The overall quality of evidence of recommendations surrounding nutrition and diet in inflammatory bowel disease.

BACKGROUND AND AIM: Recently, there has been an increased focus on the role nutrition and diet play in maintaining health in inflammatory bowel disease (IBD). We aimed to assess the overall quality, strength, and transparency of conflicts among guidelines on nutrition/diet in IBD. METHODS: A systematic search was performed on multiple databases from inception until January 1, 2021, to identify guidelines pertaining to nutrition or diet in IBD. All guidelines were reviewed for disclosure of conflicts of interest (COI) and funding, recommendation quality and strength, external document review, patient representation, and plans for update-as per Institute of Medicine (IOM) standards. In addition, recommendations and their quality were compared between guidelines/societies.​ RESULTS: Seventeen distinct societies and a total of 228 recommendations were included. Not all guidelines provided recommendations on key aspects of diet-such as the role of supplements or the appropriate micro/macro nutrition in IBD. Fifty-nine percent of guidelines reported on COI, 24% underwent external review, and 41% included patient representation. 18.4%, 25.9%, and 55.7% of recommendations were based on high-, moderate-, and low-quality evidence, respectively. 10.5%, 24.6%, and 64.9% of recommendations were strong, weak/conditional, and did not provide a strength, respectively. The proportion of high-quality evidence (p = 0.12) and strong recommendations (p = 0.83) did not significantly differ across societies. CONCLUSIONS: Many guidelines do not provide recommendations on key aspects of diet/nutrition in IBD. As over 50% of recommendations are based on low-quality evidence, further studies on nutrition/diet in IBD are warranted to improve the overall quality of evidence.

Molecular docking study on vitamin D supplements to understand their interaction with VDR-RXRα heterodimer and VDRE of TAGAP gene.

The expression level of T cell activation Rho GTPase activating protein (TAGAP) gene is higher in rheumatoid arthritis (RA) patients compared to healthy individuals. Vitamin D receptor element (VDRE) sequences present in the regulatory region of TAGAP gene are targeted by vitamin D dependent Vitamin D receptor (VDR) - retinoic acid X receptor (RXR) heterodimer complex to regulate the TAGAP gene expression. Reduction in the expression of the TAGAP gene can prevent different severity of RA disease conditions. Calcitriol is a proven vitamin D supplement prescribed to patients with RA. However, it is involved in causing hypercalcemia. Maxacalcitol, an analog of vitamin D, is shown to have less hypercalcemic activity when compared to calcitriol. This study was done to analyze and compare the binding modes of calcitriol and maxacalcitol with VDR. We also studied the interactions of these compounds with the VDR-RXRα heterodimer complex. In addition, the binding of the ligand-activated heterodimer complexes with VDREs of the TAGAP gene was also analyzed to comprehend the binding affinities of calcitriol and maxacalcitol to the gene. The current work utilizes in silico molecular docking and simulation analysis to understand the mechanism in each complex formation.Communicated by Ramaswamy H. Sarma.

An attempt to unravel the association of TAGAP gene SNPs with rheumatoid arthritis in the Indian population using high-resolution melting analysis.

Rheumatoid arthritis (RA) is an inflammatory disease that causes inflammation of the synovium, cartilage, and deformity of the bones. Single nucleotide polymorphism (SNPs) at 5'UTR rs1738074 (A/G) and a novel candidate non-synonymous single nucleotide polymorphism (nsSNP) rs759674898 (G/A) of TAGAP gene were studied for its association with RA in the Indian population. Real-time PCR coupled with High Resolution Melting analysis technique was employed to detect SNPs. The resulting outcomes were confirmed using the "traditional" Sanger's sequencing method. From this study, we identified that rs1738074 SNP was associated with RA. The odds ratio (OR) obtained for the AG genotype was 3.3379 (Confidence Interval (C.I) 1.7881 to 6.2350); for AA genotype 0.5510 (C.I 0.3043 to 0.9979) and GG genotype 0.5609 (CI 0.3062 to 1.0275). The study also confirmed that AG heterozygous condition had more significant association with RA than AA and GG genotypes. The obtained relative risk (RR) for the AA genotype was 0.676; for AG genotype (RR = 2.253) and GG genotype (RR = 0.6741). The novel candidate nsSNP rs759674898 had only the G allele, and the A allele was not detected in the population studied. In conclusion, this study emphasizes that the rs1738074 SNP in the TAGAP gene's 5'UTR is substantially linked to RA in the Indian population.

