Rotavirus serotypes and electropherotypes identified among hospitalised children in São Luís, Maranhão, Brazil

During June 1997-June 1999 rotavirus infection was screened in infants aged up to 2 years and hospitalised with acute diarrhoea in S?o Luís, Northeastern Brazil. Altogether, 128 stool samples were collected from diarrhoeic patients and additional 122 faecal specimens from age and temporal matched inpatients without diarrhoea were obtained; rotavirus positivity rates for these groups were 32.0% (41/128) and 9.8% (12/122), respectively (p < 0.001). Both electropherotyping and serotyping could be performed in 42 (79.2%) of the 53 rotavirus-positive stool samples. Long and short electropherotypes were detected at similar rates--38.1% and 40.5% of specimens, respectively. Overall, a G serotype could be assigned for 35 (83.3%) of specimens, the majority of them (66.7%) bearing G1-serotype specificity. Taking both electropherotypes and serotypes together, G1 rotavirus strains displaying long and short RNA patterns accounted for 30.9% and 19.0% of tested specimens, respectively; all G2 strains had short electropherotype. Rotavirus gastroenteritis was detected year-round and, in 1998, the incidence rates tended to be higher during the second semester than in the first semester: 45.2% and 26.1% (p = 0.13), respectively. Rotavirus infections peaked at the second semester of life with frequencies of 30.1% and 13.5% for diarrhoeic children and controls, respectively. While the six rotavirus strains bearing G2-type specificity were circulating throughout the whole study period, G1 serotypes (n = 27) emerged as from June 1998 onwards, 20 (74.1%) of which clustering in 1998. These data underscore the importance of rotaviruses in the aetiology of severe infantile gastroenteritis in Northeastern Brazil and sustain the concept that a future vaccine should confer protection against more than one serotype.     

Adiponutrin: A new gene regulated by energy balance in human adipose tissue

Adiponutrin is a newly identified nonsecreted adipocyte protein regulated by changes in energy balance in rodents. We documented the influence of energy balance modification on adiponutrin gene expression in humans. We investigated the mRNA expression in sc adipose tissue of nonobese women and in obese women during 2-d very low-calorie diet (VLCD) and subsequent refeeding as well as before and after a VLCD of 3 wk (21-d VLCD). The adiponutrin mRNA levels of the nonobese and obese women were not different (P > 0.05). Two-day VLCD reduced the average level of adiponutrin mRNA expression by 36% (P = 0.0016), whereas refeeding elevated the mRNA level by 31% (P = 0.004). The 3-wk VLCD caused a dramatic 58% fall of the adiponutrin mRNA expression level (P = 0.001). The mRNA level was negatively correlated with fasting glucose (Rho = -0.62; P < 0.0001), and subjects with high adiponutrin mRNA level had an increased insulin sensitivity. Compared with other adipocyte proteins such as leptin and adiponectin, adiponutrin mRNA did not show correlation with either adiposity indexes or with leptin or adiponectin mRNAs. These results indicate that adiponutrin gene expression in humans is highly regulated by changes in energy balance.     

Effectiveness of intervention led by a community pharmacist for improving recognition of sleep apnea in primary care – a cohort study

Despite its high prevalence and major public health ramifications, obstructive sleep apnea syndrome (OSAS) remains underdiagnosed. The aim of this study was to determine whether the involvement of a community pharmacist (CP) in the care pathway of a patient at risk of OSAS, through the implementation of a community pharmacist (CP) intervention, was effective, i.e. increased the use of diagnostic tests in this population. We compared a cohort of patients included in a research protocol (exposed to a CP intervention) with patients having the same characteristics taken from a general population database who did not receive the intervention (unexposed group). The aim of the CP intervention was to educate patients about the risk of untreated OSAS, encouraging them to consult their general practitioner, and urging the doctor to continue investigations. We included 782 patients at risk of OSAS, i.e. taking one or more anti‐hypertensive drugs, being overweight (body mass index >25) and snoring almost every night (88 in the exposed group and 694 in the unexposed group). After a 6‐month follow‐up, the number of patients who underwent an OSAS diagnostic test was significantly higher in the exposed group compared to the unexposed group (22.7 versus 11.4%, P = 0.003). Being exposed to the pharmacist intervention was associated with a higher chance of undergoing a diagnostic test for OSAS, adjusted odds ratio: 2.24 (1.25–4.01). In conclusion, these findings provide arguments for the implementation of a CP OSAS screening intervention in CP routine practice.     

