Discovering key residues of dengue virus NS2b-NS3-protease: New binding sites for antiviral inhibitors design.

The NS2B-NS3 protease is essential for the Dengue Virus (DENV) replication process. This complex constitutes a target for efficient antiviral discovery because a drug could inhibit the viral polyprotein processing. Furthermore, since the protease is highly conserved between the four Dengue virus serotypes, it is probable that a drug would be equally effective against all of them. In this article, a strategy is reported that allowed us to identify influential residues on the function of the Dengue NS2b-NS3 Protease. Moreover, this is a strategy that could be applied to virtually any protein for the search of alternative influential residues, and for non-competitive inhibitor development. First, we incorporated several features derived from computational alanine scanning mutagenesis, sequence, structure conservation, and other structure-based characteristics. Second, these features were used as variables to obtain a multilayer perceptron model to identify defined groups (clusters) of key residues as possible candidate pockets for binding sites of new leads on the DENV protease. The identified residues included: i) amino acids close to the beta sheet-loop-beta sheet known to be important in its closed conformation for NS2b ii) residues close to the active site, iii) several residues evenly spread on the NS2b-NS3 contact surface, and iv) some inner residues most likely related to the overall stability of the protease. In addition, we found concordance on our list of residues with previously identified amino acids part of a highly conserved peptide studied for vaccine development.

Diabetes nutrition therapy and dietary intake among individuals with Type 1 diabetes in China.

AIMS: To describe the contribution of diabetes nutrition therapy to disease self-management among individuals with Type 1 diabetes in China and to estimate the association of diabetes nutrition therapy with dietary intake. METHODS: The 3C Study was an epidemiological study of the coverage, cost and care of Type 1 diabetes in China. The data reported in the present study are from the 3C Nutrition Ancillary Study, a follow-up study conducted a mean ± sd of 1.6 ± 0.2 years later. Diabetes nutrition therapy was assessed by an interviewer-administered questionnaire. Dietary intake was assessed using three 24-h recalls. The association of diabetes nutrition therapy with dietary intake was estimated using ancova. RESULTS: Participants (n = 100; 54% male) had a mean ± sd age of 41.7 ± 16.3 years and a mean ± sd diabetes duration of 11.8 ± 9.7 years. Fewer than half of the participants reported that they had 'ever' met with a dietitian. While 64% of participants were taught carbohydrate counting, only 12% 'ever' use this tool. Participants on insulin pumps and those testing ≥ 1 time/day reported greater dietary flexibility and higher fruit intakes compared with participants on other insulin regimens and testing less frequently. After adjustment for confounding by age and occupation, there were no consistent differences in dietary intake across subgroups of diabetes nutrition therapy. CONCLUSIONS: In this sample of individuals with Type 1 diabetes in China there is little dietitian involvement or carbohydrate counting. Increased frequency of nutrition education in conjunction with intensified self-monitoring of blood glucose is needed to improve care.

Meningoencephalitis caused by Non-O1, non-O139 Vibrio cholerae in an infant from southern Spain.

Non-O1, non-O139 Vibrio cholerae (NOVC) is an emergent pathogen that mainly causes gastroenteritis. Also, it causes ear, wound infections, and bacteremia but the nervous system is rarely affected. We report on a case of NOVC meningoencephalitis in an infant that recovered after antimicrobial therapy but later presented neurologic sequelae.

Phylogenetic and phylogeographic analysis of the genus Orestias (Teleostei: Cyprinodontidae) in the southern Chilean Altiplano: the relevance of ancient and recent divergence processes in speciation.

This study presents phylogenetic molecular data of the Chilean species of Orestias to propose an allopatric divergence hypothesis and phylogeographic evidence that suggests the relevance of abiotic factors in promoting population divergence in this complex. The results reveal that diversification is still ongoing, e.g. in the Ascotán salt pan, where populations of Orestias ascotanensis restricted to individual freshwater springs exhibit strong genetic differentiation, reflecting putative independent evolutionary units. Diversification of Orestias in the southern Altiplano may be linked to historical vicariant events and contemporary variation in water level; these processes may have affected the populations from the Plio-Pleistocene until the present.

