Effects of Probiotics at the Interface of Metabolism and Immunity to Prevent Colorectal Cancer-Associated Gut Inflammation: A Systematic Network and Meta-Analysis With Molecular Docking Studies.

Background: Dysbiosis/imbalance in the gut microbial composition triggers chronic inflammation and promotes colorectal cancer (CRC). Modulation of the gut microbiome by the administration of probiotics is a promising strategy to reduce carcinogenic inflammation. However, the mechanism remains unclear. Methods: In this study, we presented a systematic network, meta-analysis, and molecular docking studies to determine the plausible mechanism of probiotic intervention in diminishing CRC-causing inflammations. Results: We selected 77 clinical, preclinical, in vitro, and in vivo articles (PRISMA guidelines) and identified 36 probiotics and 135 training genes connected to patients with CRC with probiotic application. The meta-analysis rationalizes the application of probiotics in the prevention and treatment of CRC. An association network is generated with 540 nodes and 1,423 edges. MCODE cluster analysis identifies 43 densely interconnected modules from the network. Gene ontology (GO) and pathway enrichment analysis of the top scoring and functionally significant modules reveal stress-induced metabolic pathways (JNK, MAPK), immunomodulatory pathways, intrinsic apoptotic pathways, and autophagy as contributors for CRC where probiotics could offer major benefits. Based on the enrichment analyses, 23 CRC-associated proteins and 7 probiotic-derived bacteriocins were selected for molecular docking studies. Results indicate that the key CRC-associated proteins (e.g., COX-2, CASP9, PI3K, and IL18R) significantly interact with the probiotic-derived bacteriocins (e.g., plantaricin JLA-9, lactococcin A, and lactococcin mmfii). Finally, a model for probiotic intervention to reduce CRC-associated inflammation has been proposed. Conclusion: Probiotics and/or probiotic-derived bacteriocins could directly interact with CRC-promoting COX2. They could modulate inflammatory NLRP3 and NFkB pathways to reduce CRC-associated inflammation. Probiotics could also activate autophagy and apoptosis by regulating PI3K/AKT and caspase pathways in CRC. In summary, the potential mechanisms of probiotic-mediated CRC prevention include multiple signaling cascades, yet pathways related to metabolism and immunity are the crucial ones.

Outcomes of Left Bundle Branch Area Pacing for Cardiac Resynchronization Therapy: An Updated Systematic Review and Meta-analysis.

BACKGROUND: Real-world data on the use of left bundle branch area pacing (LBBAP) as an alternative novel pacing strategy to biventricular pacing (BVP) for cardiac resynchronization therapy (CRT) remains scarce. We aimed to investigate the outcomes of LBBAP as an alternative to BVP as a method of CRT. METHODS: Electronic databases were searched for studies on the use of LBBAP as CRT and studies that compared LBBAP with BVP. The main outcomes examined were changes in New York Heart Association classification, left ventricular end-diastolic diameter, left ventricular ejection fraction, and paced QRS duration post CRT device implantation. RESULTS: Our meta-analysis included 8 nonrandomized studies with a total of 527 patients who underwent LBBAP as CRT. In studies with a BVP comparison group, patients with LBBAP had a greater reduction in paced QRS (mean difference [MD], 27.91 msec; 95% confidence interval [CI], 22.33-33.50), and a greater improvement in New York Heart Association class (MD, 0.59; 95% CI, 0.28-0.90) and left ventricular ejection fraction (MD, 6.77%; 95% CI, 3.84-9.71). Patients with underlying left bundle branch block appeared to benefit the most from LBBAP compared with patients without underlying left bundle branch block. CONCLUSIONS: LBBAP might be a reasonable option for patients who meet indications for CRT, particularly in those who have limited anatomy or do not benefit from CRT. Randomized trials are needed to compare LBBAP with BVP for CRT and to identify which populations might benefit the most from LBBAP.

