T cells exhaustion, inflammatory and cellular activity markers in PBMCs predict treatment outcome in pulmonary tuberculosis patients.

BACKGROUND: Pulmonary tuberculosis (PTB) is a well-known disease caused by Mycobacterium tuberculosis. Its pathogenesis is premised on evasion of the immune system and dampened immune cells activity. METHODS: Here, the transcription pattern of immune cells exhaustion, inflammatory, and cellular activity markers were examined in peripheral blood mononuclear cells (PBMCs) from PTB patients at various stages of treatment. PBMCs were isolated, and RNA extracted. cDNA synthesis was performed, then amplification of genes of interest. RESULTS: The T cell exhaustion markers (PD-L1, CTLA4, CD244 and LAG3) showed varied levels of expressions when comparing 0 T and 1 T to the other treatment phases, suggesting their potential roles as markers for monitoring TB treatment. IL-2, IFN-g and TNF-a expression at the gene level returned to normal at completion of treatment, while granzyme B levels remained undetectable at the cured stage. At the cured stage, the cellular activity monitors Ki67, CD69, GATA-3, CD8 and CD4 expressions were comparable to the healthy controls. Correlation analysis revealed a significantly strong negative relationship with CD244 expression, particularly between 1 T and 2 T (r = -0.94; p = 0.018), and 3 T (r = -0.95; p = 0.013). Comparing 0 T and 3 T, a genitive correlation existed in PD-L1 (r = -0.74) but statistically not significant, as seen in CTLA4 and LAG-3 expressions. CONCLUSION: Collectively, the findings of the study suggest that T-cells exhaustion marker particularly CD244, inflammatory markers IL-2, IFN-g and TNF-a, and cellular activity indicators such as Ki67, CD69, GATA-3, CD8 and CD4 are promising markers in monitoring the progress of PTB patients during treatment.

Seroprevalence and Risk Factors of Sexually Transmitted Blood-Borne Infections among Pregnant Women Attending Antenatal Care in Jirapa, Upper West Region of Ghana.

Background: Sexually transmitted blood-borne infections (STBBIs) contribute to negative outcomes of pregnancy. Hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), and syphilis infections in pregnancy contribute significantly to maternal and child morbidities and mortalities. This study assessed the prevalence, knowledge, and risk factors of STBBIs (HBV, HCV, HIV, and syphilis) among pregnant women attending antenatal clinics in Jirapa. Methods: A cross-sectional study design involving 246 pregnant women was employed for the study. A structured questionnaire was used to solicit information about the knowledge, prevalence, and risk factors of STBBIs. Results: The overall prevalence of STBBIs was 11.4%; HBV prevalence was 9.8% and 0.8% each for HCV, HIV, and syphilis. About 66% of mothers were aware of mother-to-child transmission of infections during pregnancy. Knowledge of transmission of HIV (93.9%), hepatitis (67.1%), and syphilis (53.7%) in pregnancy was relatively high. Knowledge of risk factors for HIV, hepatitis, and syphilis was 97.6%, 74.4%, and 76.0%, respectively. More than 98% of respondents knew about the prevention of HIV, hepatitis, and syphilis. Significant risk factors associated with and predictive of STBBIs were female genital mutilation (FGM) and gravidity. Conclusion: The occurrence of STBBIs among pregnant women was strongly associated with FGM and gravidity. Public health education should be directed at stopping the practice of FGM and improving reproductive health in the study area.

TRIM32 Deficiency Impairs Synaptic Plasticity by Excitatory-Inhibitory Imbalance via Notch Pathway.

Synaptic plasticity is the neural basis of physiological processes involved in learning and memory. Tripartite motif-containing 32 (TRIM32) has been found to play many important roles in the brain such as neural stem cell proliferation, neurogenesis, inhibition of nerve proliferation, and apoptosis. TRIM32 has been linked to several nervous system diseases including autism spectrum disorder, depression, anxiety, and Alzheimer's disease. However, the role of TRIM32 in regulating the mechanism of synaptic plasticity is still unknown. Our electrophysiological studies using hippocampal slices revealed that long-term potentiation of CA1 synapses was impaired in TRIM32 deficient (KO) mice. Further research found that dendritic spines density, AMPA receptors, and synaptic plasticity-related proteins were also reduced. NMDA receptors were upregulated whereas GABA receptors were downregulated in TRIM32 deficient mice, explaining the imbalance in excitatory and inhibitory neurotransmission. This caused overexcitation leading to decreased neuronal numbers in the hippocampus and cortex. In summary, this study provides this maiden evidence on the synaptic plasticity changes of TRIM32 deficiency in the brain and proposes that TRIM32 relates the notch signaling pathway and its related mechanisms contribute to this deficit.