Presence of Hypoderma lineatum stage I larvae in the esophagus of cattle slaughtered in Chihuahua, Chih., Mexico.

In order to detect the presence of Hypoderma lineatum stage I larvae within the esophagus of cattle slaughtered in Chihuahua, Chihuahua, Mexico, a total of five samplings were carried out between July and November 2000. In each instance, a random sample was taken from 10% of the animals slaughtered in a single work shift in each of the two slaughterhouses included in this study. The esophagus were cut longitudinally in order to carry out visual inspection and detect the presence of H. lineatum stage I larvae in the submucosa. The larvae were separated and counted. We identified the presence of H. lineatum stage I larvae in the esophagus for all sampling dates, nevertheless, within the last sampling only one esophagus had them. For all sampling dates the prevalence ranged between 11 and 33%; the latter corresponded to the sampling in October. A total of 287 esophagus was inspected of which 54 were positive with one or more larvae (19%); 233 larvae were obtained from these cases. The number of larvae recovered per sampling ranged from 46 to 74 between July and October, the highest number was found in September's sampling. The largest amount of stage I larvae per esophagus was 22 in the months of July and August. Larvae were always located in the submucosa of the esophagus and all were oriented longitudinally.

Photothermal techniques applied to the determination of the water vapor diffusion coefficient and thermal diffusivity of edible films.

Water vapor diffusion coefficient (WVDC) and thermal diffusivity (alpha) were determined in gelatin-starch films through photothermal techniques. The effect of different variables in the elaboration of these films, such as starch and glycerol concentrations and pH, were evaluated through the response surface methodology. The results indicated that an increase in the glycerol concentration and pH favored the WVDC of the films. On the other hand, alpha was influenced principally by the starch content and pH of the film-forming solution. The minimum alpha value was 4.5 x 10(-4) cm2/s, which is compared with alpha values reported for commercial synthetic polymers.

Shifting brain inhibitory balance and connectivity of the prefrontal cortex of adults with autism spectrum disorder.

Currently, there are no effective pharmacologic treatments for the core symptoms of autism spectrum disorder (ASD). There is, nevertheless, potential for progress. For example, recent evidence suggests that the excitatory (E) glutamate and inhibitory (I) GABA systems may be altered in ASD. However, no prior studies of ASD have examined the 'responsivity' of the E-I system to pharmacologic challenge; or whether E-I modulation alters abnormalities in functional connectivity of brain regions implicated in the disorder. Therefore, we used magnetic resonance spectroscopy ([1H]MRS) to measure prefrontal E-I flux in response to the glutamate and GABA acting drug riluzole in adult men with and without ASD. We compared the change in prefrontal 'Inhibitory Index'-the GABA fraction within the pool of glutamate plus GABA metabolites-post riluzole challenge; and the impact of riluzole on differences in resting-state functional connectivity. Despite no baseline differences in E-I balance, there was a significant group difference in response to pharmacologic challenge. Riluzole increased the prefrontal cortex inhibitory index in ASD but decreased it in controls. There was also a significant group difference in prefrontal functional connectivity at baseline, which was abolished by riluzole within the ASD group. Our results also show, for we believe the first time in ASD, that E-I flux can be 'shifted' with a pharmacologic challenge, but that responsivity is significantly different from controls. Further, our initial evidence suggests that abnormalities in functional connectivity can be 'normalised' by targeting E-I, even in adults.

Green chemistry synthesis of nano-cuprous oxide.

Green chemistry and a central composite design, to evaluate the effect of reducing agent, temperature and pH of the reaction, were employed to produce controlled cuprous oxide (Cu2O) nanoparticles. Response surface method of the ultraviolet-visible spectroscopy is allowed to determine the most relevant factors for the size distribution of the nanoCu2O. X-ray diffraction reflections correspond to a cubic structure, with sizes from 31.9 to 104.3 nm. High-resolution transmission electron microscopy reveals that the different shapes depend strongly on the conditions of the green synthesis.

