Nicolau Syndrome (Embolia Cutis Medicamentosa): A Rare and Poorly Recognized Iatrogenic Cause of Cutaneous Thrombotic Vasculopathy.

Nicolau syndrome is a rare form of iatrogenic cutaneous necrosis which affects injection sites. Although classically associated with intramuscular injections, it may develop after subcutaneous or other routes of parenteral drug administration. Clinically, it manifests as necrotic ulcers that often develop in a background of erythematous and livedoid reticular patches. The histopathologic characteristics of Nicolau syndrome are poorly documented in the dermatopathology literature and features only rarely as one of the obscure causes of cutaneous thrombotic vasculopathy. We report a case of Nicolau syndrome developing secondary to subcutaneous injection of cyclizine to familiarize the clinicians and pathologists to this unusual condition. Given that it is potentially avoidable, pathologists should alert the clinicians to the possibility of Nicolau syndrome when a skin biopsy from an injection site shows signs of extensive thrombotic vasculopathy.

An analysis of Pseudomonas genomic diversity in take-all infected wheat fields reveals the lasting impact of wheat cultivars on the soil microbiota.

Manipulation of the soil microbiota associated with crop plants has huge promise for the control of crop pathogens. However, to fully realize this potential we need a better understanding of the relationship between the soil environment and the genes and phenotypes that enable microbes to colonize plants and contribute to biocontrol. A recent 2 years of investigation into the effect of wheat variety on second year crop yield in the context of take-all fungal infection presented the opportunity to examine soil microbiomes under closely defined field conditions. Amplicon sequencing of second year soil samples showed that Pseudomonas spp. were particularly affected by the wheat cultivar grown in year one. Consequently, 318 rhizosphere-associated Pseudomonas fluorescens strains were isolated and characterized across a variety of genetic and phenotypic traits. Again, the wheat variety grown in the first year of the study was shown to exert considerable selective pressure on both the extent and nature of Pseudomonas genomic diversity. Furthermore, multiple significant correlations were identified within the phenotypic/genetic structure of the Pseudomonas population, and between individual genotypes and the external wheat field environment. The approach outlined here has considerable future potential for our understanding of plant-microbe interactions, and for the broader analysis of complex microbial communities.

Effect of additional vitamin E and zinc supplementation on immunological changes in peripartum Sahiwal cows.

This study was conducted to exploit ameliorative effect of additional vitamin E and/or zinc supplementation on immune response of peripartum Sahiwal cows. Thirty-two pregnant dry Sahiwal cows were blocked into four treatment groups (n = 8), namely control, zinc (Zn), vitamin E (Vit E) and zinc + vitamin E (Zn + Vit E). Feeding regimen was same in all the groups except that the Sahiwal cows in the zinc-, vitamin E- and zinc + vitamin E-fed groups were additionally supplemented with 60 mg Zn/kg DM, 1000 IU vitamin E and 60 mg/kg + 1000 IU Zn + vitamin E, respectively, from day 60 pre-partum to day 90 post-partum. Blood samples were collected on days -60, -45, -30, -15, -7, -3, 0, 3, 7, 15, 30, 45, 60, 90 and 120 with respect to day of parturition and analysed for total immunoglobulin (TIG), immunoglobulin G (IgG), interleukin-2 (IL-2), vitamin E (Vit E) and zinc (Zn) status. Before calving, cows showed a decrease in plasma TIG, IgG, IL-2, Vit E and Zn levels. However, increased levels of plasma TIG, IgG, IL-2, Vit E and Zn were observed after calving. After calving, Sahiwal cows supplemented with Zn + Vit E had higher plasma TIG, IgG and IL-2 in comparison with cows of control and Zn + Vit E-fed groups. In the present study, plasma vitamin E level was higher in Vit E-fed and Zn + Vit E-fed cows; however, zinc level was higher in Zn- and Zn + Vit E-supplemented cows. In conclusion, a reduced immune response during peripartum period in Sahiwal cows was ameliorated by dietary vitamin E and zinc supplementation.

Population genetic structure of malaria vector Anopheles stephensi using mitochondrial cytochrome oxidase II gene in Indian populations.