Secophragmalin-Type Limonoids from Trichilia connaroides: Isolation, Semisynthesis, and Their Cytotoxic Activities.

Two new secophragmalin-type limonoids, secotrichagmalins B (1) and C (2) along with two known compounds, were isolated from the fruits of Trichilia connaroides. The structures of the new compounds were elucidated by analysis of spectroscopic (IR, MS, and 2D NMR) data and single crystal X-ray diffraction studies. In addition, semisynthetic derivatives (2a-2l) were efficiently synthesized and evaluated for their in vitro cytotoxicity along with the isolated limonoids against a panel of human cancer cell lines. The results indicated that new analogues 2a, 2d, and 2e showed cytotoxicity on the DU145 cell line with IC50 values of 3.6, 4.2, and 5.2 µM, respectively. Flow cytometric analysis revealed that these analogues arrested the cell cycle in the G0/G1 phase and markedly induced apoptosis.

Distinguishing Specific and Nonspecific Complexes of Alkyladenine DNA Glycosylase.

Human alkyladenine DNA glycosylase (AAG) recognizes many alkylated and deaminated purine lesions and excises them to initiate the base excision DNA repair pathway. AAG employs facilitated diffusion to rapidly scan nonspecific sites and locate rare sites of damage. Nonspecific DNA binding interactions are critical to the efficiency of this search for damage, but little is known about the binding footprint or the affinity of AAG for nonspecific sites. We used biochemical and biophysical approaches to characterize the binding of AAG to both undamaged and damaged DNA. Although fluorescence anisotropy is routinely used to study DNA binding, we found unexpected complexities in the data for binding of AAG to DNA. Systematic comparison of different fluorescent labels and different lengths of DNA allowed binding models to be distinguished and demonstrated that AAG can bind with high affinity and high density to nonspecific DNA. Fluorescein-labeled DNA gave the most complex behavior but also showed the greatest potential to distinguish specific and nonspecific binding modes. We suggest a unified model that is expected to apply to many DNA binding proteins that exhibit affinity for nonspecific DNA. Although AAG strongly prefers to excise lesions from duplex DNA, nonspecific binding is comparable for single- and double-stranded nonspecific sites. The electrostatically driven binding of AAG to small DNA sites (∼5 nucleotides of single-stranded and ∼6 base pairs of duplex) facilitates the search for DNA damage in chromosomal DNA, which is bound by nucleosomes and other proteins.

Development of a Genetic Map for Onion (Allium cepa L.) Using Reference-Free Genotyping-by-Sequencing and SNP Assays.

Single nucleotide polymorphisms (SNPs) play important roles as molecular markers in plant genomics and breeding studies. Although onion (Allium cepa L.) is an important crop globally, relatively few molecular marker resources have been reported due to its large genome and high heterozygosity. Genotyping-by-sequencing (GBS) offers a greater degree of complexity reduction followed by concurrent SNP discovery and genotyping for species with complex genomes. In this study, GBS was employed for SNP mining in onion, which currently lacks a reference genome. A segregating F2 population, derived from a cross between 'NW-001' and 'NW-002,' as well as multiple parental lines were used for GBS analysis. A total of 56.15 Gbp of raw sequence data were generated and 1,851,428 SNPs were identified from the de novo assembled contigs. Stringent filtering resulted in 10,091 high-fidelity SNP markers. Robust SNPs that satisfied the segregation ratio criteria and with even distribution in the mapping population were used to construct an onion genetic map. The final map contained eight linkage groups and spanned a genetic length of 1,383 centiMorgans (cM), with an average marker interval of 8.08 cM. These robust SNPs were further analyzed using the high-throughput Fluidigm platform for marker validation. This is the first study in onion to develop genome-wide SNPs using GBS. The resulting SNP markers and developed linkage map will be valuable tools for genetic mapping of important agronomic traits and marker-assisted selection in onion breeding programs.

