Host markers in Quantiferon supernatants differentiate active TB from latent TB infection: preliminary report

<p>Abstract</p> <p>Background</p> <p>Interferon gamma release assays, including the QuantiFERON^®TB Gold In Tube (QFT) have been shown to be accurate in diagnosing <it>Mycobacterium tuberculosis </it>infection. These assays however, do not discriminate between latent TB infection (LTBI) and active TB disease.</p> <p>Methods</p> <p>We recruited twenty-three pulmonary TB patients and 34 household contacts from Cape Town, South Africa and performed the QFT test. To investigate the ability of new host markers to differentiate between LTBI and active TB, levels of 29 biomarkers in QFT supernatants were evaluated using a Luminex multiplex cytokine assay.</p> <p>Results</p> <p>Eight out of 29 biomarkers distinguished active TB from LTBI in a pilot study. Baseline levels of epidermal growth factor (EGF) soluble CD40 ligand (sCD40L), antigen stimulated levels of EGF, and the background corrected antigen stimulated levels of EGF and macrophage inflammatory protein (MIP)-1β were the most informative single markers for differentiation between TB disease and LTBI, with AUCs of 0.88, 0.84, 0.87, 0.90 and 0.79 respectively. The combination of EGF and MIP-1β predicted 96% of active TB cases and 92% of LTBIs. Combinations between EGF, sCD40L, VEGF, TGF-α and IL-1α also showed potential to differentiate between TB infection states. EGF, VEGF, TGF-α and sCD40L levels were higher in TB patients.</p> <p>Conclusion</p> <p>These preliminary data suggest that active TB may be accurately differentiated from LTBI utilizing adaptations of the commercial QFT test that includes measurement of EGF, sCD40L, MIP-1β, VEGF, TGF-α or IL-1α in supernatants from QFT assays. This approach holds promise for development as a rapid diagnostic test for active TB.</p

Long-Term Impact of Neonatal Intake of Oleanolic Acid on the Expression of AMP-Activated Protein Kinase, Adiponectin and Inflammatory Cytokines in Rats Fed with a High Fructose Diet

Abstract: AMP-activated protein kinase (AMPK) is known to regulate both glucose and lipid metabolism, which play vital roles in the development of metabolic syndrome. One way of regulating AMPK is through hormonal activation using adiponectin. Patients diagnosed with type-2 diabetes (T2D) and obesity exhibit low adiponectin concentration levels in their blood. Moreover, studies have also shown that inflammatory processes play a significant role in the etiology of these metabolic diseases. In this study, the long-term effects of neonatal intake of oleanolic acid (OA) on the AMPK gene, genes associated with glucose transport and lipid metabolism, adiponectin levels, and inflammatory biomarkers in rats fed with a high fructose diet were investigated. Seven day old pups were randomly divided into five groups and treated as follows; 0.5% dimethylsulphoxide v/v in distilled water vehicle control (CON), oleanolic acid (OA, 60 mg/kg), high fructose diet (HF, 20% w/v), high fructose diet combined with oleanolic acid (HF+OA), and high fructose diet combined with metformin (HF+MET, 500 mg/kg)..

Vindoline effectively ameliorated diabetes-induced hepatotoxicity by docking oxidative stress, inflammation and hypertriglyceridemia in type 2 diabetes-induced male Wistar rats

