The anti-inflammatory potential of eight indigenous probiotic Lactobacillus isolates was evaluated in vitro in terms of modulating the expression of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in human acute monocytic leukemia (THP-1) cells under inflammatory conditions. Amongst these, Lactobacillus plantarum Lp91 was the most potent anti-inflammatory strain as it evoked a significant (P < 0.001) down-regulation of TNF-α by −1.45-fold relative to the control in THP-1 cells. However, in terms of IL-6 expression, all the strains could up-regulate its expression considerably at different levels. Hence, based on in vitro expression of TNF-α, Lp91 was selected for in vivo study in lipopolysaccharide (LPS)-induced mouse model to look at the expression of TNF-α, IL-6, monocyte chemotactic protein-1 (MCP-1), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule (ICAM-1) and E-selectin in mouse aorta. In LPS challenged (2 h) mice group fed with Lp91 for 10 days, TNF-α, IL-6, MCP-1, VCAM-1, ICAM-1 and E-selectin expressions were significantly down-regulated by 3.10-, 10.02-, 4.22-, −3.14-, 2.28- and 5.71-fold relative to control conditions. In conclusion, Lp91 could serve as a candidate probiotic strain to explore it as a possible biotherapeutic anti-inflammatory agent against inflammatory diseases including cardiovascular disease.
Electronic supplementary material
The online version of this article (doi:10.1007/s12263-013-0347-5) contains supplementary material, which is available to authorized users.
Probiotic; Lipopolysaccharide; Cardiovascular diseases; Intercellular adhesion molecule; Monocyte chemotactic protein-1; Vascular cell adhesion molecule-1
Five strains of Streptomyces (CAI-24, CAI-121, CAI-127, KAI-32 and KAI-90) were earlier reported by us as biological control agents against Fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceri (FOC). In the present study, the Streptomyces were characterized for enzymatic activities, physiological traits and further evaluated in greenhouse and field for their plant growth promotion (PGP) of sorghum and rice. All the Streptomyces produced lipase, β-1-3-glucanase and chitinase (except CAI-121 and CAI-127), grew in NaCl concentrations of up to 6%, at pH values between 5 and 13 and temperatures between 20 and 40°C and were highly sensitive to Thiram, Benlate, Captan, Benomyl and Radonil at field application level. When the Streptomyces were evaluated in the greenhouse on sorghum all the isolates significantly enhanced all the agronomic traits over the control. In the field, on rice, the Streptomyces significantly enhanced stover yield (up to 25%; except CAI-24), grain yield (up to 10%), total dry matter (up to 18%; except CAI-24) and root length, volume and dry weight (up to 15%, 36% and 55%, respectively, except CAI-24) over the control. In the rhizosphere soil, the Streptomyces significantly enhanced microbial biomass carbon (except CAI-24), nitrogen, dehydrogenase (except CAI-24), total N, available P and organic carbon (up to 41%, 52%, 75%, 122%, 53% and 13%, respectively) over the control. This study demonstrates that the selected Streptomyces which were antagonistic to FOC also have PGP properties.
Antagonistic Streptomyces; Plant growth promotion; Field evaluation; Fusarium oxysporum f. sp. ciceri; Sorghum; Rice
IABPs are frequently used to provide hemodynamic support during high risk percutaneous coronary intervention (PCI), but clinical evidence to support their use is mixed. We examined hospital variation in IABP use among high risk PCI patients, and determined the association of IABP use on mortality in this population.
Methods and Results
We analyzed data submitted to the CathPCI Registry® between January 2005 and December 2007. High risk PCI was defined as having at least one of the following features: unprotected left main artery as the target vessel, cardiogenic shock, severely depressed left ventricular function, or ST segment elevation myocardial infarction. Hospitals were categorized into quartiles by their proportional use of IABP. We examined differences in in-hospital mortality across hospital quartiles using a hierarchical logistic regression model to adjust for differences in patient and hospital characteristics across hospital quartiles of IABP use. IABPs were used in 18,990 (10.5%) of 181,599 high risk PCIs. Proportional use of IABP varied significantly across hospital quartiles: Q1: 0.0%–6.5%; Q2: 6.6% to 9.2%; Q3: 9.3% to 14.1%; and Q4: 14.2% to 40.0%. In multivariable analysis, after adjustment for differences in patient and hospital characteristics, in-hospital mortality was comparable across quartiles of hospital IABP usage (Q1: Ref; Q2: Odds Ratio (OR) 1.11, 95% CI 0.99–1.24; Q3: OR 1.03, 95% CI 0.92–1.15; Q4: OR 1.06, 95% CI 0.94–1.18).
