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The authors describe the case of a forty-two-year-old patient with acute myocardial infarction (MI) on the third day of puerperium, who underwent direct angioplasty. The authors review the main physiological cardiovascular and hemostatic changes in pregnancy, which are usually well tolerated but which increase the probability of cardiovascular events when risk factors or previous cardiac disease are associated. Blood volume and cardiac output increase by around 50% during pregnancy, hemoglobin concentration falls, and reduced peripheral vascular resistance leads to a fall in arterial blood pressure during the first half of pregnancy. End-diastolic pressure, stroke volume and cardiac output increase in the immediate puerperium, and cardiovascular risk is highest in the perinatal period. The incidence of MI during pregnancy and puerperium is low; however, it is on the increase due to the growing number of pregnancies in women close to the end of their reproductive age.

Arterial complications after liver transplantation are frequent. Hepatic artery thrombosis (HAT) is usually associated with biliary complications. Herein we have reported a case of a patient who was admitted for jaundice, itch, and elevated aspartate aminotransferase and alanine aminotransferase levels at 6 weeks after liver transplantation. HAT associated with a biloma was diagnosed and an urgent operation performed requiring a new biliodigestive anastomosis technique. Fourteen months after the first transplant, the patient was retransplanted. The operation performed may be an alternative to treat biliary complications due to late HAT.

In a previous study, we showed that the Polybia paulista wasp venom causes strong myonecrosis. This study was undertaken to characterize the myotoxic potency of mastoparan (Polybia-MPII) isolated from venom (0.25 microg/microl) and injected in the tibial anterior (TA) muscle (i.m.) of Balb/c mice. The time course of the changes was followed at muscle degenerative (3 and 24h) and regenerative (3, 7, and 21 days) periods (n=6) after injection and compared to matched controls by calculation of the percentage of cross-sectional area affected and determination of creatine kinase (CK) activity (n=10). The results showed that although MP was strongly myotoxic, its capacity for regeneration was maintained high. Since the extent of tissue damage was not correlated with the CK serum levels, which remained very low, we raised the hypothesis that the enzyme underwent denaturation by the peptide. Evidence suggested that MP induced the death of TA fibers by necrosis and apoptosis and had the sarcolemma as its primordial target. Given its amphiphilic polycationic nature and based on the vast spectrum of functions attributed to the peptide, we suggest that MP interaction with cell membrane impaired the phosphorylation of dystrophin essential for sarcolemma mechanical stability, and disturbed Ca2+ mobilization with obvious implications on sarcoplasmic reticulum and mitochondrial functioning.

Background: The AZFc region of the human Y chromosome is a highly recombinogenic locus containing multi-copy male fertility genes located in repeated DNA blocks (amplicons). These AZFc gene families exhibit slight sequence variations between copies which are considered to have functional relevance. Yet, partial AZFc deletions yield phenotypes ranging from normospermia to azoospermia, thwarting definite conclusions on their real impact on fertility., Results: The amplicon content of partial AZFc deletion products was characterized with novel amplicon-specific sequence markers. Data indicate that partial AZFc deletions are a male infertility risk [odds ratio: 5.6 (95% CI: 1.6-30.1)] and although high diversity of partial deletion products and sequence conversion profiles were recorded, the AZFc marker profiles detected in fertile men were also observed in infertile men. Additionally, the assessment of rearrangement recurrence by Y-lineage analysis indicated that while partial AZFc deletions occurred in highly diverse samples, haplotype diversity was minimal in fertile men sharing identical marker profiles., Conclusion: Although partial AZFc deletion products are highly heterogeneous in terms of amplicon content, this plasticity is not sufficient to account for the observed phenotypical variance. The lack of causative association between the deletion of specific gene copies and infertility suggests that AZFc gene content might be part of a multifactorial network, with Y-lineage evolution emerging as a possible phenotype modulator.

