Exploring the resistome, virulome, and mobilome of multidrug-resistant Klebsiella pneumoniae isolates: deciphering the molecular basis of carbapenem resistance.

BACKGROUND: Klebsiella pneumoniae, a notorious pathogen for causing nosocomial infections has become a major cause of neonatal septicemia, leading to high morbidity and mortality worldwide. This opportunistic bacterium has become highly resistant to antibiotics due to the widespread acquisition of genes encoding a variety of enzymes such as extended-spectrum beta-lactamases (ESBLs) and carbapenemases. We collected Klebsiella pneumoniae isolates from a local tertiary care hospital from February 2019-February 2021. To gain molecular insight into the resistome, virulome, and genetic environment of significant genes of multidrug-resistant K. pneumoniae isolates, we performed the short-read whole-genome sequencing of 10 K. pneumoniae isolates recovered from adult patients, neonates, and hospital tap water samples. RESULTS: The draft genomes of the isolates varied in size, ranging from 5.48 to 5.96 Mbp suggesting the genome plasticity of this pathogen. Various genes conferring resistance to different classes of antibiotics e.g., aminoglycosides, quinolones, sulfonamides, tetracycline, and trimethoprim were identified in all sequenced isolates. The highest resistance was observed towards carbapenems, which has been putatively linked to the presence of both class B and class D carbapenemases, blaNDM, and blaOXA, respectively. Moreover, the biocide resistance gene qacEdelta1 was found in 6/10 of the sequenced strains. The sequenced isolates exhibited a broad range of sequence types and capsular types. The significant antibiotic resistance genes (ARGs) were bracketed by a variety of mobile genetic elements (MGEs). Various spontaneous mutations in genes other than the acquired antibiotic-resistance genes were observed, which play an indirect role in making these bugs resistant to antibiotics. Loss or deficiency of outer membrane porins, combined with ESBL production, played a significant role in carbapenem resistance in our sequenced isolates. Phylogenetic analysis revealed that the study isolates exhibited evolutionary relationships with strains from China, India, and the USA suggesting a shared evolutionary history and potential dissemination of similar genes amongst the isolates of different origins. CONCLUSIONS: This study provides valuable insight into the presence of multiple mechanisms of carbapenem resistance in K. pneumoniae strains including the acquisition of multiple antibiotic-resistance genes through mobile genetic elements. Identification of rich mobilome yielded insightful information regarding the crucial role of insertion sequences, transposons, and integrons in shaping the genome of bacteria for the transmission of various resistance-associated genes. Multi-drug resistant isolates that had the fewest resistance genes exhibited a significant number of mutations. K. pneumoniae isolate from water source displayed comparable antibiotic resistance determinants to clinical isolates and the highest number of virulence-associated genes suggesting the possible interplay of ARGs amongst bacteria from different sources.

Comprehensive insights into Klebsiella pneumoniae: unravelling clinical impact, epidemiological trends and antibiotic-resistance challenges.

Klebsiella pneumoniae, a challenging opportunistic bacterium, became a notable global health concern owing to its clinical impact, widespread epidemiology and escalating antibiotic resistance. This comprehensive review delves into the multifaceted dimensions of K. pneumoniae, with a focus on its clinical implications, epidemiological patterns and the critical issue of antibiotic resistance. The review also emphasizes the implications of K. pneumoniae in the context of antimicrobial stewardship and infection control. Epidemiological aspects are scrutinized, shedding light on the global distribution and prevalence of K. pneumoniae. Factors influencing its transmission and persistence in healthcare facilities and communities are examined, with patient demographics, healthcare practices and geographical variations. The review centres on antibiotic resistance, a critical issue in the era of bacteria displaying resistance to multiple drugs. The mechanisms of resistance used by K. pneumoniae against various classes of antibiotics are elucidated, along with the alarming rise of carbapenem-resistant strains. It also highlights ongoing research efforts and innovative strategies aimed at addressing this critical public health issue. This comprehensive review offers a holistic understanding of K. pneumoniae, emphasizing its clinical significance, global epidemiology and the immediate necessity for effective strategies to combat antibiotic resistance. It serves as a valuable resource for healthcare practitioners, researchers and policymakers seeking to manage better and mitigate the impact of this pathogen on public health.