IABP use varied significantly across hospitals for high risk PCI. However, this variation in IABP use was not associated with differences in in-hospital mortality.
Angioplasty; Atherosclerosis; Heart assist device
HapR has been given the status of a high cell density master regulatory protein in Vibrio cholerae. Though many facts are known regarding its structural and functional aspects, much still can be learnt from natural variants of the wild type protein. This work aims at investigating the nature of functional inertness of a HapR natural variant harboring a substitution of a conserved glutamate residue at position 117 which participates in forming a salt bridge by lysine (HapRV2G-E117K). Experimental evidence presented here reveals the inability of this variant to interact with various cognate promoters by in vitro gel shift assay. Furthermore, the elution profiles of HapRV2G-E117K protein along with the wild type functional HapRV2G in size-exclusion chromatography as well as circular dichroism spectra did not reflect any significant differences in its structure, thereby indicating the intactness of dimer in the variant protein. To gain further insight into the global shape of the proteins, small angle X-ray scattering analysis (SAXS) was performed. Intriguingly, increased radius of gyration of HapRV2G-E117K of 27.5 Å in comparison to the wild type protein from SAXS data analyses implied a significant alteration in the global shape of the dimeric HapRV2G-E117K protein. Structure reconstruction brought forth that the DNA binding domains were substantially “parted away” in this variant. Taken together, our data illustrates that substitution of the conserved glutamate residue by lysine in the dimerization domain induces separation of the two DNA binding domains from their native-like positioning without altering the dimeric status of HapR variant.
Factors affecting somatic embryogenesis from immature cotyledon of gum arabic tree [Acacia senegal (L.) Willd.] were investigated. Induction of somatic embryogenesis was influenced by plant growth regulator concentrations and addition of amino acids in medium. Best induction of somatic embryogenesis was obtained on MS medium supplemented with 0.45 μM 2, 4-D, 2.32 μM Kin and 15 mM L-glutamine. L-glutamine plays a significant role in the maturation of somatic embryos and most of embryos attained maturity only on L-glutamine (15 mM) containing medium. Maximum percent (75.0 ± 2.5) germination of somatic embryos was recorded on medium containing 0.22 μM BAP.
A. senegal; L-glutamine; Plant growth regulators; Somatic embryo induction
The peptide hormone cholecystokinin (CCK) exerts a wide range of digestive and CNS-related physiological signaling via CCK receptors in brain and gut. There is very limited information available on these receptors in Atlantic salmon. The aim of this study was to characterize CCK receptors in gut and brain of salmon. We have identified and cloned one CCK-1 receptor and duplicates of CCK-2 receptor in salmon. The phylogenetic analysis indicates the existence of one common ancestor gene for all CCK receptors. CCK-1R mRNA is highly expressed in pancreas followed by midgut, hindgut, gallbladder, and stomach indicating an involvement in pancreatic regulation and gallbladder contractions. CCK-2R1/gastrin mRNA is expressed at high levels in midgut and at relatively low levels in stomach, gallbladder, and pancreas. We postulate CCK-2R1/gastrin receptor to have gastrin-related functions because of its distribution and abundance in gastro-intestinal (GI) tissues. CCK-2R2 is relatively abundant in brain but has low expression levels in gut tissues supporting the hypothesis for involvement in the gut-brain signaling. Major functional motifs and ligand interaction sites in salmon are conserved with that of mammals. This information will be instrumental for comparative studies and further targeting receptor activation and selectivity of biological responses of CCK in salmon.
