A Novel Diagnostic Target in the Hepatitis C Virus Genome

BackgroundDetection and quantification of hepatitis C virus (HCV) RNA is integral to diagnostic and therapeutic regimens. All molecular assays target the viral 5'-noncoding region (59-NCR), and all show genotype-dependent variation of sensitivities and viral load results. Non-western HCV genotypes have been under-represented in evaluation studies. An alternative diagnostic target region within the HCV genome could facilitate a new generation of assays.Methods and FindingsIn this study we determined by de novo sequencing that the 3'-X-tail element, characterized significantly later than the rest of the genome, is highly conserved across genotypes. To prove its clinical utility as a molecular diagnostic target, a prototype qualitative and quantitative test was developed and evaluated multicentrically on a large and complete panel of 725 clinical plasma samples, covering HCV genotypes 1-6, from four continents (Germany, UK, Brazil, South Africa, Singapore). To our knowledge, this is the most diversified and comprehensive panel of clinical and genotype specimens used in HCV nucleic acid testing (NAT) validation to date. The lower limit of detection (LOD) was 18.4 IU/ml (95% confidence interval, 15.3-24.1 IU/ml), suggesting applicability in donor blood screening. The upper LOD exceeded 10(-9) IU/ml, facilitating viral load monitoring within a wide dynamic range. In 598 genotyped samples, quantified by Bayer VERSANT 3.0 branched DNA (bDNA), X-tail-based viral loads were highly concordant with bDNA for all genotypes. Correlation coefficients between bDNA and X-tail NAT, for genotypes 1-6, were: 0.92, 0.85, 0.95, 0.91, 0.95, and 0.96, respectively; X-tail-based viral loads deviated by more than 0.5 log10 from 5'-NCR-based viral loads in only 12% of samples (maximum deviation, 0.85 log10). The successful introduction of X-tail NAT in a Brazilian laboratory confirmed the practical stability and robustness of the X-tail-based protocol. The assay was implemented at low reaction costs (US$8.70 per sample), short turnover times (2.5 h for up to 96 samples), and without technical difficulties.ConclusionThis study indicates a way to fundamentally improve HCV viral load monitoring and infection screening. Our prototype assay can serve as a template for a new generation of viral load assays. Additionally, to our knowledge this study provides the first open protocol to permit industry-grade HCV detection and quantification in resource-limited settings.

Laparoscopic management of iatrogenic lesions

Purpose: To present our series of patients who underwent laparoscopic correction of iatrogenic lesions and a review of the literature.Patients and Methods: We evaluated 23 patients who underwent laparoscopic correction of iatrogenic lesions. Thirteen patients had open surgery, 6 had an endoscopic procedure, and 4 had a laparoscopic approach as the first surgical procedure. Vesicovaginal fistulas (VVF) developed in seven patients after open abdominal hysterectomies, and 1 patient presented with a VVF after ureterolithotripsy. A urethral cutaneous fistula developed in one patient after a laparoscopic resection of endometriosis nodules, and 1 patient presented with a ureterovaginal fistula after a perineoplasty. Three patients presented with encrusted ureteral stents after ureterolithotripsy. Ureteral stenosis developed in seven patients: three after open abdominal surgery, three after ureteroscopy, and one after pyeloplasty. One patient had a ureteral injury during laparoscopic partial nephrectomy, and two patients had bowel injuries after a tension-free vaginal tape procedure and a laparoscopic radical prostatectomy.Results: All patients underwent laparoscopic correction of the iatrogenic injuries. One patient had an early recurrence of a VVF, and one patient had a recurrence of a ureteral stenosis. There was one conversion to open surgery because of technical difficulties and one major bleeding event that necessitated blood transfusion. A lower limb compartmental syndrome developed in one patient.Conclusion: Despite the small number of patients and different types of surgeries performed, laparoscopic management of iatrogenic lesions seems to be feasible and safe in experienced hands. Its precise role in the management of this stressful condition still needs to be determined.