Distribution characteristics of reactive silicon in six water bodies in the Yangtze River Basin in China.

Terrestrial silicon (Si) from biogeochemically weathered rocks and soils into oceans must pass through several water bodies, resulting in some Si immobilized. Hence, the knowledge on Si distribution characteristics in different water bodies at a basin scale is helpful to understand Si immobilization. A total of 65 surface sediments and corresponding overlying water samples were sampled from six water bodies (Dianchi Lake, DL; Dadu River, DR; Tuojiang River, TR; Honghu Lake, HL; Donghu Lake, DhL; Taihu Lake, TL) in the Yangtze River Basin of China, total dissolved Si (TDSi) in overlying water and exchangeable Si (Ex-Si), active non-biogenic Si (NBSi), and total acid dissolved Si (TADSi) in sediments were analyzed. Water chemical parameters (pH, EC, and TDP) and sediment components (LOI, TN, TP, and TADFe) showed that the water environment characteristics of six water bodies differed. TDSi differed among regions and between lakes and rivers, significantly higher in water bodies in the upper reaches and rivers than the middle or lower reaches and lakes (p < 0.05), respectively. Ex-Si in sediments in the upper reaches was significantly higher than in the middle or lower reaches (p < 0.05), except for DhL, whose Ex-Si was the highest. Mean TADSi and active NBSi were significantly higher in lakes than rivers (p < 0.05). Oxidation of sediments significantly increased TDSi in overlying water and active NBSi in sediments (p < 0.01). Si forms in six water bodies significantly depended on components of the sediments (e.g. active Ca2+, Mg2+, Fe, and Al3+) and water chemical parameters (p < 0.05). Our results suggest that immobilization of Si in water bodies in the Yangtze River Basin depends on the types of water bodies and sediments, lakes and Fe-Al dominated sediments have a high potential to immobilize Si, but anthropogenic interference should not be ignored.

A conserved guided entry of tail-anchored pathway is involved in the trafficking of a subset of membrane proteins in Plasmodium falciparum.

Tail-anchored (TA) proteins are defined by the absence of N-terminus signal sequence and the presence of a single transmembrane domain (TMD) proximal to their C-terminus. They play fundamental roles in cellular processes including vesicular trafficking, protein translocation and quality control. Some of the TA proteins are post-translationally integrated by the Guided Entry of TA (GET) pathway to the cellular membranes; with their N-terminus oriented towards the cytosol and C-terminus facing the organellar lumen. The TA repertoire and the GET machinery have been extensively characterized in the yeast and mammalian systems, however, they remain elusive in the human malaria parasite Plasmodium falciparum. In this study, we bioinformatically predicted a total of 63 TA proteins in the P. falciparum proteome and revealed the association of a subset with the P. falciparum homolog of Get3 (PfGet3). In addition, our proximity labelling studies either definitively identified or shortlisted the other eligible GET constituents, and our in vitro association studies validated associations between PfGet3 and the corresponding homologs of Get4 and Get2 in P. falciparum. Collectively, this study reveals the presence of proteins with hallmark TA signatures and the involvement of evolutionary conserved GET trafficking pathway for their targeted delivery within the parasite.

Biosynthesis and Characterization of Silver Nanoparticles Using Tribulus terrestris Seeds: Revealed Promising Antidiabetic Potentials.

