A multimodality therapeutic application on Toxoplasma gondii encephalitis utilizing Spiramycin and 'de novo' Ferula asafetida in immunodeficient mice.

This study investigated a 'de Novo' medicinal herb, Ferula asafetida (FA), against toxoplasma encephalitis either alone or combined with spiramycin (SP). Female Swiss-Webster mice (n = 72) were divided into three batches. Batch-I received no DMS to serve as an immunocompetent control, batch-II was immune-suppressed with the DMS (0.25 mg/g/day) for 14 days pre-infection, whilst batch-III was immune-suppressed with the DMS on the same day of infection. All experimental mice were inoculated with Toxoplasma gondii ME49 cysts (n = 75). Each batch was split into four subgroups: Mono-SP, mono-FA, combined drug (SP + FA), or neither. Therapies were administered on day zero of infection in batches (I and II) and 35 days post-infection in batch (III). Treatments lasted for 14 days, and mice were sacrificed 60 days post-infection. Histopathological changes, cysts load, and CD4 and CD8 T-cells were counted in brain tissues. The cyst-load count in mice receiving SP + FA was significantly (p < .0001) the least compared to the mono treatments in all protocols. Interestingly, the combined therapy demolished the T-cell subsets to zero in immunocompetent and immunocompromised infected mice. In conclusion, F. asafetida might be a powerfully natural, safe vehicle of SP in the digestive system and/or across the brain-blood barrier to control toxoplasmosis even through immunodeficient conditions.

Regulatory roles of SP-A and exosomes in pneumonia-induced acute lung and kidney injuries.

Introduction: Pneumonia-induced sepsis can cause multiple organ dysfunction including acute lung and kidney injury (ALI and AKI). Surfactant protein A (SP-A), a critical innate immune molecule, is expressed in the lung and kidney. Extracellular vesicles like exosomes are involved in the processes of pathophysiology. Here we tested one hypothesis that SP-A regulates pneumonia-induced AKI through the modulation of exosomes and cell death. Methods: Wild-type (WT), SP-A knockout (KO), and humanized SP-A transgenic (hTG, lung-specific SP-A expression) mice were used in this study. Results: After intratracheal infection with Pseudomonas aeruginosa, KO mice showed increased mortality, higher injury scores, more severe inflammation in the lung and kidney, and increased serum TNF-α, IL-1ß, and IL-6 levels compared to WT and hTG mice. Infected hTG mice exhibited similar lung injury but more severe kidney injury than infected WT mice. Increased renal tubular apoptosis and pyroptosis in the kidney of KO mice were found when compared with WT and hTG mice. We found that serum exosomes from septic mice cause ALI and AKI through mediating apoptosis and proptosis when mice were injected intravenously. Furthermore, primary proximal tubular epithelial cells isolated from KO mice showed more sensitivity than those from WT mice after exposure to septic serum exosomes. Discussion: Collectively, SP-A attenuates pneumonia-induced ALI and AKI by regulating inflammation, apoptosis and pyroptosis; serum exosomes are important mediators in the pathogenesis of AKI.

Is Salvage of a Kidney Graft Possible as a Result of Hyperacute Rejection Immediately After Kidney Transplantation?

Hyperacute rejection is a rare complication of renal transplantation. It is mainly caused by preformed human leukocyte antigen antibodies and can lead to the loss of the transplanted kidney. Renal transplantation is a highly beneficial treatment for people with end-stage renal disease, greatly improving their quality of life. However, antibody-mediated rejection is a significant challenge for the long-term survival of transplanted kidneys. An 18-year-old male with nephrotic syndrome, who underwent bilateral renal nephrectomy due to severe proteinuria, received a living donor kidney. Pretransplant panel reactive antibodies were low. Cytotoxic T- and B-cell and non-HLA cross-match was negative. The graft became cyanotic and mottled within half an hour of transplantation. Allograft was quickly extracted, and a biopsy showed hyperacute rejection. The patient was treated with plasmapheresis, intravenous immunoglobulin, and eculizumab. The graft was successfully re-implanted after 18 hours. Further treatment included additional sessions of plasmapheresis, intravenous immunoglobulin, eculizumab, T-cell-depleting agent, and immunosuppressive therapy. Serum creatinine became stable, and renal biopsy after one month demonstrated intact parenchyma with no inflammation or fibrosis. This case highlights the critical importance of promptly removing the transplanted kidney and using aggressive immunotherapy to save renal allografts in cases of hyperacute rejection.