Evaluation of different protocols for anatomical studies in Parmeliaceae (lichenized Ascomycota)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

INFLUENCE of ENCAPSULATING MATERIALS on WATER SORPTION ISOTHERMS of VACUUM-DRIED PERSIMMON PULP POWDER

Water sorption isotherms for vacuum-dried persimmon pulp (PP) powder were determined in the temperature range of 20-50C, and the effects of maltodextrin (MD) or gum arabic (GA) addition on the water sorption behavior of the dried powders were analyzed. Several models were evaluated to fit the experimental data and the Guggenheim-Anderson-de Boer model was selected as the most adequate to describe the observed behavior. Addition of encapsulants affected the isotherms: at the same water activity, PP powder with added GA (PP + GA) or MD (PP + MD) presented lower equilibrium water content than pure PP and were less affected by temperature variations. Samples of PP + MD presented lower equilibrium moisture content than those of PP + GA. The isosteric heats of sorption of pulp powders with encapsulants were higher (less negative) than those of PP, suggesting that there are more active polar sites in PP than in pulp powder containing encapsulants.PRACTICAL APPLICATIONSThe choice of persimmon to carry out this work was due to the large persimmon production available in Brazil; moreover, persimmon pulp is rich in vitamin C, vitamin A and iron, as well as in phenolic compounds. Drying of fruit pulps with high sugar content presents technical difficulties because the hygroscopicity and thermoplasticity of the resulting powders when exposed to high temperature and relative humidity. For this reason, addition of high-molar-mass biopolymers, such as maltodextrin or gum arabic, is a strategy to aid drying and to improve storage stability. Knowledge of water sorption isotherms and net isosteric heats of sorption is important to various food processing operations, including drying, storage and packaging. They are useful in calculating time and energy consumptions during drying, modeling moisture changes during storage and predicting shelf life of food products.

Mesocosm experiment on the impact of invertebrate predation on zooplankton of a tropical lake

Invertebrate predation on zooplankton was investigated in mesocosms in the shallow tropical Lake Monte Alegre, Sauo Paulo State, Brazil, in the summer of 1999. Two treatments were applied: one with natural densities of prey and the predators Chaoborus brasiliensis and the water mite Krendowskia sp. (Pr+), and another without predators (Pr-). Three enclosures (volume: 6.6 m(3) of water per enclosure) per treatment were installed in the sediment of the deepest area of the lake (5.0 m). At the beginning, Chaoborus larvae were present in Pr- enclosures, because of technical difficulties in preventing their entrance, but they virtually disappeared in the course of the experiment. Water mites were almost absent in Pr- enclosures. Chaoborus predation negatively influenced the Daphnia gessneri population, but not the populations of the copepods Tropocyclops prasinus and Thermocyclops decipiens and the rotifers Keratella spp. Death rates of Daphnia were generally significantly higher in the Pr+ treatment; Daphnia densities increased after the disappearance of Chaoborus in Pr-. Copepod losses to predation in the experiment may be compensated by higher fecundity, shorter egg development time, and lower pressure on egg-bearing females, resulting in a lower susceptibility to Chaoborus predation. The predation impact of water mite on microcrustaceans and rotifers in the experiment was negligible.

ESTENOSE SUBGLOTICA

In the present study we report the experience of the Department of Otolaryngology-Head and Neck Surgery, University Hospital, Botucatu School in 19 cases of laryngotracheal stenosis. Fifteen cases had subglottic stenosis, one had stenosis of inferior tracheal and subglottic, one had stenosis of trachea and two others had extensive involvement of larynx and trachea due to chronic inflammatory disease. In eleven patients a laryngotracheal reconstrutive technique with costal cartilagem graft was performed with good results. The major technical difficulties, occurring mainly in children, were the withdrawal of the cannula of the tracheostomy due to granulation tissue formation and the subglottic partial stenosis after surgical reconstruction.

Cytogenetic studies in Diplopoda

The Diplopoda have received little attention from cytogeneticists owing mostly to technical difficulties in obtaining mitotic chromosomes, restricting the studies to meiosis and eventual spermatogonial metaphases, which limits the use of modern cytogenetical techniques. A literature search shows that only about 0.1% of all known species have been cytogenetically studied. There are 80,000 species estimated for this group, making it the 3rd. larger class in Arthropoda, after Insecta and Arachnida. The diploid chromosomal number in diplopods varies from 2n=8 to 2n=30 and the sex determination mechanism commonly found is XY/XX. In meiotic prophase, the bouquet formation and the diffuse state in pachytene are typical events. The few works performed on Brazilian fauna add up to 16 species, out of an estimated number of 2000 to 3000 species. The present review reports all the species of diplopods that have been cytogenetically studied so far, each with its chromosome number and sex determination system.