IL-34 modulates rheumatoid synovial fibroblasts proliferation and migration via ERK/AKT signalling pathway.

OBJECTIVES: The interface between pro-inflammatory cytokines and rheumatoid synovial fibroblast (sFLS) has central effects on rheumatoid arthritis (RA). The present study aimed to explore the role of IL-34 expression as one of major cytokine implicated in RA. METHODS: We examined the expression of IL-34 after RA sFLS stimulated by IL-1ß and TGF-ß1 separately by reverse transcription polymerase chain reaction (RT-PCR). Transwell and wound closure techniques were used to detect whether IL-34 is involved in promoting cell migration. Cellular viability was determined via CCK-8 and cultural morphology assays between IL-34 downregulated group and non-transfected counterpart. We also tested the expression of VEGF gene with RT-PCR analysis and activation of the major signalling pathways by western blot in IL-34 down-regulated group, IL-1ß or TGF-ß1 treated groups. Propidium iodide (PI) staining and fluoresceine isothiocyanate (FITC) Annexin V and propidium iodide apoptosis assay were used to analyse cell cycle arrest and apoptosis separately in IL-34 down-regulated cells. RESULTS: We found that IL-1ß significantly enhanced IL-34 expression, while contrarily, TGF-ß1 restrained IL-34 gene expression. Transwell and wound closure techniques showed that IL-34 was involved considerably in promoting cell migration. However, IL-34 knock-down restricted sFLS migration possibly through the diminishing of MMP2 and MMP9 expression. Interestingly, IL-34 down-regulated cells exhibited significantly low cellular viability compared with the non-transfected counterpart via CCK-8 and cultural morphology assays. We found that IL-34 down-regulated cells have low VEGF gene expression compared with treated cells. PI staining showed a G0/G1 cell cycle arrest in IL-34 down-regulated cells. FITC Annexin V and propidium iodide apoptosis assay verified that IL-34 down-regulated cells induced massive apoptosis through apoptotic signalling caspase3, while IL-1ß treated cells presented termination of cellular apoptosis signalled by BCL-2. Furthermore, we observed IL-34 induced activation of ERK1/2 and AKT pathways while IL-34 down-regulation significantly decreased the activation of these pathways. CONCLUSIONS: Our data add novel insights into the pathogenesis of RA and we suggest that IL-34 plays a dominant role in controlling migration and proliferation of sFLS. Consequently, therapeutic strategies targeting IL-34 could be a potent therapy for RA.

GDC-0941 and CXCL8 (3-72) K11R/G31P combination therapy confers enhanced efficacy against breast cancer.

Aim: Herein is presented the combined effect of PI3K inhibitor (GDC-0941) and CXCR1/2 analogue (G31P) in breast cancer. Materials & methods: Breast cancer cell lines and xenograft model were employed to test the efficacy of the combination therapy. Results: GDC-0941+G31P treatment substantially inhibited multiplication of all the breast cancer cell lines used in this study (BT474, HCC1954 and 4T1). Even though single therapies caused a meaningful S-phase cell cycle arrest, the inhibition effect was more potent with the combined treatment. Similarly, enhanced apoptosis accompanied GDC-0941+G31P treatment. Furthermore, the migration ability of the breast cancer cell lines were significantly curtailed by the combination therapy compared with the single treatments. Conclusion: The findings suggest that combination treatment involving PI3K inhibitor and CXCR1/2 analogue (G31P) could be a potent therapeutic option for breast cancer treatment.

Assessing the knowledge of expectant mothers on mother-to-child transmission of viral hepatitis B in Upper West region of Ghana.

BACKGROUND: Viral Hepatitis B is of a major public health concern globally, especially in developing countries. Expectant mothers' knowledge of Mother-To-Child Transmission (MTCT) of the disease is significant in preventing the spread from an infected mother to her child. This study sought to assess the expectant mothers' knowledge of Mother-To-Child Transmission of viral hepatitis B in the Wa Municipality and Lawra District of Upper West Region, Ghana. METHODS: A descriptive cross-sectional study with a multi-stage sampling technique was employed to select a total of 450 study respondents (expectant mothers), and a semi-structured questionnaire was used for the data collection. Respondents were interviewed using face-to-face interview technique. RESULTS: Majority (54.0%) of the respondents were aged between 25 and 35 years and the results were similar in both districts. Overall, 62.4% (281/450) of the respondents had at least Junior High level education, and 76.2% (343/450) were multigravida. Educational levels among respondents in the two areas were above 50.0% and considered relatively high. Respondents' general knowledge of hepatitis B infection and disease was 46.0% (208/450). However, there was a slight difference between the two districts (40.1% in Lawra District and 51.6% in Wa Municipality). The overall knowledge level on MTCT of viral hepatitis B among the respondents was 34.7% (156/450): the Wa Municipality recorded higher knowledge (43.3%) compared to 24.8% in Lawra District. CONCLUSION: The knowledge level of the expectant mothers on MTCT of viral hepatitis B is relatively low in Upper West Region, Ghana. Majority of the respondents had some form of formal education. The age, marital status, education, occupation, gravity and family setup were found to be associated with knowledge of Hepatitis B infection and MTCT. Thus, there is urgent need to intensify efforts of health staff to educate expectant mothers. In addition, home education and outreach activities should be intensified on HBV infection as well as MTCT. Consequently, planning, implementation and execution of preventive activities, especially in the antenatal clinics should critically consider the social and demographic variations of mothers.

The use of antibiotics during pregnancy: A cross-sectional study of knowledge, attitude, and practices among antenatal care attendees in Northern Ghana.

Background and aims: The promotion of rational use of antibiotics among pregnant women is eminent not only for the risk of teratogenicity in the developing fetus but also the risk of drug resistance with its concomitant high cost of health care. Studies on antibiotic self-medication among pregnant women in Northern Ghana are rare. Improving the knowledge and awareness among the vulnerable groups about the appropriate use of antibiotics can help in limiting the antibiotic resistance menace. We, therefore, conducted this study to assess the knowledge, attitude, and practice (KAP) toward antibiotic use among pregnant women attending an antenatal clinic at a primary health care in Tolon, Northern Region, Ghana. Method: We conducted a cross-sectional study using an interviewer-administered questionnaire to assess the KAP of 702 pregnant women on antibiotic use. This study was conducted in the Tolon Health Center (THC) from March 2021 and ended in October 2021. Results: In this study, 55.6% of pregnant women had good knowledge and 45.3% of them had engaged in self-medication with antibiotics while pregnant. There were statistically significant associations between participants' background and obstetric characteristics and knowledge of antibiotic use and antibiotic resistance, except for age, marital status, and parity. Also, there was a significant association between pregnant women's knowledge and self-medication or over-the-counter purchase of antibiotics. Conclusion: We concluded that higher education level, monthly income, good practice, and good knowledge were significantly associated with a reduced likelihood of self-medication with antibiotics. A well-structured education that could be easily accepted and understood by pregnant women on the risks of antibiotic self-medication should be included in the routine education at the antenatal clinics.

Pregnancy and the risk of NICU admissions in Nandom Municipality of Ghana: A cross-sectional retrospective study.

Background: Neonatal intensive care units (NICU) are specialized units that provide medical attention to neonates, and thus have become a vital aspect in the provision of critical care to infants who are faced with special challenges following birth. Aim: To determine antepartum and intrapartum factors that predispose to NICU admissions in the Nandom Municipal of the Upper West Region of Ghana. Method: This was a cross-sectional retrospective study, spanning from January 1, 2021 to December 31, 2021. Records covering 1777 women who were delivered or had their babies referred to the St. Theresa's Hospital in the Nandom Municipality were involved in the study. Descriptive statistics and multinomial logistic regression analysis were used to compare variables, and statistical significance was determined where the p-value was less than 0.05. Results: From the study, the rate of NICU admission was 10.4%. There was a significant association between mothers who attended less than four antenatal sessions (p = 0.004) and admission to NICU. Nulliparous mothers (p = 0.027) and mothers who presented with multiple pregnancy (p < 0.001) were more likely to have their babies sent to NICU. Both preterm delivery (p < 0.001) and post-term delivery (p < 0.001) were prone to admission to NICU. Also, instrumental delivery (p < 0.001), cesarean section (p < 0.001), low birth weight (p < 0.001), and male infants (p = 0.003) had an increased risk of being admitted to NICU. Furthermore, severe (p < 0.001) and moderate (p < 0.001) birth asphyxia in the first minute following delivery were significantly associated with NICU admission whereas severely asphyxiated babies at 5 min (p < 0.001) were associated with NICU admission. Conclusion: The study revealed a relatively high NICU admission rate in the study area, and the predictors are multifaceted. Tailored intervention programs aimed at curbing these predictors will be required to reduce the rate of NICU admissions in the Nandom Municipality of Ghana.

Drivers of household antibiotic use in urban informal settlements in Northern Ghana: Implications for antimicrobial resistance control.

Background: Urban informal settlements have been described as the epicenters of frequent antibiotic misuse, which has local and global consequences on the goals of antimicrobial stewardship. The aim of this study was to assess the relationship between knowledge, attitude, and practices of antibiotic use among households in urban informal settlements in the Tamale metropolis of Ghana. Method: This study was a prospective cross-sectional survey of the two major informal settlements in the Tamale metropolis, namely Dungu-Asawaba and Moshie Zongo. In all, 660 households were randomly selected for this study. Households with an adult and at least a child under 5 years old were randomly chosen. An adult with knowledge of household healthcare practices was selected  to respond to a structured questionnaire. Results: In all, 291 (44.1%) of the 660 households reported taking at least one type of antibiotic within the last month before the study and 30.9% (204/660) had used antibiotics without a prescription. Information on which antibiotics to use was obtained mostly from friends/family members 50 (24.5%) and were commonly purchased from a medical store or a pharmacy 84 (41.2%), saved up from a previously used antibiotic 46 (22.5%), a friend/family members 38 (18.6%), and drug hawkers 30 (14.7%). Amoxicillin 95 (26.0%) was the most frequently used antibiotic and the commonest indication for antibiotics use was diarrhea 136 (37.9%). Female respondents (odds ratio [OR] = 3.07; 95% confidence interval [CI] = 2.199-4.301; p < 0.0001), larger households (OR = 2.02; 95% CI = 1.337-3.117; p = 0.0011) and those with higher monthly household income (OR = 3.39; 95% CI = 1.945-5.816; p < 0.0001) were more likely to have good knowledge of appropriate antibiotic use and antibiotic resistance. Furthermore, bad attitudes influenced participants' use of antibiotics without prescription (OR = 2.41; 95% CI = 0.432-4.05; p = 0.0009). Conclusion: This study exposes the drivers of inappropriate use of antibiotics at the household level, particularly in urban informal settlements. Policy interventions aimed at controlling the indiscriminate use of antibiotics in such settlements could improve the responsible use of antibiotics. Keywords: antibiotic resistance, informal settlements, Tamale, Ghana.

Transfusion transmissible infections among blood donors in Ghana: A 3-year multicentered health facility-based retrospective study.

Transfusion transmissible infections (TTIs) remain a major health challenge particularly in developing countries. Here, we present a multicentered hospital-based retrospective study on the prevalence, distribution, and risk factors of TTIs in Ghana. Data on blood donors from four health facilities, namely Nkwanta South Municipal Hospital (Oti region), Weija-Gbawe Municipal Hospital (Greater Accra region), SDA Hospital (Northern region) and Wa Municipal Hospital (Upper West region) were extracted and analyzed. Descriptive statistics and multinomial logistic regression were applied to compare sociodemographic data with TTI status. A total of 6094 blood donors were included in this study, and 2% were females. The overall prevalence of TTIs was 21.0% (1232/5868). Specifically, the prevalence of HBV, HCV, HIV, and Syphilis was 6.6% (385/5868), 4.9% (286/5830), 2.9% (168/5867), and 6.8% (393/5739), respectively. Wa dominated in all the viral agents considered in this study, while the Oti region recorded the highest prevalence in T. pallidum. The odds of HBV infection was 3.1 (p = 0.008) among first-time donors, while that for HCV was 2.8 (p = 0.042). For rural dwellers, donors significantly had T. pallidum (p < 0.001; OR = 2.8), HCV (p < 0.001; OR = 2.9), and HIV (p = 0.028; OR = 1.5) infections. Generally, the recipients of transfused blood were predominantly pregnant mothers, followed by children and accident victims. This study has revealed significant disparities and relatively high prevalence of TTIs in Ghana, specifically HBV, HCV, HIV and T. pallidum infections. The variations suggest the presence of unique health challenges per study area, hence the need for a tailored intervention for each study site.

In vitro antimicrobial activity of ethanolic fractions of Cryptolepis sanguinolenta.