CONTEXTE: On dispose de peu de données obtenues en contexte réel sur l'utilisation de la stimulation de la branche gauche (SBG) comme nouvelle stratégie remplaçant la stimulation biventriculaire (SBV) dans le cadre d'une thérapie de resynchronisation cardiaque (TRC). Nous avons voulu étudier les résultats de la SBG à titre de solution de rechange à la SBV comme méthode de TRC. MÉTHODOLOGIE: Nous avons cherché dans les bases de données électroniques les études examinant l'utilisation de la SBG comme TRC, et les études comparant la SBG à la SBV. Les principaux résultats examinés étaient les changements dans les classes de la New York Heart Association (NYHA), le diamètre télédiastolique du ventricule gauche, la fraction d'éjection du ventricule gauche (FEVG) et la durée du QRS stimulé après l'implantation du dispositif de TRC. RÉSULTATS: Notre méta-analyse portait sur huit études sans répartition aléatoire, portant sur un total de 527 patients ayant subi une SBG comme TRC. Dans les études comportant un groupe témoin ayant subi une SBV, les patients ayant subi une SBG présentaient une réduction plus importante du QRS stimulé (différence moyenne [DM] : 27,91 ms; intervalle de confiance [IC] à 95 % : 22,33-33,50), ainsi qu'une amélioration plus importante des classes de la NYHA (MD : 0,59; IC à 95 % : 0,28-0,90) et de la FEVG (MD : 6,77 %; IC à 95 % : 3,84-9,71). Les patients avec un bloc de la branche gauche (BBG) sous-jacent ont semblé bénéficier davantage de la SBG que les patients sans BBG sous-jacent. CONCLUSIONS: La SBG peut être une option raisonnable pour les patients chez qui la TRC est indiquée, en particulier ceux qui ont des restrictions sur le plan de l'anatomie ou qui ne bénéficient pas de la TRC. Des essais randomisés sont nécessaires pour comparer la SBG à la SBV comme TRC, et pour déterminer les populations qui pourraient bénéficier le plus de la SBG.

Systematic Network and Meta-analysis on the Antiviral Mechanisms of Probiotics: A Preventive and Treatment Strategy to Mitigate SARS-CoV-2 Infection.

With the alarming rise of infected cases and deaths, COVID-19 is a pandemic, affecting 220 countries worldwide. Until now, no specific treatment is available against SARS-CoV-2. The causal virus SARS-CoV-2 primarily infects lung cells, leading to respiratory illness ranging in severity from the common cold to deadly pneumonia. This, with comorbidities, worsens the clinical outcome, particularly for immunosuppressed individuals with COVID-19. Interestingly, the commensal gut microbiota has been shown to improve lung infections by modulating the immune system. Therefore, fine-tuning of the gut microbiome with probiotics could be an alternative strategy for boosting immunity and treating COVID-19. Here, we present a systematic biological network and meta-analysis to provide a rationale for the implementation of probiotics in preventing and/or treating COVID-19. We have identified 90 training genes from the literature analysis (according to PRISMA guidelines) and generated an association network concerning the candidate genes linked with COVID-19 and probiotic treatment. The functional modules and pathway enrichment analysis of the association network clearly show that the application of probiotics could have therapeutic effects on ACE2-mediated virus entry, activation of the systemic immune response, nlrp3-mediated immunomodulatory pathways, immune cell migration resulting in lung tissue damage and cardiovascular difficulties, and altered glucose/lipid metabolic pathways in the disease prognosis. We also demonstrate the potential mechanistic domains as molecular targets for probiotic applications to combat the viral infection. Our study, therefore, offers probiotics-mediated novel preventive and therapeutic strategies for COVID-19 warfare.

Hypothyroidism in Pancreatic Cancer: Role of Exogenous Thyroid Hormone in Tumor Invasion-Preliminary Observations.

According to the epidemiological studies, about 4.4% of American general elderly population has a pronounced hypothyroidism and relies on thyroid hormone supplements daily. The prevalence of hypothyroidism in our patients with pancreatic cancer was much higher, 14.1%. A retrospective analysis was performed on patients who underwent pancreaticoduodenectomy (Whipple procedure) or distal pancreatectomy and splenectomy (DPS) at Thomas Jefferson University Hospital, Philadelphia, from 2005 to 2012. The diagnosis of hypothyroidism was correlated with clinicopathologic parameters including tumor stage, grade, and survival. To further understand how thyroid hormone affects pancreatic cancer behavior, functional studies including wound-induced cell migration, proliferation, and invasion were performed on pancreatic cancer cell lines, MiaPaCa-2 and AsPC-1. We found that hypothyroid patients taking exogenous thyroid hormone were more than three times likely to have perineural invasion, and about twice as likely to have higher T stage, nodal spread, and overall poorer prognostic stage (P < 0.05). Pancreatic cancer cell line studies demonstrated that exogenous thyroid hormone treatment increased cell proliferation, migration, and invasion (P < 0.05). We conclude that exogenous thyroid hormone may contribute to the progression of pancreatic cancer.

Conserved Metabolic Changes in Nondiabetic and Type 2 Diabetic Bariatric Surgery Patients: Global Metabolomic Pilot Study.

The goal of this study was to provide insight into the mechanism by which bariatric surgical procedures led to weight loss and improvement or resolution of diabetes. Global biochemical profiling was used to evaluate changes occurring in nondiabetic and type 2 diabetic (T2D) patients experiencing either less extreme sleeve gastrectomy or a full gastric bypass. We were able to identify changes in metabolism that were affected by standard preoperation liquid weight loss diet as well as by bariatric surgery itself. Preoperation weight-loss diet was associated with a strong lipid metabolism signature largely related to the consumption of adipose reserves for energy production. Glucose usage shift away from glycolytic pyruvate production toward pentose phosphate pathway, via glucose-6-phosphate, appeared to be shared across all patients regardless of T2D status or bariatric surgery procedure. Our results suggested that bariatric surgery might promote antioxidant defense and insulin sensitivity through both increased heme synthesis and HO activity or expression. Changes in histidine and its metabolites following surgery might be an indication of altered gut microbiome ecology or liver function. This initial study provided broad understanding of how metabolism changed globally in morbidly obese nondiabetic and T2D patients following weight-loss surgery.

Osteopontin (OPN) isoforms, diabetes, obesity, and cancer; what is one got to do with the other? A new role for OPN.

Alternative splicing of osteopontin (OPN) produces three isoforms: OPNa, OPNb, and OPNc. The aims of this study were to examine the expression profile of OPN isoforms in sera from patients with pancreatic lesions and to determine their correlation with the presence of comorbid systemic inflammatory conditions, such as diabetes and/or obesity. Sera from 90 patients undergoing pancreatic surgery and 29 healthy volunteers were analyzed. Seventeen patients were diabetics, 17 were obese, and 6 had both diabetes and obesity. In patients with pancreatic lesions, OPNb was expressed in 48% of the patients' sera, OPNc in 34%, and both in 5%. The presence of diabetes and/or obesity was associated with complete disappearance of OPNb and expression of only OPNc. OPNc presence was significantly associated with diabetes and obesity (OR = 7.06 [95% CI 1.97-23.3]; p = 0.003). No OPNb or OPNc was detected in the normal sera. Overexpression of OPNb and OPNc isoforms in PDA cells significantly (p < 0.05) increased their activity in soft-agar colony formation and wound healing assays, induced the transcription of interleukin (IL)-6, and reduced tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), and IL-10. Our data show for the first time the significant association between serum OPNc and diabetes and/or obesity. Unraveling the functional role of OPN isoforms in systemic inflammation is essential to understanding their significance as therapeutic targets in diabetes and obesity, and during metastasis development in PDA.

Tubeless tracheotomy for survival airway surgery in the leporine model.

OBJECTIVES/HYPOTHESIS: The ideal animal experimental tracheostomy technique is one that is 1) safe and easy to perform, 2) requires no tracheostomy tube, and 3) requires minimal cleaning or suctioning to maintain patency. The leporine model for airway injury has been well established and offers an inexpensive and practical animal model for experimental evaluation. However, previous research has demonstrated a high mortality rate with survival airway surgery in rabbits. This study demonstrates the feasibility of airway management in the leporine model using a simple maturing suture tracheostomy that avoids a tracheostomy tube. STUDY DESIGN: Tracheostomy was performed in six New Zealand white rabbits in the setting of survival surgery over a 2-week study period. METHODS: A vertical tracheal incision was made from the second to the sixth tracheal ring. The anterior portion of the tracheal rings was removed and the skin surrounding the stoma was sutured down to the tracheal wall. The lateral tracheal wall was then suspended to the soft tissue in the lateral neck. RESULTS: All six rabbits survived the study period with minimal care and maintained stoma patency until sacrifice. Granulation tissue and edema were noted during the first week and largely resolved by the second week. An average of 5-mm-diameter stoma was measured 14 days after surgery without intraluminal stenosis or laryngeal edema. CONCLUSIONS: This method meets the defined criteria for the ideal experimental tracheostomy, demonstrating potential benefit in a laryngotracheal stenosis model and a rabbit model of evoked phonation.