Comprehensive strategy for the design of precision drugs and identification of genetic signature behind proneness of the disease-a pharmacogenomic approach.

The proneness of diseases and susceptibility towards drugs vary from person to person. At present, there is a strong demand for the personalization of drugs. The genetic signature behind proneness of the disease has been studied through a comprehensive 'octopodial approach'. All the genetic variants included in the approach have been introduced. The breast cancer associated with BRCA1 mutation has been taken as the illustrative example to introduce all these factors. The genetic variants associated with the drug action of tamoxifen have been fully illustrated in the manuscript. The design of a new personalized anti-breast cancer drug has been explained in the third phase. For the design of new personalized drugs, a metabolite of anti-cancer drug chlorambucil has been taken as the template. The design of drug has been made with respect to the protein 1T15 of BRCA1 gene corresponding to the genetic signature of rs28897696.

Industry-Led Access-To-Medicines Initiatives In Low- And Middle-Income Countries: Strategies And Evidence.

Global biopharmaceutical companies are increasingly establishing access-to-medicines (AtM) initiatives in low- and middle-income countries. We reviewed the initiatives of twenty-one research-based global biopharmaceutical companies to assess their strategies for improving access and the quality of evidence on the impact of their initiatives. The number of operating initiatives increased from 17 in 2000 to 102 in 2015. Of the 120 different AtM initiatives identified, 48 percent used a medicine donation strategy, and 44 percent used a price reduction strategy. While companies have frequently claimed that their initiatives have had positive impacts, we found published evaluations for only seven initiatives, and nearly all of the evaluations were of low (62 percent) or very low (32 percent) quality. It is clear that the biopharmaceutical industry has increased its commitment to improving access to medicines in low- and middle-income countries. However, companies should do more to generate high-quality evidence on their initiatives, and the global health community should do more to assist the developing of evidence about the initiatives.

Concurrent production of cellulase and xylanase from Trichoderma reesei NCIM 1186: enhancement of production by desirability-based multi-objective method.

Application of multiple response optimizations using desirability function in the production of microbial metabolites improves economy and efficiency. Concurrent production of cellulase and xylanase in Trichoderma reesei NCIM 1186 using an agricultural weed, Prosopis juliflora pods, was studied. The main aim of the study was to optimize significant medium nutrient parameters for maximization of cellulase and xylanase by multi-objective optimization strategy using biomass. Process parameters such as the nutrient concentrations (pods, sucrose, and yeast extract) and pH were investigated to improve cellulase and xylanase activities by one factor at a time approach, single response optimization and multi-objective optimization. At the corresponding optimized process parameters in single response optimization, the maximum cellulase activity observed was 3055.65 U/L where xylanase highest activity was 422.16 U/L. Similarly, the maximum xylanase activity, 444.94 U/L, was observed with the highest cellulase activity of 2804.40 U/L. The multi-objective optimization finds a tradeoff between the two objectives and optimal activity values in between the single-objective optima were achieved, 3033.74 and 439.13 U/L for cellulase and xylanase, respectively.

Withanolides against TLR4-Activated Innate Inflammatory Signalling Pathways: A Comparative Computational and Experimental Study.

Innate inflammations are dominant causes of poor health and high mortality. The pathogen-associated molecular pattern and lipopolysaccharide (LPS) are sensed by immune cells through activation of toll-like receptor 4 leading to mitogen-activated protein kinases (MAPKs) and NF-κB activations. Controlled MAPK and Nf-κB inhibitors have been proposed as potential antiinflammatory drugs. Withania somnifera is an important medicinal herb with known antiinflammatory activity. In this study, the selected Withania somnifera extracts and withanolides were analysed on LPS-induced macrophages comparatively. Molecular docking analysis revealed withaferin A, withanone and withanolide A as effective withanolides against inflammatory target molecules. In experiments, withaferin A and withanone treatment had prominent suppressions on LPS-induced expression of pro-inflammatory cytokines in bone marrow-derived macrophages. Withaferin A regulated all the major four pathways (MAPKs and NF-κB) involved in innate inflammations. Similarly among the Withania extracts analysed, the in vitro propagated leaf and field grown root extracts containing high withaferin A content suppressed the inflammatory molecules through NF-κB and MAPK pathways. Withaferin A was found to be best in suppressing the activated inflammatory pathways among all the analysed withanolides. Therefore, withaferin A and extracts with high withaferin A content can be used as promising drug candidates against innate inflammations. Copyright © 2016 John Wiley & Sons, Ltd.

