussia are characteristic of hyperendemic territory. The geometric mean of the antibody titer has been noted to increase with age, which is probably due to repeated infections of persons who have already had the disease. The use of this value for the characterization of collective immunity and epidemiological situation has been proposed.


[Antibody prevalence for hepatitis C and other parenterally transmissible viral diseases in i. v. drug dependent patients]. Serum samples of 151 intravenous drug users were tested for markers of hepatitis B, hepatitis C, HIV and HTLV-I infection to estimate the prevalence of blood born virus infection in this high risk group. Anti HCV antibodies were found in 75% of sera. Seroprevalence for HIV was found 13%, for hepatitis B markers 68% and none for HTLV-I. Multiple infections have been very frequent.


Selectivity and minimal androgenicity of norgestimate in monophasic and triphasic oral contraceptives. The contraceptive progestin norgestimate (NGM) has a high affinity for uterine progestin receptors and a lack of affinity for androgen receptors similar to that of natural progesterone. NGM's selectivity results in excellent efficacy, cycle control, and minimal androgenicity when it is combined with ethinyl estradiol (EE). Clinical studies of a monophasic regimen of NGM/EE indicate a positive impact on lipid metabolism, revealing an increase in serum levels of high-density lipoprotein cholesterol with a concomitant and significant decrease in the low-density lipoprotein/high-density lipoprotein cholesterol ratio. Little impact on carbohydrate metabolism was noted. Serum levels of sex hormone binding globulin, an indicator of androgen-estrogen balance, also increased significantly with NGM/EE in accordance with its low androgenic activity. A significant between-regimen difference in SHBG was seen in a comparison study of NGM/EE and LNG/EE triphasic formulations (a mean rise of 68.6% with NGM/EE vs a decrease of 6.1% with LNG/EE). NGM's lack of estrogenicity was evidenced by unchanged prolactin levels and absence of effect on the coagulation system. In a large study of the monophasic formulation in 59,701 women, some improvement in acne was reported as well as minimal weight gain. An overview of clinical data is provided from United States and European trials as well as some preclinical data relevant to NGM's selectivity.


Cycle control with triphasic norgestimate and ethinyl estradiol, a new oral contraceptive agent. Effective cycle control was demonstrated based on two multicenter, 2-year studies of the triphasic oral contraceptive (OC) agent containing the new progestin norgestimate. The estrogen in this OC is ethinyl estradiol. These open-label Phase III studies were conducted in the United States by 33 investigators at 33 sites who treated a total of 1,783 subjects, healthy women 17 to 38 years of age with menstrual cycle characteristics considered to be within the normal range. The norgestimate/ethinyl estradiol preparation was taken for up to 24 cycles. Follow-up information was collected 3 to 4 months post-treatment. Bleeding pattern analyses were based on 27,970 valid cycles. Normal cyclic bleeding patterns were experienced by most of the women during the study; only minimal and statistically and clinically insignificant variations in menstrual flow, dysmenorrhea, and premenstrual tension occurred. There was a low incidence of failed withdrawal bleeding in single cycles (less than 1.0% after cycle 6). There were no cases of amenorrhea, defined as two consecutive cycles of missed withdrawal flow. The incidence of breakthrough bleeding or spotting was highest during the initial treatment cycles and diminished with continued use of the formulation. The mean incidence of breakthrough bleeding was 2.36% in cycles 13 to 24. Apart from somewhat higher initial percentages among women new to oral contraception, the pattern of midcycle bleeding or spotting was similar to that of all women studied. Effective long-term cycle control was demonstrated in women who used this OC agent.


Clinical evaluation of a new triphasic oral contraceptive: norgestimate and ethinyl estradiol. The safety and efficacy of the triphasic oral contraceptive agent containing norgestimate and ethinyl estradiol were evaluated in a 12-month study of 661 women. Excellent contraceptive efficacy was achieved, with two pregnancies ascribed to product failure in a total of 6,511 treatment cycles. The life-table predicted pregnancy rate was 0.57 per 100 woman-years of use. The overall and theoretical Pearl indexes were 0.55 and 0.37, respectively. Good cycle control was maintained in patterns similar to those noted in previous studies. The incidence of dysmenorrhea and premenstrual syndrome was sharply reduced. Side effects reported were typical of those associated with use of low-dose oral contraceptive agents. Acceptability was high compared with agents used previously by the subjects. Total cholesterol did not change but high-density lipoprotein cholesterol was significantly elevated at 3 and 12 months. There were no clinically significant changes in the parameters of hematology or blood chemistry tested.