Contrast-enhanced ultrasound-guided Sentinel lymph node biopsy of the ocular conjunctiva.

OBJECTIVES/HYPOTHESIS: Sentinel lymph node biopsy (SLNB) has been utilized for cutaneous melanoma and other malignancies arising from the eye and ocular adnexa. Currently, SLNB requires blue dyes and/or radiopharmaceuticals; both of which have significant shortcomings. This study sought to evaluate the feasibility of SLNB with the use of real-time, contrast-enhanced ultrasound (CEUS) as an alternative technique for tumors arising in the conjunctiva. STUDY DESIGN: Prospective feasibility study in a porcine model. METHODS: Twelve experiments were performed on six non-tumor-bearing Yorkshire swine. An ultrasound contrast agent, Sonazoid (GE Healthcare, Oslo, Norway), (99m) technetium ((99m) Tc), and methylene blue (MB) (Covidien, Mansfield, MA) were injected in the ocular conjunctiva. Sentinel lymph nodes (SLNs) were localized with CEUS and findings were compared to that of MB and (99m) Tc. Fisher exact test was used. RESULTS: Contrast-enhanced SLNs were identified within an average of 6.2 minutes from time of injection of Sonazoid. A total of 17 SLNs were identified by at least one of the three techniques. Correlation between Sonazoid and (99m) Tc was 94.1% (16/17 SLNs). Correlation between (99m) Tc and MB was 88.2% (15/17). One SLN that was positive for (99m) Tc but negative for Sonazoid and was considered to be a false positive (1/17); findings were similar for MB (1/17). Differences between the three techniques were not significant (P = .886). CONCLUSIONS: CEUS-guided injection of conjunctiva for SLNB is technically feasible and correlates well with standard detection techniques. This technique shows promise for rapid, real-time, intraoperative imaging for SLNB, using a widely available imaging modality and avoiding the need for radiopharmaceuticals. LEVEL OF EVIDENCE: NA

Overexpressing TNF-alpha in pancreatic ductal adenocarcinoma cells and fibroblasts modifies cell survival and reduces fatty acid synthesis via downregulation of sterol regulatory element binding protein-1 and activation of acetyl CoA carboxylase.

The effect of tumor necrosis factor-alpha (TNF-α) gene delivery has been suggested as a potentially useful therapeutic approach to improve the chemotherapeutic treatment of patients with pancreatic ductal adenocarcinoma (PDA), but the exact mechanism of its action is not clearly understood. In this study, we analyzed the expression profile of TNF-α in PDA tissue and explored its potential role in fatty acid synthase (FAS) regulation in PDA cells and in fibroblasts. Quantitative real-time polymerase chain reaction was used to examine the expression of TNF-α in PDA, matching adjacent tissues, and benign lesions. Logistic regression models with robust variance were used to analyze the gene expression levels, and Kaplan-Meier survival curves were generated. In vitro, we overexpressed the TNF-α gene in PDA cells and fibroblasts and analyzed its effect on cell survival, migration, and on members of the FAS signaling pathway. We also evaluated TNF-α effects on a panel of inflammation-, angiogenesis-, and metastasis-related markers. In the tumor tissue of PDA patients, compared with their matched adjacent tissue, expression levels of TNF-α were not statistically different and did not correlate with survival or any other examined clinicopathological features. Overexpression of TNF-α significantly (p < 0.05) reduced PDA and fibroblast cell migration. In PDA cells that highly overexpress TNF-α, this was associated with a significant reduction of FAS mRNA and protein expression levels and significant (p < 0.05) reduction of SREBP-1 and ACC mRNA. Reduction of FAS by TNF-α was inhibited when either SREBP-1 or ACC was knocked down by siRNA. PDA cells and fibroblasts that overexpress TNF-α displayed differential regulation of several inflammation-related markers and reduced levels of metastasis-related genes. Our data demonstrate a previously unknown multi-targeted involvement of TNF-α in PDA lipogenesis and inflammation and metastasis and suggest that intratumoral introduction of TNF-α may have the potential as a novel therapeutic approach in human PDA.

