Cellular and molecular biology最新文献

Increasing evidence suggests that inhibition of receptor-interacting serine/threonine-protein kinase (RIPK) 1/RIPK3/mixed lineage kinase domain-like pseudokinase (MLKL) necrosome has protective effects in vivo models of painful conditions seen in humans associated with inflammation and demyelination in the central nervous system. However, the contribution of RIPK1-driven necroptosis to inflammatory pain remains unknown. Therefore, this study aims to determine the effect of necrostatin (Nec) -1s, a selective RIPK1 inhibitor, on lipopolysaccharide (LPS)-induced inflammatory pain and related underlying mechanisms. In the saline-, LPS-, and/or Nec-1s-injected male mice, thermal hyperalgesia was evaluated by hot plate test. Alterations in the expression of proteins involved in the RIPK1, toll-like receptor (TLR) 4, myeloid differentiation factor (MyD) 88/toll-interleukin (IL)-1 receptor domain-containing adapter-inducing interferon-β (TRIF)/nuclear factor (NF)-kB, nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain containing (NLRP) 3/apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC)/pro-caspase-1, and caspase-11/gasdermin D (GSDMD) signaling pathways, as well as proteins related to demyelination and remyelination in the brain and spinal cord were determined by the immunoblotting method. The LPS-induced alleviation of thermal hyperalgesia was prevented by necrostatin-1s. Necrostatin-1s reversed (1) increased activity of RIPK1, RIPK3, MLKL, and NF-kB p65, (2) enhanced expression of TLR4, MyD88, TRIF, NF-kB p65, HMGB1, NLRP3, ASC, caspase-1 p20, IL-1β, caspase-11 p20, p30-GSDMD, and semaphorin 3A, and (3) diminished myelin PLP expression induced by LPS. These findings suggest that the use of RIPK1 inhibitors could be a therapeutic approach in the management of inflammatory pain associated with necroptosis, pyroptosis, and demyelination.

The rising global incidence of syphilis underscores the risk of transmission through blood transfusions. Treponema pallidum, the pathogen responsible for syphilis, represents a major public health challenge. Accurate detection is essential for controlling the disease, particularly in asymptomatic blood donors. This study aimed to evaluate the seroprevalence of specific antibodies against T. pallidum in blood donors, confirmed by DNA testing for seropositivity. The goal was to enhance our understanding of syphilis exposure and improve the safety of blood donations. A total of 1,260 HIV, HCV, and HBsAg-negative blood donors were screened for T. pallidum-specific antibodies using enzyme-linked immunosorbent assay (ELISA). Initially, reactive samples were re-evaluated, and those repeatedly reactive were classified as seropositive for syphilis. ELISA-positive samples were further tested for T. pallidum DNA using real-time polymerase chain reaction (RT-PCR). Data analysis was done using SPSS with a level of significance p< 0.05 Of 1,260 blood donors, the seroprevalence of anti-T. pallidum antibodies was 0.158%, with both positive cases confirmed by PCR. The prevalence was 0.2% in males and 0.00% in females, with no significant gender differences (P > 0.05). The highest prevalence was in the 31-40 age group (0.5%), but this was not statistically significant (P > 0.05). There were no significant differences by donation type or marital status. Significant associations were observed with educational level (P < 0.05), with higher prevalence among high school graduates Our results confirm syphilis in Iraqi blood donors, highlighting the need for routine T. pallidum ELISA screening at transfusion centers. Positive cases should be discarded and affected donors treated. ELISA is an effective primary screening method, consistent with WHO guidelines for low-prevalence settings, and is essential for preventing transfusion transmission.

Rice salt tolerance is highly anticipated to meet global demand in response to decreasing farmland and soil salinization. Therefore, dissecting the genetic loci controlling salt tolerance in rice for improving productivity is of utmost importance. Here, we evaluated six salt-tolerance-related traits of a biparental mapping population comprising 280 F2 rice individuals (Oryza sativa L.) at the seedling and reproductive stages. We performed a genome-wide association study (GWAS) to identify marker-trait associations under artificially induced salt stress using the 1K RICA chip (Agriplex Genomics, Cedar Avenue, Suite 250, Cleveland, 011444106, USA). We have identified 8 single nucleotide polymorphisms (SNPs) representing eight genomic regions on chromosomes 5, 8, 9, and 10. These were significantly associated with the six salt-tolerance-related traits, no. of tillers per plant (TPP), effective tillers per plant (ETP), spikelet fertility percentage (SFP), field grain number (FGN), grain length breadth ratio (LBR) and thousand-grain weight (TGW).  FGN has two significant SNPs (SNP0758 and SNP0759) on Chromosome 9, whereas SFP on chromosomes 8 and 12 (SNP1127 and SNP0966, respectively). Similarly, for TPP (SNP0796), a significant SNP was detected on chromosome 10, and for ETP (SNP0414) on chromosome 5.  Two significant SNPs were found in chromosome 12 for LBR (SNP0920) and TGW (SNP0976). Based on all loci, we screened 3 possible candidate genes in chromosomes 8, 9, and 12 between the genomic region of SNP0920 and SNP1127 under salt stress. Interestingly, these genes were involved in protein coding, none of which was previously reported as being involved in plant salt tolerance. Further, the genetic relationship between the mapping population and population structure was classified by STRUCTURE v 2.3. Genotypes with ≥ 80% of shared ancestry were explained into two major clusters (I and II), and < 80% of shared ancestry were categorized as admixtures. An unrooted alpha was developed by TASSEL 5.0, dividing the genotypes into three major groups where 97 individuals were in Cluster 1, cluster 2 consisted of 93 individuals, and the remaining Cluster 3 included 90 individuals. A kinship matrix developed from 860 SNPs indicated group formation and more substantial relatedness among the genotypes with a red zone. Our findings provide valuable information for enhancing the understanding of complicated salt tolerance mechanisms in rice seedlings and the identified candidates potentially used for breeding salt-tolerant genotypes.