Immobilization of levan-xylanase nanohybrid on an alginate bead improves xylanase stability at wide pH and temperature.

Despite the sustainable availability, levan, a fructose based natural polysaccharide has not received significant attention in the development of enzyme immobilization technology. Herein, we prepared levan-xylanase (LXy) nanohybrid and characterized by scanning electron microscopy, particle size analyzer and zeta potential. To prevent the enzyme leakage from the nanohybrid, LXy was immobilized onto an alginate beads (NaAlg). Immobilization yield was optimized using a statistical method, central composite design. A maximum immobilization yield of 95.3% was achieved at 2.13% (w/v) of sodium alginate, 2.14% (w/v) of calcium chloride, 64min of curation time and 1.4mm bead size. Immobilized LXy retains nearly 80% of the enzyme activity at a wide range of temperature (20-90°C) and pH (3-10). Immobilization of LXy onto NaAlg increases the activation energy from 28.50Jmol-1K-1 to 39.38Jmol-1K-1. Collectively, this result implies that LXy immobilized onto NaAlg increases the enzyme stability and retains its activity.

Minimally Invasive Surgery in Patients With Adolescent Idiopathic Scoliosis: Is it Better than the Standard Approach? A 2-Year Follow-up Study.

STUDY DESIGN: This is a retrospective controlled study. OBJECTIVE: To compare the safety and efficacy of minimally invasive surgery (MIS) for the surgical management of adolescent idiopathic scoliosis (AIS) to the standard open posterior approach (PSF). SUMMARY OF BACKGROUND DATA: MIS approaches offer the potential to reduce soft-tissue trauma, intraoperative blood loss, and surgical-site infection. Thus far, MIS has been successfully utilized for the surgical correction of multilevel spine pathology in adults. It is not yet known if these results can be replicated in the surgical management of AIS. MATERIALS AND METHODS: Seven MIS patients were compared with 15 PSF patients using minimum 2-year follow-up data. Parameters studied included preoperative patient and deformity characteristics, perioperative details, extent of deformity correction, and complications. Data were compared using Mann-Whitney tests for continuous variables and Fisher exact tests for categorical variables. RESULTS: The MIS and PSF groups were similar for all preoperative characteristics collected (P>0.05). MIS patients had fewer fixation points (P=0.015), but a longer median operative time (P=0.011). There was no significant difference in estimated blood loss (EBL) (P=0.051), EBL/fixation point (P=0.204), or amount of fluids administered (P=0.888). Postoperative recovery did not differ between the 2 groups in number of intensive care unit days (P=0.362), length of hospital stay (P=0.472), time to mobilization (P=1.00), Visual Analogue Scale pain scores (P=0.698), or patient-controlled analgesia (P=1.00). The MIS technique had similar deformity correction, screw placement accuracy, and fusion status when compared with the PSF group. MIS patients had lower blood transfusion rate (P=0.02), shorter fusion (P=0.046) and fewer pedicle screws (P=0.015). CONCLUSIONS: The short-term advantages seen in MIS for adult scoliosis were not as obvious in our series. We found similar deformity correction and adequate fusion, however shortcomings related to learning curve, and instrumentation persist. MIS surgery is an innovative treatment for AIS that is technically feasible with significantly lower transfusion rate, shorter fusion lenghts and lesser pedicle screw fixation. Despite these advanatges, its role in AIS is currently difficut to define.

Design, Synthesis, and Evaluation of Novel 1,2,3-Triazole-Tethered Glycolipids as Vaccine Adjuvants.

