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Molecular investigations of pathogenesis-related Bet v 1 homologues in Passiflora (Passifloraceae).

The major birch pollen allergen, Bet v 1, responsible for allergic reactions in many areas of the world, is homologous to a large number of pathogenesis-related proteins (PRs), identified as PR10. As part of a long-range investigation of these types of proteins and of evolution in Passiflora, DNA sequences from eight Bet v 1 homologue isoforms were obtained from five species of this genus in Brazil, and their sequences compared among themselves and with 30 others from 8 different species, classified in different taxonomic units. The objective was a first characterization of these PRs in wild passionflowers, and their use for evolutionary and applied investigations. High interspecific, but low intraspecific variability was observed, as expected from multigenic families subjected to concerted evolution. The relationships obtained both within Passiflora and between it and seven other genera probably best reflect functional similarities than evolutionary history.

Allergens↗

Effect of temperature on ppp(A2'p)nA binding protein activities in rabbit reticulocyte lysates and other mammalian extracts.

Lysates of rabbit reticulocytes and other mammalian cells are known to contain an activity which binds with high specificity ppp(A2'p)3A,3'-[32P]pCp. The binding activity shows a marked dependence on preincubation of lysates at different temperatures (4 degrees C - 45 degrees C). For example, binding was increased 50% by preincubation of rabbit reticulocyte lysates at 37 degrees C for 60 minutes. An identical preincubation of mouse brain extracts results in a greater than 90% loss of binding activity. Fractionation of rabbit reticulocyte lysates into the postribosomal supernatant (PRS) and the ribosomal salt wash (RSW), followed by heparin-agarose column chromatography, showed that with the PRS fraction, most of the binding activity is eluted with 600 mM KCl. With the RSW fraction, more than 50% of the binding activity is eluted with 250 mM KCl. These data suggest that multiple ppp(A2'p)nA binding protein activities exist in mammalian cells.

Adenine Nucleotides↗

Pulvinar and lateral geniculate neuronal activity in the cat during operantly conditioned appetitive behavior.

Cats were trained to press a lever for 0.5--1.0 ml of milk reward both in the presence and absence of ambient light. 'Floating' microelectrodes were implanted in the pulvinar and lateral geniculate nuclei to allow chronic recording from single or multiple neurons over a period of 1--16 days. Thirty-one of 36 pulvinar neurons (86%) showed a consistent periodicity in the firing rate that was clearly related to certain phases of operant behavior. In the presence of ambient light, the firing rate was maximal shortly prior to or immediately after a rewarded bar press, particularly during the animal's initial contact with reward (the first lap). This burst of activity was followed by an abrupt and strong inhibition of firing which was, in most instances, associated with high voltage slow wave (6--9 c/sec) postreinforcement synchronization (PRS) of the electroencephalogram over the visual and association cortices as well as in the pulvinar. Postreinforcement inhibition of neuronal activity was also observed in the lateral geniculate. Neuronal firing in both nuclei dropped significantly below the respective mean rate observed for each nucleus during relaxed wakefulness. In the dark, however, the reward-induced inhibition was abolished in both nuclei. Moreover, some pulvinar neurons in the dark showed a conspicuous and significant increase in firing during the consummatory response (dark reversal pattern). These findings suggest that a transient but powerful inhibitory process triggered by positive reinforcement depends on visual input. Firing patterns of some pulvinar neurons were noted to be entrained not only by the rhythm of PRS oscillations (inhibitory phasing and postinhibitory discharges) but also by the much slower 3--4 c/sec rhythm of lapping and licking. Furthermore, the patterns of entrainment by initial lapping appear to be different from those observed during the later phase of the consummatory response, thus indicating that the pulvinar may receive information regarding the content of the delivery cup. No entrainment of neuronal activity was observed in the lateral geniculate. These findings reveal remarkable plasticity in the pulvinar neurons that is consistent with their role in polysensory integration.

Action Potentials↗

Molecular cloning and sequencing of human cDNA for phosphoribosyl pyrophosphate synthetase subunit II.

cDNA clones for human phosphoribosyl pyrophosphate synthetase subunit II (PRS II) were isolated. The five overlapping clones contained 2457 base pairs (bp) covering a 954-bp complete coding region for 318 amino acid residues. Homologies between human and rat PRS II were 99% of the amino acids and 88% of the nucleotides in the coding region. This amino acid homology seems to be the highest so far reported for enzymes involved in nucleotide metabolism and glycolysis. The highly conserved structure may be required for unique catalysis and rigid regulation of this enzyme.