BACKGROUND: Following claims that some plants have antimicrobial activities against infectious microbes, the in vitro antimicrobial activities of different solvent fractions of ethanolic extract of Cryptolepis sanguinolenta were evaluated against eight standard bacteria and clinical isolates. METHODS: The solvent partitioning protocol involving ethanol, petroleum ether, chloroform, ethyl acetate and water, was used to extract various fractions of dried pulverized Cryptolepis sanguinolenta roots. Qualitative phyto-constituents screening was performed on the ethanol extract, chloroform fraction and the water fraction. The Kirby Bauer disk diffusion method was employed to ascertain the antibiogram of the test organisms while the agar diffusion method was used to investigate the antimicrobial properties of the crude plant extracts. The microplate dilution method aided in finding the MICs while the MBCs were obtained by the method of Nester and friends. The SPSS 16.0 version was used to analyze the percentages of inhibitions and bactericidal activities. RESULTS: The phytochemical screening revealed the presence of alkaloids, reducing sugars, polyuronides, anthocyanosides and triterpenes. The ethanol extract inhibited 5 out of 8 (62.5%) of the standard organisms and 6 out of 8 (75%) clinical isolates. The petroleum ether fraction inhibited 4 out of 8 (50%) of the standard microbes and 1 out of 8 (12.5%) clinical isolates. It was also observed that the chloroform fraction inhibited the growth of all the organisms (100%). Average inhibition zones of 14.0 ± 1.0 mm to 24.67 ± 0.58 mm was seen in the ethyl acetate fraction which halted the growth of 3 (37.5%) of the standard organisms. Inhibition of 7 (87.5%) of standard strains and 6 (75%) of clinical isolates were observed in the water fraction. The chloroform fraction exhibited bactericidal activity against all the test organisms while the remaining fractions showed varying degrees of bacteriostatic activity. CONCLUSION: The study confirmed that fractions of Cryptolepis sanguinolenta have antimicrobial activity. The chloroform fraction had the highest activity, followed by water, ethanol, petroleum ether and ethyl acetate respectively. Only the chloroform fraction exhibited bactericidal activity and further investigations are needed to ascertain its safety and prospects of drug development.

EnDuo, a novel derivative of Endostar, inhibits the migration of colon cancer cells, suppresses matrix metalloproteinase-2/9 expression and impedes AKT/ERK activation.

BACKGROUND/AIMS: Colon cancer remains a life-threating disease with increasing morbidity and mortality worldwide despite the advancement in modern medical treatment. Therefore, novel and effective anti-colon cancers drugs are urgently needed. In this study, we investigated the anti-metastatic property EnDuo, a modified version of Endostar, and the underlying mechanisms. METHODS: Colon cancer cells were treated with different concentrations of EnDuo (50 µg/mL, 100 µg/mL, 200 µg/mL), and Endostar (100 µg/mL) as positive control. Cell Counting Kit-8 assay was performed to test the effect of EnDuo on cell viability. A scratch wound assay and transwell assay were employed to evaluate the relocation and motility of malignant colon cells following treatment with EnDuo. Western blot analysis was used to determine inhibitory effects of EnDuo by detecting the phosphorylation level of AKT and ERK proteins, and the expression of MMP-2 and MMP-9 proteins. RESULTS: Our results showed that EnDuo impedes the migration of colon cancer cells in a dose-dependent manner. At the molecular level, EnDuo induced a significant reduction in the phosphorylation of AKT and ERK proteins, and inhibited the expression of MMP-2 and MMP-9 proteins. CONCLUSIONS: Collectively, these results demonstrate that EnDuo exhibits a comparable anti-metastatic effect by suppressing the migration of colon cancer cells. Possibly, EnDuo interrupts the PI3K/AKT/ERK signaling pathway to arrest cell migration. Our study provides a novel insight to the potential clinical applications of EnDuo against colon cancers in the future.

TL1A/TNFR2 Axis Enhances Immunoregulatory Effects of Bone Marrow Derived Mesenchymal Stem Cell by Indian Hedgehog Signaling Pathway.