Green synthesis is the most effective and environmentally friendly way to produce nanoparticles. The present research aimed at the biosynthesizing of silver nanoparticles (AgNPs) using Tribulus terrestris seed extract as the reducing and stabilizing agent and investigating their anti-diabetic properties. Fourier transformation infrared (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and UV-Vis spectroscopy were used to analyze the synthesized silver nanoparticles from Tribulus terrestris (TT-AgNPs). The spectroscopic characterization revealed a surface Plasmon resonance band at 380 nm, which verified the development of TT-AgNPs. The transmittance peaks were observed at 596, 1450, 1631, 2856, 2921, and 3422 cm-1 through the FTIR spectrophotometer. The XRD spectrum showed four distinct diffraction peaks in the 2θ range at 20° to 60°. Intense peaks were at 26.32°, 30.70°, 44.70°, 56.07°, 53.75°, 66.28°, and 75.32°. The SEM analysis revealed that the prepared TT-AgNPs were clustered loosely with a smooth and spherical structure and were of relatively uniform size. The in vitro antidiabetic potential of TT-AgNPs was assessed by using glucose yeast uptake, glucose adsorption, and alpha-amylase assays. TT-AgNPs showed the highest activity (78.45 ± 0.84%) of glucose uptake by yeast at 80 µg/mL. In the glucose adsorption assay, the highest activity of TT-AgNPs was 10.40 ± 0.52% at 30 mM, while in the alpha-amylase assay, TT-AgNPs exhibited the maximum activity of 75.68 ± 0.11% at 100 µg/mL. The results indicate a substantial anti-diabetic effect of the TT-AgNPs. Furthermore, the in vivo antidiabetic study was performed on TT-AgNPs in streptozotocin-induced diabetic mice. After receiving TT-AgNPs treatment for 30 days, the mice were sacrificed for biochemical and histological analyses of pancreatic and liver samples, which demonstrated a good improvement when compared to the control group. Mice treated with TT-AgNPs showed a significant drop in blood sugar levels, showing that the biosynthesized TT-AgNPs have effective anti-diabetic properties.

The efficiency of boric acid as an alternate to NaCl for the preservation of cattle hides.

The fresh buffalo hides (n = 6) were cut into two equal parts and categorized into three equal groups. The first group was treated with 50% NaCl; the second group hides were treated with 5% of boric acid (BA), and the third group hides were with NaCl + BA (10:1). Hair loss was seen at the sample margins of hides treated with 50% NaCl, with a slight odor. In the second group, there was neither hair loss nor a pungent smell was felt. The nitrogen content of the preserved hide was measured at different durations during the experimental period, i.e., 0 h, 24 h on day 7th, and day 14th. The nitrogen level reduced significantly (P < 0.05) from 0 h to day 14th of the experiment in hides treated with 50% of NaCl and 5% of boric acid, while this trend was found non-significant (P > 0.05) in hides treated with the combination of NaCl + BA. At 0 h, the moisture content for 50% of NaCl-treated hides was 64.82 ± 0.38% moisture content for 5% of boric acid was 63.89 ± 0.59, while for the NaCl + BA combination 61.69 ± 1.09 was observed. Moisture content for 50% of NaCl on day 14th was 38.87 ± 0.42; for boric acid, it was 37.76 ± 1.12, and for the combination of both, the moisture content was 34.56 ± 0.41%. A similar decreasing trend of moisture contents was found in hides treated with different preservatives. After 14 days of treatment, the bacterial count for 50% of NaCl was 2 × 109; for boric acid, it was 1 × 109, and for the combination of both, the bacterial count was 3 × 109. The lowest pollution load was observed hides treated with the combination of NaCl + BA (10:1). Total solids (TS) were 21.69 ± 0.57 while total dissolved solids (TDS) were 21.10 ± 0.57, and total suspended solids were 0.60 ± 0.57 mg/l. It is concluded from the present study that boric acid alone or in combination with NaCl efficiently reduced nitrogen content and bacterial count and can reduce water pollution in tannery and hence could be used as a preservative for the hide in the tannery industry.

Impact of graded levels of coated calcium butyrate on growth performance and serological indices during pre-weaning stage in Holstein calves.

The study aimed to analyze the impact of calcium butyrate supplementation in calf starter on growth performance indices associated with early rumen development to decrease the volume of milk or milk replacer feeding and enhance early starter intake in Holstein calves. For this purpose, twelve Holstein calves were randomly assigned into three treatments (n = 4/treatment); a control without coated calcium butyrate, T1, and T2 treatments supplemented with coated calcium butyrate 3 g and 6 g per day/head, respectively. Body weight was measured at days 7, 14, 21, 28, 35, 42, 49, and 56 of the trial, and the average daily weight gain and feed conversion ratio were determined. Blood samples were collected at 14, 28, 42, and 56 days of trial for serological parameters. Gut morphometry was performed at the end of trial at slaughtering by collecting duodenal samples. Furthermore, the meat was also evaluated for its quality parameters including pH and tenderness after slaughtering. The results indicated that the feed intake, average daily weight gain, feed conversion ratio, and gut morphometric parameters involving villus height and crypts depth of calves were improved in coated calcium butyrate-supplemented groups. Furthermore, the supplementation of calf starter with coated calcium butyrate significantly enhanced serum concentrations of glucose and total protein. Besides, Beta hydroxy butyrate (BHBA) levels of blood were also found to be elevated in both treatment groups. However, it was revealed that coated calcium butyrate supplementation had no significant effect on meat quality parameters. In conclusion, the supplementation of calf starter with coated calcium butyrate could improve calf performance.

Association of blood lead level with vitamin D binding protein, total and free 25-hydroxyvitamin D levels in middle-school children.

A negative association between blood Pb level (BPbL) and vitamin D metabolites in occupationally exposed populations has been reported, but data from the general population are scarce. Furthermore, the association between BPbL and vitamin D binding protein (DBP) and free 25-hydroxyvitamin D (25(OH)D) has not been reported. We investigated the association of BPbL with DBP, total and free 25(OH)D in healthy adolescents (n 1347; age range 11-16 years) cross-sectionally selected from all Governorates of Kuwait, utilising multi-stage cluster random sampling. Pb in whole blood was analysed by inductively coupled plasma MS, and DBP with ELISA. Plasma 25(OH)D was analysed by LC-MS/MS, and free 25(OH)D was calculated utilising the levels and binding affinities of DBP and albumin for 25(OH)D. DBP was positively associated with BPbL (ß = 0·81; 95 % CI 0·14, 0·22; P < 0·001). A negative association between BPbL and total 25(OH)D was non-significant (P = 0·24) when BPbL was used as a continuous variable but was significant when used as quartiles (P = 0·02). The negative association between BPbL and free 25(OH)D was significant whether BPbL was used as continuous, as quartiles or as cut-off point of <5 µg/dl (0·24 µmol/l). In multinomial logistic regression, the odds of vitamin D insufficiency and deficiency were more than two-fold higher in the upper quartiles of BPbL compared with the lowest quartile. The negative correlation of BPbL with free 25(OH)D was more robust than its correlation with total 25(OH)D. Future studies must consider the levels of DBP when assessing the association between Pb and vitamin D metabolites.

Transcriptional regulation of adipogenic marker genes for the improvement of intramuscular fat in Qinchuan beef cattle.

The intramuscular fat content plays a crucial role in meat quality traits. Increasing the degree of adipogenesis in beef cattle leads to an increase in the content of intramuscular fat. Adipogenesis a complex biochemical process which is under firm genetic control. Over the last three decades, the Qinchuan beef cattle have been extensively studied for the improvement of meat production and quality traits. In this study, we reviewed the literature regarding adipogenesis and intramuscular fat deposition. Then, we summarized the research conducted on the transcriptional regulation of key adipogenic marker genes, and also reviewed the roles of adipogenic marker genes in adipogenesis of Qinchuan beef cattle. This review will elaborate our understanding regarding transcriptional regulation which is a vital physiological process regulated by a cascade of transcription factors (TFs), key target marker genes, and regulatory proteins. This synergistic action of TFs and target genes ensures the accurate and diverse transmission of the genetic information for the accomplishment of central physiological processes. This information will provide an insight into the transcriptional regulation of the adipogenic marker genes and its role in bovine adipogenesis for the breed improvement programs especially for the trait of intramuscular fat deposition.

Increased LRG1 Levels in Overweight and Obese Adolescents and Its Association with Obesity Markers, Including Leptin, Chemerin, and High Sensitivity C-Reactive Protein.