Amino Acid Sequence↗

Solid phase synthesis of the proteinase of bovine leukemia virus. Comparison of its specificity to that of HIV-2 proteinase.

The 126-residue proteinase (PR) of bovine leukemia virus (BLV) was synthesized by solid-phase peptide synthesis and its activity was shown using various oligopeptide substrates representing cleavage sites in BLV, human T-cell leukemia virus type 1 (HTLV-1), murine leukemia virus (MuLV) and human immunodeficiency virus type 1 (HIV-1). The specificity of the BLV PR was also compared to that of chemically synthesized human immunodeficiency virus type 2 (HIV-2) PR. Many of the peptides were cleaved at the expected site, however, 6 out of 15 were hydrolyzed only by one of the PRs. Furthermore, one BLV peptide was processed differently by the two enzymes. These results, together with the relative activities and the lack of inhibition of BLV PR by two HIV-1 PR inhibitors, suggest that the BLV PR specificity is substantially different from that of HIV PRs.

Amino Acid Sequence↗

A randomized comparison of intrauterine and intraperitoneal insemination in the treatment of infertility.

OBJECTIVE: To compare the pregnancy rates (PRs) between intrauterine insemination (IUI) and intraperitoneal insemination (IPI) in women receiving comparable ovarian stimulation in couples who were infertile for various infertility conditions. METHOD: Fifty-four couples were treated. Of these, 25 were diagnosed as having unexplained infertility, 21 as having male infertility and 8 as having cervical mucus hostility. RESULT: Nine pregnancies occurred with 89 cycles of IUI and 6 pregnancies with 85 cycles of IPI, giving an overall PR of 28%. Twelve of the 15 pregnancies occurred during the first treatment cycle and three during the second treatment cycle. There was no significant difference in the (PRs) between the IUI and IPI (P > 0.25). CONCLUSION: Both procedures combined with superovulation proved to be simple, inexpensive safe and worthwhile for infertile couples with unexplained infertility, male infertility and cervical mucus hostility.

Adult↗

Relationship between reversible acetylcholinesterase inhibition and efficacy against soman lethality.

Carbamate pretreatment (45% inhibition, reversible), combined with therapy, protected rats from soman-induced lethality [The Pharmacologist 23, 224 (1981)]. The present study was done to see if less than 45% inhibition protects and to see if reversible acetylcholinesterase (AChE) inhibition and efficacy against soman lethality are correlated. At 30 min pre-soman, guinea pigs and rats received (im) either pyridostigmine (Py) or physostigmine (Ph) to inhibit whole blood AChE from 10 to 70%; at 1 min post-soman (sc), they received (im) atropine (16 mg/kg)/2-PAMCl (50 mg/kg) and mecamylamine (0.8 mg/kg)/atropine (16 mg/kg), respectively. Protective ratios (PRs) were computed and they ranged from 3.1 to 7.7 for guinea pigs and from 1.8 to 2.4 for rats. In guinea pigs the PRs for Py + therapy were roughly similar to those of Ph + therapy. In both species at 30 min after im injection of Py and Ph, a linear relationship was found between percentage of whole blood AChE inhibition and ln dosage of carbamate. Positive correlation (p less than 0.05) was found between the degree of reversible AChE inhibition by pretreatment, coupled with therapy, and efficacy against soman lethality. The present data indicate that inhibition levels as low as 10% may provide some protection.

Acetylcholinesterase↗

Dynamic respiratory mechanics in monkeys measured by forced oscillations.

We measured impedances of the lungs (ZL) and respiratory system (Zrs) by discrete frequency (f) forced oscillations in adult bonnet monkeys at two lung volumes, FRC and at transrespiratory system pressure (Prs) of -5 cm H2O. Measurements were made from 2 to 32 Hz to study f dependence of effective resistances (Reff) and reactances, and from 1 to 8 Hz to study f dependence of effective pulmonary compliance (Ceff,L). Estimates of resistances (R), inertances (I) and compliances (C) of the lungs (L), chest wall (W) and respiratory system (rs) were obtained by fitting the 2-32 Hz data to a series RIC network model. Reff,L, Reff,W and Reff,rs were found to be f dependent. The magnitude of ZL was small relative to that of Zrs at all f with RL only 30% of Rrs and CL six times Crs. Decreasing lung volume from FRC to Prs = -5 cm H2O increased RL, RW and Rrs, slightly decreased CL, CW and Crs, slightly increased resonance frequencies, and increased the f dependent behavior of Ceff,L. Our results indicate both similarities and differences between the respiratory system of these primates and man.

