Optimization of A(H1N1)pdm09 vaccine seed viruses: The source of PB1 and HA vRNA as a major determinant for antigen yield.

Vaccination prevents and reduces the severity of influenza virus infections. Continuous evolution of influenza hemagglutinin (HA) and neuraminidase (NA) supports the virus to evade pre-existing immunity, which demands vaccines to be reformulated every year. Incorporation of polymerase basic protein 1 (PB1) viral RNA (vRNA) of the same origin of HA and NA vRNA has been observed in previous pandemic viruses and occasionally reported for influenza A vaccine prototype strains of prior seasons. At this point, it remains to be explored whether this phenomenon translates into an improved growth phenotype. In this work, we showed that the HA vRNA of A(H1N1)pdm09 is generally incorporated with the PB1 vRNA of the same origin, establishing the beneficial effect of the presence of PB1 and the pattern of the PB1-HA co-incorporation in the A(H1N1)pdm09 model. We further investigated the putative interplay between PB1 and antigenic proteins regarding the vRNA composition of the progeny and observed that vRNA segregation does not appear to be mainly determined by protein-protein interactions; while vRNA-vRNA interactions can be suggested as the main driving force. Our data also indicate an increase in the hemagglutination capacity and neuraminidase activity due to incorporation of PB1, HA and NA from A(H1N1)pdm09, in comparison with the recombinant virus incorporating only HA and NA from A(H1N1)pdm09 - which have the potential to improve current limitations regarding antigenicity and immunogenicity of influenza vaccines. Further knowledge of the complex vRNA-vRNA interaction network between PB1 and HA will additionally contribute to improve current vaccine formulation, and to gradually optimize the production of A(H1N1)pdm09 reverse genetics vaccine seed virus towards a higher cost-effectiveness.

An update of posterior retroperitoneoscopic adrenalectomy - Case series.

INTRODUCTION: Posterior retroperitoneoscopic adrenalectomy (PRA) has advantages over transperitoneal approach. A second group of 10 patients is analyzed and compared with the first 10 procedures. Conclusions on feasibility, safety and learning curve are taken. MATERIAL AND METHODS: A retrospective analysis of a second group of 10 patients submitted to PRA was conducted. All patients with functioning and non-functioning adrenal tumors <6-8 cm and without features of malignancy were included. A comparison with the previous 10 cases was conducted, and the results of all 20 cases were compared with other surgeons. RESULTS: Pre-operative diagnoses: Conn's syndrome - 8 (80%); Pheochromocytoma - 1 (10%); Non-functioning tumor (≥ 4 cm) - 1 (10%). Mean size of adrenal tumors was 2,9 cm. Mean operative time for first group was 46,7 min and 31,1 min for the second (p = 0,036). Postoperative in-hospital days decreased in the second group (p = 0,01). Conversion rate was equal (10%). Morbidity and mortality were similar. DISCUSSION: Comparing the evolution of operative time in both groups, a constant and faster operative time was noted for the second group and a decreasing linear tendency was noted as more cases were being performed. Postoperative in-hospital days lowered in the second group, because with experience we started discharging patients earlier. Outcomes are stable between both groups. Our results match other authors data. CONCLUSION: These results are consistent with our first report and support the small learning curve for PRA, which is technically feasible and safe. Operative time and in-hospital days are influenced by surgeon's experience. More cases need to be collected so that these results can be validated.

To hit or not to hit: Large-scale sequence analysis and structure characterization of influenza A NS1 unlocks new antiviral target potential.

Influenza NS1 protein is among the most promising novel druggable anti-influenza target, based on its structure; multiple interactions; and global function in influenza replication and pathogenesis. Notwithstanding, drug development guidance based on NS1 structural biology is lacking. Here, we design a promising strategy directed to highly conserved druggable regions as a result of an exhaustive large-scale sequence analysis and structure characterization of NS1 protein across human-infecting influenza A subtypes, over the past 100 years. We have identified 3 druggable pockets and 8 new potential hot spot residues in the NS1 protein, not described before, additionally to other 16 sites previously identified, which represent attractive targets for pharmacological modulation. This study provides a rationale towards structure-function studies of NS1 druggable sites, which have the potential to accelerate the NS1 target validation. This research also contributes to a deeper comprehension and insight into the evolutionary dynamics of influenza A NS1 protein.