Acanthoscelides obtectus is a devastating storage insect pest capable of causing severe bean crop losses. In order to maintain their own development, insect pest larvae feed continuously, synthesizing efficient digestive enzymes. Among them, cysteine proteinases (CPs) are commonly produced as inactive precursors (procysteines), requiring a cleavage of the peptide proregion to become active. The proregion fits tightly into the active site of procysteines, efficiently preventing their activity. In this report, a CP cDNA (cpao) was isolated from A. obtectus midgut larvae. In silico studies indicated that the complete CP sequence contains a hydrophobic signal peptide, a prodomain and a conserved catalytic region. Moreover, the encoding cDNA contains 963bp translating into a 321 residue protein, CPAo, which was expressed in E. coli, fused with thioredoxin. Enzymatic assays using the recombinant protein revealed that the enzyme was catalytically active, being able to cleave the synthetic substrate Z-Phe-Arg-7-AMC. Additionally, this report also focuses the cpao propeptide (PCPAo) subcloning and expression. The expressed propeptide efficiently inhibited CPAo, as well as digestive CP of other bean bruchids. Little or no activity was found against proteolytic enzymes of two other coleopterans: Rhyzopertha dominica and Anthonomus grandis. The data reported here indicate the possibility of endogenous propeptides as a novel strategy on bruchids control, which could be applicable to bean improvement programs.

The first complete mitochondrial DNA (mtDNA) sequences (approximately 16,569 bp) in 20 patients with asthenozoospermia and a comparison with 23 new complete mtDNA sequences in teratoasthenozoospermic individuals, confirmed no sharing of specific polymorphisms or specific mitochondrial lineages between these individuals. This is strong evidence against the accepted claim of a major role played by mtDNA in male fertility, once supported by haplogroup association studies based on the screening of hypervariable region I. The hypothesis of maternally driven selection acting in male reproductive success must thus be treated with caution.

An integrated framework for ventricular arrhythmias (VA) assessment, composed of two levels, is proposed in this work. The first level consists of four independent neural networks (NN), designed for specific detection tasks: signal quality, premature ventricular contractions (PVC), ventricular tachycardia (VT) and ventricular fibrillation (VF). Time and frequency domain features, obtained from the electrocardiogram (ECG) and selected through a correlation analysis procedure, form the inputs to the neural modules. The outputs feed the second layer, which consists of a global classifier (ANFIS structure), returns the global result for the VA assessment scheme. Sensitivity and specificity values, evaluated from public MIT-BIH databases, show the effectiveness of the proposed strategy.

A mitochondrial DNA (mtDNA) haplogroup association study carried out in 101 southern Portuguese males with oligozoospermia showed to be negative when comparing with a geographically matching control sample. Misleading positive association signs were however obtained when using other control samples from the same country. This shows that mtDNA population substructure can also introduce spurious signs in haplogroup association studies, as previously reported for Y-chromosome. However, our data do not exclude the probability that a particular mtDNA mutation contributes significantly to the reduction of sperm production, but indeed precludes the hypothesis of a significant association between such a mutation and specific haplogroup.

Objective: The aim of the study was to investigate the impact of the climacterium (before and after menopause) on platelet activation., Background: Platelet activation has been associated to the risk of cardiovascular disease. There is much speculation about the relationship between platelet function and sex steroids, due to peculiarities of platelet action between the genders, including concerns about the influence of low estradiol status in menopausal women., Methods: By means of a cross-sectional study design, 37 female patients divided into two groups were compared. Group A consisted of ten women, mean age 43.9 years, in the premenopausal period, with normal estrogen levels; and Group B comprised 27 patients, mean age 53.0 years, who had all reached menopause. Platelet activation markers, namely P-selectin and glycoprotein IIb-IIIa complex (GPIIb-IIIa), were evaluated by flow cytometry with monoclonal antibodies. A binding index was calculated for both parameters (percentage of positive platelets x mean fluorescence of positive platelets). Also, thromboxane A2 was quantified by means of its main plasma metabolite, thromboxane B2, by enzyme immunoassay., Results: P-selectin and GPIIb-IIIa expression results revealed lower platelet activation status after menopause, as there was a decrease in both the percentage of P-selectin+ platelets and of GPIIb-IIIa mean fluorescence of positive platelets, lowering both binding indices. P-selectin binding index differed significantly between Group A (12.3 +/- 3, n = 10) and Group B (6.2 +/- 2.9, n = 27; mean +/- standard deviation (SD), p < 0.001). GPIIb-IIIa binding index also differed significantly between both groups (Group A: 18.8 +/- 2.3, n = 10 vs. Group B: 16.2 +/- 3.1, n = 27; mean +/- SD, p < 0.0018). Plasma concentration of thromboxane B2 was 1.07 +/- 0.5 pg/well before menopause (Group A, n = 10) and 1.9 +/- 4.1 pg/well after menopause (Group B, n = 27), not significantly different (mean +/- SD, baseline x therapy, p = 0.85)., Conclusions: After the menopause, climacteric women--whose estradiol status is low--have a decreased activation platelet status compared with premenopausal women. Nevertheless, further studies on a larger sample are necessary for conclusive data regarding cardiovascular disease.

