Production and Characterization of a Human Recombinant Monoclonal Fab Fragment Specific for Influenza A Viruses

A human recombinant monoclonal Fab fragment that specifically recognizes all the influenza A virus strains tested was produced in transformed Escherichia coli using the phage display technique. No strain of influenza B virus reacted with it. It was purified after four cycles of panning and by a single passage through an immunoaffinity column. About 1 mg of pure monoclonal antibody was obtained from 1 liter of culture medium in 3 working days. The Fab fragment reacted with a viral 27-kDa protein, which could reasonably be a matrix protein. Indirect immunofluorescence tests performed on virus-infected MDCK cells showed that this Fab fragment was at least equally efficient as other commercial monoclonal antibody-based systems in detecting influenza A viral infections. The potential advantages of human recombinant Fabs on murine monoclonal antibodies are discussed.     

Hemodynamic changes during continuous infusion with dobutamine in candidates for lung transplantation. Lung Transplantation Group

BACKGROUND: The aim of this study is to analyze the effects of dobutamine (DBT) on pulmonary and systemic hemodynamics and oxygenation in lung transplant candidates. METHODS: Forty-five patients (21M, 24F) to be introduced in waiting list for lung transplantation were studied (14 pulmonary fibrosis, 15 COPD, and 16 cystic fibrosis). They were studied awake, while spontaneously breathing in two different phases: baseline--O2 100%; DBT phase--O2 100% after 10 minutes of DBT continuous infusion (10 mcg/Kg/min). Blood gas samples and hemodynamic data were collected during right heart catheterization. Data were statistically analyzed with Student's "t" test and values for p < 0.05 were considered as significant. RESULTS: During DBT phase, a significant increase of cardiac output with a decreasing in systemic and pulmonary vascular resistance was observed. Since the fall in pulmonary vascular resistance (PVRI) was not proportional to the increase of cardiac output, mean pulmonary artery pressure and transpulmonary gradient increased. The prevalent role of vascular recruitment as mechanism in PVRI reduction during DBT is supported by the concomitant fall in PaO2/FiO2. This strongly suggests a worsening of regional Va/Qc due to an increased perfusion of poorly ventilated areas. CONCLUSIONS: DBT reduces PVRI through a recruitment of vessels due to an increase of pulmonary flow. Dobutamine has a favorable hemodynamic effect in mild-to-moderate pulmonary hypertension in lung transplant candidates.     

Ecotoxicological effects of bisphenol A and nonylphenol on the freshwater cladocerans Ceriodaphnia silvestrii and Daphnia similis

Toxicities of bisphenol A (BPA) and nonylphenol (NP) to the neotropical freshwater cladocerans Ceriodaphnia silvestrii and Daphnia similis were studied under laboratory conditions. Acute exposures to BPA generated mean 48-h EC₅₀ values of 14.44 (6.02–22.85) mg L^?1 for C. silvestrii and 12.05 (1.73–22.37) mg L^?1 for D. similis. When the organisms were exposed to acute doses of NP, mean 48-h EC₅₀ values were 0.055 (0.047–0.064) mg L^?1 (C. silvestrii) and 0.133 (0.067–0.200) mg L^?1 (D. similis). Ceriodaphnia silvestrii was also tested in chronic bioassays, which resulted in mean 8-d IC₂₅ values of 2.43 (2.16–2.69) mg L^?1 BPA [no observed effect concentration (NOEC): 1.38?mg L^?1] and 0.020 (0.015–0.026) mg L^?1 NP (NOEC: 0.015?mg L^?1). These laboratory tests are valuable to broaden the understanding of the environmental threat posed by BPA and NP in aquatic ecosystems, and to increase the knowledge about the sensitivity of neotropical indigenous species to these contaminants. In addition to the laboratory bioassays, species sensitivity distributions were used to suggest protective concentrations of BPA and NP to prevent adverse effects on freshwater organisms. According to the obtained results, concentrations lower than 36.47?µg L^?1 BPA and 1.39?µg L^?1 NP are not expected to adversely impact aquatic organisms in natural ecosystems.     

Molecular Investigation of Canine Parvovirus-2 (CPV-2) Outbreak in Nevis Island: Analysis of the Nearly Complete Genomes of CPV-2 Strains from the Caribbean Region