CITATION: Gobozaa, M., et al. 2019. Vindoline effectively ameliorated diabetes-induced hepatotoxicity by docking oxidative stress, inflammation and hypertriglyceridemia in type 2 diabetes-induced male Wistar rats. Biomedicine and Pharmacotherapy, 112:108638, doi:10.1016/j.biopha.2019.108638.The original publication is available at https://www.sciencedirect.comENGLISH ABSTRACT: Vindoline, an indole alkaloid present in the leaves of Catharanthus roseus plant, has been recently reported to have insulotropic effects. This present study evaluated the possible hepatoprotective effects of vindoline in a type 2 diabetes mellitus rat model. Diabetes mellitus was induced by exposing rats to 10% fructose water for two weeks followed by a single intraperitoneal injection of 40 mg/kg body weight of streptozotocin (STZ). Rats were randomly divided into six groups (n = 8) and treated daily for 6 weeks with the vehicle via oral gavage, vindoline (20 mg/kg) or glibenclamide (5 mg/kg). Weekly fasting blood glucose (FBG) levels and body weight were measured and recorded. Administration of vindoline significantly (p < 0.05) reduced FBG by 15% when compared to the diabetic controls. Vindoline significantly (p < 0.05) decreased diabetes-induced hepatic injury shown by decreased levels of serum alanine transferase (ALT) (-42%), aspartate aminotransferase (AST) (-42%) and alkaline phosphatase (-62%) compared to the diabetic controls. The oxygen radical absorbance capacity and the activities of superoxide dismutase (SOD) and catalase (CAT) were also improved following treatment with vindoline. The results also showed decreased levels of pro-inflammatory cytokines such as TNF-ɑ by (-41%) and IL-6 (-28%) which may have also contributed to the reduction of serum triglycerides (-65%) in the diabetic group treated with vindoline. Histopathological findings showed improvement of both the hepatic and pancreatic tissues following vindoline treatment. Overall, these findings suggest that vindoline may protect the diabetic hepatic tissue from injury via antioxidant, anti-inflammatory and anti-hypertriglyceredemia mechanisms thereby retarding the development of diabetic complications.https://www.sciencedirect.com/science/article/pii/S0753332218374663?via%3DihubPublisher's versio

Reproductive Parameters in Streptozotocin-Induced Diabetic Male Wistar Rats: Beneficial Role of Basella Alba Aqueous Leave Extract

Background: Diabetes mellitus (DM) is widely reported to have adverse effect on most systems of the body. This study investigated the effects of DM on male reproductive parameters and possible role of aqueous leave extract from Basella alba in ameliorating such effects. Methods: Male rats (n = 40) aged 8-10 weeks were randomly divided into 4 equal groups: Healthy Control (HC), Diabetic Control (DC), Healthy Treatment (HT) and Diabetic Treatment (DT). Diabetes was induced in animal by using (STZ single intraperitoneal injection of streptozotocin (STZ), (55 mg/kg). Treatment was oral via gavage HC and DC groups received 0.5 ml/100 g normal saline daily via oral gavage, HT and DT groups received 200 mg/kg plant extract daily for 4 weeks. Fasting blood sugar and body weights were recorded weekly. Animals were sacrificed upon completion of treatments and tissues were collected for further analysis. Results: There was significant decrease in weights of the body, testis, and epididymis in DC compared to HC rats (P < 0.05). Relative testicular weight was significantly increased in both DC and DT compared to HC groups (P < 0.0001). Sperm concentration, viability and morphology, were all significantly reduced in DC versus HC rats (P<0.0001), but improved in DT versus DC (P < 0.05). Histopathological examination showed degenerative changes in DC specimens that were alleviated in DT rats. Conclusion: Aqueous extract of Basella alba plays a major role in ameliorating male reproductive complications due to streptozotocin-induced diabetes mellitus

Administration of S-allyl cysteine to neonatal rats modulates inflammatory biomarkers in high-fructose-fed rats in adulthood

PURPOSE : To investigate the potential prophylactic effect of S-allyl cysteine (SAC), found in garlic (Allium sativum), against the development of apro-inflammatory status induced by diet in neonatal rats later on in adulthood. METHODS : Suckling Wistar rat pups (4-day-old; male = 21 and female = 21) were randomly allocated to either of 3 groups and orally gavaged daily with the following treatments from postnatal day (PND) 6 – 20: group 1 (control) - 10 mL/kg distilled water; group 2 - 10 mL/kg of 20 % w/v fructose solution (FS) and group 3 - 10 mL/kg FS + SAC. The rat pups were weaned on PND 21, and given ad libitum access to standard rat chow and plain drinking up to PND 115. The rats were euthanized on PND 116 and plasma was collected for the determination of interleukins (IL-1β, IL-4, IL-5, IL-10), vascular endothelial growth factor (VEGF) and monocyte chemotactic protein-1 (MCP-1)] using Bio-Plex Pro magnetic beadbased assays on Bio-Plex platform. RESULTS : Oral administration of FS during suckling increased significantly (p < 0.05) plasma concentrations of IL-5, MCP-1 and VEGF in adult male rats, and plasma MCP-1 in adult female rats. Neonatal oral administration of SAC prevented FS-programmed increase in pro-inflammatory cytokines (p < 0.05) later on in adulthood. CONCLUSION : Oral administration of SAC during the neonatal period protected against FS-induced proinflammatory status and thus, could possibly be exploited as a prophylactic or intervention agent against a pro-inflammatory status induced by a high fructose diet.https://www.tjpr.org/homeam2021Physiolog

Reproductive Parameters in Streptozotocin-Induced Diabetic Male Wistar Rats: Beneficial Role of Basella Alba Aqueous Leave Extract

Background: Diabetes mellitus (DM) is widely reported to have adverse effect on most systems of the body. This study investigated the effects of DM on male reproductive parameters and possible role of aqueous leave extract from Basella alba in ameliorating such effects. Methods: Male rats (n = 40) aged 8-10 weeks were randomly divided into 4 equal groups: Healthy Control (HC), Diabetic Control (DC), Healthy Treatment (HT) and Diabetic Treatment (DT). Diabetes was induced in animal by using (STZ single intraperitoneal injection of streptozotocin (STZ), (55 mg/kg). Treatment was oral via gavage HC and DC groups received 0.5 ml/100 g normal saline daily via oral gavage, HT and DT groups received 200 mg/kg plant extract daily for 4 weeks. Fasting blood sugar and body weights were recorded weekly. Animals were sacrificed upon completion of treatments and tissues were collected for further analysis. Results: There was significant decrease in weights of the body, testis, and epididymis in DC compared to HC rats (P < 0.05). Relative testicular weight was significantly increased in both DC and DT compared to HC groups (P < 0.0001). Sperm concentration, viability and morphology, were all significantly reduced in DC versus HC rats (P<0.0001), but improved in DT versus DC (P < 0.05). Histopathological examination showed degenerative changes in DC specimens that were alleviated in DT rats. Conclusion: Aqueous extract of Basella alba plays a major role in ameliorating male reproductive complications due to streptozotocin-induced diabetes mellitus

Modulatory effects of oleanolic acid on cardiac anti-oxidant status and inflammatory response in high fructose-fed neonatal Sprague-Dawley rats

This present study investigated the antioxidant and inflammatory properties of oleanolic acid (OA) on neonatal rats administered with high fructose diet (HFD). Neonatal rats (24) were assigned at random to four (4) groups namely: Group A (control) which had distilled water only; Group B was administered with OA only; Group C was administered with HFD; Group D received HFD and OA. Animals were administered orally using orogastric gavage at a dosage of 10 ml/kg for 7 days (postnatal day 7-14. The antioxidant status of the hearts such as TEAC, Ferric Reducing Anti-oxidant Power, FRAP, Trolox Equivalence Antioxidant Capacity and oxidative stress biomarkers (MDA, Malondialdehyde and GSH, Glutathione) were evaluated using standard procedures. The levels of inflammatory cytokines in the hearts were determined using magnetic bead-based assays procedure. The TEAC values were significantly decreased in HFD+OA treatment (p < 0.05) in comparison with HFD group. Glutathione concentration in the HFD group had significant increase (p < 0.05) following treatment with oleanolic acid. FRAP values and MDA level were significantly (p < 0.01) elevated post exposure to HFD and treatment with oleanolic acid insignificantly decreased MDA level when compared with HFD group. The pro-inflammatory cytokines (IL-1β, IL-6, IL-12, IFN-γ, TNF-a and MCP-1) were significantly (p < 0.05) increased HFD group when compared to the control. Oleanolic acid administration significantly reduced inflammation in postexposure to HFD. Neonatal intake of oleanolic acid may help to prevent inflammation and oxidative damage in the progression of cardiovascular related diseases.https://www.tjnpr.orgam2023Physiolog

Chronic obstructive pulmonary disease (COPD) : neutrophils, macrophages and lymphocytes in patients with anterior tuberculosis compared to tobacco related COPD

CITATION: Guiedem, E., et al. 2018. Chronic obstructive pulmonary disease (COPD): neutrophils, macrophages and lymphocytes in patients with anterior tuberculosis compared to tobacco related COPD. BMC Research Notes, 11:192, doi:10.1186/s13104-018-3309-6.The original publication is available at https://bmcresnotes.biomedcentral.comObjective: The inflammatory profile of chronic obstructive pulmonary disease (COPD) related to tobacco is known in certain studies while that of the post tuberculosis form is not yet known. This study aimed to evaluate the levels of neutrophils, macrophages and lymphocytes cells in sputum of COPD patients with history of smoking or anterior tuberculosis. Enumeration of cells in samples was analyzed using standard microscopy. Results: We enrolled 92 participants, 46 (50%) were COPD subjects comprising 22 (47.83%) smokers and 24 (52.17%) with anterior tuberculosis while 46 (50%) healthy persons constituted the control group. The levels of neutrophils, lymphocytes and monocytes were statistically higher in COPD patients compared to the control group with p-values of 0.0001 respectively. Neutrophils levels were higher in COPD patients with history of tobacco than in COPD patients with anterior tuberculosis with a mean rate of 4.72 × 106/ml and 2.48 × 106/ml respectively (p = 0.04). The monocytes and lymphocytes levels were not statistically different between the two sub-groups of COPD patients with p-value of 0.052 and 0.91 respectively. Neutrophils are the only inflammatory cells that were significantly higher in COPD patients with history of smoking as compared to COPD patients with anterior tuberculosis.https://bmcresnotes.biomedcentral.com/articles/10.1186/s13104-018-3309-6Publisher's versio