Airway Resistance↗

Management of upper airway obstruction in the Pierre Robin syndrome.

Eight patients with Pierre Robin syndrome (PRS), were treated according to our management protocol. All our patients initially were given a trial of conservative, positional treatment, with high caloric gavage feeding. In five patients this therapeutic approach resulted in a good weight gain, with no significant respiratory distress. The remaining three patients showed no improvement, failed to thrive and therefore underwent the tongue to lip adhesion (TLA) procedure. Two patients then improved dramatically; whereas one continued with respiratory distress and failure to thrive and required tracheostomy. We conclude that when symptoms of respiratory distress and failure to thrive coexist in patients with PRS despite conservative management, surgical intervention to the airway is mandatory. TLA should be the first surgical procedure considered and if the ultimate goals of weight gain and respiratory comfort are still not achieved then tracheostomy seems inevitable.

Airway Obstruction↗

A phase II study of oral weekly 4-demethoxydaunorubicin in advanced breast cancer.

Thirty-eight patients with advanced breast cancer were treated with oral 4-demethoxydaunorubicin in a continuous weekly schedule at a dose of 15 mg/m2/week. Subjective toxicity consisted of mild nausea (grade 1) in 52% with more severe GI side effects (grade 2) in 15%. Three patients developed grade 1 alopecia and there were no episodes of cardiac failure. Significant neutropenia (grade 2/3) only occurred in patients with marrow involvement or widespread bone disease. There was one CR and 5 PRs, an overall response rate of 15.7% (95% confidence limits 6-31%). In addition 6 patients had disease stabilization for at least 6 months. Fourteen patients progressing on 4-demethoxydaunorubicin have received adriamycin 60 mg/m2 21 days. There have been 5 PRs in this group indicating possible non-cross-resistance between these two agents.

Adult↗

Influence of selenium deficiency on glutathione disulfide metabolism in isolated perfused rat heart.

Selenium deficiency causes a fall in rat cardiac glutathione peroxidase activity. As a consequence, isolated perfused selenium-deficient heart does not release increased amounts of GSSG when hydroperoxide is infused. However, the total amount of glutathione measured as intracellular GSH, intracellular GSSG and GSSG released from the heart when hydroperoxide is infused does not equal the total glutathione measured in these pools in untreated hearts (Xia, Y., Hill, K.E. and Burk, R.F. (1985) J. Nutr. 115, 733-742). GSSG can react with protein sulfhydryl groups to form glutathione-protein mixed disulfides (PrS-SG). PrS-SG were measured in perfused selenium-deficient and control hearts infused with t-butylhydroperoxide and were found to account for the previously unmeasured glutathione. The ability of the selenium-deficient heart to transport GSSG was also examined. GSSG was produced non-enzymatically by infusing diamide. The diamide-treated selenium-deficient heart formed GSSG and released it at the same rate as similarly-treated control heart. Thus although selenium deficiency decreases GSSG formation by glutathione peroxidase, it does not affect cardiac GSSG transport.

Animals↗

Heat, drugs, and radiation given in combination is palliative for unresectable esophageal cancer.

From April 1966 to April 1986, 101 men and women with unresectable squamous cell carcinoma of the esophagus were treated in our clinic. Since 1983, 21 were treated with a combination of hyperthermia, chemotherapy, and radiotherapy (group I). Before 1983, for another 80 patients, radiation plus chemotherapy had been prescribed (group II). Nine of 21 patients in group I had an unresectable carcinoma due to an advanced tumor, 9 had an associated severe clinical status, and 3 refused surgery. Out of 80 in group II, 50 had a far advanced tumor, 21 had a poor clinical condition, and 9 refused operation. With regard to staging, for 21 in group I, 6 were classified as Stage I, 5 as Stage II, 7 as Stage III, and 3 as Stage IV. As to the 80 in group II, those in Stage I, II, III, and IV accounted for 8, 22, 39, and 11, respectively. The median doses of each modality, for patients in group I, were 6 times of hyperthermia at 42-45 degrees C for 30 minutes, 40 Gy of X ray and 30 mg of bleomycin. For patients in group II, a median dose of 56 Gy of X ray was given. Response rates determined by esophagograms and endoscopies for the patients in groups I and II were 76.2% (16/21; 4 CRs, 12 PRs) and 39.2% (31/79; 2 Crs, 29 PRs), respectively (p less than 0.001). The effective rates determined by improvement in quality of life (relief of pain and dysphagia) for groups I and II were 61.9% and 37.2%, respectively. A longer survival was obtained for patients in group I (median survival: 9 months vs 6 months). Especially for the patients classified as Stage I, a significantly longer survival was obtained with a combination of hyperthermia, chemotherapy, and radiotherapy (p less than 0.01).