To date, there is a dearth of information on canine parvovirus-2 (CPV-2) from the Caribbean region. During August–October 2020, the veterinary clinic on the Caribbean island of Nevis reported 64 household dogs with CPV-2-like clinical signs (hemorrhagic/non-hemorrhagic diarrhea and vomiting), of which 27 animals died. Rectal swabs/fecal samples were obtained from 43 dogs. A total of 39 of the 43 dogs tested positive for CPV-2 antigen and/or DNA, while 4 samples, negative for CPV-2 antigen, were not available for PCR. Among the 21 untested dogs, 15 had CPV-2 positive littermates. Analysis of the complete VP2 sequences of 32 strains identified new CPV-2a (CPV-2a with Ser297Ala in VP2) as the predominant CPV-2 on Nevis Island. Two nonsynonymous mutations, one rare (Asp373Asn) and the other uncommon (Ala262Thr), were observed in a few VP2 sequences. It was intriguing that new CPV-2a was associated with an outbreak of gastroenteritis on Nevis while found at low frequencies in sporadic cases of diarrhea on the neighboring island of St. Kitts. The nearly complete CPV-2 genomes (4 CPV-2 strains from St. Kitts and Nevis (SKN)) were reported for the first time from the Caribbean region. Eleven substitutions were found among the SKN genomes, which included nine synonymous substitutions, five of which have been rarely reported, and the two nonsynonymous substitutions. Phylogenetically, the SKN CPV-2 sequences formed a distinct cluster, with CPV-2b/USA/1998 strains constituting the nearest cluster. Our findings suggested that new CPV-2a is endemic in the region, with the potential to cause severe outbreaks, warranting further studies across the Caribbean Islands. Analysis of the SKN CPV-2 genomes corroborated the hypothesis that recurrent parallel evolution and reversion might play important roles in the evolution of CPV-2.     

Post-thoracotomy analgesia: epidural vs intravenous morphine continuous infusion

BACKGROUND: We compared thoracic morphine epidural analgesia (TEA) and I.V. analgesia (IVA) with morphine, in respect to the time to extubation, the quality of postoperative analgesia, side effects, complications, postoperative hospital length of stay in patients having thoracotomy lung resection. METHODS: We prospectively studied 563 consecutive patients, undergoing thoracotomy (lobectomy, bilobectomy or pneumonectomy), randomized in two groups: TEA 286 patients and IVA 277 patients. In the epidural group, before the induction of anesthesia, continuous infusion of 15 mg of morphine in 250 mL of normal saline at 5 mL/h was started. In the IVA group a continuous infusion of 30 mg of morphine associated with 180 mg ketorolac in 250 mL of normal saline at 5 mL/h was started before the induction of anesthesia. The pain degree was evaluated on an analogic scale by Keele modified at 1 (end of anesthesia) 6, 12, 24, and 48 postoperative hours, at rest and after movements. Data obtained were analysed by means of the analysis of variance for repeated measures. RESULTS: The time from the end of surgery to tracheal extubation was similar in both groups. Significantly lower numeric verbal pain scores at rest and after movements were found in the epidural group (p<0.001). Postop complications, nausea and vomiting were higher in the IVA group (p<0.05). Postoperative mean hospital length of stay was 9+/-4 days in TEA and 11+/-4 in the IVA group (p<0.05). CONCLUSIONS: In our study the epidural root was superior in terms of analgesia, side effects, length of stay and postoperative complications after thoracotomy.     

Preprotachykinin A gene expression after administration of 3,4-methylene dioxymethamphetamine (Ecstasy)

This study tested the effects of 8 days of subchronic administration of 3,4-methylene dioxymethamphetamine (MDMA) (5 mg/kg b.w.) on preprotachykinin A mRNA levels in discrete rat brain regions. In situ hybridization examined preprotachykinin A mRNA levels in the core and shell of the nucleus accumbens, the islands of Calleja, the olfactory tubercle, the dorsal and ventral caudate–putamen, the bed nucleus of the stria terminalis, the medial preoptic area, the medial habenular nucleus and in the postero-dorsal part of the medial amygdala. Higher levels of preprotachykinin A mRNA were found in the core and shell of the nucleus accumbens, in the islands of calleja, in the olfactory tubercle, in the bed nucleus of the stria terminalis, in the medial habenular nucleus and the postero-dorsal part of the medial amygdala, compared to control animals. Conversely, increased preprotachykinin A mRNA levels were observed in the dorsal and ventral caudate–putamen in MDMA treated when compared to control rats. In the social memory test, MDMA significantly impaired rats' short-term working memory. These results show that chronic exposure to MDMA strongly affects preprotachykinin A mRNA levels in discrete rat brain regions. These changes occur in experimental conditions in which working memory is markedly reduced, suggesting that changes in gene expression of tachykinin mechanisms may contribute to the effects of MDMA on memory function.     

A quantitative model of cellular senescence influence on cancer and longevity

Contrary to the paradigm that cancer incidence increases indefinitely with age, significant data now suggest cancer incidence may markedly reduce beyond age 80 years for humans and beyond 800 days for mice, and is not inevitable. We show that increasing cellular senescence with age is a possible cause of this reduction, since senescent cells are removed from the pool of cells that retain proliferative ability necessary for cancer. We further show that animal interventions appearing to alter senescence, p53 mutation and melatonin dosing, support the prediction that increasing senescence rate reduces cancer while reducing lifespan, and vice versa. Studies of environmental agents associated with increased cancer might be re-examined to find if there is an association with longevity increases, which may markedly alter our view of such agents. We also show that if an agent functions by slowing both senescence and carcinogenesis, longevity is increased while reducing cancer. Dietary restriction is the only known intervention that accomplishes this, but there may be others.