Mutation in FBXO32 causes dilated cardiomyopathy through up-regulation of ER-stress mediated apoptosis.

Endoplasmic reticulum (ER) stress induction of cell death is implicated in cardiovascular diseases. Sustained activation of ER-stress induces the unfolded protein response (UPR) pathways, which in turn activate three major effector proteins. We previously reported a missense homozygous mutation in FBXO32 (MAFbx, Atrogin-1) causing advanced heart failure by impairing autophagy. In the present study, we performed transcriptional profiling and biochemical assays, which unexpectedly revealed a reduced activation of UPR effectors in patient mutant hearts, while a strong up-regulation of the CHOP transcription factor and of its target genes are observed. Expression of mutant FBXO32 in cells is sufficient to induce CHOP-associated apoptosis, to increase the ATF2 transcription factor and to impair ATF2 ubiquitination. ATF2 protein interacts with FBXO32 in the human heart and its expression is especially high in FBXO32 mutant hearts. These findings provide a new underlying mechanism for FBXO32-mediated cardiomyopathy, implicating abnormal activation of CHOP. These results suggest alternative non-canonical pathways of CHOP activation that could be considered to develop new therapeutic targets for the treatment of FBXO32-associated DCM.

Role of RONS and eIFs in Cancer Progression.

Various research works have piled up conflicting evidence questioning the effect of oxidative stress in cancer. Reactive oxygen and nitrogen species (RONS) are the reactive radicals and nonradical derivatives of oxygen and nitrogen. RONS can act as a double-edged weapon. On the one hand, RONS can promote cancer initiation through activating certain signal transduction pathways that direct proliferation, survival, and stress resistance. On the other hand, they can mitigate cancer progression via their resultant oxidative stress that causes many cancer cells to die, as some recent studies have proposed that high RONS levels can limit the survival of cancer cells during certain phases of cancer development. Similarly, eukaryotic translation initiation factors are key players in the process of cellular transformation and tumorigenesis. Dysregulation of such translation initiation factors in the form of overexpression, downregulation, or phosphorylation is associated with cancer cell's altering capability of survival, metastasis, and angiogenesis. Nonetheless, eIFs can affect tumor age-related features. Data shows that alternating the eukaryotic translation initiation apparatus can impact many downstream cellular signaling pathways that directly affect cancer development. Hence, researchers have been conducting various experiments towards a new trajectory to find novel therapeutic molecular targets to improve the efficacy of anticancer drugs as well as reduce their side effects, with a special focus on oxidative stress and initiation of translation to harness their effect in cancer development. An increasing body of scientific evidence recently links oxidative stress and translation initiation factors to cancer-related signaling pathways. Therefore, in this review, we present and summarize the recent findings in this field linking certain signaling pathways related to tumorigeneses such as MAPK and PI3K, with either RONS or eIFs.

Impact of the factors shaping gut microbiota on obesity.