Clinical significant of serum chromogranin A levels for diagnosing pheochromocytoma in hypertensive patients

OBJECTIVE: CMA is a widespread glycoprotein located in the secretory vesicles of neuroendocrine cells and is co-released with peptides and biogenic amines into the circulation. The present study set out to investigate the clinical utility of assessing serum CGA levels in comparison with the urinary KTCO and their urinary metabolites concentrations, which are to date the gold standard validated diagnostic test. METHODS: From January 2000 to June 2001, 202 consecutive patients, aged 53 +/- 12.7, 102 males, were admitted to our department for a hypertension evaluation. Blood samples for measurements of plasma concentrations of chromogranin A were collected and serum CGA levels were quantified by RIA technique (RIACT). This radioimmunometric technique consisted in using 2 monoclonal antibodies directed to 2 specific antigenic domains of the middle portion of the CGA. The fixed threshold value for identifying positive results was, set at 100 ng/ml according to previous studies. RESULTS: No pheochomocytoma was diagnosed by conventional urinary KTCO essay. Of the 202 CGA blood samples, 32 turned out to be positive, due to commonly encountered false positive causes (inhibitor of the pump with protons, corticotherapy, hypergastrinemia, chronic renal insufficiency, respectively, in 11, 2, 1, 18 cases). The CGA plasma concentration averaged 77 +/- 77 mg/ml and 203 +/- 125 ng/ml in the CGA subgroup over the threshold value. CONCLUSION: The reliability of immunoradiometric serum CGA concentrations appeared according to this work to be comparable to that of the urinary KTCO levels and their urinary metabolites in hypertensives. Moreover, it solely requires a simple, easily done blood taking, less expensive than urinary KTCO collection. Besides, no antihypertensive drugs interfered with the analysis of CGA levels. However, some false positive results have to be mentioned in the presence of renal impairment, hypergastrinemia, corticotherapy, inhibitor of the pump with protons.     

Insulin secretion and insulin sensitivity in diabetic and non-diabetic subjects with hepatic nuclear factor-1alpha (maturity-onset diabetes of the young-3) mutations

OBJECTIVE: To evaluate insulin secretion and sensitivity in affected (diabetes mellitus or impaired glucose tolerance; n=7) and in unaffected (normal glucose tolerance; n=3) carriers of hepatocyte nuclear factor-1alpha (maturity-onset diabetes of the young-3 (MODY3)) gene mutations. METHODS: Insulin secretion was assessed by an i.v. glucose tolerance test (IVGTT), hyperglycemic clamp and arginine test, and insulin sensitivity by an euglycemic hyperinsulinemic clamp. Results were compared with those of diabetic MODY2 (glucokinase-deficient) and control subjects. RESULTS: The amount of insulin secreted during an IVGTT was decreased in affected MODY3 subjects (46+/-24 (s.d.) pmol/kg body weight (BW)) as compared with values in MODY2 (120+/-49pmol/kg BW) and control (173+/-37pmol/kg BW; P=0.0004) subjects. The amount of insulin secreted during a 10mmol/l glucose clamp was decreased in affected MODY3 subjects (171+/-78pmol/kg BW) and MODY2 subjects (302+/-104pmol/kg BW) as compared with control subjects (770+/-199pmol/kg BW; P=0.0001). Insulin secretion in response to arginine was decreased in affected MODY3 subjects. Milder and heterogeneous defects were observed in the unaffected MODY3 subjects; the amount of insulin secreted during the hyperglycemic clamp was 40-79% of that of controls. The response to arginine was abnormally delayed. Insulin sensitivity was decreased in diabetic but not in non-diabetic MODY3 subjects. CONCLUSIONS: Beta-cell dysfunction in response to glucose and arginine is observed in affected and unaffected MODY3 subjects. The MODY3 and MODY2 subtypes present different insulin secretion profiles. Secondary insulin resistance might contribute to the chronic hyperglycemia of MODY3 patients and modulate their glucose tolerance.     

Platelet aggregation impacts thrombin generation assessed by calibrated automated thrombography

A calibrated automated thrombogram (CAT) is performed usually with human platelet-free plasma (PFP) but may be more relevant with platelet-rich plasma (PRP). In this case, platelets are not stimulated by subendothelial molecules like collagen. Our aim was to assess the consequence of strong (collagen) or weak (ADP) induction of platelet release and aggregation on thrombin generation. Platelet aggregation in PRP was triggered with 10 µg/mL collagen or 10 µM ADP using a lumi-aggregometer. Thrombin generation curves were monitored by CAT in different conditions: PRP, PRP with activated platelets (actPRP), aggregated PRP (agPRP), aggregated platelets resuspended in autologous PFP (resPRP), PFP and PFP obtained after aggregation (agPFP). We found a 3-fold shortening of the lag time and time to peak and a marked increase in velocity and thrombin peak without changes in endogenous thrombin potential (ETP) in agPRP with both agonists compared with PRP. The same holds true in agPFP but with a marked increase in ETP compared with PFP. Similar changes in the kinetics of thrombin generation were observed with actPRP-collagen and to a lesser extent in resPRP-collagen compared with PRP. By contrast, there were no modifications of the thrombin generation curves in actPRP-ADP. Alpha-2-macroglobin-thrombin complexes were unchanged in the different PRP conditions but were increased in PFP prepared from agPFP compared to control PFP. Platelet aggregation during activation by agonists other than thrombin did not increase thrombin generation but accelerated its kinetics mainly via platelet content release and platelet-derived extracellular vesicules formation. In diseases characterized by altered platelet granule content or release as well as altered platelet activation, a platelet aggregation step prior to CAT analysis may be clinically relevant to improve laboratory estimation of the bleeding/thrombotic balance.