Leucine-rich α-2 glycoprotein1 (LRG1) is a member of the leucine-rich repeat (LRR) family that is implicated in multiple diseases, including cancer, aging, and heart failure, as well as diabetes and obesity. LRG1 plays a key role in diet-induced hepatosteatosis and insulin resistance by mediating the crosstalk between adipocytes and hepatocytes. LRG1 also promotes hepatosteatosis by upregulating de novo lipogenesis in the liver and suppressing fatty acid ß-oxidation. In this study, we investigated the association of LRG1 with obesity markers, including leptin and other adipokines in adolescents (11−14 years; n = 425). BMI-for-age classification based on WHO growth charts was used to define obesity. Plasma LRG1 was measured by ELISA, while other markers were measured by multiplexing assay. Median (IQR) of LRG1 levels was higher in obese (30 (25, 38) µg/mL) and overweight (30 (24, 39) µg/mL) adolescents, compared to normal-weight participants (27 (22, 35) µg/mL). The highest tertile of LRG1 had an OR [95% CI] of 2.55 [1.44, 4.53] for obesity. LRG1 was positively correlated to plasma levels of high sensitivity c-reactive protein (HsCRP) (ρ = 0.2), leptin (ρ = 0.2), and chemerin (ρ = 0.24) with p < 0.001. Additionally, it was positively associated with plasma level of IL6 (ρ = 0.17) and IL10 (ρ = 0.14) but not TNF-α. In conclusion, LRG1 levels are increased in obese adolescents and are associated with increased levels of adipogenic markers. These results suggest the usefulness of LRG1 as an early biomarker for obesity and its related pathologies in adolescents.

Magnesium and Zinc Oxide Nanoparticles from Datura alba Improve Cognitive Impairment and Blood Brain Barrier Leakage.

Epilepsy is a neurological disorder involving persistent spontaneous seizures and uncontrolled neuronal excitability that leads to cognitive impairments and blood-brain barrier (BBB) disruption. Currently available antiepileptic drugs present side effects and researchers are trying to discover new agents with properties to overcome these drawbacks. The aim was to synthesize magnesium oxide (MgO) and zinc oxide (ZnO) nanoparticles from Datura alba fresh leaf extracts and evaluate their anti-epileptic potential in mice kindling or a repetitive seizures model. The phytoassisted synthesized nanoparticles were characterized using spectroscopy; FT-IR, XRD, SEM, and EDX. Analysis of the NPs confirmed the crystalline pleomorphic shape using the salts of both zinc and magnesium possibly stabilized, functionalized and reduced by bioactive molecules present in plant extract. By using several characterization techniques, NPs were confirmed. UV-Vis spectroscopy of biologically produced ZnO and MgO revealed distinctive peaks at 380 nm and 242 nm, respectively. Our findings categorically demonstrated the reductive role of biomolecules in the formation of ZnO and MgO NPs. The mice kindling model was induced using seven injections of Pentylenetetrazole (PTZ, 40 mg/kg, i.p) for 15 days alternatively. The results showed that mice post-treated with either ZnO or MgO nanoparticles (10 mg/kg, i.p) significantly improved in respect of behavior and memory as confirmed in the Morris water maze (MWM), open field (OF), novel object recognition (NOR) test compared with PTZ treated mice. Furthermore, the ZnO and MgO nanoparticle treatment also maintained the integrity of the BBB, reducing the leakage, as confirmed by Evans blue dye (EBD) compared with PTZ treated mice only. In summary, the current finding demonstrates that green synthesized ZnO and MgO nanoparticles have neuroprotective, ant-epileptic potential, molecular mechanisms, and clinical implications need to be further explored.

Low Doses of Thymoquinone and Ferulic Acid in Combination Effectively Inhibit Proliferation of Cultured MDA-MB 231 Breast Adenocarcinoma Cells.

Thymoquinone (TQ) and Ferulic Acid (FA) are natural ingredients from Nigella sativa and Ferula asafetida, respectively. Individually both TQ and FA have shown anticancer properties in a variety of cancer cell lines. We investigated the combination effect of lower doses of TQ and FA on proliferation, apoptosis, and cell cycle of a breast cancer cell line MDA-MB 231. Cells were treated with various concentrations of TQ, FA and various combinations of TQ + FA for 48 hrs. Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, whereas cell cycle and apoptosis were measured with flow-cytometry after propidium iodide staining and Annexin-V/FITC staining, respectively. TQ (50 and 100 µM) and FA (450 µM), and all the doses of TQ and FA in combination significantly decreased cell proliferation. 25 µM TQ and 250 µM FA individually did not affect cell proliferation but in combination significantly reduced cell proliferation. TQ at 50 µM caused significant apoptosis, whereas, FA did not affect apoptosis. These in vitro data suggest that in combination, relatively lower doses of TQ and FA are effective in decreasing cancer cell proliferation, and thus have anticancer therapeutic potential. Further research is needed to corroborate these findings in an animal model of breast cancer.

Risk factors of the severity of COVID-19: A meta-analysis.

OBJECTIVE: We intend to identify some probable risk factors that are responsible for the severity of COVID-19 using a meta-analysis. METHODS: The literature exploration lasted up to 18 April 2020 and through PubMed, Google Scholar, EMBASE and Cochrane Library we have identified 10 pertinent publications. To paraphrase the outcomes of autonomous researches, we have performed a random-effect meta-analysis. RESULTS: A total of 2272 patients' information was extracted from the selected literature. We have found gender (male) (Risk ratio [RR] = 1.29, 95% Confidence Interval [CI] 1.07 to 1.54), hypertension (RR = 1.79, 95% CI 1.57 to 2.04), diabetes (RR = 1.57, 95% CI 1.25 to 1.98), fatigue or myalgia (RR = 1.17, 95% CI 1.02 to 1.35), and smoking history (RR = 1.71, 95% CI 1.25 to 2.35) are potential risk factors for the severity of COVID-19. We found fever (RR = 1.21, 95% CI 0.66 to 2.22), cough (1.13, 95% CI 0.98 to 1.30) and diarrhoea (RR = 1.14, 95% CI 0.93 to 1.40) as insignificant risk factors for COVID-19 severity. CONCLUSIONS: The findings of this research may be beneficial to identify patients with higher risks to provide additional medical attention from the very beginning of the treatment.

Prevalence of lead toxicity in adolescents in Kuwait.

BACKGROUND: Elevated blood lead level (EBLL) is a public health problem in both developing and industrialized countries. Being a petrochemical-based economy, lead (Pb) levels are expected to be high in Kuwait, but systematic data on population exposure are lacking. This study aimed at determining the prevalence of EBLL in adolescents in Kuwait. METHODS: Adolescents (N = 1385; age range 11-16 years) were cross-sectionally selected from public middle schools from all Governorates of Kuwait, utilizing multistage cluster random sampling. Pb in whole blood was analyzed by inductively coupled plasma mass spectrometry. Distribution of blood Pb levels (BLL) among Governorates and sexes were compared by non-parametric tests and the prevalence of EBLL (defined as BLL above the CDC reference level of ≥5 µg/dL) was estimated by χ2 test. Binary logistic regression was used for assessing the association between EBLL and Governorate. RESULTS: Median (IQR) BLL was 5.1(3.6-7.1) µg/dL [4.9 (3.8-6.5) µg/dL in males and 5.4 (3.3-7.6) µg/dL in females; p = 0.001]. In the overall sample, 51% had BLL ≥5 µg/dL; 13% had ≥10 µg/dL and 3% > 20 µg/dL. Prevalence of EBLL was 47% in males and 56% in females (p < 0.001). EBLLs were clustered in Al-Asima, Al-Ahmadi (in both sexes); Al-Jahra (in males) and Mubarak Al-Kabeer (in females) Governorates. CONCLUSIONS: EBLL is a significant public health problem in adolescents in Kuwait. Urgent public health intervention is required in areas with EBLL, and the sources of exposure need to be identified for prevention.

Bioinformatics analysis and transcriptional regulation of TORC1 gene through transcription factors NRF1 and Smad3 in bovine preadipocytes.

Intramuscular fat is the an important factor that defines meat quality; however, enhancing its deposition without increasing the other three adipose depots (subcutaneous, visceral, and intermuscular fat) is a challenge for animal science and the meat industry. The TORC1 is a key regulator of adipogenesis and its regulation in bovine intramuscular preadipocytes has not been studied. The TORC1 is a member of the gene family that codes for a binding proteins which regulate transcription of cAMP which, is a key regulator of adipogenesis. In the present study, expression and sub-cellular localization of the TORC1 gene was analyzed in bovine preadipocytes. Bioinformatics tools were applied to characterize TORC1. To investigate the molecular mechanism of bovine TORC1 gene regulation, we cloned a 1008 bp of the 5'UTR regulatory region into a luciferase reporter vector. Different fragments were amplified using 5'UTR unidirectional deletion of the TORC1 promoter. Site directed mutation, dual luciferase reporter assay, RNAi interference and DNA-protein interaction (EMSA) were used to validate the regulatory roles of Smad3 and NRF1 in the regulation of TORC1 gene in bovine preadipocytes. The core promoter region of the TORC1 gene was identified at a location -410 to -155 bp upstream of transcription start site. Different vectors were constructed through serial deletion of the 5'UTR flanking region and in combination with site directed mutations and transcription interference through siRNA or shRNA, two transcription factors of NRF1 and Smad3 were found to be repressors in the promoter of the TORC1 gene. These findings were further confirmed through Electrophoretic Mobility Shift Assay (EMSA) within nuclear extracts of bovine adipocytes. The core promoter region of the TORC1 gene, spanning from -410 to -155 bp upstream of the transcription start site was specified in this study and this information will provide opportunity for the improvement of intramuscular fat in cattle.