Rare case of symmetrical peripheral gangrene due to septic shock, disseminated intravascular coagulation and inotropic use.

INTRODUCTION: Symmetrical peripheral gangrene (SPG) is a rare syndrome defined by the peripheral ischemic lesion of two or more extremities in the absence of major vascular obstructive disease. PRESENTATION OF CASE: A 45yo woman, admitted in intensive care unit due to urinary septic shock, in need of high doses of amines, developed cold extremities with acrocyanosis that rapidly progressed to gangrene. Laboratory analysis revealed increased inflammatory parameters, liver shock, thrombocytopenia, prolonged coagulation times, increased D-Dimers and isolation of Acinetobacter baumanni in urine culture. An intravenous vasodilator was initiated with clinical benefits. After improvement and delimitation of the lesions, the patient underwent the amputation of the distal phalanges of the 2nd, 3rd and 4th fingers of the right hand and the toes of both feet. DISCUSSION/CONCLUSION: Even though there is no consensus regarding SPG treatment, consequences should be mitigated, particularly when vasodilators are used, in order to avoid major amputation.

Posterior retroperitoneoscopic adrenalectomy-Case series.

INTRODUCTION: Posterior retroperitoneoscopic adrenalectomy has advantages over transperitoneal technique. However many surgeons prefer the transperitoneal technique because they get a familiar and wider working space. MATERIAL AND METHODS: A retrospective analysis of the first 10 patients submitted to posterior retroperitoneoscopic adrenalectomy was conducted. Data collected included: diagnosis, size, operation time, blood loss, conversion rate, morbidity and mortality, in-hospital length of stay. Compare our outcomes with worldwide bigger series, and take conclusions on the feasibility of the technique was the objective. RESULTS: We included 2 pheochromocytomas, 1 giant cystic pheochromocytoma, 4 Conn's, 2 Cushing's, 1 non-functioning tumor with 4 cm. Mean operation time was 46,7 min for lesions ranging from 1,8 to 14 cm. Blood loss was negligible. One patient (10%) was converted to laparotomy because of a past clinical history of dorsal and lumbar trauma. No morbidity and no mortality. Mean hospital length of stay was 2,2 days. DISCUSSION: Mean operation time found in bigger series published in worldwide literature is 40-105,6 min. Complication rate reported ranges from 0 to 14,4%. No mortality has been ever reported. Blood loss reported in other series is 10-50 ml. The data found in our study matches other studies data. Since the same surgeon who had never performed the technique before operated all patients, makes us believe the technique is safe and feasible. CONCLUSION: Posterior retroperitoneoscopic adrenalectomy has a small learning curve. It is technically safe and feasible. More patients will be collected to validate these results.

Oral and dental anomalies in purebred, brachycephalic Persian and Exotic cats.

OBJECTIVE To determine the prevalence of dental anomalies in brachycephalic cats from various geographic regions and analyze potential relationships with oral disease. DESIGN Prospective multicenter cross-sectional study. ANIMALS 50 purebred Persian (n = 42) and Exotic (8) cats. PROCEDURES Cats were anesthetized, and a complete dental examination, dental charting, 3-view oral photography, and full-mouth dental radiography were performed. RESULTS Malocclusions were observed in 36 (72%) cats. Crowding of the teeth was evident in 28 (56%) cats, with the incisors being most commonly affected. Malpositioned teeth were noted in 32 (64%) cats, with abnormal orientation being the most common anomaly followed by rotation and impaction. Numerical abnormalities were present in 38 (76%) cats, including 6 (12%) with hyperdontia and 32 (64%) with hypodontia. Periodontal disease was reported in 44 (88%) cats, and cats with periodontal disease were older than cats without. Tooth resorption was evident in 35 (70%) cats. Overall, 123 of 1,349 (8.7%) teeth had external inflammatory resorption and 82 (6.1%) had external replacement resorption. The premolar teeth were the teeth most commonly affected with inflammatory resorption, whereas the canine teeth were the teeth most commonly affected with replacement resorption. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that because of their brachycephaly, Persian and Exotic cats have unique oral and dental features that may predispose them to dental disease (eg, tooth resorption and periodontal disease). Knowledge of the particular dental anomalies common in brachycephalic cats could aid in early detection and mitigation of dental disease in these breeds.