A comparison study of lipid and androgen metabolism with triphasic oral contraceptive formulations containing norgestimate or levonorgestrel. The effects of norgestimate triphasic (Ortho Tri-Cyclen, Tri-Ciles) and levonorgestrel triphasic (Triphasi) formulations on lipid and androgen metabolism were assessed in a study of 66 healthy women treated through six menstrual cycles. Levels of the following were measured: cholesterol and its subfractions, triglycerides, carrier lipoproteins, estradiol, testosterone, and sex hormone binding globulin (SHBG). Comparison of baseline values with values after 3 and 6 months of treatment indicated that both regimens influenced lipid and androgen metabolism. There was a statistically significant between-regimen difference in levels of high-density lipoprotein, which were favorably increased with norgestimate triphasic but reduced with levonorgestrel triphasic. Related data on SHBG showed that plasma levels of this marker of estrogen/androgen balance were increased significantly more in the norgestimate triphasic group, providing additional evidence of low androgenicity. Both regimens inhibited follicular growth to the same extent, as evidenced by low mean levels of estradiol in all on-therapy cycles; and both decreased free testosterone. Side effects in both groups were minor and characteristic of those observed with low-dose oral contraceptive agents. The results of the study support the reported safety and positive effects of norgestimate on lipid and androgen metabolism, in comparison with a levonorgestrel-containing combined oral contraceptive.


Lipid and carbohydrate effects of a new triphasic oral contraceptive containing norgestimate. Data are reported from the combined results of two studies assessing the lipid and carbohydrate effects of a triphasic preparation of norgestimate and ethinyl estradiol (Ortho TriCyclen, Tri-Cilest) over a 2-year period in 1,783 healthy women. Mean values for serum levels of high-density lipoprotein cholesterol (HDL-C) were increased significantly, with a percent change at 24 months of 13.2. Values for the ratio of low-density lipoprotein cholesterol to HDL-C were reduced throughout the study period (mean change of -6.4% at cycle 24). There were no clinically significant changes in fasting blood glucose levels or insulin levels or in values for glycosylated hemoglobin. These results are consistent with those of previous studies and indicate that the triphasic preparation of norgestimate and etinyl estradiol is a selective and minimally androgenic oral contraceptive agent. Long-term therapeutic benefit may accrue from the favorable influences on the lipid profile.


Comparative contraceptive efficacy and mechanism of action of the norgestimate-containing triphasic oral contraceptive. Norgestimate (NGM), a derivative of 19-nortestosterone with very specific affinity for the progesterone receptor, has been used in combination with ethinyl estradiol (EE) at low doses in both monophasic and triphasic oral contraceptives (OCs). An open-label comparative clinical trial was conducted with 4,234 healthy women using comparative clinical trial was conducted with 4,234 healthy women using triphasic levonorgestrel (LUG)/EE and NGM/EE through a total of 22,312 menstrual cycles. Contraceptive (LUG)/EE and NGM/EE through a total of 22,312 menstrual cycles. Contraceptive efficacy was excellent with both preparations, with no statistically significant between-regimen differences in pregnancy rates. The theoretical Pearl index was the NGM/EE triphasic, and 0.34 for the LNG/EE triphasic. Adverse experiences in groups were typical of those that may occur among women taking low-dose OC agents. was similar with the two preparations: 8.6% for the NGM/EE triphasic and 6.8% for the LNG/EE triphasic. In a separate mechanism of action study, specific endocrine parameters were investigated in 20 subjects using the NGM/EE triphasic for 4 cycles. Ovulation suppression was demonstrated in statistically significant decreases from pretreatment values in serum levels of luteinizing hormone, follicle-stimulating hormone, progesterone, and estradiol. Significant on-treatment increases in serum levels of sex hormone binding globulin evidenced minimal androgenicity. All hormonal values returned to or toward normal in the post-treatment cycle. The study results support those obtained in large noncomparative studies of the NGM/EE triphasic. This phased-dose combination suppresses ovulation and is a very effective, minimally androgenic contraceptive agent with a good safety profile.


T cell activation and T cell receptor variable region gene usage in measles. T cell activation and T cell receptor variable (V) regions were studied with monoclonal antibodies in peripheral blood lymphocytes from 22 patients with measles. Increased (greater than 5%) activated T cells (HLA-DR+ CD3+ cells) were noted in 14 of the 22 patients. Elevations of V beta 5+ and V beta 8+ T cells were observed in two and four patients, respectively, and appeared to be associated with T cell activation. The duration of fever was significantly prolonged in those with increased (greater than 10%) activated T cells (p less than 0.01). These results suggest that T cell activation and the preferential expansion of V beta 8+ and V beta 5+ T cells are associated with the pathogenic process of measles.


Human herpesvirus 6 (HHV-6) infection in the central nervous system. Human herpesvirus 6 (HHV 6) was isolated from patients with exanthema subitum (ES) with a high frequency, and it is now believed that this virus causes ES as a primary infection in childhood. HHV 6 infection is highly prevalent in early childhood and this virus may infect infants through the saliva mainly from mother to child. HHV 6 has a tropism to CD4+ cells and destroys cells in vitro. Although children recover from ES without any sequelae, neurological symptoms associated with exanthema subitum are often observed, and we could detect HHV 6 in the cerebrospinal fluid of ES patients. This result suggests that HHV 6 may invade the central nervous system and cause neurological symptoms.