O uso do pneumoperitônio progressivo no pré-operatório das hérnias volumosas da parede abdominal

Context - Correction of voluminous hernias and large abdominal wall defects is a big challenge in surgical practice due to technical difficulties and the high incidence of respiratory and cardiovascular complications. Objectives - To present the authors experience with inducing progressive pneumoperitoneum preoperative to surgical treatment of voluminous hernias of the abdominal wall. Methods - Retrospective study of six patients who presented voluminous hernias of the abdominal wall and were operated after installation of a pneumoperitoneum. The procedure was performed by placing a catheter in the abdominal cavity at the level of the left hypochondrium with ambient air insufflation for 10 to 15 days. Results - Four of the six patients were female and two male. Ages ranged from 42 to 62 years. Hernia duration varied from 5 to 40 years. Four patients had incisional, one umbilical, and one inguinal hernias. Mean pneumoperitoneum time was 11.6 days. There were no complications related to pneumoperitoneum installation and maintenance. All hernias were corrected without technical difficulties. The Lichtenstein technique was used to correct the inguinal hernia, peritoneal aponeurotic transposition for one of the incisional hernias, with the rest corrected using polypropylene mesh. One death and one wall infection were observed post operatively. No recurrences were reported until now, in 4 to 36 months of follow-up. Conclusion - Preoperative progressive pneumoperitoneum is a safe and easy executed procedure, which simplifies surgery and reduces post-operative respiratory and cardiovascular complications. It is indicated for patients with hernias that have lost the right of domain in the abdominal cavity.

Land parceling and neighborhood impacts in Brazil

The proposal and implementation of parceling real estate into smaller lots in Brazil is done according to legal and technical formalities. However, these instruments have proved inefficient in reducing the resulting environmental impacts. The ambiguities of the federal, state and municipal laws and regulations have limited the effectiveness of the actions of urban administrators. Law 10257/2001 emerged as an alternative to overcome these difficulties, proposing the adoption of neighborhood impact studies as an instrument to evaluate new proposals of urban occupation for purposes of environmental licensing. Thus, the purpose of this law is to provide the foundations for municipal public authorities to establish criteria for the assessment, mitigation and compensation of impacts resulting from new occupations. However, the very vagueness of the generic nature of this federal law and its incorrect application in the municipal sphere has posed the greatest obstacles to the good use of this instrument of urban environmental management. These deficiencies are classified herein in the categories of philosophical, technical and operational problems. The problems of a philosophical nature lead to technical difficulties, which in turn trigger operational deficiencies. This article discusses these deficiencies and points out ways to reduce them. © 2009 Journal of Urban and Environmental Engineering (JUEE). All rights reserved.

Apexification with white MTA in an immature permanent tooth with dens invaginatus

Dens invaginatus, also known as dens in dente, is a developmental dental anomaly resulting in an invagination of the enamel organ into the dental papilla. These cases present technical difficulties to the root canal treatment. Apexification using an apical plug of mineral trioxide aggregate (MTA) has been indicated as an alternative to long-term intracanal use of calcium hydroxide in immature permanent teeth. It is considered as a simple and rapid technique. This paper reports a case of Oehlers' Type 1 dens invaginatus in an immature permanent maxillary right lateral incisor, which presented pulp necrosis secondary to dental trauma and was treated by apexification with white MTA apical plugging followed by conventional root canal therapy. The operative procedures are described and the technique is discussed. The physical and biological properties of MTA, associated with appropriate instrumentation and obturation techniques, make this material an excellent option in the endodontic therapy of immature permanent teeth with dens invaginatus.