A pathological condition in the peripheral nerve tissue, which provides the connection between the organism and the external environment, negatively affects the standard of living. The nerve tissue histotechnology is of serious importance both for scientific studies and for clinical diagnosis. The fixation, which is one of the leading procedures for histological examination of tissues, aims to preserve tissue morphology. Another essential part of the histological examination is staining process. This study, it was aimed to determine the fixative that provides optimal histological appearance in peripheral nerve tissue. Therefore, various histochemical stainings of tissues fixed with some fixatives used in practice were compared. Sciatic nerves from each rat (n=7) used in the study were fixed with different fixatives and histochemical staining was performed. In histological examination, cellular (nucleus-cytoplasm) and intercellular morphological details, staining intensity and distribution were evaluated. At the end of the study, formaldehyde was found to be the most ideal fixing agent for all stains. Although Bouin and Carnoy fixatives differed according to the staining type, their fixation quality was similar in general. Glutaraldehyde did not give as good results as other fixatives in all stainings. This study is an important technical reference for clinical and experimental studies.

Today, methamphetamine (METH) is being used by adolescents and young adults. Our previous research demonstrated that intrauterine exposure to METH induces apoptosis in testicles and seminiferous tubes. However, based on available literature, the mechanism of this effect remains unidentified. This study aimed to investigate proteins involved in sperm growth and development pathways, such as testis-specific serine/threonine kinases (TSSK) and receptor-interacting protein kinases 2 (RIPK2), and to study the serine-threonine kinase pathway in the testes of rats whose mothers received intraperitoneal METH during pregnancy. In the present study, female rats during pregnancy received either 5 or 10 mg/kg of METH or normal saline for ten days. After reaching maturity, their testes were isolated and examined for histopathological and immunohistochemical mechanisms. Results were analyzed and reported using statistical software. Results revealed that following intrauterine exposure to METH, TSSK protein expression reduced from 52.68±2.4% in the control group to 48.04±2.29% in the 2 mg/kg/day group and 12.83±3.35% in the 5 mg/kg/day group with P=0.0029 and F=72.63. In addition, RIPK2 protein expression increased from 8.34±2.69% in the control group to 31.17±3.69% in the 2 mg/kg/day group and 98.49±4.66% in the 5 mg/kg/day group, with p=0.0037 and F=61.14. Histopathological findings indicated a reduction in the thickness of germ layers following intrauterine exposure to METH, with the seminiferous tubule's thickness decreasing. Inflammatory cell populations increased, and the number of vessels decreased due to intrauterine exposure to METH. Our study suggests intrauterine exposure to METH increases the prevalence of inflammatory cell populations, enhances RIPK2 protein expression, reduces the number of vessels, reduces the diameter of seminiferous tubes, decreases TSSK protein expression, and reduces the thickness of germ layers in testicular tissue. Apoptosis of spermatid cells observed in our previous study may be related to the signaling pathways of TSSK and RIPK2 proteins.

Chronic kidney disease (CKD) is often complicated by diabetes, impacting various biochemical and immunological markers. This study aimed to investigate the relationship between irisin, apelin-13, and immunological markers IL-1α and IL-1β in diabetic patients with CKD. This cross-sectional study was conducted from January to June 2023 in a tertiary care hospital in Tikrit City, Iraq. This study included 120 CKD patients and a control group including 20 healthy individuals. Patients were included in the study by convenience sampling method. Participants were evaluated using ELISA kits for irisin, apelin-13, and cytokines, with blood samples analyzed for relevant biochemical markers. Patients had irisin levels of 10.98 ± 2.5 ng/mL, significantly different from non-diabetic patients (12.40 ± 3.54 ng/mL) and controls (5.36 ± 1.06 ng/mL) (p<0.001). Apelin-13 was higher in diabetic patients (537.71 ± 124.78 pg/mL) compared to controls (181.26 ± 29.98 pg/mL) (p<0.001). IL-1α levels in diabetic patients were 715.30 ± 392.48 pg/mL, significantly higher than in control patients (206.27 ± 26.49 pg/mL) (p<0.001). IL-1β levels were 351.50 ± 81.82 pg/mL in diabetics, also higher than in control (145.79 ± 38.49 pg/mL) (p<0.001). The study highlights significant associations between biochemical markers and CKD in diabetic patients. Elevated levels of irisin, apelin-13, IL-1α, and IL-1β may serve as potential biomarkers for diabetes-related CKD complications.