Aged↗

Thermoradiotherapy in the treatment of locally advanced nonsmall cell lung cancer.

PURPOSE: To improve the treatment results of locally advanced non-small cell lung cancer (NSCLC), we have been conducting a clinical trial using regional hyperthermia combined with radiotherapy. METHODS AND MATERIALS: Between 1985 and 1990, 19 patients were treated. All cases except one were regarded as initially unresectable. There were 10 Stage IIIA cases and nine Stage IIIB cases. In 10 cases thermoradiotherapy was used definitively, and in the other nine cases preoperatively. Radiotherapy was administered with conventional fractionation. Total dose ranged from 42 to 80 Gy (mean 62.9 Gy) for definitive treatment cases, and 38 to 47 Gy (mean 40.6 Gy) for preoperative cases. Radiofrequency (RF) capacitive hyperthermia was administered twice weekly, immediately after radiotherapy. Total sessions of hyperthermia ranged from 5 to 16 times (mean 9.0) for definitive treatment cases and 3 to 8 times (mean 6.7) for preoperative cases. RESULTS: The results of thermoradiotherapy group (HTRT group) were compared with our historical control group (RT group); initially unresectable Stage III NSCLC irradiated definitively with 50 Gy or more (26 cases), or became resectable after radiotherapy and operated (4 cases). As for initial response, there were 5 complete responses (CRs), 13 partial responses (PRs), and 1 no change (NC) (CR rate 26%, response rate 95%) in the HTRT group, whereas there were no CR, 21 PRs, and 9 NCs in the RT group (CR rate 0%, p < 0.005, response rate 70%, p < 0.05). Overall 3-year local relapse-free survival and survival rate for the HTRT group was 73% and 37%, respectively, and 20% and 6.7%, respectively, for the RT group (p < 0.01, p < 0.01). The rate of death from uncontrolled primary disease for the HTRT group was significantly lower than for the RT group (21% vs. 53%, p < 0.03). CONCLUSION: Although the number of cases is rather small, thermoradiotherapy in the treatment of locally advanced NSCLC is promising in raising resectability, local control, and, thus, long-term survival.

Adolescent↗

Virulence factors associated with F165-positive Escherichia coli strains isolated from piglets and calves.

In this study, 91 F165-positive Escherichia coli isolated from calves and piglets with diarrhea or septicemia were characterized with respect to receptor binding specificity, presence of the aerobactin system, production of colicin V, resistance to the bactericidal effects of serum. Although most F165-positive isolates shared similar DNA sequences with pap operon sequences, less than half of these isolates demonstrated MRHA to P antigen of human red blood cells and Forssman antigen of sheep red blood cells recognized by P and F (or Prs) adhesins respectively. Certain F165-positive isolates sharing similar DNA sequences with both pap and sfa operon sequences demonstrated mannose-resistant hemagglutination of sheep erythrocytes, as observed in human uropathogenic E. coli possessing the prs operon. Most isolates caused mannose-resistant, neuraminidase-resistant hemagglutination of human, equine, feline, and bovine erythrocytes. Thus, F165-positive isolates express one or more adhesins with different receptor binding specificities. An association was observed between the various receptor binding specificities and serogroup. Most F165-positive isolates possessed the aerobactin system and were resistant to the bactericidal effects of serum, but only 38.5% isolates produced colicin V.

Adhesins, Escherichia coli↗

Technical considerations in the selection of a personal emergency response system.

It is the purpose of this report to provide an internal review of the personal response system (PRS) in our hospital. Our system is coordinated by volunteers without emergency physician supervision. It uses antiquated equipment in the client's homes and emergency response center. Calls are answered by individuals who have not been trained as dispatchers, and who do not record pertinent performance data. The technical considerations involved in the selection and operation of an ideal PRS system are described.

Aged↗

De novo insG619 mutation in PAX2 gene in a Japanese patient with papillorenal syndrome.