Appetite regulation; Atlantic salmon; cholecystokinin receptors; digestive physiology; gene expression; ligand binding; protein structure
Congenital eyelid imbrication syndrome (CEIS) is an extremely rare, benign, transient, self-limiting eyelid malposition disorder. The classic triad of signs in patients with a CEIS consists of bilateral upper eyelids overriding the lower eyelids when child was in sleep, bilateral medial and lateral canthal tendon laxity and tarsal conjunctival hyperemia. We report a third case of congenital combined eyelid imbrication and floppy eyelid syndrome in healthy neonate that was resolved within a week with conservative treatment.
Congenital ectropion; congenital eyelid imbrication syndrome; congenital floppy eyelid syndrome; congenital lax upper eyelid syndrome; down syndrome
Ryanodine receptor 1 (RyR1) is well-known to be expressed in systemic and pulmonary vascular smooth muscle cells (SMCs); however, its functional roles remain largely unknown. In the present study, we attempted to determine the potential importance of RyR1 in membrane depolarization-, neurotransmitter-, and hypoxia-induced Ca2+ release and contraction in pulmonary artery SMCs (PASMCs) using RyR1 homozygous and heterozygous gene deletion (RyR1−/− and RyR1+/−) mice. Our results indicate that spontaneous local Ca2+ release and caffeine-induced global Ca2+ release are significantly reduced in embryonic RyR1−/− and adult RyR+/− cells. An increase in [Ca2+]i following membrane depolarization with high K+ is markedly attenuated in RyR1−/− and RyR1+/− PASMCs in normal Ca2+ or Ca2+-free extracellular solution. Similarly, muscle contraction evoked by membrane depolarization is reduced in RyR1+/− pulmonary arteries in the presence or absence of extracellular Ca2+. Neurotransmitter receptor agonists and inositol 1,4,5-triphosphate elicit a much smaller increase in [Ca2+]i in both RyR1−/− and RyR1+/− cells. We have also found that neurotransmitter-evoked muscle contraction is significantly inhibited in RyR1+/− pulmonary arteries. Hypoxia-induced increase in [Ca2+]i and contraction are largely blocked in RyR1−/− and/or RyR1+/− PASMCs. Collectively, our findings provide genetic evidence for the functional importance of RyR1 in spontaneous local Ca2+ release, and membrane depolarization-, neurotransmitter-, as well as hypoxia-induced global Ca2+ release and attendant contraction in PASMCs.
Ryanodine receptor; Calcium release; Membrane depolarization; Hypoxia; Pulmonary artery smooth muscle cell
Breath holding spells are a common and dramatic form of syncope and anoxic seizure in infancy. They are usually triggered by an emotional stimuli or minor trauma. Based on the color change, they are classified into 3 types, cyanotic, pallid, and mixed. Pallid breath holding spells result from exaggerated, vagally-mediated cardiac inhibition, whereas the more common, cyanotic breathholding spells are of more complex pathogenesis which is not completely understood. A detailed and accurate history is the mainstay of diagnosis. An EKG should be strongly considered to rule out long QT syndrome. Spontaneous resolution of breath-holding spells is usually seen, without any adverse developmental and intellectual sequelae. Rare cases of status epilepticus, prolonged asystole, and sudden death have been reported. Reassurance and education is the mainstay of therapy. Occasionally, pharmacologic intervention with iron, piracetam; atropine may be of benefit. Here we present 2 cases, one of each, pallid and cyanotic breath holding spells.
The aim of this study was to compare the accuracy of cone beam CT (CBCT) with intraoral radiographs for detection of occlusal caries.
A set of 60 extracted teeth were imaged using a Sirona Galileos CBCT system (Sirona Dental Systems, Bensheim, Germany) and an intraoral Planmeca® system (Planmeca OY, Helsinki, Finland). Six observers looked at both modalities and used a five-point confidence scale to evaluate presence or absence of occlusal caries. Histology was used as the gold standard. Receiver operating characteristic analysis and weighted kappa statistics were used for statistical analysis. Differences in the area under the curve (AUC) values between observers and modalities were analysed using analysis of variance (ANOVA). Differences in sensitivity and specificity were analysed using the Wilcoxon test. Interobserver and intraobserver reliability was assessed by weighted kappa scores.