We investigate the angular distribution of photoionization fragments at low photon energies (12-40 eV) in an open shell atom, by synchrotron radiation recoil ion momentum spectroscopy in a laser cooled and trapped sample. For cesium atoms, for which relativistic effects play an important role and the ion recoil is relatively small, we could determine large and rapid changes of the asymmetry parameter beta from two, observed for s electrons outside resonances and far from the Cooper minimum. They can be explained by relativistic effects and interchannel coupling arising from final state configuration mixing.

A bacteriological survey of kidneys from 145 abattoir horses was performed, which resulted in the isolation of two Leptospira strains. The isolates were serologically typed as belonging to serogroups Australis and Pomona, and REA identified them as L. interrogans serovar Bratislava and L. kirschneri serovar Tsaratsovo, respectively. These are the first Leptospira isolates obtained from horses in Portugal and the Bratislava strain is the first serogroup Australis strain to be isolated in this country. The 145 horses were also serologically tested for leptospiral antibodies, and 37% had MAT titres #10878;1:10.

Aims: To assess whether feeding bioassays using culture-supernatant proteins could be combined with PCR into a first-tier screening strategy for Vip3A-like genes efficient against tropical Spodoptera frugiperda., Methods and Results: Out of 12 Bacillus thuringiensis strains studied, the total protein concentrated from the culture supernatant of only the strain HD125 yielded a significantly increased armyworm mortality and an intense band of the predicted size for VIP3A protein in SDS-PAGE. However, PCR and sequencing data indicated Vip-like genes are ubiquitous in tropical B. thuringiensis isolates. Interestingly, the HD125 strain was also the only one displaying a single-band amplification pattern and the highest sequence identity to the reported Vip3A(a) gene., Conclusions: Results suggest the insecticidal effectiveness of putative VIPs in B. thuringiensis isolates can be preliminarily estimated by the use of supernatant-derived total protein in feeding experiments, though only in a limited manner., Significance and Impact of the Study: A simple and cost-effective first-tier screening strategy for VIP-derived activities in B. thuringiensis collections can be developed by combining PCR and feeding bioassays. Moreover, the employed primers showed to be useful as a tool for strains differentiation at DNA level, and for characterization and isolation of Vip-like genes in tropical B. thuringiensis germplasm.

Background: To determine the prevalence of different modes of death and to identify therapeutic limitations in patients admitted to the Pediatric Intensive Care Unit (PICU) at a teaching hospital., Methods: A retrospective study was carried out. Information was collected from the medical records of patients who died at the PICU of Hospital de Clínicas de Porto Alegre between July 1st, 1996 and June 30th, 1997. Four modes of death were considered: patient not responding to cardiopulmonary resuscitation methods (NRCPR), brain death (BD), withdrawal / withhold life-support measures (W/WLS) and decision not to resuscitate (DNR). For cause of death we employed the criterion of organ failure., Results: Out of 61 deaths occurred during the period under analysis, 44 patients were included in the study. Patient age median was 28 months. The cause of death for all patients was multiple organ failure. Twenty-six patients (59%) were classified in group I (NRCPR and BD) and 18 (41%) in group II (W/WLS and DNR). Among patients of group II, 83% had a chronic and/or debilitating disease (p = 0.017; chi(2)). The prevalent reason for patient admission was the need for organ support (55%), both respiratory and cardiovascular. The median for duration of patient stay at the PICU was 5 days and at the hospital was 11 days. There was no statistically significant difference between the two groups in terms of reason for patient admission or duration of PICU/hospital stay., Conclusions: There was a high prevalence of W/WLS and DNR deaths among ICU patients, thus suggesting therapeutic limitation for them. We were unable to evaluate the level of participation by the medical team and by the family in this decision making process.