Complex visual disturbances in Alzheimer's disease.

Although Alzheimer's disease (AD) involves visual association cortex, previous studies have not systematically investigated complex visual disturbances in AD. We examined 30 community-based AD patients, 13 (43%) of whom had complex visual complaints, and compared them with 30 controls on 7 types of complex visual tasks. Despite preserved visual acuity and color recognition, the AD patients were impaired in the visual evaluation of common objects, famous faces, spatial locations, and complex figures. In the AD patients, we found that all 30 had disturbances in figure-ground analysis; 17 (57%) had difficulties visually recognizing actual objects ("agnosia"); those with worse dementia disability had the most complex visual disturbances; and a subgroup (6) with Balint's syndrome performed the most poorly on the complex visual tasks. This study demonstrates that a range of complex visual disturbances are common in AD and suggests that they may result from the known neuropathology in the visual association cortex.

Rapid sustained recovery after cardiac operations.

After successful cardiac operations in the early 1980s the most common causes of prolonged hospitalizations were noncardiac disorders. We prevented or quickly corrected these noncardiac disorders after operations in succeeding patients and observed in the following 2 years that the shortest postoperative stays in the hospital were followed by the fewest rehospitalizations. In 240 consecutive patients the median length of hospital stay after operation was 4 days. The operations included coronary artery bypass procedures, aortic valve replacements, and mitral valve operations. Six patients (2.5%) were rehospitalized within 6 months after discharge and five patients (2.1%) were rehospitalized 6 to 24 months after discharge: Longer initial hospitalizations would not have prevented rehospitalizations. Forty of the 240 patients were discharged on the third postoperative day or earlier (one patient). None died or were rehospitalized in the following 2 years. Prevention or quick correction of noncardiac disorders allowed rapid recovery after heart operations, and rapid recovery indicated that health would be maintained.

Malignant pleural effusion from prostatic adenocarcinoma resolved with hormonal therapy.

A 73-year-old man presented with dyspnea, right-sided pleural effusion, and bilateral pulmonary infiltrates. The pleural fluid revealed adenocarcinoma cells that stained positively for prostatic specific antigen (PSA), which confirmed this uncommon metastatic involvement from prostate cancer. The dyspnea, effusion, and infiltrates disappeared after therapy with flutamide and leuprolide was started. This report demonstrates both the usefulness of immunocytochemical staining for PSA in ascertaining the origin of malignant pleural effusion in men and the effectiveness of the aforementioned endocrine therapy in such setting.

Surgical treatment of mitral insufficiency secondary to coronary artery disease.

From 1970 to 1978, 61 patients were operated upon for mitral insufficiency secondary to coronary artery disease. These patients were between 44 and 71 years of age and all were in Class III or IV of the New York Heart Association Classification. The left ventricular end-diastolic pressure was 15 mm Hg or more in 32 of the 39 patients in whom it was measured. Twenty-four of 31 patients in whom right heart catheterization was performed had a systolic pulmonary artery pressure of 50 mm Hg or greater. All 61 patients had myocardial revascularization, 52 had repair of the mitral valve, and nine had mitral valve replacement. There were five hospital deaths in these 61 patients. Among the nine patients with a preoperative ejection fraction of 0.1 to 0.2, there were two hospital deaths; among the 20 patients with a preoperative ejection fraction of 0.25 to 0.40, there were two hospital deaths; and among the 32 patients with a preoperative ejection fraction of 0.45 to 0.70, there was only 1 hospital death. For those patients with repair and revascularization, the survivability was 81% at 7 years. In the patients with repair and myocardial revascularization, the incidence of peripheral embolization was 0.5% per patient-year.

A ribosomal RNA gene probe differentiates member species of the Anopheles gambiae complex.