A Cu-mediated azide-alkyne click chemistry protocol was employed for the synthesis of a focused library of novel 1,2,3-triazolyl conjugates bearing various carbohydrate-steroid/triterpenoid entities. The immunogenicity of these compounds was examined initially by ex vivo assays. The lead compound 15g was further subjected to in vivo evaluation in BALB/c mice immunized with ovalbumin. These in vivo biological studies revealed an increase in B cell-mediated proliferation, higher expression levels of IL-2, TNF-α, IL-12, and IFN-γ indicating Th1 activation, together with an enhanced OVA-specific antibody (IgG) response compared to alum, affirming adjuvanticity of these glycolipids. The primary indications of response skewed toward Th1 immunity induced by the new triazoyl analogs indicate the potential of these molecules for possible application as adjuvants.

CT-Based Anatomical Evaluation of Pre-Vertebral Structures With Respect to Vertebral Body Using a Clock-Face Analogy.

STUDY DESIGN: Retrospective Chart and CT Scan Review. OBJECTIVE: To define the relationship of the pre-vertebral structures for each level to assist in easier intraoperative visualization. SUMMARY OF BACKGROUND DATA: Vascular and visceral injuries from pedicle screws are well-known. This study will define the relationship of the pre-vertebral structures for each level to assist in avoiding potential complications. METHODS: Pre- and post-operative CT scans were reviewed to define the pre-vertebral structures in relation to a clock-face. On reformatted axial slices, a clock-face was superimposed so that the left transverse process (TP) represented 8 o'clock and the right TP represented 4 o'clock. The positions of the TP on the clock-face did not change with rotation of the vertebra. RESULTS: 108 patients had pre-operative CT scans. 78 had post-operative CT scans. Median age was 15 years, median Cobb angle was 50°, fused were 12, with 21 fixation points. 6324 axial CT slices were reformatted and analyzed. The trachea was located at 12 o'clock at T1, 1 o'clock at T2-T4, and between 12 and 1 o'clock at T5. The esophagus starts as a midline structure at 12 o'clock from T1-T2, moves to 11 o'clock from T3-T6, and further to 10 o'clock from T7-T9. The aorta starts at 10 o'clock at T5-T6, moves left at T7-T8 to 9 o'clock, and returns to 10 o'clock from T9-T11. It appears at 11'clock at T12, and at 12 o'clock from L1-L4. In about a third of cases, it is at 1 o'clock from L1 to L4, where it bifurcates. CONCLUSIONS: This CT-based anatomical study provides a simple reference frame to help surgeons visualize the vital structures at each level. This three-dimensional visualization is facilitated by fixing the position of TP on the clock-face. Knowledge of this anatomical relationship can help avoid direct injury, and is easier to recall intra-operatively. LEVEL OF EVIDENCE: 3.

Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis.

BACKGROUND: The production of metabolites via in vitro culture is promoted by the availability of fully defined metabolic pathways. Withanolides, the major bioactive phytochemicals of Withania somnifera, have been well studied for their pharmacological activities. However, only a few attempts have been made to identify key candidate genes involved in withanolide biosynthesis. Understanding the steps involved in withanolide biosynthesis is essential for metabolic engineering of this plant to increase withanolide production. RESULTS: Transcriptome sequencing was performed on in vitro adventitious root and leaf tissues using the Illumina platform. We obtained a total of 177,156 assembled transcripts with an average unigene length of 1,033 bp. About 13% of the transcripts were unique to in vitro adventitious roots but no unique transcripts were observed in in vitro-grown leaves. A putative withanolide biosynthetic pathway was deduced by mapping the assembled transcripts to the KEGG database, and the expression of candidate withanolide biosynthesis genes -were validated by qRT PCR. The accumulation pattern of withaferin A and withanolide A varied according to the type of tissue and the culture period. Further, we demonstrated that in vitro leaf extracts exhibit anticancer activity against human gastric adenocarcinoma cell lines at sub G1 phase. CONCLUSIONS: We report here a validated large-scale transcriptome data set and the potential biological activity of in vitro cultures of W. somnifera. This study provides important information to enhance tissue-specific expression and accumulation of secondary metabolites, paving the way for industrialization of in vitro cultures of W. somnifera.

Pedicle screws adjacent to the great vessels or viscera: a study of 2132 pedicle screws in pediatric spine deformity.

STUDY DESIGN: A retrospective study. OBJECTIVE: To determine the incidence of pedicle screws close to vital structures and to identify patient or curve characteristics that increase the risk of screw misplacement. SUMMARY AND BACKGROUND: Most pedicle screw misplacements are asymptomatic, thus they are frequently undetected. This study identifies the rate of screw placement in proximity to vital structures using postoperative computed tomography scans. METHODS: A total of 2132 screws in 101 patients, who underwent posterior spinal fusion for spinal deformity, were reviewed. Screws adjacent to great vessels and viscera were identified and evaluated. Patients with screws at risk (group B) were compared with patients without screws at risk (group A). Patient and curve characteristics were analyzed to determine whether a correlation with screw misplacement exists. RESULTS: A total of 40 at risk screws (∼2%) were identified in 25 patients (∼25%). These 40 screws were in proximity to the aorta (31), left subclavian artery (1), esophagus (3), trachea (3), pleura (1), and diaphragm (1). Of the 31 screws close to the aorta, 10 screws in 6 patients were impinging or distorting the aortic wall. One hundred percent of misplaced screws were in the thoracic spine, 50% were misplaced laterally, 50% were 35 mm long, 57.5% were in pedicles with normal morphology, and 75% were in curves between 40 and 70 degrees. Median screw misplacement rate was 10% in group A and 13% in group B. Adjusted for age and preoperative Cobb angle, patients with a higher misplacement rate were more likely to have screws adjacent to vital organs [adjusted odds ratio: 1.06 (95% confidence interval, 1.01-1.13), P=0.033]. CONCLUSIONS: Although only a small number of screws were at risk, they occurred in a large percentage of patients (25%). A single at-risk screw causes a significant complication for the patient. Postoperative imaging beyond routine x-rays may be needed to detect at-risk screws in asymptomatic patients.

Testosterone and insulin resistance in the metabolic syndrome and T2DM in men.

Obesity, type 2 diabetes mellitus and the metabolic syndrome are major risk factors for cardiovascular disease. Studies have demonstrated an association between low levels of testosterone and the above insulin-resistant states, with a prevalence of hypogonadism of up to 50% in men with type 2 diabetes mellitus. Low levels of testosterone are also associated with an increased risk of all-cause and cardiovascular mortality. Hypogonadism and obesity share a bidirectional relationship as a result of the complex interplay between adipocytokines, proinflammatory cytokines and hypothalamic hormones that control the pituitary-testicular axis. Interventional studies have shown beneficial effects of testosterone on components of the metabolic syndrome, type 2 diabetes mellitus and other cardiovascular risk factors, including insulin resistance and high levels of cholesterol. Biochemical evidence indicates that testosterone is involved in promoting glucose utilization by stimulating glucose uptake, glycolysis and mitochondrial oxidative phosphorylation. Testosterone is also involved in lipid homeostasis in major insulin-responsive target tissues, such as liver, adipose tissue and skeletal muscle.

Invivo comparative evaluation of tertiary dentin deposit to three different luting cements a histopathological study.

The aim of the study is to evaluate histopathologically the amount of tertiary dentin deposit stimulated by three different luting cements. With the informed consent for fifteen patients crown preparation was done for maxillary and mandibular premolar teeth which were scheduled for orthodontic extraction. Copings were cemented with three different luting cements zinc oxide eugenol, glass ionomer and zinc polycarboxylate which were classified as Groups A, B and C respectively. The teeth were later extracted and histopathologically analysed for pulpodentinal reactions using a control study group. Statistically Tukey-HSD procedure was used to identify the significant group and one way ANNOVA was used to analyse the thickness of tertiary dentin among the study group. Tertiary dentin was seen in most of the specimens. When the three groups were compared zinc oxide eugenol helps in stimulation of tertiary dentin formation.

Eventration of the diaphragm and associations.

Eventration of the diaphragm, most often an isolated entity and detected incidentally, has been known to be associated with several genetic syndromes. Authors report their experience of seeing diaphragmatic eventration in association with Poland syndrome and wandering spleen syndrome and briefly discuss the literature.