Concurrent evaluation of cytokines improves the accuracy of antibodies against Mycobacterium tuberculosis antigens in the diagnosis of active tuberculosis.

BACKGROUND: Antibodies against mycobacterial proteins are highly specific, but lack sensitivity, whereas cytokines have been shown to be sensitive but not very specific in the diagnosis of tuberculosis (TB). We assessed combinations between antibodies and cytokines for diagnosing TB. METHODS: Immuoglubulin (Ig) A and IgM antibody titres against selected mycobacterial antigens including Apa, NarL, Rv3019c, PstS1, LAM, "Kit 1" (MTP64 and Tpx)", and "Kit 2" (MPT64, Tpx and 19 kDa) were evaluated by ELISA in plasma samples obtained from individuals under clinical suspicion for TB. Combinations between the antibody titres and previously published cytokine responses in the same participants were assessed for diagnosing active TB. RESULTS: Antibody responses were more promising when used in combination (AUC of 0.80), when all seven antibodies were combined. When anti-"Kit 1"-IgA levels were combined with five host cytokine biomarkers, the AUC increased to 97% (92-100%) with a sensitivity of 95% (95% CI, 73-100%), and specificity of 88.5% (95% CI, 68.7-97%) achieved after leave-one-out cross validation. CONCLUSION: When used in combination, IgA titres measured with ELISA against multiple Mycobacterium tuberculosis antigens may be useful in the diagnosis of TB. However, diagnostic accuracy may be improved if the antibodies are used in combination with cytokines

Analysis of host responses to Mycobacterium tuberculosis antigens in a multi-site study of subjects with different TB and HIV infection states in sub-Saharan Africa.

BACKGROUND: Tuberculosis (TB) remains a global health threat with 9 million new cases and 1.4 million deaths per year. In order to develop a protective vaccine, we need to define the antigens expressed by Mycobacterium tuberculosis (Mtb), which are relevant to protective immunity in high-endemic areas. METHODS: We analysed responses to 23 Mtb antigens in a total of 1247 subjects with different HIV and TB status across 5 geographically diverse sites in Africa (South Africa, The Gambia, Ethiopia, Malawi and Uganda). We used a 7-day whole blood assay followed by IFN-γ ELISA on the supernatants. Antigens included PPD, ESAT-6 and Ag85B (dominant antigens) together with novel resuscitation-promoting factors (rpf), reactivation proteins, latency (Mtb DosR regulon-encoded) antigens, starvation-induced antigens and secreted antigens. RESULTS: There was variation between sites in responses to the antigens, presumably due to underlying genetic and environmental differences. When results from all sites were combined, HIV- subjects with active TB showed significantly lower responses compared to both TST(-) and TST(+) contacts to latency antigens (Rv0569, Rv1733, Rv1735, Rv1737) and the rpf Rv0867; whilst responses to ESAT-6/CFP-10 fusion protein (EC), PPD, Rv2029, TB10.3, and TB10.4 were significantly higher in TST(+) contacts (LTBI) compared to TB and TST(-) contacts fewer differences were seen in subjects with HIV co-infection, with responses to the mitogen PHA significantly lower in subjects with active TB compared to those with LTBI and no difference with any antigen. CONCLUSIONS: Our multi-site study design for testing novel Mtb antigens revealed promising antigens for future vaccine development. The IFN-γ ELISA is a cheap and useful tool for screening potential antigenicity in subjects with different ethnic backgrounds and across a spectrum of TB and HIV infection states. Analysis of cytokines other than IFN-γ is currently on-going to determine correlates of protection, which may be useful for vaccine efficacy trials