Vitamin D levels in relation to low back pain during adolescence.

This study aimed to investigate the association between 25-hydroxyvitamin D (25(OH)D) level and low back pain (LBP) among adolescents while adjusting for potential confounders pertinent to this age group including the weight of school bags, BMI and physical activity. A cross-sectional study was conducted on 760 randomly selected adolescents in middle schools. Data on LBP and the risk factors for LBP were collected from parents by a self-administered questionnaire and from adolescents by face-to-face interview. Blood samples were tested in an accredited laboratory; and 25(OH)D was measured using liquid chromatography-tandem MS. The lifetime prevalence and the 6-month prevalence of LBP were 32·28 (95 % CI 28·97, 35·73) % and 21·26 (95 % CI 18·40, 24·33) %, respectively. There was no difference in the geometric mean of 25(OH)D between those with and without LBP in the past 6 months (28·50 nmol/l and 30·82 nmol/l, respectively; P = 0·122). There was no association between 25(OH)D and LBP in the univariable or multivariable analysis whether 25(OH)D fitted as a continuous or as a categorical variable. We found no association between vitamin D level and LBP in adolescents in an area with high prevalence of vitamin D deficiency. Although it is important to have sufficient vitamin D levels during adolescence for several other health benefits, we concluded that vitamin D is not a major determinant for LBP among adolescents in our setting.

Effects of dietary organic acids on performance, cecal microbiota, and gut morphology in broilers.

Organic acids have exhibited great potential as an antibiotic replacement and as an additive work tremendously for health maintenance of broiler chicken. To explore more about organic acids, a total of 900 day-old broiler chicks (Cobb-500) were procured from a local hatchery and distributed into 9 treatment groups having 5 replicates of 20 birds each; duration of the biological trial was of 35 days. Group T1 served as control group without any dietary supplementation. Other groups T2 and T3 were boosted with different levels (125 g/ton and 250 g/ton) of enramycin (antibiotic), T4, T5, and T6 were supplemented with different levels (2 kg/ton, 3 kg/ton, and 4 kg/ton) of ammonium formate and ammonium propionate, and T7, T8, and T9 were fed with different levels (2 kg/ton, 3 kg/ton, and 4 kg/ton) of calcium formate and calcium propionate. The findings declared significant improvement (P < 0.05) in body weight gain and FCR in groups T3, T5, and T9 while feed intake was not affected. Carcass evaluation depicted significantly better (P < 0.05) dressed and eviscerated weight along with carcass yield (T5, T7, T8, T9). Broilers fed organic acid supplemented diet had significantly lower (P < 0.05) total bacterial count (T3, T5, T8, T9) and positively improved (P < 0.05) villi length (T5, T6, T9) as compared with control group. However, total protein, globulin, HDL, and LDL levels were determined to be non-significant (P > 0.05) among different organic acids treatments. Hence, organic acids can be utilized as a better replacement for antibiotics. Supplementation of organic acids at a dose rate of 3 kg/ton and 4 kg/ton is recommended for efficient performance of broilers.

Memantine Is Protective against Cytotoxicity Caused by Lead and Quinolinic Acid in Cultured Rat Embryonic Hippocampal Cells.