Diretriz para o gerenciamento integrado de resíduos sólidos urbanos em Estiva Gerbi (SP)

Pós-graduação em Geociências e Meio Ambiente - IGCE

Avaliação das alterações funcionais e histológicas na isquemia renal, comparando-se duas técnicas de hipotermia: estudo experimental em suínos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ovariohisterectomia por videocirurgia (via NOTES vaginal híbrida), celiotomia ou miniceliotomia em cadelas

The elective ovariohysterectomy (OH) is the most frequent procedures performed in dogs. In this study was used three groups of seven animals each (GI, GII, and GIII) that was undergone to three elective OH techniques: (i) mini-celiotomy (Snook-hook technique), (ii) hybrid Natural Orifice Translumenal Endoscopic Surgery (NOTES), and (iii) celiotomy (conventional surgery). The surgical techniques were compared considering the surgery time, trans and postoperative complications, technical difficulties, postoperative pain, surgical bleedind and some vital parameters as: heart rate (FC), respiratory rate (), rectal temperature, invasive blood pressure (PVI) and central venous pressure (PVC). The OH by hybrid vaginal NOTES was the technique with the lowest post-surgical discomfort score and the lowest surgical bleeding, although its surgical time was higher compared to the conventional and the hook (mini-celiotomy) modalities.

Two-stage rumenostomy in buffaloes

Background: Rumenostomy may be performed for therapeutic and digestibility research purposes in bovines, small ruminants and camelids. Several studies requires romenostomy in buffaloes in order to sample ruminal content for laboratorial assays. However, complications and outcome of rumenostomy was poorly studied in buffaloes. Therefore, the aim of the current study was to describe a two-stage rumenostomy technique in buffaloes, focused on intra and post-operative period.Materials, Methods & Results: Nine Murrah buffaloes were submitted to a 36-h and 12-h of food and water fastening. The animals were given acepromazine and maintained in standing position. Flank local anesthesia was carried out. A circular skin incision was carried out in the center of the left flank, followed by divulsion of the external and internal obliques and transversus abdominus muscles, and incision of the peritoneum. Subsequently, a segment of the dorsal aspect of the rumen was grasped and pulled through the flank incision. The rumen was attached to the peritoneum and skin incision margins in four points (dorsal, ventral, cranial and caudal). Additional simple interrupted sutures attaching the rumen serosa to the skin were applied subsequently. Four additional interrupted horizontal mattress sutures were applied equidistantly, taking bites only in the skin and rumen serosa. Following 12 h, the second stage was carried out. The buffaloes were prepared and restrained as performed for the first stage. A circular flap was excised from the exteriorized rumen and the silicone romenostomy cannula was placed. Clinical parameters, postoperative recovery, weight and behavioral pain scale were assessed. Positioning and anesthesia regimen were adequate for the achievement of the procedure. However, two animals fell in the restraint chute during the first surgical stage. Mild ischemia of the exteriorized rumen segment was observed on the second surgical stage, which resulted in less hemorrhage and enhanced cannula positioning. Complete cicatrization and permanent adhesion of the rumen to the skin were achieved. No ruminal leakage to the abdominal cavity occurred. No signs of pain were reported. There were few cases of laxity of the romenostomy opening leading to drop of cannula, myiasis on the margin of the stoma site and few cases of mild ruminal content leakage on the long-term assessment.Discussion: Restraint in standing position was considered adequate, although lateral recumbence constitutes another option. However, higher risk of contamination and technical difficulties in placing the cannulas are expected if lateral recumbence is considered. In other trials using acepromazine, no accidental recumbence occurred. Xylazine was also indicated for chemical restraint of buffaloes. It is known that flexible cannulas provide better anatomic adjustment and adaptation as well as being effective for sampling ruminal content, as seen in the current study. Ruminal leakage is one of the most frequent complications of romenostomy, which may affect animal's welfare. The animals in the current study presented no variations on the body score, even though on those presenting cannula loosening or ruminal content leakage. Moreover, no significant changes of the ruminal content parameters were noticed. Myiasis was also reported following ruminal surgical interventions, which were mainly attributed to extensive breeding. Loss of the cannula, subcutaneous emphysema and suture dehiscence are common complications of romenostomy. Nonetheless, none of those complications were found on the current study. Thus, romenostomy was feasible and efficient for sampling and performing assays of the ruminal content in buffaloes.

Estudo comparativo entre ressecção e eletrocoagulação endometrial em paciebtes com sangramento uterino anormal

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB