HydroCalc Proteome: a tool to identify distinct characteristics of effector proteins.

Bacterial pathogenicity is associated with secretion of effector proteins into intra- and extracellular spaces. These proteins interfere with cellular processes such as inhibition of phagosome-lysosome fusion, induction of apoptosis and autophagy, activation and suppression of kinases, regulation of receptor activity, and modulation of transcription factors. Knowledge regarding the characteristics of these proteins would assist in pathogenicity studies, and help to identify possible and novel targets for antibacterial drugs. Amino acid hydropathy is a property that can affect behavior patterns in effector proteins. The HydroCalc Proteome tool analyzes total hydropathy, average hydropathy, C-terminal hydropathy, C-terminal load, and basic polar amino acids at the C-terminus. These five properties could contribute to the identification of proteins with an effector potential. HydroCalc Proteome is a web tool that provides a simple interface for the analysis of hydropathy properties in proteins. This tool permits the analysis of a single protein or even the complete proteome, which cannot be achieved by using other hydropathy tools. The tool displays the result of five properties related to effector proteins in a single table. The HydroCalc Proteome (www.gmb.bio.br/hydrocalc) is a powerful tool for protein analysis, and can contribute to the study of effector proteins.

Hidden prevalence of chronic kidney disease in hypertensive patients: the strategic role of primary health care.

OBJECTIVE: To identify the hidden prevalence of chronic kidney disease (CKD) in hypertensive patients. STUDY DESIGN: Cross-sectional study of individuals with systemic arterial hypertension (SAH) who were registered for primary health care (PHC). METHODS: In total, 293 individuals participated. Data were collected through interviews, as well as biochemical and anthropometric assessments. The CKD-EPI formula was used to identify the occurrence of CKD. Pearson's chi-squared test or Fisher's exact test were used to compare proportions. Prevalence ratios were estimated with a confidence interval of 95% for associations between the explanatory variables and CKD. RESULTS: Most of the individuals assessed were female (74%), elderly (69%), with a low income (90%), low education levels (84%) and overweight (66.9%). A CKD prevalence of 38.6% (95% CI: 33.0-44.2) was found and approximately 14% were at an advanced stage of the disease. Upon comparison of the variables in the different stages of CKD, statistically significant association could be suggested between CKD and age, education, alcohol intake, overweight individuals, cardiovascular risk, abnormal creatinine and abnormal microalbuminuria. When the prevalence ratio was assessed, association could be suggested between CKD and age, and CKD and creatinine. CONCLUSION: The high hidden prevalence of CKD confirms the need to train health professionals involved in the treatment of SAH through PHC, enabling the prevention and diagnosis of CKD in its early stages.

Purine nucleoside phosphorylase activity and expression are upregulated in sites affected by periodontal disease.

BACKGROUND AND OBJECTIVE: Purine nucleoside phosphorylase (PNP) is an enzyme that catalyzes the reversible phosphorolysis of purine nucleosides, playing a key role in the purine salvage pathway. Activated T cells seem to rely heavily on PNP to remain functionally active and are particularly sensitive to PNP deficiency. The role of PNP in periodontal tissues has not been characterized thus far. The aim of this study therefore was to assess the activity and expression of PNP in the gingival tissues of periodontitis patients. MATERIAL AND METHODS: Ten patients consecutively admitted for treatment had their periodontal clinical variables recorded and their gingival crevicular fluid collected. After periodontal treatment the patients were seen once a month for plaque and bleeding control, and had their periodontal variables recorded and gingival crevicular fluid collected at 90 and 180 d. Purine nucleoside phosphorylase-specific activity was assessed using a spectrophotometer through the addition of the PNP substrate analog 2-amino-6mercapto-7-methyl purine riboside to the gingival crevicular fluid. In parallel, PNP expression was assessed by immunohistochemistry and real-time PCR in gingival biopsies and cell culture. RESULTS: Purine nucleoside phosphorylase activity was higher in the gingival crevicular fluid of periodontally diseased sites, which was positively correlated with improvements of the clinical variables. Treatment of periodontal disease induced a striking decrease of PNP activity in periodontally diseased sites. Expression of PNP was more pronounced in mononuclear cells and endothelial cells of the gingiva, and the mRNA levels were 5.7-fold higher in inflamed tissues compared with control samples. CONCLUSION: Purine nucleoside phosphorylase activity and expression are upregulated in periodontally diseased sites and can be detected in the gingival crevicular fluid.

Intussusception in adults: a retrospective study.

OBJECTIVE: Intestinal intussusception in adult patients is rare. In contrast with paediatric patients, it is usually secondary to a definable lesion, often malignant. The purpose of this study was to determine the causes and the management of intussusception in adult patients. METHOD: A retrospective review was performed looking at patients over 18 years with intestinal intussusception who were admitted to a tertiary university hospital from 1997 to 2007. RESULT: There were 16 patients (out of whom 10 were female subjects) of mean age 49 years (range 19-76). All presented with abdominal pain and in seven (46.6%) patients, this was acute. The diagnosis of intussusception was correctly made preoperatively in eight (50%) patients. Six (37.5%) patients had the lead point for the intussusception at the ileocaecal valve, five (31.25%) in the small bowel and five (31.25%) had a colonic lead point. An anatomical cause was found in 14 (87.5%). In two (12.5%), the intussusception occurred in the postoperative period without any definable lesion. Half the patients had a malignant neoplasm. All patients underwent surgery. In 14 (87.5%) patients, this was by resection and in two (12.5%), a reduction with no resection was carried out. CONCLUSION: The features of intussusception may be nonspecific and the diagnosis is often made only during laparotomy. An identifiable organic lesion is present in most cases. En bloc resection is recommended for ileocaecal and colocolic intussusception.

Enrofloxacin assay validation and pharmacokinetics following a single oral dose in chickens.

The pharmacokinetics of enrofloxacin (ENRO), a fluoroquinolone antimicrobial agent, was studied in male broiler chickens (Cobb) after single oral administration of 10 mg of ENRO/kg b.w. A high-performance liquid chromatography-photodiode array detector (DAD) (HPLC-DAD) method was developed and validated and used for quantitation of ENRO and its major metabolite ciprofloxacin in plasma. The HPLC analyses were carried out using a cationic-octadecyl mixed column and 0.05 mol/L phosphate buffer (pH 2.5)/acetonitrile as mobile phase. The sample preparation of plasma consisted of the precipitation of proteins followed by solid phase extraction on cationic-octadecyl mixed cartridges. The method was validated considering linear range, linearity, selectivity, sensitivity, limit of detection (LOD), limit of quantitation (LOQ), intra- and inter-day precisions and accuracy. The LOD and LOQ for both fluoroquinolones were 60 and 200 ng/mL for plasma. The plasma concentration vs. time graph was characteristic of a two-compartment open model. The maximal plasma concentration of 1.5 +/- 0.2 mg/mL was achieved at 9 +/- 2 h. The elimination half-life and the mean residence time of ENRO were 1.5 +/- 0.2 and 15.64 h, respectively. The area under the concentration-time curve was calculated as 35 +/- 4 mgxh/mL.

Pseudomyxoma peritonei arising from colonic polyps.

AIMS: Pseudomyxoma peritonei may have as its primary site a mucinous gastrointestinal adenoma or carcinoma that gains access to the peritoneal cavity. This manuscript describes this disease arising from a benign or malignant colonic polyp. METHODS: From a database of over 1000 pseudomyxoma peritonei patients and colorectal carcinomatosis patients, three cases were identified in which the primary tumor site was a colonic polyp. The clinical history and course of these patients were studied. RESULTS: In a review of the clinical management of these patients, all three had an event whereby neoplastic cells from the surface of the colonic polyp could have gained access to the free peritoneal cavity. The patients developed the characteristic pseudomyxoma peritonei syndrome. All three patients were treated with cytoreductive surgery plus perioperative hyperthermic intraperitoneal chemotherapy. CONCLUSIONS: Colonic polyps can serve as a source of dysplastic cells whereby pseudomyxoma peritonei can result. Caution to prevent seeding to the free peritoneal cavity during surgery for colonic polyps should be observed. If pseudomyxoma peritonei develops, cytoreductive surgery and perioperative intraperitoneal chemotherapy should be considered for treatment.

Late postoperative follow-up of patients submitted to subtotal splenectomy.

Subtotal splenectomy preserving the upper splenic pole vascularized only by the splenogastric vessels has been performed by us in 91 patients since 1984. This procedure was combined with central splenorenal shunt or portal variceal disconnection to treating schistosomatic portal hypertension (n = 67); it was combined with distal pancreatectomy to treat pancreatic cystadenoma (n = 1), and it was carried out for severe splenic trauma (n = 14), and to control severe Gaucher's disease (n = 4) and myeloid metaplasia due to myelofibrosis (n = 5). All patients are alive. In the present paper, we present the late postoperative follow up of 32 patients operated in a period that ranges from 1 to 12 years. All patients are satisfied with the operation. Increased counts of white blood cell (n = 10) and platelets (n = 7) were the only hematological alterations. Esophageal varices were present in 23 of 27 patients operated due to portal hypertension, but all of them were shorter than in the preoperative period. The ultrasound confirmed the presence of the splenic remnant (30 of 32) without significant size changes. Splenic scintigraphies were positive in 30 of 32 patients. Subtotal splenectomy seems to be a safe procedure that can be useful in portal hypertension, some metabolic and hematological diseases, and benign growth of the distal pancreas.

[Delayed (modified) oral cholecystography in the study of biliary dyskinesia]. / Colecistografia oral retardada (modificada) no estudo da discinesia biliar.

Biliary dyskinesia is difficult to be either diagnosed or treated. A modified oral cholecystography technique has been tested to make it suitable for studying such gallbladder disease. Routine oral cholecystography was performed and then two other roentgenograms were taken both 24 and 48 hours after it. In our opinion, further studies should be performed whenever gallbladder visualization lasts longer than 48 hours even in order to indicate surgery.

Effect of air temperature and humidity on ingestive behaviour of sheep.

Thirty-two Polwarth ewes, of ages up to 1 year, were observed in a climatic chamber (24 to 45 degrees C) for eight periods of 5 h each. The observations were made through a window in the chamber wall. All animals were observed four times, then shorn and observed four times again. The animals were given weighed quantities of water and feed consisting of commercial concentrate plus Rhodes grass (Chloris gayana) hay. The water and feed remaining after 5 h of observation were weighed. The following traits were analysed: time eating hay (TEH), time eating concentrate (TEC), time drinking water (TDW), weight of hay eaten (WHE), weight of concentrate eaten (WCE), volume of ingested water (VIW), ruminating time standing up (RTS), ruminating time lying down (RTL), idling time standing up (ITS), and idling time lying down (ITL). Shearing had a significant effect for all traits except ITS. Shearing resulted in higher values for all traits except for ITS and ITL. Ingestion of hay (TEH and WHE) decreased with increased air temperature and humidity, while the ingestion of concentrate (TEC) and WHE) and water (TDW and VIW) increased. Rumination decreased with increased air temperature and humidity, and was higher in shorn than in unshorn sheep.

Influence of hot environments on some blood variables of sheep.

Thirty-two Polwarth sheep of ages up to 1 year were observed under temperatures varying from 10.5 to 46.5 degrees C. The following blood cell counts were made: erythrocyte (RBC), leucocyte (WBC), eosinophil (EOS), neutrophil (NEU), lymphocyte (LYM) and monocyte (MON). Other traits measured were: haemoglobin (HB), haematocrit (HT), blood glucose (GLU) and serum protein (PROT). Multivariate analysis of variance was used and the results showed a significant (P < 0.001) effect for the interaction of shearing and temperature treatment. Under temperatures > 25 degrees C, sheep presented a decrease of RBC, WBC, HB and HT, these differences being greater in the shorn than in the unshorn animals. Unshorn animals presented higher variations in EOS, NEU, LYM, MON and GLU. Blood glucose increased under high temperatures in the shorn animals (from 56.36 +/- 0.65 mg/100 ml to 60.52 +/- 0.69 mg/100 ml) as in the unshorn animals (from 54.72 +/- 0.74 mg/100 ml to 57.56 +/- 0.77 mg/100 ml).

Dependence of the C-6 sulfate of the glucosamine moiety and 1----4 glycosidic linkage of heparin disaccharides for production of hemorrhage: reversal of the antihemostatic activity of heparin and their fragments by adenosine triphosphate and myosin.

Topical application or intraperitoneal injection of heparin and heparin oligosaccharides produces a potent inhibition of skin hemostasis. Studies conducted with disaccharides derived from heparin, heparan sulfate, and chondroitin sulfates have shown that delta-4,5-uronyl-(1----4)-glucosamine, bearing a sulfate at the C-6 position of the glucosamine residue, is the minimum structure for the antihemostatic activity. The disaccharides with this basic structure produce uncontrollable hemorrhage from small blood vessels, similar to that observed for heparin. The finding that other sulfated disaccharides, with the same sulfate to hexosamine to uronic acid ratios but with the sulfate at a different position (C-2) or with a different glycosidic linkage (1----3), were inactive as inhibitors of hemostasis indicates that a specific structure is needed to produce the effect. The inhibitory activity of the normal hemostatic process produced by heparin and its products could be reversed either by ATP or myosin. Molecular models show that part of the disaccharide inhibitors and ATP have a similar structural conformation.