Obesity is considered as a risk factor for chronic health diseases such as heart diseases, cancer and diabetes 2. Reduced physical activities, lifestyle, poor nutritional diet and genetics are among the risk factors associated with the development of obesity. In recent years, several studies have explored the link between the gut microbiome and the progression of diseases including obesity, with the shift in microbiome abundance and composition being the main focus. The alteration of gut microbiome composition affects both nutrients metabolism and specific gene expressions, thereby disturbing body physiology. Specifically, the abundance of fibre-metabolizing microbes is associated with weight loss and that of protein and fat-metabolizing bacteria with weight gain. Various internal and external factors such as genetics, maternal obesity, mode of delivery, breastfeeding, nutrition, antibiotic use and the chemical compounds present in the environment are known to interfere with the richness of the gut microbiota (GM), thus influencing weight gain/loss and ultimately the development of obesity. However, the effectiveness of each factor in potentiating the shift in microbes' abundance to result in significant changes that can lead to obesity is not yet clear. In this review, we will highlight the factors involved in shaping GM, their influence on obesity and possible interventions. Understanding the influence of these factors on the diversity of the GM and how to improve their effectiveness on disease conditions could be keys in the treatment of metabolic diseases.

Using the Food Skills Questionnaire (FSQ) to Evaluate a Cooking Intervention for University Students: A Pilot Study.

Purpose: To pilot test the Food Skills Questionnaire (FSQ) to evaluate a cooking intervention.Methods: Students attending Western University were invited to participate in 3 cooking classes over a 3-month period. All participants were asked to complete the FSQ pre- and post-intervention. The FSQ evaluated food skills in 3 domains-Food Selection and Planning, Food Preparation, and Food Safety and Storage-with a maximum score of 100 per domain. Domain scores were then computed as a weighted average for the Total Food Skills Score out of 100. Open-ended questions assessed participants' perceptions of the classes.Results: Forty-four students participated. There was a significant increase in food planning (70.6 ± 13.5-77.6 ± 14.3, P < 0.01), food preparation (67.5 ± 14.0-74.9 ± 12.9, P < 0.01), food safety (78.0 ± 9.9-80.8 ± 13.0, P = 0.04), and total food skills (71.9 ± 8.9-77.8 ± 10.6, P < 0.01) post-intervention. Content analysis of open-ended questions indicated that participants enjoyed healthy recipes, supportive Peer Educators, discussions, the cooking experience, socializing, and the safe environment.Conclusions: The FSQ shows strong potential for evaluating basic (e.g., peeling, chopping, slicing) to intermediate (e.g., meal planning) food skills in an effective and feasible manner. It can also capture changes in specific domains, allowing the development of more focused nutrition education and skills-based interventions.

Berberine chloride ameliorated PI3K/Akt-p/SIRT-1/PTEN signaling pathway in insulin resistance syndrome induced in rats.

The liver is the main organ involved in lipid metabolism process and it helps in drug detoxification. Insulin resistance is considered one of risk reasons which lead to several metabolic diseases. Currently, berberine (BER) occupies a huge challenge against multiple diseases with no toxic effect. The present work was aimed to identify, does BER-chloride has a poisonous influence on the liver? and investigating the outcome of BER-chloride on PI3K/Akt-p/SIRT-1/PTEN pathway during insulin resistance syndrome. The insulin resistance model was achieved in experimental female rats via high-fat diet (HFD). Glucose, insulin, lipid profiles, and hepatic oxidative stress parameters were assessed. PI3K, AKt-p, SIRT-1, and PTEN levels in hepatic tissue were determined at genome and protein levels. Further adiponectin concentration was performed in serum, hepatic, and white adipose tissues. Molecular study of fold alteration in insulin, insulin receptor, and retinol binding protein-4 (RBP4) in liver was done. PRACTICAL APPLICATIONS: Obesity syndrome causes multiple obstacles in modern years. The current results revealed elevation the body weight of rats, plasma glucose, homeostatic model assessment, glycated hemoglobin, insulin, and lipid profiles concentrations in a group of rats, which nourished HFD for 8 weeks and this rise, was diminished after 2 weeks from BER-chloride administration. Further, BER-chloride improved transaminases enzymes, pro-oxidant, and antioxidant defense system, PI3K, AKt-p, SIRT-1, and PTEN in the liver, with downregulation of hepatic RBP4. Hence, these data provide a crucial message that BER-chloride enhanced both hepatic function and insulin signaling pathways that might be of therapeutic importance to insulin resistance with no harmful effect on the liver. BER-chloride is predicted to be a drug of choice for obesity complications cure.