The effect produced by the nitric oxide donor SIN-1 on monosynaptic reflexes was examined. Experiments were performed on anesthetized, paralyzed and spinalized cats. Lumbar monosynaptic reflexes were produced by stimulation of Ia afferents. I.v. application of SIN-1 (500 microg/kg) produced a mean marked potentiation of 704% of pre-drug control (100%) in the amplitude of monosynaptic reflexes. In addition, in other experiments a concentration-dependent effect on the amplitude of monosynaptic reflexes was observed after microinjections of SIN-1 into the ventral horn (1 microl; 10(-12) - 10(-3) M), with a mean facilitatory effect of 355%. In both cases, the potentiation was reversible 45 min after i.v. or local application of SIN-1. These results provide the first evidence that monosynaptic reflexes can be potentiated by nitric oxide.

Medical records and biopsy specimens were retrospectively reviewed from 25 dogs diagnosed with unresectable urinary bladder carcinoma and treated with chemotherapy. Our intention was to identify clinical, histologic, and immunohistochemical indicators of prognosis. Immunohistochemical stains for P-glycoprotein, glutathione-S-transferase pi, and factor VIII-related antigen were applied to archived tissue. There were more spayed female dogs than castrated male dogs (76% versus 24%). Transitional cell carcinoma was the most common tumor (88%, n = 22), followed by undifferentiated carcinoma (8%, n = 2) and squamous cell carcinoma (4%, n = 1). Overall median survival was 251 days. Histologic diagnosis and immunohistochemical characteristics did not correlate with prognosis. Spayed females survived significantly longer than castrated males (358 days versus 145 days, P = .042). Dogs that received either doxorubicin or mitoxantrone in addition to a platinum-based chemotherapeutic (either cisplatin or carboplatin) lived significantly longer than those that received only a platinum compound (358 days versus 132 days, P = .042).

A total of 56 weekly samples from Oporto and 40 from Vienna were collected and analysed, for 23 n-alkanes and 16 polycyclic aromatic hydrocarbons (PAH), by GC-MS after extraction with a toluene/methanol mixture. Total and elemental carbon were in the same range of values for both sampling sites. Although parts of the spectrum of species in both sampling sites were constant over the sampling period there is no evidence for suggesting a universal tracer for alkanes. For both Oporto and Vienna, Principal Component Analysis (PCA) identified two PCs and in both cases the first PC contained only PAHs while the second contained only alkanes. This separation between alkanes and PAHs and the observation of an alternating pattern with higher concentrations of odd carbon numbered from C27 to C30 (natural emissions from biological origin) is believed to result from a separation between anthropogenic and biological contributions associated with the first and second PC, respectively.

With the purpose of studying data on spontaneous customary changes in diabetic rats, we induced diabetes in 28 Wistar rats with streptozotocin. The animals were observed for 27 weeks in an attempt to characterize spontaneous customary changes that could suggest signs of chronic pain. Morphine, as a central-acting potent analgesic and its specific antagonist naloxone, were used. Our results evidenced in the animals a clinical syndrome similar to human diabetes. Long-term customary analysis revealed a significant (p < 0.05) increase of scratching and resting/sleeping behaviors, but diminished motor, eating and grooming customs. Moreover, the thermal tests revealed hyperalgesia in 43% of the animals, what may corroborate the meaning of scratching as a sign of pain. Pharmacological tests with morphine showed a significant (p < 0.05) inhibition of scratch, with concomitant increase of motor and eating activities and diminished rest/sleep capacity. Naloxone antagonized the effects induced by morphine. Such results suggest that these animals exhibit evoked behavior of hyperalgesia and that scratch may possibly be a spontaneous manifestation of chronic pain also in Wistar rats with this experimental model of painful diabetic neuropathy.