The genetic differentiation in A. stephensi based on haplotype diversity using Restriction Fragment Length Polymorphism and bysequencing of CO II gene across different localities in India has been analyzed. The presence of only one DraI restriction site in CO II gene conferred to haplotype B indicating that the gene is very much conserved and the gene flow is not affected even by a major geographical distance barrier. The sequencing and analysisof various population parameters revealed seven haplotypes in all populations. The West Bengal population was found to be more genetically diverse than others. The geographic distance between populations was found to be contributing to the genetic differentiation. The sign of demographic expansion were found in three of the five populations. The local geographic barriers were found to be ineffective in prevention of gene flow.

[Evaluation of reference genes for quantitative real-time PCR normalization in cotton bollworm, Helicoverna armigera].

Reverse-transcription quantitative real-time PCR (RT-qPCR), a sensitive technique is being extensively employed in quantification of gene expression. However this requires normalization with suitable reference gene (RG) which is crucial in minimizing inter sample variations. Information regarding suitable RG is scarce in general and more so in insects, including the cotton bollworm, Helicoverpa armigera, an economically important pest. In management of this pest RNA interference (RNAi), is perceived as a potential tool, which is achieved by double-stranded RNA (dsRNA) delivery. These studies demand accurate quantification of gene silencing. In this study we assessed the suitability of five RGs viz. ß-actin (ACTB), 18S rRNA (18S), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ß-tubulin (TUB) and elongation fator-1-alfa (EF1-α) for gene expression studies in dsRNA treatment and across different developmental stages of H. armigera and ranked using geNorm, NormFinder and BestKeeper software programs. Data analysis revealed that best ranked RGs were varied in dsRNA treatment and in developmental stages. Under dsRNA treatment, 18S and GAPDH were more stable whereas, TUB and GAPDH were more stable across developmental stages. We also demonstrate that inappropriate selection of RG led to erroneous estimation of the target gene, chymotrypsin, expression. These results facilitate accurate quantification of gene expression in H. armigera.

A comparative evaluation of intranasal dexmedetomidine, midazolam and ketamine for their sedative and analgesic properties: a triple blind randomized study.

OBJECTIVES: To evaluate and compare the efficacy and safety of Intranasal (IN) Dexmedetomidine, Midazolam and Ketamine in producing moderate sedation among uncooperative pediatric dental patients. STUDY DESIGN: This randomized triple blind comparative study comprises of eighty four ASA grade I children of both sexes aged 4-14 years, who were uncooperative and could not be managed by conventional behavior management techniques. All the children were randomized to receive one of the four drug groups Dexmedetomidine 1 microg/ kg (D1), 1.5 microg/kg (D2), Midazolam 0.2 mg/kg (M1) and Ketamine 5 mg/kg (K1) through IN route. These drug groups were assessed for efficacy and safety by gauging overall success rate and by monitoring vital signs, respectively. RESULTS: The onset of sedation was significantly rapid with M1 and K1 as compared to D1 and D2 (p = < 0.001). The overall success rate was highest in D2 (85.7%) followed by D1 (81%), K1 (66.7%) and M1 (61.9%), however, the difference among them was not statistically significant (p = > 0.05). Even though all the vital signs were within physiological limits, there was significant reduction in pulse rate (PR) (p = < 0.001) and systolic blood pressure (SBP) (p = < 0.05) among D1 and D2 as compared to M1 and K1. D1, D2 and K1 produced greater intra- and post-operative analgesia as compared to M1. There were no significant adverse effects with any group. CONCLUSION: Dexmedetomidine, Midazolam and Ketamine, all the three drugs evaluated in the present study can be used safely and effectively through IN route in uncooperative pediatric dental patients for producing moderate sedation.

Isolated megalourethra: A rare case report and review of literature.

Megalourethra is a rare congenital disorder of anterior urethra and erectile tissue of penis. It mainly appears in two types-a milder scaphoid type and severe fusiform type. Fusiform type is commonly associated with congenital anomalies of various systems of the body. Isolated megalourethra without other congenital anomalies is extremely rare. We report one such case which was detected postnatally and successfully treated by reduction urethroplasty.