BACKGROUND AND OBJECTIVES: The immunomodulatory potential of mesenchymal stem cells (MSCs) can be regulated by a variety of molecules, especially cytokines. The inflammatory cytokine, TNF-like ligand 1A (TL1A), has been reported as an inflammation stimulator in-multiple autoimmune diseases. Here, we studied the effects of TL1A/TNF-receptor 2 (TNFR2) pathway on the therapeutic potency of bone marrow-derived MSCs (BMSCs). METHODS AND RESULTS: BMSCs, fibroblast-like synoviocytes (FLSs), and H9 and jurkat human T lymphocytes were used in this study. BMSCs paracrine activities, differentiation, proliferation, and migration were investigated after stimulation with TL1A, and intervened with anti-TNFR2. Additionally, the effects of TL1A on BMSCs therapeutic potency were evaluated by treating RA-FLSs, and H9 and jurkat T cells with TL1A-stimulated BMSCs conditioned medium (CM). Indian hedgehog (IHH) involvement was determined by gene silencing and treatment by recombinant IHH (rIHH). TL1A induced BMSCs stemness-related genes, COX-2, IL-6, IDO, TGF-ß and HGF through TNFR2. Also, TL1A corrected biased differentiation and increased proliferation, and migration through TNFR2. Meanwhile, CM of TL1A-stimulated BMSCs decreased the inflammatory markers of RA-FLSs and T cells. Moreover, TL1A-stimulated BMSCs experienced IHH up-regulation coupled with NF-κB and STAT3 signaling up-regulation, while p53 and oxidative stress were down-regulated. Furthermore, treatment of BMSCs by rIHH increased their anti-inflammatory effects. More importantly, knockdown of IHH decreased the ability of TL1A-stimulated BMSCs to alleviating the inflammation in RA-FLSs and T cells. CONCLUSIONS: This study reports the effects of TL1A/TNFR2 pathway on the biological behaviors and therapeutic potency of BMSCs through IHH. These findings could introduce novel procedures to increase the stemness of MSCs in cellular therapy.

Aquaporin 5 promotes tumor migration and angiogenesis in non-small cell lung cancer cell line H1299.

Non-small cell lung cancer (NSCLC) constitutes the majority of all lung-cancer cases. Aquaporin 5 (AQP5) may be involved in NSCLC by promoting lung-cancer initiation and progression. The present study aimed to determine the role of AQP5 in migration and angiogenesis using NSCLC cells and HUVECs. AQPs 1, 3, 4, 5, 8 and 9 were screened in the NSCLC cell line H1299, and the present results showed that AQP5 mRNA was upregulated compared with the other AQP genes. At the protein level, AQP5 was significantly increased in H1299 cells compared with 16HBE cells. AQP5 knockdown in H1299 cells significantly decreased cell migration compared with untransfected cells, as demonstrated by both Transwell and wound closure assays. The present study further investigated H1299 ability to promote HUVEC vascularisation. The supernatants of both transfected and untransfected H1299 cells were used as conditioned medium for HUVECs, and tube formation was measured. The supernatant of AQP5-downregulated cells exhibited significantly low tube formation potential compared with untransfected cells. Similarly, vascular endothelial growth factor was significantly increased in control cells (si-NC) compared with cells transfected with small interfering RNA targeting AQP5. The present study found that AQP5 downregulation significantly decreased the phosphorylation level of epidermal growth factor receptor and the activity of the ERK1/2 pathway. In summary, the present study suggested that AQP5 influenced migration and angiogenesis in NSCLCs in vitro and may potentially exhibit similar in vivo effects.

TL1A/TNFR2-mediated mitochondrial dysfunction of fibroblast-like synoviocytes increases inflammatory response in patients with rheumatoid arthritis via reactive oxygen species generation.