Mitochondrial ribosomal protein S23 (MRPS23), encoded by a nuclear gene, is a well-known driver of proliferation in cancer. It participates in mitochondrial protein translation, and its expression association has been explored in many types of cancer. However, MRPS23 expression associations are rarely reported in breast cancer (BC). In this study, we explored the MRPS23 expression in BC cells compared with the non-tumoral breast cells. Overexpression and knockdown analysis of MRPS23 were performed in BC cells. Transfection efficiency was evaluated by western blot and qRT-PCR analysis. The role of MRPS23 in the malignant biological behaviors of BC cells was investigated using in-vitro experiments. Our results demonstrate that MRPS23 was aberrantly overexpressed at both the transcript and protein levels in BC cells. Additional findings reveal that deficiency of MRPS23 is associated with a decrease in cell proliferation/viability and compromised cell migration/invasion in BC cells. Relative to the sh-Ctrl group, the expression levels of cadherin, SNAI 1, and TWIST 1 decreased in the MRPS23 knockdown BC cells. We further found a significant decrease in the expression levels of Cyclin D1, Axin 2, LEF1, NKD1, and Survivin in MRPS23 knockdown cells. In conclusion, we found an association between MRPS23 knockdown and the metastasis ability of BC cells. These findings reveal that MRPS23 significantly decreases the migration and invasion of BC, thus inhibiting BC progression. We confirmed for the first time that MRPS23 expression determines the metastasis features of BC. Hence, the findings justify the key role of this protein in BC progression; therefore, it may be a potential therapeutic target for BC therapy.

In this study, the effects of histone deacetylase inhibitor CI-994 and nanotechnological drug liposomal cisplatin LipoPlatin on Luminal A breast cancer and triple-negative breast cancer were explored using agents alone and in combination. MCF-7 and MDA-MB-231 cell lines were used. Cell viability, and cell index values obtained from xCELLigence System, MI, BrdU LI and AI were evaluated in experiments. In monotherapy applications, 10 μM, 20 μM and 40 μM concentrations of CI-994 and 80 μM, 160 μM, and 250 μM concentrations of LipoPlatin were applied to both cell lines. IC50 concentration of CI-994 was 10 μM for both cell lines. While IC50 concentration of LipoPlatin was 80 μM for Luminal A breast cancer cell line, this concentration was 160 μM for triple-negative breast cancer cell line. The application of a combination of CI-994 and Lipoplatin for each cell line has shown that while there was a significant decrease in cell viability, mitotic index (MI), and BrdU labeling index (LI), there was a significant increase in AI. Consequently, it was thought that combined treatments were more efficient than single medication applications and decreased the cell prognosis with a synergistic and additive impact.

Klebsiella pneumoniae is a non-motile, encapsulated, environmental gram-negative bacterium. Once the bacteria have infiltrated the body, they can display substantial degrees of resistance to drugs and virulence. Extended Spectrum Beta-Lactamases (ESBLs) are most typically seen in K. pneumoniae. The objective of this study was to investigate the morbidity and mortality associated with ESBL K. pneumoniae infection in different albino rat administration route groups. Four cohorts of albino rats were acquired and categorized into the subsequent groups: inhalation, oral administration via food, water, and control group. Each group was infected independently and the isolate administration lasted 6 days. The clinical diagnosis revealed the presence of K. pneumoniae infection. Within one day of infection, the inhalation group exhibited the initial clinical signs and symptoms, such as red eyes, coughing, and closed eyelids. Subsequently, the infection was verified through the process of sample cultivation. Additionally, blood clinical findings, including blood tests such as CBC, lipid profile, CRP, and kidney and liver function tests, further supported the confirmation of the infection. The K. pneumoniae isolates had a severe influence on the CBC, liver, and kidney functioning causing elevated liver enzymes, and high RBC levels with impaired kidney functioning. Due to K. pneumonia's affinity for lung tissue, it had the greatest impact in the albino rat inhalation group.

One of the prevailing trends in contemporary agriculture is the application of biological control. Nevertheless, several reports suggest that biocontrol bacteria exhibit poor survival rates in host plants. Consequently, the concept of shielding biological control agents by encapsulating them in outer coatings has gained popularity. Several techniques, including extrusion, spray drying, and emulsification, have been introduced to encapsulate biocontrol bacteria. Much research has focused on the preparation of suitable synthetic hormone products capable of influencing plant growth and development in agriculture. The most effective approach to address this demand is through controlled release systems. One of these techniques involves encapsulating growth hormones. The encapsulation procedure must adhere to crucial standards such as biocompatibility, biodegradability, and provision for sustained viability and performance. Nonetheless, it is essential to conduct further research on the consequences of encapsulation and targeted release in organic farming systems. The creation of a novel composition grounded on biodegradable polymers has the potential to enhance the volume and quality of agricultural yields significantly. The current investigation endeavors to scrutinize the encapsulation of plant hormones and microencapsulation and their effectiveness in counteracting plant pathogens.