Quinolinic acid (QA) is an excitotoxic metabolite of the kynurenine pathway of tryptophan metabolism produced in response to inflammation and oxidative stress. Lead (Pb) causes oxidative stress and thus may produce neurotoxicity by increasing QA production. We investigated the in vitro cytotoxic effects of Pb and QA and the protective effects of the NMDA receptor antagonist memantine. Primary cultures of embryonic hippocampal cells from Wistar rats were treated with different concentrations of Pb, QA, and Pb + QA with and without memantine. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Apoptosis was analyzed by flow cytometry after Annexin-V/propidium iodide staining. The numbers of immunostained neurons (with ß3-Tubulin; Tuj1) and astrocytes (with glial fibrillary acidic protein) were counted. Pb at 20 µg/dL (0.97 µM) and QA at 500 nM concentrations showed significant cytotoxic effects, as evidenced by decreased cell viability, increased apoptosis, and a decrease in the number of both astrocytes and neurons. The combination of Pb and QA showed significant synergistic apoptotic effects at lower doses. Memantine (500 nM) was largely protective against the cytotoxic effects of both Pb and QA, suggesting that Pb's and QA's cytotoxicity involves NMDA receptor activation. Whereas the neuroprotection by memantine from QA-induced toxicity has been previously reported, this is the first study reporting the protection by memantine against Pb-induced cytotoxicity in cultured hippocampal cells. Protection by memantine against these neurotoxicants in vivo needs to be investigated.

Intraventricular infusion of quinolinic acid impairs spatial learning and memory in young rats: a novel mechanism of lead-induced neurotoxicity.

BACKGROUND: Lead (Pb), a heavy metal, and quinolinic acid (QA), a metabolite of the kynurenine pathway of tryptophan metabolism, are known neurotoxicants. Both Pb and QA impair spatial learning and memory. Pb activates astrocytes and microglia, which in turn induce the synthesis of QA. We hypothesized increased QA production in response to Pb exposure as a novel mechanism of Pb-neurotoxicity. METHODS: Two experimental paradigms were used. In experiment one, Wistar rat pups were exposed to Pb via their dams' drinking water from postnatal day 1 to 21. Control group was given regular water. In the second protocol, QA (9 mM) or normal saline (as Vehicle Control) was infused into right lateral ventricle of 21-day old rats for 7 days using osmotic pumps. Learning and memory were assessed by Morris water maze test on postnatal day 30 or 45 in both Pb- and QA-exposed rats. QA levels in the Pb exposed rats were measured in blood by ELISA and in the brain by immunohistochemistry on postnatal days 45 and 60. Expression of various molecules involved in learning and memory was analyzed by Western blot. Means of control and experimental groups were compared with two-way repeated measure ANOVA (learning) and t test (all other variables). RESULTS: Pb exposure increased QA level in the blood (by ~ 58%) and increased (p < 0.05) the number of QA-immunoreactive cells in the cortex, and CA1, CA3 and dentate gyrus regions of the hippocampus, compared to control rats. In separate experiments, QA infusion impaired learning and short-term memory similar to Pb. PSD-95, PP1, and PP2A were decreased (p < 0.05) in the QA-infused rats, whereas tau phosphorylation was increased, compared to vehicle infused rats. CONCLUSION: Putting together the results of the two experimental paradigms, we propose that increased QA production in response to Pb exposure is a novel mechanism of Pb-induced neurotoxicity.

Halal slaughtering, welfare, and empathy in farm animals: a review.

Pre-slaughter and slaughter stressors are considered major concerns in animal welfare. Halal slaughtering method is considered one of the slaughtering stressors in livestock. This method seems to cause fear followed by stress in animals mainly due to inhuman handling. In this review, empathy and animal welfare are discussed in light of Islamic sharia and has further linked with animal's physiology and behavioral responses during slaughtering. Islam as a religion forbids slaughtering an animal in front of another animal as through optic, olfactory, and cochlear senses animals can perceive the stress state of conspecifics. This suggests and strengthens the hypothesis that animals being slaughtered in front of each other may produce stress in them. This argument further leads to a claim that animals can experience empathy of each other through olfaction of semiochemicals (stress pheromones) emitted from animals slaughtered in the stressful condition that can be detected by other animals in abattoirs. Hence, research is needed to find out these specific stress pheromones and legislation needs to be adopted in slaughterhouses to isolate the areas of butchery from slaughtering lines to ensure proper guidelines of Halal slaughtering in slaughterhouses.