The mean value and standard deviation of AUC was 0.719 ± 0.038 for CBCT and 0.649 ± 0.062 for the intraoral radiographs. The ANOVA results demonstrated that there was no significant difference between the modalities and the observers. The interobserver kappa for pairs of observers ranged from fair to substantial for bitewings (0.244–0.543) and CBCT (0.152–0.401). Four out of six observers reported higher sensitivity but lower specificity with CBCT. The Wilcoxon exact p-value showed no difference in sensitivity (0.175) or specificity (0.573) between the two modalities.
Based on the results we conclude that the Sirona CBCT unit cannot be used for the sole purpose of looking at occlusal caries.
caries; cone beam computed; cone beam computed tomography; dental; tomography
Common carp (Cyprinus carpio) is a widely cultivated freshwater fish for human consumption, while koi carp, is a farmed colored sub species of common carp used for ornamental purposes. Since 1998, both common carp and koi carp are severely affected by a viral disease called as Koi herpes virus disease (KHVD). This disease is caused by Koi herpes virus (KHV), also known as cyprinid herpes virus-3. The virus causes interstitial nephritis and gill necrosis in carps, so it is also termed as carp interstitial nephritis and gill necrosis virus. KHV is a double stranded icosahedral DNA virus belonging to family Alloherpesviridae, with a genome size of 295 kbp, larger than any member of Herpesviridae. The viral genome encodes 156 potential protein coding open reading frames. Each virion consists of forty structural proteins, which are classified as capsid (3), envelope (13), tegument (2) and unclassified (22) structural proteins. Diagnosis of KHVD is mainly based on detection of viral DNA by polymerase chain reaction amplification using specific primers or loop mediated isothermal amplification. Temperature dependent latent infection is unique to KHV; and carrier fish are often not detected, thereby possibly resulting in spread of this pathogen to newer areas. The disease is now known to occur in, or has been recorded from at least 26 different countries of the world. Fortunately, KHVD has not been reported from India or from Indian major carps. To monitor the disease status of the country, a total of 254 fish samples collected from different parts of India were screened by PCR for the presence of KHV. None of the tested samples were found to be positive for KHV. These results demonstrate that tested samples from different parts of India were apparently free from KHV. Preliminary risk assessment of KHV suggest that in the event of unrestricted importation of koi carps into our country, there is a higher probability of risk to aquaculture as compared to natural waters. So there is strong need to develop diagnostic capabilities and launch surveillance programmes for KHV in India.
Koi carp; Common carp; Koi herpes virus
Previous studies have described an “obesity paradox” with heart failure, whereby higher body mass index (BMI) is associated with lower mortality. However, little is known about the impact of obesity on survival after acute myocardial infarction.
Data from 2 registries of patients hospitalized in the United States with acute myocardial infarction between 2003–04 (PREMIER) and 2005–08 (TRIUMPH) were used to examine the association of BMI with mortality. Patients (n=6359) were categorized into BMI groups (kg/m2) using baseline measurements. Two sets of analyses were performed using Cox proportional hazards regression with fractional polynomials to model BMI as categorical and continuous variables. To assess the independent association of BMI with mortality, analyses were repeated adjusting for 7 domains of patient and clinical characteristics.
Median BMI was 28.6. BMI was inversely associated with crude 1-year mortality (normal, 9.2%; overweight, 6.1%; obese, 4.7%; morbidly obese; 4.6%; p<0.001), which persisted after multivariable adjustment. When BMI was examined as a continuous variable, the hazards curve declined with increasing BMI and then increased above a BMI of 40. Compared with patients with a BMI of 18.5, patients with higher BMIs had a 20% to 68% lower mortality at 1 year. No interactions between age (p=0.37), gender (p=0.87) or diabetes mellitus (p=0.55) were observed.
There appears to be an “obesity paradox” among acute myocardial infarction patients such that higher BMI is associated with lower mortality, an effect that was not modified by patient characteristics and was comparable across age, gender, and diabetes subgroups.
body mass index; mortality; myocardial infarction; fractional polynomials; obesity paradox
Despite the favorable outcome of most pediatric patients with Hodgkin lymphoma (HL), there is rising concern about risks of carcinogenesis from both diagnostic and therapeutic radiation exposure for patients treated on study protocols. Although previous studies have investigated radiation exposure during treatment, radiation from post-treatment surveillance imaging may also increase the likelihood of secondary malignancies. All diagnostic imaging examinations involving ionizing radiation exposure performed for surveillance following completion of therapy were recorded for 99 consecutive pediatric patients diagnosed with HL from 2000 to 2010. Cumulative radiation dosage from these examinations and the frequency of relapse detection by these examinations were recorded. In the first 2 years following completion of therapy, patients in remission received a median of 11 examinations (range 0–26). Only 13 of 99 patients relapsed, 11 within 5 months of treatment completion. No relapse was detected by 1- or 2-view chest radiographs (n = 38 and 296, respectively), abdomen/pelvis computed tomography (CT) scans (n = 211), or positron emission tomography (PET) scans alone (n = 11). However, 10/391 (2.6%) of chest CT scans, 4/364 (1.1%) of neck CT scans, and 3/47 (6.4%) of PET/CT scans detected relapsed disease. Thus, only 17 scans (1.3%) detected relapse in a total of 1358 scans. Mean radiation dosages were 31.97 mSv for Stage 1, 37.76 mSv for Stage 2, 48.08 mSv for Stage 3, and 51.35 mSv for Stage 4 HL. Approximately 1% of surveillance imaging examinations identified relapsed disease. Given the very low rate of relapse detection by surveillance imaging stipulated by current protocols for pediatric HL patients, the financial burden of the tests themselves, the high cure rate, and risks of second malignancy from ionizing radiation exposure, modification of the surveillance strategy is recommended.
Hodgkin disease; late effects; radiology
Due to the functional defects in apoptosis signaling molecules or deficient activation of apoptosis pathways, leukemia has become an aggressive disease with poor prognosis. Although the majority of leukemia patients initially respond to chemotherapy, relapse is still the leading cause of death. Hence targeting apoptosis pathway would be a promising strategy for the improved treatment of leukemia. Hydantoin derivatives possess a wide range of important biological and pharmacological properties including anticancer properties. Here we investigated the antileukemic activity and mechanism of action of one of the potent azaspiro hydantoin derivative, (ASHD).
Materials and Methods
To investigate the antileukemic efficacy of ASHD, we have used MTT assay, cell cycle analysis by FACS, tritiated thymidine incorporation assay, Annexin V staining, JC1 staining and western blot analysis.
Results showed that ASHD was approximately 3-fold more potent than the parent compounds in inducing cytotoxicity. Tritiated thymidine assay in conjunction with cell cycle analysis suggests that ASHD inhibited the growth of leukemic cells. The limited effect of ASHD on cell viability of normal cells indicated that it may be specifically directed to cancer cells. Translocation of phosphatidyl serine, activation of caspase 3, caspase 9, PARP, alteration in the ratio of BCL2/BAD protein expression as well as the loss of mitochondrial membrane potential suggests activation of the intrinsic pathway of apoptosis.
These results could facilitate the future development of novel hydantoin derivatives as chemotherapeutic agents for leukemia.
Reconstitution of GLUT4 vesicle fusion in a defined fusion system shows that the C2-domain factor Doc2b activates the SNARE-dependent fusion reaction by a calcium- and membrane bending–dependent mechanism. Of importance, certain features of Doc2b function appear to be distinct from how synaptotagmin-1 promotes synaptic release.