Response surface and simplex methods were used for optimizing the determination of total sulfur present in gaseous samples using a quartz crystal microbalance. The optimization methodology increased the analytical signal from three to five times. The simplex method showed the highest efficiency in the search for the maximum sensor response. However, the response surface allows ranking the variables according to their effects and highlights the existence of a plateau in the optimal region.

This paper presents a review of Leptospira infection in farm animals in Portugal which is based mainly on serological results obtained in the National Veterinary Research Laboratory between January 1987 and December 1993. Serum samples were tested by the microscopic agglutination test, at a minimum dilution of 1:100. Positive titres were obtained in 15.3% of the 9,543 bovine samples examined. Sejroe, Pomona, Hebdomadis, Tarassovi and Icterohaemorrhagiae were the principal serogroups which reacted in the tests. A total of 3,195 pigs were tested, of which 20.2% showed positive reactions. The main serogroups which reacted were Australis, Pomona, Cynopteri, Icterohaemorrhagiae and Autumnalis. Field observations on outbreaks of leptospirosis in pigs, along with data obtained from an experimental infection with serovar mozdok in pregnant gilts suggest that this serovar, rather than serovar pomona, may be causing Pomona group infections in pigs. Serum samples from 5,298 sheep were tested and 3.3% gave positive results. The predominant serogroups involved were Canicola, Pomona, Cynopteri, Sejroe and Icterohaemorrhagiae. From the 1,631 goats examined serologically, 5.0% gave positive results, mainly to serogroups Grippotyphosa, Canicola, Pomona, Icterohaemorrhagiae and Pyrogenes. Only 83 serum samples from horses were obtained, of which 43.4% showed positive titres. Serogroups Australis, Autumnalis, Cynopteri and Pyrogenes were those most commonly found. Serological evidence of leptospirosis in farm animals is widespread in Portugal, particularly in cattle and pigs. Leptospirosis in horses needs to be studied further. In an attempt to provide a general view on the occurrence of leptospirosis in these animal species in Portugal, the present results are compared with results obtained in previous studies and are complemented with both previous and recent bacteriological findings.

HTLV-I infection and associated myelopathy has been reproduced experimentally in vitro and in vivo and these studies have shown the possibility of creating several lines of infective cells and of detecting minor and major clinical expressions of HTLV-I associated myelopathy in rabbits and rats.

A new method for the analysis of both total and bound SO(2) in wine is proposed, based on a quartz crystal microbalance (QCM), and it is compared with the widely used Ripper method. The proposed method is faster than the Ripper's, and the instrumentation is either home-made or widely available. When both methods are applied to the same sample, the results obtained using the QCM method are bracketed in an interval less than one-tenth the size of that obtained using the Ripper method. Although the SO(2) concentrations found using the QCM method correlate well with the ones obtained with the Ripper method, the results are systematically higher, which can be explained as due to the absence of interferences known to affect the Ripper method.

High expression and secretion of recombinant ovine beta-lactoglobulin has been achieved in the yeast Kluyveromyces lactis. The yield of beta-lactoglobulin is 40-50 mg per litre of culture supernatant and accounts for approx. 72% of the total secreted protein. Constitutive expression is under the control of the Saccharomyces cerevisiae phosphoglycerate kinase promoter from an intronless version of the beta-lactoglobulin gene. Secretion is specified by the ovine protein's own signal sequence. this system, coupled to an efficient and novel recovery protocol, allows 30 mg of pure protein to be isolated from a typical 1 litre culture. The protein is virtually indistinguishable from beta-lactoglobulin conventionally purified from sheep milk by its behaviour in native PAGE and SDS/PAGE, reactivity to antibodies, CD, fluorescence spectroscopy and N-terminal sequencing. Attempts to achieve a similar expression and secretion system in the yeast S. cerevisiae met with only limited success, although it was found that heat-shock treatment modestly increased the yield up to approx. 3-4 mg per litre of culture supernatant. Site-directed mutagenesis showed that secretion in S. cerevisiae depended upon correct formation of the two disulphide bonds present in beta-lactoglobulin.