PURPOSE: To describe a Japanese patient with papillorenal syndrome (PRS) and to identify the genetic defect responsible for the disease. DESIGN: Interventional case report. METHODS: Complete ophthalmologic and systemic examinations were performed, and direct genomic sequencing of the PAX2 gene. RESULTS: Fundus examination of a 3-year-old Japanese girl showed atypical coloboma bilaterally. At 6 years of age, she presented with proteinuria, and renal ultrasonography showed hypoplastic kidneys bilaterally. Molecular genetic analysis of the PAX2 gene revealed a de novo heterozygous insertion of a G at position 619. CONCLUSIONS: Our findings suggest that an abnormal development of the optic stalk led to the optic disk dysplasia in PAX2-associated PRS. This indicates that we should consider renal abnormalities when an atypical round coloboma is present. Molecular genetic analysis of the PAX2 gene in combination with renal ultrasonography can help in making an earlier diagnosis of the disease.

Abnormalities, Multiple↗

Anesthetic effects of progesterone are undiminished in progesterone receptor knockout mice.

Progesterone has sedative and anesthetic effects but the underlying molecular mechanisms remain unclear. The two possible mechanisms by which progesterone affects the function of the brain include binding to intracellular progesterone receptors (PR) and metabolism to GABA(A) receptor-modulating neurosteroids. In this study, PR knockout (PRKO) mice were used as model to study the role of PRs in the anesthetic activity of progesterone. The progesterone-induced anesthetic activity was undiminished in female PRKO mice (ED50, 172 mg/kg) as compared to their wild-type littermates (ED50, 167 mg/kg). The progesterone-induced anesthetic activity was highly correlated with increased plasma allopregnanolone levels. Pretreatment of PRKO mice with the 5alpha-reductase inhibitor finasteride significantly reduced the progesterone-induced anesthetic activity. Allopregnanolone also evoked dose-dependent anesthetic activity in PRKO mice, which was similar to those of wild-type mice. Thus, the anesthetic activity of progesterone is not mediated by its interaction with PRs. The neurosteroid allopregnanolone partially mediates the anesthetic activity of progesterone by potentiation of GABA(A) receptor function.

Analysis of Variance↗

Hepatic disorders in patients treated with COX-2 selective inhibitors or nonselective NSAIDs: a case/noncase analysis of spontaneous reports.

BACKGROUND: Hepatic adverse events associated with the use of nonaspirin drugs and NSAIDs are uncommon, but the widespread use of these drugs may impact public health. OBJECTIVE: We conducted a case/noncase analysis of spontaneous reports to compare the hepatic safety profile of cyclooxygenase (COX)-2 selective inhibitors with that of nonselective NSAIDs. METHODS: This case/noncase analysis was conducted using the US Food and Drug Administration Freedom of Information (FDA/FOI) database (through quarter 1, 2003) and the World Health Organization Uppsala Monitoring Centre (WHO/UMC) database (through quarter 3, 2003). Council for International Organizations of Medical Sciences and WHO Adverse Reaction Terminology preferred terms were used to classify hepatic disorders with broad and specific case definitions. After reports involving established hepatotoxic drugs (bromfenac, nimesulide, sulindac) were excluded, the proportion of reports (PRs) of each case definition was calculated for each NSAID. Crude and adjusted reporting odds ratios (RORs) were used to compare the overall proportions of hepatic disorders and hepatic failure of celecoxib and rofecoxib versus nonselective NSAIDs. RESULTS: A total of 158,539 and 185,253 reports of NSAIDs were identified in the FDA/FOI and WHO/UMC databases and 25% and 16%, respectively, involved other hepatotoxic drugs. The PRs of hepatic disorders for all COX-2 selective inhibitors and non-selective NSAIDs were 3.0% in the FDA/FOI database and 2.7% in the WHO/UMC database. In the FDA/FOI and WHO/UMC databases, respectively, mmesulide (16.7% and 14.4%), bromfenac (12.0% and 20.7%), diclofenac (8.1% and 4.7%), and sulindac (6.1 % and 9.9%) were reported to be associated with higher proportions of overall hepatic disorders compared with those of other NSAIDs. Crude and adjusted RORs for the prevalences of overall hepatic disorders and hepatic failure with celecoxib and rofecoxib versus the other NSAIDs were <1 (indicating that the proportion was not higher than that of the comparator) in both databases. The interpretation of the results was unchanged when bromfenac, nimesulide, and sulindac were excluded from the analysis. CONCLUSIONS: In this case/noncase analysis, bromfenac, nimesulide, sulindac, and diclofenac had higher proportions of reports of hepatic disorders compared with those of other NSAIDs in the FDA/FOI and WHO/UMC databases. The analysis did not raise a safety concern for celecoxib or rofecoxib versus NSAIDs for overall hepatic disorders and hepatic failure.

Anti-Inflammatory Agents, Non-Steroidal↗