The glucose transporter GLUT4 plays a central role in maintaining body glucose homeostasis. On insulin stimulation, GLUT4-containing vesicles fuse with the plasma membrane, relocating GLUT4 from intracellular reservoirs to the cell surface to uptake excess blood glucose. The GLUT4 vesicle fusion reaction requires soluble N-ethylmaleimide–sensitive factor attachment protein receptors (SNAREs) as the core fusion engine and a group of regulatory proteins. In particular, the soluble C2-domain factor Doc2b plays a key role in GLUT4 vesicle fusion, but its molecular mechanism has been unclear. Here we reconstituted the SNARE-dependent GLUT4 vesicle fusion in a defined proteoliposome fusion system. We observed that Doc2b binds to GLUT4 exocytic SNAREs and potently accelerates the fusion kinetics in the presence of Ca2+. The stimulatory activity of Doc2b requires intact Ca2+-binding sites on both the C2A and C2B domains. Using electron microscopy, we observed that Doc2b strongly bends the membrane bilayer, and this membrane-bending activity is essential to the stimulatory function of Doc2b in fusion. These results demonstrate that Doc2b promotes GLUT4 exocytosis by accelerating the SNARE-dependent fusion reaction by a Ca2+- and membrane bending–dependent mechanism. Of importance, certain features of Doc2b function appear to be distinct from how synaptotagmin-1 promotes synaptic neurotransmitter release, suggesting that exocytic Ca2+ sensors may possess divergent mechanisms in regulating vesicle fusion.
Post-transcriptional gene silencing is commonly observed in polyploid species and often poses a major limitation to plant improvement via biotechnology. Five plant viral suppressors of RNA silencing were evaluated for their ability to counteract gene silencing and enhance the expression of the Enhanced Yellow Fluorescent Protein (EYFP) or the β-glucuronidase (GUS) reporter gene in sugarcane, a major sugar and biomass producing polyploid. Functionality of these suppressors was first verified in Nicotiana benthamiana and onion epidermal cells, and later tested by transient expression in sugarcane young leaf segments and protoplasts. In young leaf segments co-expressing a suppressor, EYFP reached its maximum expression at 48–96 h post-DNA introduction and maintained its peak expression for a longer time compared with that in the absence of a suppressor. Among the five suppressors, Tomato bushy stunt virus-encoded P19 and Barley stripe mosaic virus-encoded γb were the most efficient. Co-expression with P19 and γb enhanced EYFP expression 4.6-fold and 3.6-fold in young leaf segments, and GUS activity 2.3-fold and 2.4-fold in protoplasts compared with those in the absence of a suppressor, respectively. In transgenic sugarcane, co-expression of GUS and P19 suppressor showed the highest accumulation of GUS levels with an average of 2.7-fold more than when GUS was expressed alone, with no detrimental phenotypic effects. The two established transient expression assays, based on young leaf segments and protoplasts, and confirmed by stable transgene expression, offer a rapid versatile system to verify the efficiency of RNA silencing suppressors that proved to be valuable in enhancing and stabilizing transgene expression in sugarcane.
To determine the burden of trachoma and its related risk factors amongst the native population of Car-Nicobar Island in India.
Rapid assessment for trachoma was conducted in ten villages of Car- Nicobar Island according to standard WHO guidelines. An average of 50 children aged 1–9 years were assessed clinically for signs of active trachoma and facial cleanliness in each village. Additionally, all adults above 15 years of age in these households were examined for evidence of trachomatous trichiasis and corneal opacity. Environmental risk factors contributing to trachoma like limited access to potable water & functional latrine, presence of animal pen and garbage within the Nicobari hut were also noted in all villages.
Out of a total of fifteen villages in Car-Nicobar Island, ten villages were selected for trachoma survey depending on evidence of socio-developmental indicators like poverty and decreased access to water, sanitation and healthcare facilities. The total population of the selected clusters was 7277 in the ten villages. Overall, 251 of 516 children (48.6%;CI: 46.5–55.1) had evidence of follicular stage of trachoma and 11 children (2.1%;CI:1.0–3.4) had evidence of inflammatory stage of trachoma. Nearly 15%(CI:12.1–18.3) children were noted to have unclean faces in the ten villages. Trachomatous trichiasis was noted in 73 adults (1.0%;CI:0.8–1.2). The environmental sanitation was not found to be satisfactory in the surveyed villages mainly due to the co-habitance of Nicobari people with domestic animals like pigs, hens, goats, dogs, cats etc in most (96.4%) of the households.
Active trachoma and trachomatous trichiasis was observed in all the ten villages surveyed, wherein trachoma control measures are needed.
