The impact of diabetes mellitus on the emergence of multi-drug resistant tuberculosis and treatment failure in TB-diabetes comorbid patients: a systematic review and meta-analysis

BackgroundThe existence of Type 2 Diabetes Mellitus (DM) in tuberculosis (TB) patients is very dangerous for the health of patients. One of the major concerns is the emergence of MDR-TB in such patients. It is suspected that the development of MDR-TB further worsens the treatment outcomes of TB such as treatment failure and thus, causes disease progression.AimTo investigate the impact of DM on the Emergence of MDR-TB and Treatment Failure in TB-DM comorbid patients.MethodologyThe PubMed database was systematically searched until April 03, 2022 (date last searched). Thirty studies met the inclusion criteria and were included in this study after a proper selection process.ResultsTuberculosis-Diabetes Mellitus patients were at higher risk to develop MDR-TB as compared to TB-non-DM patients (HR 0.81, 95% CI: 0.60–0.96, p < 0.001). Heterogeneity observed among included studies was moderate (I2 = 38%). No significant change was observed in the results after sub-group analysis by study design (HR 0.81, 95% CI: 0.61–0.96, p < 0.000). In the case of treatment failure, TB-DM patients were at higher risk to experience treatment failure rates as compared to TB-non-DM patients (HR 0.46, 95% CI: 0.27–0.67, p < 0.001).ConclusionThe results showed that DM had a significant impact on the emergence of MDR-TB in TB-diabetes comorbid patients as compared to TB-non-DM patients. DM enhanced the risk of TB treatment failure rates in TB-diabetes patients as compared to TB-non-DM patients. Our study highlights the need for earlier screening of MDR-TB, thorough MDR-TB monitoring, and designing proper and effective treatment strategies to prevent disease progression

Effect of Temperature and Storage Time on DNA Quality and Quantity from Normal and Diseased Tissues

DNA extraction and purification is an initial step for authentic results in advance molecular biology, therefore DNA degradation is unavoidable. The aim of present study is to investigate the DNA quality and quantity in terms of shorter time preservation with normal and diseased tissue, therefore tissues of normal (n = 18) and diseased (n = 18) liver, lung and heart was collected from goat after slaughtered. For DNA extraction Gene JET Genomic DNA Purification Kit protocol was followed, then stored at -20 oC and -04 oC temperatures for 24hrs and 48hrs period of time. The concentration and purity of the extracted DNA were measured with Spectrophotometer and purity confirmed at an absorbance ratio of 260 or 280. It was observed that at a -20 oC temperature for 24hours the concentration of DNA yield was numerically higher than at -04 oC temperature for tissue stored at 48hrs, whereas absorbance was higher, however in normal tissues in contrast with diseased the concentration and absorbance of DNA was somehow same at -20 and -04 oC but different in storage time. On the basis of these findings, it was concluded that time elapsed between sampling with the storage condition and with normal or diseased samples for DNA extraction will largely depend on the experiment. If tissue preservative conditions and sampling are appropriate, storage time will not be a factor at least for short storage periods

Effect of Temperature and Storage Time on DNA Quality and Quantity from Normal and Diseased Tissues

DNA extraction and purification is an initial step for authentic results in advance molecular biology, therefore DNA degradation is unavoidable. The aim of present study is to investigate the DNA quality and quantity in terms of shorter time preservation with normal and diseased tissue, therefore tissues of normal (n = 18) and diseased (n = 18) liver, lung and heart was collected from goat after slaughtered. For DNA extraction Gene JET Genomic DNA Purification Kit protocol was followed, then stored at -20 oC and -04 oC temperatures for 24hrs and 48hrs period of time. The concentration and purity of the extracted DNA were measured with Spectrophotometer and purity confirmed at an absorbance ratio of 260 or 280. It was observed that at a -20 oC temperature for 24hours the concentration of DNA yield was numerically higher than at -04 oC temperature for tissue stored at 48hrs, whereas absorbance was higher, however in normal tissues in contrast with diseased the concentration and absorbance of DNA was somehow same at -20 and -04 oC but different in storage time. On the basis of these findings, it was concluded that time elapsed between sampling with the storage condition and with normal or diseased samples for DNA extraction will largely depend on the experiment. If tissue preservative conditions and sampling are appropriate, storage time will not be a factor at least for short storage periods

A Time-Saving Strategy to Generate Double Maternal Mutants by an Oocyte-Specific Conditional Knockout System in Zebrafish