Rheumatoid arthritis (RA) is the major autoimmune destructive disease of joints with a complicated pathogenesis. The contribution of tumor necrosis factor-like ligand 1A (TL1A) in RA pathogenesis, especially on fibroblast-like synoviocytes (FLS), has been suggested clinically. The present study investigated the role of TL1A in mitochondrial dysfunction, induced oxidative stress in mitochondria, apoptosis resistance and the inflammatory response in FLS obtained from RA patients (RA-FLS). RA-FLS were incubated with TL1A and tumor necrosis factor receptor 2 (TNFR2) antagonist. Respiratory function, mitochondrial membrane potential and respiration associated genes of mitochondria were measured in both TL1A stimulated and non-stimulated RA-FLS. Additionally, the effects of TL1A on reactive oxygen species (ROS) production in mitochondria, apoptosis and the inflammatory response in RA-FLS were also assessed. The role of TL1A in association between ROS generation, especially mitochondrial type and the inflammatory response, was evaluated by measuring inflammation-related cytokines and signaling pathways using ROS inhibitors, diphenyleneiodonium chloride and Mito-TEMPO (Sigma-Aldrich, Miamisburg, OH, USA). We found that TL1A induced mitochondrial dysfunction by weakening mitochondrial respiration and membrane potential, which was blocked by a TNFR2 antagonist. Increased ROS synthesis in impaired mitochondria was observed with MitoSOX (Invitrogen, CA, USA) immunofluorescence staining in TL1A-stimulated RA-FLS but inhibited by a TNFR2 antagonist. TL1A influenced apoptosis resistance and inflammatory mediators via TNFR2. Inhibition of mitochondria-derived ROS compromised the production of inflammatory factors in TL1A-stimulated RA-FLS, suggesting that mitochondrial dysfunction mediated by the TL1A/TNFR2 axis might amplify the inflammatory response via regulation of mitochondria-derived ROS generation. Collectively, our results reveal that TL1A might be involved in making FLS more aggressive in RA pathogenesis via cell respiration interruption.

Indian Hedgehog regulates senescence in bone marrow-derived mesenchymal stem cell through modulation of ROS/mTOR/4EBP1, p70S6K1/2 pathway.

Premature senescence of bone marrow-derived mesenchymal stem cells (BMSC) remains a major concern for their application clinically. Hedgehog signaling has been reported to regulate aging-associated markers and MSC skewed differentiation. Indian Hedgehog (IHH) is a ligand of Hedgehog intracellular pathway considered as an inducer in chondrogenesis of human BMSC. However, the role of IHH in the aging of BMSC is still unclear. This study explored the role IHH in the senescence of BMSC obtained from human samples and senescent mice. Isolated BMSC were transfected with IHH siRNA or incubated with exogenous IHH protein and the mechanisms of aging and differentiation investigated. Moreover, the interactions between IHH, and mammalian target of rapamycin (mTOR) and reactive oxygen species (ROS) were evaluated using the corresponding inhibitors and antioxidants. BMSC transfected with IHH siRNA showed characteristics of senescence-associated features including increased senescence-associated ß-galactosidase activity (SA-ß-gal), induction of cell cycle inhibitors (p53/p16), development of senescence-associated secretory phenotype (SASP), activation of ROS and mTOR pathways as well as the promotion of skewed differentiation. Interestingly, BMSC treatment with IHH protein reversed the senescence markers and corrected biased differentiation. Moreover, IHH shortage-induced senescence signs were compromised after mTOR and ROS inhibition. Our findings presented anti-aging activity for IHH in BMSC through down-regulation of ROS/mTOR pathways. This discovery might contribute to increasing the therapeutic, immunomodulatory and regenerative potency of BMSC and introduce a novel remedy in the management of aging-related diseases.

IL-8 antagonist, CXCL8(3-72)K11R/G31P coupled with probiotic exhibit variably enhanced therapeutic potential in ameliorating ulcerative colitis.

Inflammatory bowel disease (IBD) remains a major health challenge due in part to unsafe and limited treatment options, hence there is the need for alternatives. CXCL8/interleukin 8 (IL-8) is elevated in inflammation, and binds preferentially to G protein-couple receptors (GPCRs) CXCR1/2 of the CXC chemokine family to initiate cascades of downstream inflammatory signals. A mutant CXCL8 protein, CXCL8(3-72)K11R/G31P (G31P), competitively and selectively binds to CXCR1/2, making CXCL8 redundant. We explore the therapeutic potential of G31P in dextran sulfate sodium (DSS) induced ulcerative colitis (UC), and the corresponding effect if G31P treatment is augmented with Lactobacillus acidophilus (LACT). The treatment options administered significantly reduced TNF-α, IFN-γ, IL-1ß, IL-6, and IL-8, but maintained elevated levels of IL-10. CD68 and F4/80 expressions were down-regulated and showed restricted infiltration to inflamed colon, while IL-17F levels were insignificantly different from the DSS treated mice. Also, we observed up-regulation of IL-17A in G31P + LACT but not G31P treated mice if compared with Control group. The treatments ameliorated colonic fibrosis by reducing VEGF, TGF-ß, MMP-2 and MMP-9. In addition, we observed elevated levels of E-cadherin, and marginal up-regulation of occludin, suggesting the role of the treatments in regulating tight intestinal junction and adherence proteins. Mechanism-wise, G31P interferes with AKT and ERK signaling pathways. Our study suggests that G31P confers protection in IBD, particularly UC, and when G31P treatment is augmented with Lactobacillus acidophilus, the protection is variably enhanced.