Considerable attention has been devoted to the effect of social support on patient outcomes after acute myocardial infarction (AMI). However, little is known about the relation between patient living arrangements and outcomes. Thus, we used data from PREMIER, a registry of patients hospitalized with AMI at 19 US centers between 2003-04, to assess the association of living alone with post-AMI outcomes. Outcome measures included 4-year mortality, 1-year readmission, and 1-year health status, using the Seattle Angina Questionnaire (SAQ) and Short Form-12 physical health component (SF-12 PCS) scales. Patients who lived alone had higher crude 4-year mortality (21.8% vs. 14.5%, P<0.001), but comparable rates of 1-year readmission (41.6% vs. 38.3%, p=0.79). Living alone was associated with lower unadjusted quality of life (mean SAQ −2.40 (95% confidence interval [CI] −4.44, −0.35), p=0.02), but had no impact on SF-12 PCS (−0.45 (95% CI −1.65, 0.76), p=0.47) compared with patients who did not live alone. After multivariable adjustment, patients who lived alone had a comparable risk of mortality (hazard ratio [HR] 1.35, 95% CI: 0.94-1.93) and readmission (HR 0.99, 95% CI: 0.76-1.28) as patients who lived with others. Mean quality of life scores remained lower among patients who lived alone (SAQ −2.91 (95% CI −5.56, −0.26), p=0.03). Living alone may be associated with poorer angina-related quality of life one year post-MI, but is not associated with mortality, readmission, or other health status measures after adjusting for other patient and treatment characteristics.
Living alone; acute myocardial infarction; social support
Okra (Abelmoschus esculentus (L) Moench) is an important vegetable crop of India. Dried okra pods have wide use in snacks and are in great demand for domestic as well as export market. Hence, effect of four slice sizes (1, 2, 3 and 4 cm) and four drying temperatures (50, 60, 70 and 80 °C) on quality of hot air dried okra were studied. Okra pods were dried in the form of slices cut across the length at different temperatures. Quality assessment of okra was done on the basis of protein, ascorbic acid and fibre content. Okra slice sizes and drying temperatures affected all the quality parameters significantly (p < 0.05). Maximum retention of protein, ascorbic acid and fibre content were found in 2 cm long slices dried at 60 °C temperature.
Okra; Drying; Protein; Ascorbic acid; Fibre
Ophthalmomyiasis is the infestation of human eye by the larvae of certain flies. Sheep botfly commonly manifests as Ophthalmomyiasis externa when there is conjunctival involvement or rarely as Opthalmomyiasis interna when there is larval penetration into the eyeball. It appears to be more common than what has been indicated by previously published reports. We present a report of seven cases of Ophthalmomyiasis by Oestrus ovis, from central India who presented with features of conjunctivitis varying between mild to severe. The larvae were seen in bulbar and palpebral conjunctiva and also entangled in lashes with discharge. Since the larvae are photophobic, it is prudent to look for them in the fornices and also in discharge. Prompt removal of the larvae from the conjunctiva helps in relieving the symptoms and also prevents serious complications. Taxonomic identification of the species is important to estimate the risk of globe penetration by the larvae.
Central India; conjunctivitis; Oestrus ovis; Ophthalmomyiasis
Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of healthcare-associated infections and significant contributor to healthcare cost. Community-associated-MRSA (CA-MRSA) strains have now invaded healthcare settings. A convenience sample of 97 clinical MRSA isolates was obtained from seven hospitals during a one-week period in 2010. We employed a framework integrating Staphylococcus protein A typing and full-genome next-generation sequencing. Single nucleotide polymorphisms were analyzed using phylodynamics. Twenty-six t002, 48 t008, and 23 other strains were identified. Phylodynamic analysis of 30 t008 strains showed ongoing exponential growth of the effective population size the basic reproductive number (R0) ranging from 1.24 to 1.34. No evidence of hospital clusters was identified. The lack of phylogeographic clustering suggests that community introduction is a major contributor to emergence of CA-MRSA strains within hospitals. Phylodynamic analysis provides a powerful framework to investigate MRSA transmission between the community and hospitals, an understanding of which is essential for control.
To evaluate the relationship between A1C and glucose therapy intensification (GTI) in patients with diabetes mellitus (DM) hospitalized for acute myocardial infarction (AMI).