Maternal products are those mRNAs and proteins deposited during oogenesis, which play critical roles in controlling oocyte formation, fertilization, and early embryonic development. However, loss-of-function studies for these maternal factors are still lacking, mainly because of the prolonged period of transgenerational screening and technical barriers that prevent the generation of maternal (M) and maternal and zygotic (MZ) mutant embryos. By the transgenic expression of multiple sgRNAs targeting a single gene of interest in the background of a transgenic line Tg(zpc:zcas9) with oocyte-specific cas9 expression, we have successfully obtained maternal or maternal–zygotic mutant for single genes in F1 embryos. In this work, we tandemly connected a maternal GFP marker and eight sgRNA expression units to target dvl2 and dvl3a simultaneously and introduced this construct to the genome of Tg(zpc:zcas9) by meganuclease I-Sce I. As expected, we confirmed the existence of Mdvl2;Mdvl3a embryos with strong defective convergence and extension movement during gastrulation among outcrossed GFP positive F1 offspring. The MZdvl2;MZdvl3a embryos were also obtained by crossing the mutant carrying mosaic F0 female with dvl2+/−;dvl3a−/− male fish. This proof-of-principle thus highlights the potential of this conditional knockout strategy to circumvent the current difficulty in the study of genes with multiple functionally redundant paralogs

LIPOPHAGY: a novel form of steroidogenic activity within the LEYDIG cell during the reproductive cycle of turtle

Abstract Background Steroidogenesis is an indispensable process that is indirectly associated with spermatogenesis in the Leydig cell (LC) to utilize the lipid droplets (LDs) that are critical to maintaining normal testosterone synthesis. The regulation of LD mobilization, known as lipophagy, in the LC is still largely unknown. Method In the present study, the LC of the Chinese soft-shelled turtle was investigated to identify the steroidogenic activity and lipophagy during the annual reproductive cycle by light microscopy, immunohistochemistry (IHC), immunofluorescence (IF), and transmission electron microscopy (TEM). Results The LC showed a dynamic steroidogenic function with strong activity of 3β-HSD, vimentin and tubular ER during hibernation by IHC and TEM. The tubulo-vesicular ER had a weak immunopositive reaction for 3β-HSD in the LC during reproductive phase, suggesting persistent steroidogenic activity. ORO staining and TEM demonstrated that a larger number of LDs had accumulated in the LC during hibernation than in the reproductive phase. These LDs existed in close association with mitochondria and lysosomes by being dynamically surrounded by intermediate filaments to facilitate LD utilization. Lysosomes were found directly attached to large LDs, forming an autophagic tube and engulfing LDs, suggesting that micro-lipophagy occurs during hibernation. Furthermore, the IHC of ATG7 (Autophagy Related Gene 7) and the IF of the LC3 (Microtubule-associated protein light chain 3), p62 (Sequestosome-1 (SQSTM1) and LAMP1(Lysosomal-associated membrane protein 1) results demonstrated strong expression, and further confirmation by TEM showed the existence of an autophagosome and an autolysosome and their fusion during the hibernation season. Conclusion In conclusion, the present study provides clear evidence of LD consumption in the LC by lipophagy, lysosome and mitochondria during the hibernation period, which is a key aspect of steroidogenesis in the Chinese soft-shelled turtle

Therapeutic strategy of biological macromolecules based natural bioactive compounds of diabetes mellitus and future perspectives: A systematic review

High blood glucose levels are a hallmark of the metabolic syndrome known as diabetes mellitus. More than 600 million people will have diabetes by 2045 as the global prevalence of the disease continues to rise. Contemporary antidiabetic drugs reduce hyperglycemia and its consequences. However, these drugs come with undesirable side effects, so it's encouraging that research into plant extracts and bioactive substances with antidiabetic characteristics is on the rise. Natural remedies are preferable to conventional anti-diabetic drugs since they are safer for the body, more affordable and have fewer potential adverse effects. Biological macromolecules such as liposomes, niosomes, polymeric nanoparticles, solid lipid nanoparticles, nanoemulsions and metallic nanoparticles are explored in this review. Current drug restrictions have been addressed, and the effectiveness of plant-based antidiabetic therapies has enhanced the merits of these methods. Plant extracts' loading capacity and the carriers' stability are the primary obstacles in developing plant-based nanocarriers. Hydrophilic, hydrophobic, and amphiphilic drugs are covered, and a brief overview of the amphipathic features of liposomes, phospholipids, and lipid nanocarriers is provided. Metallic nanoparticles' benefits and attendant risks are highlighted to emphasize their efficiency in treating hyperglycemia. Researchers interested in the potential of nanoparticles loaded with plant extracts as antidiabetic therapeutics may find the current helpful review