One-step DNA fragment assembly for expressing intron-containing hairpin RNA in plants for gene silencing.

Double-stranded RNA-mediated RNA interference in plants involves generating a construct expressing intron-containing hairpin RNA (ihpRNA), which usually is a cumbersome, multistep process. Here, we describe a simplified method involving single steps of PCR, restriction, ligation, and transformation for assembling an ihpRNA construct for plant transformation. Our method has several advantages over the currently available ones, viz., wider choice of restriction sites and facility for rapid screening of positive clones, among others. We demonstrate the utility of this approach in assembling the tomato phytoene desaturase gene. This simplified DNA fragment assembly strategy for ihpRNA construction facilitates high-throughput gene silencing in plants.

Effect of diet delivered various concentrations of double-stranded RNA in silencing a midgut and a non-midgut gene of Helicoverpa armigera.

Ribonucleic acid interference (RNAi) is a sequence-specific gene silencing mechanism induced by double-stranded RNA (dsRNA). Recently, RNAi has gained popularity as a reverse genetics tool owing to its tremendous potential in insect pest management, which includes Helicoverpa armigera. However, its efficiency is mainly governed by dsRNA concentration, frequency of application, target gene, etc. Therefore, to obtain a robust RNAi response in H. armigera, we evaluated various concentrations of dsRNA and its frequency of applications delivered through diet in silencing a midgut gene, chymotrypsin and a non-midgut gene, juvenile hormone acid methyl transferase (jhamt) of H. armigera. The extent of target gene silencing was determined by employing reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Our study revealed four significant findings: (i) single application of dsRNA elicited a delayed and transient silencing, while multiple applications resulted in early and persistent silencing of the above genes; (ii) silencing of the non-midgut gene (jhamt) through diet delivered dsRNA revealed prevalence of systemic silencing probably due to communication of silencing signals in this pest; (iii) the extent of silencing of chymotrypsin was positively correlated with dsRNA concentration and was negatively correlated with jhamt; (iv) interestingly, over-expression (15­18 folds) of an upstream gene, farnesyl diphosphate synthase (fpps), in juvenile hormone (JH) biosynthetic pathway at higher concentrations of jhamt dsRNA was the plausible reason for lesser silencing of jhamt. This study provides an insight into RNAi response of target genes, which is essential for RNAi design and implementation as a pest management strategy.

Effect of vitamin e and zinc supplementation on energy metabolites, lipid peroxidation, and milk production in peripartum sahiwal cows.

The study was conducted to evaluate the effect of vitamin E and zinc supplementation on energy metabolites, lipid peroxidation, and milk production in peripartum Sahiwal cows. For this, thirty-two pregnant dry Sahiwal cows were selected at sixty days prepartum and divided into four groups viz control, T1, T2, and T3 of eight each. Group T1 were supplemented with zinc at 60 ppm/d/cow, group T2 were supplemented with vitamin E at 1,000 IU/d/cow and group T3 were supplemented with combination of vitamin E at 1,000 IU/d/cow and zinc at 60 ppm/d/cow during d 60 prepartum to d 90 postpartum. Blood samples were collected on d -60, -45, -30, -15, -7, -3, 0, 3, 7, 15, 30, 45, 60, 90, and 120 with respect to day of parturition and analysed for glucose, non esterified fatty acid, and thiobarbituric acid reactive substance. Body condition score was maintained significantly better (p<0.05) in T3 than in the control, T1 and T2 groups. Overall glucose level was higher (p<0.05) in T3 than control, T1, and T2 groups. Levels of nonesterified fatty acid, and thiobarbituric acid reactive substance were lower (p<0.05) in T3 than control, T1, and T2 groups. Milk yield was higher (p<0.05) in T3 than control, T1, and T2 groups. In conclusion, the present study indicated that the supplementation of vitamin E and zinc in peripartum Sahiwal cows enhanced milk production by reducing negative energy balance.

Modified competing polymerase chain reaction primer for single tube quantitative PCR.

We developed a modified polymerase chain reaction (PCR) primer with 3' phosphate instead of hydroxyl group for single-tube accurate transcript quantification. 18S ribosomal RNA (rRNA) reference gene-specific modified primer was used for precise single-tube quantification of two target transcripts, namely chymotrypsin and jhamt (juvenile hormone acid methyl transferase) of Helicoverpa armigera. A comparative study of 3' phosphorylated primers, 3' mismatched primers, and commercial Competimers revealed that 3' phosphorylation was more efficient than the 3' mismatch and was on par with Competimers in blocking the primer extension. Thus, the modified primers can be used in single-tube, economical, and accurate PCR quantification of the target gene using any assay-specific reference gene.

Association of promoter methylation status of NRF2 and PNPLA3 genes in alcoholic liver disease.

BACKGROUND: Alcoholic liver disease (ALD) is the leading cause of chronic liver disease. In the liver, metabolism of alcohol occurs through multiple mechanisms and it results in the generation of various toxic products. Multiple genetic causes have been identified that are associated with the development and progression of ALD. The present study assessed the promoter site methylation status of nuclear factor erythroid 2-related factor 2 (NRF2) and patatin-like phospholipase domain-containing protein-3 (PNPLA3) genes in different subgroups of ALD. METHODS: The patients recruited were cases of alcohol dependence syndrome with hepatic dysfunction, compensated cirrhosis, decompensated cirrhosis, and acute-on-chronic liver failure due to alcohol as an etiology along with healthy control subjects. Routine biochemical investigations were performed along with methylation-specific polymerase chain reaction (MS-PCR) to qualitatively assess the promoter methylation status of NRF2 and PNPLA3 in all these cases. RESULTS: There was significant difference in methylation status of NRF2 gene in ALD when compared to healthy controls but there was no such difference in PNPLA3. All biochemical and clinical parameters studied were significantly different in subgroups of ALD except the serum aspartate aminotransferase (AST) level. Subgroups of ALD did not show any significant association with NRF2 or PNPLA3 methylation status. Gamma-glutamyl transferase (GGT) and creatinine levels in serum were significantly associated with the methylation status of NRF2 gene while no such association was seen with PNPLA3 gene. Model for end-stage liver disease (MELD) score varied differentially with NRF2 methylation and PNPLA3 methylation but there was no statistical significance. CONCLUSIONS: The present study showed that methylation status of NRF2 and PNPLA3 genes could not differentiate between subgroups of alcoholic liver diseases. However, the unmethylation of NRF2 promoter is associated with higher serum levels of GGT.

A transposon insertion single-gene knockout library and new ordered cosmid library for the model organism Streptomyces coelicolor A3(2).

A simple and high-throughput transposon mediated mutagenesis system employing in vitro shuttle transposon mutagenesis has been used to systematically mutagenise the Streptomyces coelicolor genome. To achieve the highest coverage, a new ordered cosmid library was also constructed. Individual cosmids from both the existing and new libraries were disrupted using the Tn5-based mini-transposon Tn5062. A total of 35,358 insertions were sequenced resulting in the disruption of 6,482 genes (83% of the predicted open reading frames). Complete information for both the newly generated cosmids as well as all the insertions has been uploaded onto a central database, StrepDB ( http://strepdb.streptomyces.org.uk/ ). All insertions, new cosmids and a range of transposon exchange cassettes are available for study of individual gene function.

First report on embryonic and larval development of 2n/3n mosaic sterlet.

Mosaicism is frequently observed in aquaculture practices, and it adversely affects the production as well as the restoration programme of the sturgeon. The purpose of the present study was the induction of 2n/3n mosaic in sterlet, Acipenser ruthenus L., and compare their embryonic and larval development with diploid control sterlet. Microsatellite DNA loci genotyping was conducted for the identification of the genotypes and parentage analysis. Embryonic development was monitored in experimental groups at every 24 h interval. Identification of individual stages of embryonic development was recorded based on a 36-degree scale of development. Additionally, the BW and body length (LT) of experimental fishes were taken during 110 days of the rearing period. The Fulton's condition coefficient (F), length-weight parameters, and specific growth rate (SGR) coefficient were calculated. The analysis of embryonic development of the 2n/3n mosaic and the diploid control group did not show differences. However, higher mortality (88%) was observed in 2n/3n mosaic groups in comparison to the diploid control groups (55%). BW and body length of 2n/3n mosaic sterlet were slightly lower than the diploid control sterlet, but the differences were not statistically significant. F analysis did not confirm a lower growth performance of the fishes in the 2n/3n mosaic group. Microsatellite DNA loci genotyping confirmed both the incidence of polyspermy and retention of the second polar body. This paper presents the first report on embryonic development and growth performance of 2n/3n mosaic sturgeons.

Bile acids: natural ligands for an orphan nuclear receptor.

Bile acids regulate the transcription of genes that control cholesterol homeostasis through molecular mechanisms that are poorly understood. Physiological concentrations of free and conjugated chenodeoxycholic acid, lithocholic acid, and deoxycholic acid activated the farnesoid X receptor (FXR; NR1H4), an orphan nuclear receptor. As ligands, these bile acids and their conjugates modulated interaction of FXR with a peptide derived from steroid receptor coactivator 1. These results provide evidence for a nuclear bile acid signaling pathway that may regulate cholesterol homeostasis.

Reinforcing mitochondrial functions in aging brain: An insight into Parkinson's disease therapeutics.

Mitochondria, the powerhouse of the neural cells in the brain, are also the seat of certain essential gene signaling pathways that control neuronal functions. Deterioration of mitochondrial functions has been widely reported in normal aging as well as in a spectrum of age-associated neurological diseases, including Parkinson's disease (PD). Evidences accumulated in the recent past provide not only advanced information on the causes of mitochondrial bioenergetics defects and redox imbalance in PD brains, but also much insight into mitochondrial biogenesis, quality control of mitochondrial proteins, and genes, which regulate intra- and extra-mitochondrial signaling that control the general health of neural cells. The mitochondrial quality control machinery is affected in aging and especially in PD, thus affecting intraneuronal protein transport and degradation, which are primarily responsible for accumulation of misfolded proteins and mitochondrial damage in sporadic as well as familial PD. Essentially we considered in the first half of this review, mitochondria-based targets such as mitochondrial oxidative stress and mitochondrial quality control pathways in PD, relevance of mitochondrial DNA mutations, mitophagy, mitochondrial proteases, mitochondrial flux, and finally mitochondria-based therapies possible for PD. Therapeutic aspects are considered in the later half and mitochondria-targeted antioxidant therapy, mitophagy enhancers, mitochondrial biogenesis boasters, mitochondrial dynamics modulators, and gene-based therapeutic approaches are discussed. The present review is a critical assessment of this information to distinguish some exemplary mitochondrial therapeutic targets, and provides a utilitarian perception of some avenues for therapeutic designs on identified mitochondrial targets for PD, a very incapacitating disorder of the geriatric population, world over.

Drug-Drug Interaction Studies of Methadone and Antiviral Drugs: Lessons Learned.

Different views appear in the literature on the extent of specific cytochrome P450 (CYP) involvement in methadone metabolism. The aim of this work is to leverage knowledge from drug-drug interaction (DDI) studies in new drug applications between methadone and antiviral medications to better understand methadone disposition and to inform design of future DDI studies with methadone. A database of DDI studies between all FDA-approved human immunodeficiency virus and hepatitis C virus medications and methadone was constructed. The database contains data from 29 DDI studies. Sixteen of the 29 studies had statistically significant changes in methadone area under the concentration-time curve. Methadone exposure was either decreased or unchanged when it was coadministered with weak to strong CYP3A inhibitors or a moderate CYP3A4 inducer. Methadone exposure was reduced when it was coadministered with CYP2B6 inducers. The role of other enzymes (CYP2C9, CYP2C19, and CYP2D6) cannot be fully elucidated from these studies. In conclusion, CYP2B6 plays a prominent role in methadone metabolism, although methadone exposure is not sensitive to CYP3A perturbation. In designing methadone DDI studies, (1) measuring R- and S-methadone is more informative than measuring total methadone, and (2) CYP2B6 genotyping of subjects enrolled in methadone DDI studies should be considered. Finally, there is a need for the development of predictive models to determine the influence of medications on methadone disposition.