A leptospira-like spirochete was isolated from a lymphocyte culture from a HIV-I and HTLV-I/II-positive patient. On the basis of serological, biological and morphological characteristics of the cells of the isolated strain (strain Lisboa), we conclude that it is a member of the genus Leptonema.

Three pregnant gilts were experimentally infected with leptospires of the serovar mozdok, isolated from an aborted pig fetus from a Portuguese pig farm with abortion problems. All the gilts aborted dead or dying piglets on days 105 or 106 of pregnancy. Serovar mozdok was isolated from 12 of the 22 piglets in the three litters. Histological examination of the livers and kidneys of the gilts at the end of the experiment revealed evidence of disease, and leptospires were isolated from their kidneys. Their serological responses up to 42 days after inoculation were monitored by means of a microscopic agglutination test, using 21 antigens from 18 serogroups. Cross reactions to heterologous antigens belonging to the Grippotyphosa, Australis, Icterohaemorrhagiae and Cynopteri groups were observed in all of them.

The genomic region of the Anticarsia gemmatalis multiple nucleocapsid nuclear polyhedrosis virus (AgMNPV) strain 2D encoding the polyhedrin gene was cloned and mapped, and a 2085 bp SphI-PstI fragment containing the gene was sequenced. The polyhedrin polypeptide of the parental isolate AgMNPV was manually sequenced, and the amino acid sequence obtained agreed with that deduced from the DNA coding region sequence. AgMNPV and Orgyia pseudotsugata MNPV (OpMNPV) are similar in terms of promoter structure and polyhedrin primary sequence, and the polyhedrin gene of both viruses is transcribed in the anti-clockwise direction in relation to their physical maps. The region upstream from the polyhedrin gene of AgMNPV, OpMNPV, Bombyx mori NPV and Autographa californica MNPV (AcMNPV) was compared and this showed that the open reading frame (ORF) common to all four viruses (ORF 5) has sequence homology with the AcMNPV 25K gene. The sequences between ORF 5 and the polyhedrin gene were found to be variable among the polyhedrin gene loci compared. Additionally, conserved elements in the promoters of the very late genes encoding polyhedrin and granulin, and those encoding two p10 proteins were found to share sequence homology and positional similarity with consensus regions in the conserved boxes A and C, responsible for binding transcription factors to eukaryotic 5S ribosomal RNA genes, and to box C of tRNA genes.

Adults with pulmonary hypertension and polycythemia (N = 22) have low levels of plasma antithrombin III (84 +/- 18 vs 98 +/- 13% for controls, N = 35, P less than 0.005) and protein C (66 +/- 21 vs 125 +/- 30%, N = 8, P less than 0.0002) but normal levels of total protein S. Data are reported as means +/- SD and percent normal values obtained for pooled plasma from normal healthy adults. Children with the same disorder (N = 6) also had low protein C levels (66 +/- 16 vs 85 +/- 5%, P less than 0.025). Total protein S was normal for children, but free protein S was decreased (66 +/- 13 vs 91 +/- 23%, P less than 0.025). Since the levels observed in these patients are above those reported for congenital deficiencies, the reduction in plasma levels of anticoagulant proteins may be the result of chronic intravascular coagulation. Furthermore, normal levels of plasminogen and fibrin degradation products suggest a localized disorder or an acquired decrease in fibrinolytic activity.

1. Seven patients submitted to myocardial revascularization surgery with cardiopulmonary bypass were studied. Blood samples were obtained immediately before and 24 h after surgery. The parameters studied were the production of platelet activating factor (PAF-acether) and superoxide anion, cellular beta-glucuronidase activity as well as polymorphonuclear cell (PMN) and platelet counts. 2. Twenty-four h after surgery, there was a 54% decrease in platelet number (P less than 0.005), a 121% increase in PMN number (P less than 0.005), a 353% increase in PAF-acether (P less than 0.01), a 211% increase in superoxide anion (O2-) and a 104% increase in beta-glucuronidase (P less than 0.05) levels when compared with the pre-surgery levels. 3. The present results indicate that PMN are more reactive after surgery with cardiopulmonary bypass.