RESEARCH DESIGN AND METHODS
A1C was measured as part of routine care (clinical A1C) or in the core laboratory (laboratory A1C, results unavailable to clinicians). GTI predictors were identified using hierarchical Poisson regression.
Of 1,274 patients, 886 (70%) had clinical A1C and an additional 263 had laboratory A1C measured. Overall, A1C was <7% in 419 (37%), 7–9% in 415 (36%), and >9% in 315 patients (27%). GTI occurred in 31% of patients and was more frequent in those with clinical A1C both before (34 vs. 24%, P < 0.001) and after multivariable adjustment (relative risk 1.34 [95% CI 1.12–1.62] vs. no clinical A1C).
Long-term glucose control is poor in most AMI patients with DM, but only a minority of patients undergo GTI at discharge. Inpatient A1C assessment is strongly associated with intensification of glucose-lowering therapy.
The Drug Discovery Center collaborates with a wide range of academic and industrial research centers to facilitate the identification of active small molecules with high potential for use as biological probes or as starting points for drug discovery programs. The DDC operates state-of-the-art high throughput and high content screening instrumentation and a diverse 350,000 compound library. The center's personnel provide collaborators with advice in assay design, analytical technology selection, and library design via cheminformatics and/or structure-based approaches. Typical programs are exemplified by an HTS program targeting the identification of novel atypical PKC inhibitors for potential use in cancer and an HCS program targeting the identification of stimulators of the differentiation of oligodendrocyte progenitor cells to oligodendrocytes for potential use in multiple sclerosis. Activities of atypical PKCs, PKCiota and PKCzeta, were measured using both fluorescent detection of ADP production and MALDI-TOF detection of substrate phosphorylation. 30,000 compounds were screened for their effect on PKC activity using these orthogonal detection methods to identify potential inhibitors. Oligodendrocyte differentiation was measured in a High Content Screen by pairing a selective marker of oligodendrocytes with Alexa Fluor 488 secondary antibody for the detection of mature ODs. DAPI stain was used for nucleus detection. Confocal microscope images were acquired and analyzed using an algorithm for neurite outgrowth adapted for the characterization of oligodendrocyte processes. Three measurements: Mean Maximum Process Length per Cell, Mean Process Signal Intensity per Cell, and Percentage Differentiated Cells per Well, were quantified by the image analysis script and afforded a statistically significant separation of promoter controls and inhibitor controls from non-treated (neutral) controls with Mean Maximum Process Length as measurement.
Recently, mass spectrometry (MS)-based readout has been demonstrated to be a highly effective for high throughput screening (HTS) assays. The major advantages compared to the most common fluorescence readout are the paucity of false readouts, reduced reagent costs, and the ability to multiplex assays such that multiple therapeutic targets can be screened for inhibitor hits with one pass through the compound repository. Previously, we have developed MS-based methods for rapid and accurate compound screening for inhibitors to therapeutic targets. However, the limited use of MS-based methods with small test libraries has been insufficient to validate the overall utility of this readout for large screening campaigns. Thus, in this report, the MS-based readout technology was scaled to include a library of 30,400 compounds to systematically validate the reliability of MALDI-MS readout head-to-head versus a traditional methods of HTS. The target enzyme for these comparative assays is PKC-iota, which plays a role cancer cell survival, tumor growth and potentially invasion. First the MS-based assay was fully integrated into an automated workflow on a PerkinElmer Plate:Explorer HTS system in a 384-well format. Then, the primary screen of 30,400 compounds with both the MS and fluorescence-based readouts yielded a hit rate of 0.3% and 0.9% for the two methods, respectively. Only 29% of the MS-based hits confirmed in triplicate assays; however, 95% of those confirmed hits validated as concentration-dependent inhibitors with IC50 value ranging from low nM to high μM inhibitors. By contrast, 58% of the fluorescence hits were deemed as false positives since they produced fluorescence inhibition even in the absences of PKC-iota. Overall the data validate the utility of the MS-based readout in terms of sensitivity, reproducibility and minimal reagent cost. We are now investigating ways to incorporate screening technologies as an additional service and revenue stream for our core laboratory.