Efficacy of Low-level Laser Therapy, Hyaluronic Acid Gel, and Herbal Gel as Adjunctive Tools in Gingivectomy Wound Healing: A Randomized Comparative Clinical and Histological Study.

Introduction Gingival overgrowth is usually an inflammatory response to plaque present on tooth surfaces. The other causes could be drugs and other systemic conditions. When the local factors are responsible and subgingival scaling does not help, gingivectomy is performed. The gingivectomy wound is raw and heals slowly. Low-level laser therapy (LLLT), hyaluronic acid, and herbal gels aid in healing after a gingivectomy. This study compared the efficacy of LLLT, hyaluronic acid, and herbal gel when used topically after a gingivectomy. This was a single-arm, interventional trial wherein 30 patients aged between 18 and 40 years with moderate gingival overgrowth participated. The study was approved by the institutional ethical committee. (DN/0109-16). The participants signed the consent form and the study was also registered (NCT03569683). Materials and methods The samples were equally divided into three groups. Group A received LLLT immediately postop, and on the first, third, and seventh days after surgery. Group B received hyaluronic acid (Gengigel) while Group C received an herbal gel (Hiora SG) for the same time periods after surgery, respectively. Analysis of variance (ANOVA) followed by a post-hoc Bonferroni test was used to evaluate differences within groups. Intergroup comparison was performed using the independent t-test. A p-value of <0.05 was considered statistically significant. Results The plaque index (PI), gingival index (GI), and gingival enlargement index (GEI) showed good improvement postoperatively within the groups, which was statistically significant. However, on an intergroup comparison, the GEI pertaining only to the laser group showed significant changes. Also, the pain perception analyzed by the visual analog score (VAS) was the least, and histologically, the amount of mature collagen fibers laid down were more in the laser group. Conclusion Patients irradiated with laser after gingivectomy (Group A) showed superior results in the clinical, histological variables as compared to Groups B and C.

Biocontrol efficiency of odonate nymphs against larvae of the mosquito, Culex quinquefasciatus Say, 1823.

An estimation of the predatory efficiency of the nymphs of five coexisting odonate species Aeshna flavifrons, Coenagrion kashmirum, Ischnura forcipata, Rhinocypha ignipennis and Sympetrum durum using the fourth-instar larvae of Culex quinquefasciatus as prey, was made under laboratory and semi-field conditions. The daily feeding rate varied among the odonate species, at laboratory conditions. The mean number of IV instars Cx. quinquefasciatus larvae killed per day, ranged between 14 and 64 (64 mosquito larvae for I. forcipata, 57 for A. flavifrons, 45 for R. ignipennis, 25 for S. durum and 14 for C. kashmirum). The prey consumption was linearly related to the number of predators and prey available but inversely related with space. It was also noted that the feeding rates varied significantly between dark and light conditions, in all the odonate species. The presence of nymphs in semi-field conditions significantly lowered the mosquito larval density in dipper samples after 15 days from the introduction, followed by a significant increase of larval mosquito density after 15 days from the withdrawal of the nymphs. The results of the present observations are suggestive of the use of odonate nymphs in temporary pools or larger habitats where they can be a potential biological resource in regulating the larval population of the vector and pest mosquitoes.

Age composition of incriminated malaria vector in a rural foothills in West Bengal, India.

Two 1 yr surveys carried out at a gap of 10 yr revealed nine anopheline species from malaria endemic foothills area of Ayodhya - Baghmindi range of West Bengal, India, with 8.4 per cent populations of Anopheles culicifacies. An. culicifacies was incriminated as vector of Plasmodium falciparum with sporozoite rate of 1.23 per cent. Studies on age composition indicated that proportion parous and daily survival rate of An. culicifacies were assessed to be 0.50 and 0.84 respectively. The survival rate per gonotrophic cycle averaged over the two year was 0.61. Rainy season was found to be the favourable period for transmission.