Differential expression of cytochrome P450 omega-hydroxylase isoforms and their association with clinicopathological features in pancreatic ductal adenocarcinoma.

BACKGROUND: The cytochrome P450 (CYP) superfamily consists of enzymes that catalyze the oxidation of lipids, steroids, and drugs. In particular, the CYP4 family plays an essential role in lipid metabolism by the ω-hydroxylation of terminal ends of fatty acids. Disturbance of this system has been associated with increased angiogenesis, proliferation, and metastasis of several cancers. This study aimed to detect the expression of CYP4 isoforms (CYP4A11, CYP4F2, CYP4F3) in pancreatic ductal adenocarcinoma (PDA) and their association with clinicopathological features. METHODS: Pancreatic specimens were collected from 73 patients who underwent surgical resection at the Thomas Jefferson University Hospital. Quantitative polymerase chain reaction was used to examine the cytochrome P450 isoforms in PDA (n = 62), adjacent-normal (n = 30), and benign tissues (n = 11). Logistic regression models were used to analyze gene expression among tissue types. Spearman rank correlations were calculated for isoform expression and for age. Differences in expression by gender were assessed via t test. Other clinicopathological variables (diabetes, smoking, obesity, T stage, perineural invasion, nodal status) were analyzed by Wilcoxon rank sum. RESULTS: CYP4 expression for isoforms was significantly higher in PDA tissues versus matched-adjacent tissues (p < 0.01). PDA tumors expressed significantly higher levels of CYP4F2 and CYP4F3 when compared to benign lesions (p < 0.01). Significant associations were found between low levels of CYP4F2 and CYP4F3 and increased age of PDA patients. Interestingly, all isoforms were expressed at higher levels in male patients. CONCLUSIONS: Transcriptional upregulation of cytochrome P450 ω-hydroxylase suggests that these enzymes have the potential to be used as distinguishing markers in pancreatic pathology.

RAN GTPase and Osteopontin in Pancreatic Cancer.

INTRODUCTION: Pancreatic ductal adenocarcinoma (PDA) has the worst prognosis among cancers, mainly due to the high incidence of early metastases. RAN small GTPase (RAN) is a protein that plays physiological roles in the regulation of nuclear transport and microtubule spindle assembly. RAN was recently shown to mediate the invasive functions of the prometastatic protein osteopontin (OPN) in breast cancer cells. We and others have shown previously that high levels of OPN are present in PDA. In this study, we analyzed the expression and correlation of RAN with OPN in human pancreatic lesions, and explored their regulation in PDA cell lines. METHODS: Real time PCR was used to analyze RAN and OPN mRNA levels in PDA, adjacent non-malignant, and benign pancreatic tissues. Expression levels were correlated with survival and different clinicopathological parameters using different statistical methods. Transient transfection studies using OPN and RAN plasmids, and knockdown experiments using siRNA were used to examine their mutual regulation. RESULTS: OPN and RAN levels highly correlated with each other (p<0.0001). OPN or RAN levels did not correlate with venous lymphatic invasion, diabetes, obesity, T stage, BMI, or survival. However, we found a significant association between RAN levels and perineural invasion (HR=0.79, 95% CI 0.59, 1.07; p=0.0378.). OPN and RAN colocalized in PDA tissues and cell lines. Increasing RAN expression in PDA cells induced OPN transcription and RAN silencing reduced total OPN levels. OPN did not have any significant effect on RAN transcription. CONCLUSIONS: The high levels of RAN in PDA and its correlation with OPN and with perineural invasion suggest that RAN may contribute to PDA metastasis and progression through the induction of OPN. RAN's role in the regulation of OPN in PDA is unique and could provide potential novel therapeutic strategies to combat PDA aggressiveness.