A 0.59 kilobase DNA fragment cloned from an rDNA cistron of the mosquito Anopheles gambiae can be used as a probe to differentiate between A. gambiae, A. arabiensis, and A. melas, three morphologically identical sibling species in the A. gambiae complex which otherwise can be reliably distinguished only by polytene chromosome banding patterns. Although all are important (and often sympatric) African malaria vectors, their relative roles in malaria transmission have thus far been difficult to assess. The probe, an EcoRI-SalI fragment from the 3' end of the 28S beta coding region of the cistron, is present in all three species, but the species differ uniquely with respect to the location of an EcoRI site in the nontranscribed spacer (NTS) downstream of the fragment. We have routinely used the probe to identify A. gambiae complex mosquitoes to species on the basis of genomic DNA extracted from individual air dried specimens. A single mosquito abdomen provides more than sufficient DNA for the assay, and neither eggs nor a bloodmeal in the abdomen interfere with DNA yield. Moreover, the DNA extraction procedure does not degrade the bloodmeal IgG, so the residual protein pellet can be used to identify the mosquito bloodmeal source. Since the rDNA cistron organization as detected by the probe does not differ between male and female mosquitoes, the probe can be used for either sex. Preliminary experiments show that the probe is equally useful for mosquito larvae and pupae.

Thirteen years' experience with the Kay-Shiley disc valve for tricuspid replacement in Ebstein's anomaly.

From August, 1965, to November, 1974, 11 patients underwent operation for Ebstein's anomaly. In 1 patient operated on on August 16, 1965, the early Kay-Shiley disc valve was used. In the remaining 10 patients, the Kay-Shiley muscle guard valve was inserted. This valve was designed to prevent the prosthesis from encroaching on the right ventricle, thereby increasing the flow around the disc. One patient with a history of Wolff-Parkinson-White syndrome died of dysrhythmia on the first postoperative day. The other 10 patients have been followed from 4 to 13 years (mean, 6 years 2 months) after operation. Seven patients are working full-time without difficulty, 2 are housewives, and 1 attends school. All patients have improved at least one class in the New York Heart Association Functional Classification.

Limited selective posterior rhizotomy for the treatment of spasticity secondary to infantile cerebral palsy: a preliminary report.

A limited selective posterior rhizotomy was performed on 30 children suffering from spasticity secondary to infantile cerebral palsy. As opposed to standard techniques that stimulate and divide the dorsal rootlets from L2 to S1, we dissected L4, L5, and S1 dorsal roots through an L5 to S1 laminectomy. Eight to 12 rootlets from each root were electrically stimulated with two unipolar electrodes (pulse width, 50 microseconds; 10-50 V). The muscle responses were observed visually and registered by electromyography. Those rootlets associated with an abnormal motor response as evidenced by sustained muscular contraction or by prolonged electromyographic response were divided. Spasticity was scored from 0 to +. The muscular groups assessed were those involved in the flexion of the shoulder, elbow and wrist in the upper limbs, and those involved in flexion and adduction of the hip, flexion of the leg, and plantar flexion in the lower limbs. The patients were assessed 1 week before and 6 months after the operation. Reduction of spasticity was observed in all the muscular groups, and all the patients presented functional improvement of motor abilities. These preliminary results indicate that a limited procedure that reduces the extension of the laminectomy and the length of the operation could be effective for treating spasticity secondary to infantile cerebral palsy.

Pycnodysostosis: some aspects concerning kinetics of calcium metabolism and bone pathology.

A 23-year-old white woman suffering pycnodysostosis whose parents and five siblings were unaffected was investigated. Chromosomal morphology was normal. Histologic examination of a biopsy specimen obtained from the distal phalanx of the left thumb, corresponding to an area on an x-ray film of osteolysis, showed a fibrous dysplasia-like picture. Kinetic studies of calcium metabolism revealed that exchangeable pool size, turnover, and bone accretion rate were all decreased. Intestinal calcium absorption was investigated simultaneously by a double isotope technique and by deconvolution of the plasmatic specific activity curve of 47Ca given per os. Results obtained by both methods coincided in that values were found to be similarly increased. Endogenous fecal calcium was also determined and revealed a similar increase. Some physiopathological implications of these findings are discussed.

Hemoptysis, recurrent cerebrovascular accident, and emergency mitral valve replacement.