Upregulation of sulfatase-1 decreases metastatic potential of SKOV3 human ovarian cancer cells in vitro and in vivo.

AIM: Sulfatase-1 (SULF-1) is one of the genes associated with the inhibition of several signaling pathways by desulfating HSPG in cancer cells. The aim of this study is to investigate the effect of SULF-1 upregulation on SKOV3 ovarian cancer cell line and its influence on cell proliferation, migration, invasion in vitro, and lymph node metastasis in 615 inbred mice in vivo. MATERIALS AND METHODS: In in vitro study, we upregulated SULF-1 in SKOV3 cells using SULF-1 expression plasmid. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting were used to measure SULF-1 expression levels after stable upregulation. CCK-8, flow cytometry, Boyden Transwell-chamber, and scratch-wound healing assay were performed to explore the effect of SULF-1 on the proliferation, migration, and invasion. In in vivo study, immunohistochemistry and eosin stain (H and E) were used to evaluate the expression level of SULF-1 gene and to measure the lymph node metastatic rate of mice inoculated with SULF-1-SKOV3-expressed plasmid, SKOV3, and Nc-SKOV3 cells. RESULTS: qRT-PCR and western blot assay confirmed that SULF-1 was upregulated both in mRNA and protein levels. Following SULF-1 stable upregulation, the cell proliferation, migration, and invasion were significantly reduced in the SULF-1 upregulated cells (SULF-1-SKOV3) compared with the nontransfected (SKOV3) and the nonspecific sequence transfected cells (Nc-SKOV3). IHC results showed that SULF-1 was highly expressed after stably upregulation in SKOV3 cells, and H and E stain confirmed that the mice inoculated with SULF-1-SKOV3 cells decreased lymph node metastatic rate compared to the two control groups. CONCLUSIONS: Our findings showed that overexpression of SULF-1 in SKOV3 results in a decrease in ovarian cancer cell proliferation, migration, and invasion in vitro and decreased lymph node metastasis in vivo. This finding could have a potential therapeutic window in the management of ovarian cancer.

Soluble resistance-related calcium-binding protein in cancers.

Soluble resistance-related calcium binding protein (Sorcin) is an oncoprotein expressed at high levels in human cancers and confers multidrug resistance (MDR) in several tumors. Sorcin participates in a number of neoplastic processing including metastasis and apoptosis. In this review, we summarize and discuss the relationship of Sorcin with tumors as well as its regulatory mechanisms. Sorcin is increasingly considered as a potential molecular target for therapeutic intervention.

Sulfatase-1 knockdown promotes in vitro and in vivo aggressive behavior of murine hepatocarcinoma Hca-P cells through up-regulation of mesothelin.

Our previous study (Oncotarget 2016; 7:46) demonstrated that the over-expression of sulfatase-1 in murine hepatocarcinoma Hca-F cell line (a murine HCC cell with lymph node metastatic [LNM] rate of >75%) downregulates mesothelin and leads to reduction in lymphatic metastasis, both in vitro and in vivo. In current work, we investigated the effects of Sulf-1 knockdown on mesothelin (Msln) and it's effects on the in vitro cell proliferation, migration, invasion, and in vivo tumor growth and LNM rate for Hca-P cells (a murine HCC cell with LNM rate of <25%). Western blotting and qRT-PCR assay indicated that both in vitro and in vivo Sulf-1 was down-regulated by 75% and 68% and led to up regulation of Msln by 55% in shRNA-transfected-Sulf-1-Hca-P cells compared with Hca-P and nonspecific sequence control plasmid transfected Hca-P cell (shRNA-Nc-Hca-P). The in vitro proliferation, migration and invasion potentials were significantly enhanced following Sulf-1 stable down-regulation. In addition, Sulf-1 knock-down significantly promoted tumor growth and increased LNM rates of shRNA-Sulf-1-Hca-P-transplanted mice by 78.6% (11 out of 14 lymph nodes were positive of cancer). Consistent with our previous work, we confirmed that Sulf-1 plays an important role in hepatocarcinoma cell proliferation, migration, invasion and metastasis. The interaction between Sulf-1 and Msln is a potential therapeutic target in the development of liver cancer therapy.

Ezrin as a possible diagnostic and/or prognostic biomarker in mice lymphatic metastatic hepatocellular carcinoma in vivo.

Hepatocellular carcinoma (HCC) ranks in the top of cancers leading to death. Early diagnosis is the big challenge in the case of HCC. Our in vitro study showed that Ezrin expression in lymphatic metastasis hepatocellular carcinoma (LNM-HCC) was associated with the metastatic rate. Here we aim to evaluate Ezrin expression as diagnostic and/or prognostic biomarker of LNM-HCC in mice. Chinese inbred 615 mice, Hca-F and Hca-P cell lines were used in the study. Histological changes were determined by Hematoxylin and Eosin, while Ezrin expression was assessed by qRT-PCR, western blot, immunohistochemistry, and enzyme-linked immunosorbent assay. Ezrin expression in this study gives credit to our in vitro study which Ezrin expression was positively correlated with LNM-HCC and negatively with Annexin7 (A7) expression. The highest histological changes were observed in high metastatic primary/secondary tumors combined with high Ezrin expression. Ezrin and A7 are higher in total primary tumors than in total secondary tumors (P = 0.0001, P = 0.021), respectively. Ezrin expression was enhanced in Hca-P A7 down-regulated primary/secondary tumors (P = 0.004), whereas, Ezrin expression was suppressed in Hca-F A7 upregulated primary/secondary tumors. Serum ELISA indicated differential expression of Ezrin among the study groups (P ≤ 0.0001). Ezrin expression was higher in NC-Hca-F than NC-Hca-P (P ≤ 0.0001), suppressed in Hca-F A7 upregulation (P ≤ 0.0001) and in enhanced in Hca-P A7 down-regulation (P = 0.0001). In conclusion, Ezrin level may serve as a differential diagnostic and/or prognostic biomarker for high and low LNM-HCC and may be beneficial in the diagnosis of HCC disease. © 2017 BioFactors, 43(5):662-672, 2017.

Ezrin expression is altered in mice lymphatic metastatic hepatocellular carcinoma and subcellular fractions upon Annexin 7 modulation in-vitro.

Ezrin and Annexin seven (A7) have been suggested to be involved in several roles in cancers metastasis. However, the role of Ezrin and the effect of A7 on Ezrin expression in lymphatic metastatic hepatocellular carcinoma (LNM-HCC) have not been extensively explored yet. This study reports expression of Ezrin in high lymphatic metastasis (Hca-F >70%) and low metastatic metastasis (Hca-P <30%) HCC cell lines, and the effect of A7 on Ezrin expression. Real-Time PCR, Western blot, Subcellular fractionation, Immunocytochemistry and Immunofluorescence were used to investigate Ezrin expression in addition to migration and invasion behaviors of A7 up-regulated Hca-F cells, A7 down-regulated Hca-P and in their respective negative control (NC) cells. Ezrin expression was higher in high LNM-HCC than low LNM-HCC (p=0.0046). Cell fractionation analysis reveals that Ezrin was highly present in the cytoplasm, nucleus and cytoskeleton of NC-Hca-F cells. However, Ezrin was highly observed in the cell membrane, nucleus and cytoskeleton of NC-Hca-P cells. A7 up-regulation in Hca-F suppressed Ezrin expression (p=0.0248), but increase the migration and invasion, whereas Ezrin was mainly located in the cytoplasm and nucleus fractions. Down-regulation of A7 in Hca-P cells, enhanced Ezrin expression (p<0.0001) in the cytoplasm and nucleus fractions, and suppressed migration and invasion. In conclusion, Ezrin may play a role in LNM-HCC and might be inversely associated with A7 expression. The subcellular localization of Ezrin and A7 was varied according to the metastatic levels. Ezrin may thus be a potential diagnostic and/or prognostic biomarker for HCC.

Overexpression of sulfatase-1 in murine hepatocarcinoma Hca-F cell line downregulates mesothelin and leads to reduction in lymphatic metastasis, both in vitro and in vivo.

Lymphatic vessels function as transport channels for tumor cells to metastasize from the primary site into the lymph nodes. In this experiment we evaluated the effect of Sulfatase-1 (Sulf-1) on metastasis by upregulating it in murine hepatocarcinoma cell line Hca-F with high lymph node metastatic rate of >75%. The study in vitro showed that up regulation of Sulf-1 in Hca-F cells significantly reduced cell proliferation, migration and invasion (p<0.05). Also, the forced expression of Sulf-1 down regulated Mesothelin (Msln) at both the protein and mRNA levels. The experiment in vivo further showed that up-regulation of Sulf-1 with the attendant downregulation of mesothelin delayed tumor growth and decreased lymph node metastasis. In conclusion, our findings show that Sulf-1 is an important tumor suppressor gene in hepatocellular carcinoma (HCC), and its over expression downregulates Msln and results in a decrease in HCC cell proliferation, migration, invasion, and lymphatic metastasis. This functional relationship between Sulf-1 and Msln could be exploited for the development of a novel liver cancer therapy.

Adverse Long-term Metabolic and Endometrial Consequences in Women with Polycystic Ovarian Syndrome: A Report of Two Cases.

BACKGROUND: Little prospective data exist on associations between polycystic ovary syndrome (PCOS) and long-term metabolic disorders/endometrial malignancy in women. We report the cases of 2 obese, infertile women with PCOS who experienced long-term adverse metabolic and endometrial consequences. CASE: The first woman developed type 2 diabetes and hypertension after a 10-year history of PCOS and subsequent endometrioid carcinoma after having PCOS for 16 years. The second woman was first diagnosed with endometrial atypical hyperplasia, which failed to respond to high-dose progestin treatment, and then was diagnosed with PCOS after a 10-year history of oligomenorrhea. Laboratory testing revealed dyslipidemia and impaired glucose tolerance. Both women had a family history of diabetes. The second woman was administered oral contraceptives and metformin. Her endometrial atypia regressed and insulin sensitivity improved after 6 months of treatment. CONCLUSION: PCOS may be associated with long-term adverse metabolic and endometrial consequences. Timely diagnosis and appropriate monitoring and management should be emphasized in order to prevent these women from adverse long-term complications. Oral contraceptives in combination with metformin reversed endometrial atypical hyperplasia in the obese, progestin-resistant woman with PCOS. Further clinical studies may be needed to confirm the effectiveness of such treatment on endometrial atypia in obese, progestin-resistant women for fertility sparing.

Epigenetics and chromatin remodeling in adult cardiomyopathy.

The manipulation of chromatin structure regulates gene expression and the flow of genetic information. Histone modifications and ATP-dependent chromatin remodeling together with DNA methylation are dynamic processes that modify chromatin architecture and profoundly modulate gene expression. Their coordinated control is key to ensuring proper cell commitment and organ development, as well as adaption to environmental cues. Recent studies indicate that abnormal epigenetic status of the genome, in concert with alteration of transcriptional networks, contribute to the development of adult cardiomyopathy such as pathological cardiac hypertrophy. Here we consider the emerging role of different classes of chromatin regulators and how their dysregulation in the adult heart alters specific gene programs with subsequent development of major cardiomyopathies. Understanding the functional significance of the different epigenetic marks as points of genetic control may represent a promising future therapeutic tool.