TL1A mediates fibroblast-like synoviocytes migration and Indian Hedgehog signaling pathway via TNFR2 in patients with rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by joints inflammation. One of the aggressive characteristics of RA fibroblast-like synoviocytes (FLS) is the tendency for migration in the local environment, which plays a central role in the RA pathogenesis. Tumor Necrosis Factor (TNF)-like cytokine 1A (TL1A) is a member of TNF superfamily, which has a role in autoimmunity and influences the RA-FLS behavior through TNF receptor 2 (TNFR2). We investigated the effect of TNF-like cytokine 1A (TL1A) on RA-FLS migration using patients' samples. Specifically, we examined the hedgehog signaling pathway which is a key regulator in chondrocyte growth and differentiation. We found that TL1A increased significantly the hedgehog homologue Indian hedgehog (IHH) and its receptor Patched 1, 2 (PTCH 1, 2) in RA-FLS. In addition, TL1A-stimulated RA-FLS promoted significantly IHH protein expression. However, both mRNA and protein levels decreased substantially after blocking TL1A with TNFR2 antagonist. The migratory property of RA-FLS was enhanced after stimulation of RA-FLS with TL1A, but was compromised following TL1A blockage. In conclusion, our study has revealed that TL1A modulated RA-FLS migration and Indian hedgehog signaling pathway using TNFR2.

CXCR1/CXCR2 antagonist G31P inhibits nephritis in a mouse model of uric acid nephropathy.

The prevalence of gout is relatively high worldwide, and many gout patients suffer from uric acid nephropathy (UAN) concomitantly. ELR-CXC chemokines such as CXCL8 and CXCL1 have a elevated expression in UAN. In this research, a mouse UAN model was established for a 12 week duration, and uric acid-related crystals were observed. CXCL8(3-72)K11R/G31P (G31P) is a mutant protein of CXCL8/interleukin 8 (IL-8), which has been reported to have therapeutic efficacy in both inflammatory diseases and malignancies for it acts as a selective antagonist towards CXCR1/CXCR2. In this study, G31P-treated mice showed declined production of the blood urea nitrogen (BUN) level and urine volume in UAN mice compared with G31P-untreated UAN counterparts. In addition, G31P effectively improved renal fibrosis, and reduced uric acid accumulation and leukocyte infiltration in UAN kidneys. Furthermore, the expressions of CXCL1 and CXCL2 were reduced and the activation of NOD-like receptors protein 3 (NLRP3) was inhibited by G31P treatment. This study has demonstrated that G31P attenuates inflammatory progression in chronic UAN, and plays a renoprotective function.

IL-8 analogue CXCL8 (3-72) K11R/G31P, modulates LPS-induced inflammation via AKT1-NF-kß and ERK1/2-AP-1 pathways in THP-1 monocytes.

IL-8 is elevated during inflammation, and it initiates cascade of down-stream reactions. Its antagonist, CXCL8 (3-72) K11R/G31P (G31P), represses inflammatory reactions via competitive binding to CXC chemokine family, preferentially G protein-couple receptors (GPCRs) CXCR1/2. This study reports the effect of G31P on the transcription profile of lipopolysaccharide (LPS) induced inflammation in THP-1 monocytes ex-vivo. LPS (1 µg/ml) induced elevation of IL-8 was significantly reduced by G31P (20 µg/ml and 30 µg/ml), with relatively increased inhibition of CXCR2 than CXCR1. Transcription of IL-1ß, IL-6, and TNF-α were significantly inhibited, while IL-10 remained relatively unchanged. G31P treatment also had repressing effect on the inflammatory associated enzymes COX-2, MMP-2, and MMP-9. Significant restriction of c-Fos, and NF-kß mRNA expression was observed, while that of c-Jun was marginally elevated. Conversely, SP-1 mRNA expression was seen to increase appreciably by G31P treatment. While the translation of pAKT, pERK1/2, and p65- NF-kß were down-regulated by the G31P following THP-1 cells stimulation with LPS, reactive oxygen species (ROS) expression was on the positive trajectory. Collectively, the IL-8 analogue, G31P, modulates the inflammatory profile of LPS induced inflammation in THP-1 monocytes via AKT1-NF-kß and ERK1/2-AP-1 pathways.