Hemoptysis in mitral stenosis may occur frequently, but massive pulmonary hemorrhage is uncommon. We describe a patient with mitral stenosis who had severe hemoptysis and required anticoagulation for recurrent embolic cerebrovascular accidents. The patient underwent successful mitral valve replacement with a Hancock bioprosthesis. There was no evidence of pulmonary bleeding following operation. This case illustrated that mitral valve operation may be performed in acutely ill patients for the relief of hemoptysis.

Evaluation of a myoelectric hand prosthesis for children with a below-elbow absence.

A clinical evaluation of a myoelectric hand prosthesis was carried out within a multicentre trial in three centres in the United Kingdom. The aim was to assess its effectiveness as an acceptable device for young children and to evaluate its technical efficiency. The trial ran for 2 3/4 years and included 87 children. The methodology used and the results obtained are presented with indications towards future developments.

Genetic risk score and adiposity interact to influence triglyceride levels in a cohort of Filipino women.

BACKGROUND/OBJECTIVES: Individually, genetic variants only moderately influence cardiometabolic (CM) traits, such as lipid and inflammatory markers. In this study we generated genetic risk scores from a combination of previously reported variants influencing CM traits, and used these scores to explore how adiposity levels could mediate genetic contributions to CM traits. SUBJECTS/METHODS: Participants included 1649 women from the 2005 Cebu Longitudinal Health and Nutrition Survey. Three genetic risk scores were constructed for C-reactive protein (CRP), high-density lipoprotein cholesterol (HDL-C) and triglycerides (TGs). We used linear regression models to assess the association between each genetic risk score and its related trait. We also tested for interactions between each score and measures of adiposity. RESULTS: Each genetic risk score explained a greater proportion of variance in trait levels than any individual genetic variant. We found an interaction between the TG genetic risk score (2.29-14.34 risk alleles) and waist circumference (WC) (Pinteraction=1.66 × 10(-2)). Based on model predictions, for individuals with a higher TG genetic risk score (75th percentile=12), having an elevated WC (⩾80 cm) increased TG levels from 1.32 to 1.71 mmol l(-1). However, for individuals with a lower score (25th percentile=7), having an elevated WC did not significantly change TG levels. CONCLUSIONS: The TG genetic risk score interacted with adiposity to synergistically influence TG levels. For individuals with a genetic predisposition to elevated TG levels, our results suggest that reducing adiposity could possibly prevent further increases in TG levels and thereby lessen the likelihood of adverse health outcomes such as cardiovascular disease.

Glutamate/glutamine and neuronal integrity in adults with ADHD: a proton MRS study.

There is increasing evidence that abnormalities in glutamate signalling may contribute to the pathophysiology of attention-deficit hyperactivity disorder (ADHD). Proton magnetic resonance spectroscopy ([1H]MRS) can be used to measure glutamate, and also its metabolite glutamine, in vivo. However, few studies have investigated glutamate in the brain of adults with ADHD naive to stimulant medication. Therefore, we used [1H]MRS to measure the combined signal of glutamate and glutamine (Glu+Gln; abbreviated as Glx) along with other neurometabolites such as creatine (Cr), N-acetylaspartate (NAA) and choline. Data were acquired from three brain regions, including two implicated in ADHD-the basal ganglia (caudate/striatum) and the dorsolateral prefrontal cortex (DLPFC)-and one 'control' region-the medial parietal cortex. We compared 40 adults with ADHD, of whom 24 were naive for ADHD medication, whereas 16 were currently on stimulants, against 20 age, sex and IQ-matched healthy controls. We found that compared with controls, adult ADHD participants had a significantly lower concentration of Glx, Cr and NAA in the basal ganglia and Cr in the DLPFC, after correction for multiple comparisons. There were no differences between stimulant-treated and treatment-naive ADHD participants. In people with untreated ADHD, lower basal ganglia Glx was significantly associated with more severe symptoms of inattention. There were no significant differences in the parietal 'control' region. We suggest that subcortical glutamate and glutamine have a modulatory role in ADHD adults; and that differences in glutamate-glutamine levels are not explained by use of stimulant medication.