Impact of Intragastric Balloon Placement on the Stomach Wall: A Prospective Cohort Study.

PURPOSE: Endoscopic intragastric balloon (IGB) placement is a minimally invasive treatment for morbid obesity that is sometimes used as a preparatory step before surgical intervention. This study was performed to analyze the changes in the stomach wall induced by IGB placement, with particular emphasis on pathomorphology, inflammatory markers, and tissue growth factors. MATERIAL AND METHODS: In total, 30 patients with morbid obesity were prospectively analyzed. A total of 16 patients with body mass index (BMI) ≥ 53 kg/m2 underwent two-stage treatment comprising IGB placement followed by laparoscopic sleeve gastrectomy (LSG) (IGB group), while 14 patients underwent one-stage LSG (non-IGB group). The gastric specimens removed during LSG were examined. The two groups were compared regarding the surgical results, microscopic structure and inflammatory process exponents of the stomach wall, and receptors for selected tissue growth factors. RESULTS: The IGB group had a longer median hospital stay than that of the non-IGB group. Compared with the non-IGB group, the IGB group had a thicker stomach wall, more submucosal fibrosis, and increased amounts of growth factors and inflammatory markers. CONCLUSION: Patients with IGB placement before LSG showed greater changes in the stomach wall than those of patients who received LSG alone. IGB placement was associated with stomach muscle layer thickening, submucosal fibrosis, and increased levels of inflammatory markers and tissue growth factors.

Microbiome Profile and Molecular Pathways Alterations in Gastrointestinal Tract by Hydrogen Sulfide-Releasing Nonsteroidal Anti-Inflammatory Drug (ATB-352): Insight into Possible Safer Polypharmacy.

Aims: Nonsteroidal anti-inflammatory drugs, including ketoprofen, induce adverse effects within the gastrointestinal (GI)-tract. Hydrogen sulfide (H2S) is an antioxidative gaseous mediator contributing to GI-protection. We aimed to evaluate the GI safety of a novel H2S-releasing derivative of ketoprofen (ATB-352) versus classic ketoprofen and the molecular mechanisms of their activity after chronic treatment in experimental animal models. Results: Ketoprofen (10 mg/kg/day) administered intragastrically for 7 days in contrast with ATB-352 (14 mg/kg/day) reduced mucosal H2S content inducing GI damage with significantly increased injury score, altered intestinal microbiome profile, and modulation of more than 50% of 36 investigated molecular sensors (e.g., mammalian target of rapamycin or suppressor of cytokine signaling 3 [SOCS3]). Polypharmacy with aspirin (10 mg/kg/day) enhanced ketoprofen toxicity not affecting GI safety of ATB-352. Omeprazole (20 mg/kg/day) decreased ketoprofen-induced injury to the level of ATB-352 alone. Both compounds combined or not with aspirin or omeprazole maintained the ability to inhibit cyclooxygenase (COX) activity manifested by decreased prostaglandin production. Innovation and Conclusions: Ketoprofen-induced H2S-production decrease and intestinal microbiome profile alterations lead to GI toxicity observed on macro-/microscopic and molecular levels. Ketoprofen but not ATB-352 requires concomitant treatment with omeprazole to eliminate GI adverse effects. ATB-352 applied alone or in a polypharmacy setting with aspirin effectively inhibited COX and maintained GI safety due to H2S-release. Neither compound affected DNA oxidation in the GI mucosa, but ATB-352 had lower impact on molecular oxidative/inflammatory response pathways and intestinal microbiome. The GI safety of ATB-352 could be due to the involvement of heme oxygenase 1 and SOCS3 pathway activation. Antioxid. Redox Signal. 36, 189-210.

Molecular Profile of Barrett's Esophagus and Gastroesophageal Reflux Disease in the Development of Translational Physiological and Pharmacological Studies.

Barrett's esophagus (BE) is a premalignant condition caused by gastroesophageal reflux disease (GERD), where physiological squamous epithelium is replaced by columnar epithelium. Several in vivo and in vitro BE models were developed with questionable translational relevance when implemented separately. Therefore, we aimed to screen Gene Expression Omnibus 2R (GEO2R) databases to establish whether clinical BE molecular profile was comparable with animal and optimized human esophageal squamous cell lines-based in vitro models. The GEO2R tool and selected databases were used to establish human BE molecular profile. BE-specific mRNAs in human esophageal cell lines (Het-1A and EPC2) were determined after one, three and/or six-day treatment with acidified medium (pH 5.0) and/or 50 and 100 µM bile mixture (BM). Wistar rats underwent microsurgical procedures to generate esophagogastroduodenal anastomosis (EGDA) leading to BE. BE-specific genes (keratin (KRT)1, KRT4, KRT5, KRT6A, KRT13, KRT14, KRT15, KRT16, KRT23, KRT24, KRT7, KRT8, KRT18, KRT20, trefoil factor (TFF)1, TFF2, TFF3, villin (VIL)1, mucin (MUC)2, MUC3A/B, MUC5B, MUC6 and MUC13) mRNA expression was assessed by real-time PCR. Pro/anti-inflammatory factors (interleukin (IL)-1ß, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12, IL-13, tumor necrosis factor α, interferon γ, granulocyte-macrophage colony-stimulating factor) serum concentration was assessed by a Luminex assay. Expression profile in vivo reflected about 45% of clinical BE with accompanied inflammatory response. Six-day treatment with 100 µM BM (pH 5.0) altered gene expression in vitro reflecting in 73% human BE profile and making this the most reliable in vitro tool taking into account two tested cell lines. Our optimized and established combined in vitro and in vivo BE models can improve further physiological and pharmacological studies testing pathomechanisms and novel therapeutic targets of this disorder.

Ovarian Sertoli-Leydig cell tumour with α-fetoprotein-producing intestinal glandular cells. Clinical case and short review of basic literature.

Sertoli Leydig cell tumor of the ovary, is a rare neoplasm from the group of sex cord-stromal tumors of the ovary, accounting for less than 1% of all ovarian tumors. Among the Sertoli Leydig cell tumors, we distinguish a separate group of tumors secreting α-fetoprotein (AFP). The young 24-year-old woman presented to the Clinical Department of Gynaecological Endocrinology at the University Hospital in Krakow due to secondary amenorrhea, hirsutism and worsening abdominal pain for several months. During the admission draws attention was drawn to the abnormal level of testosterone, AFP and the revised structure of the ovary in the ultrasound. After a preliminary diagnosis, expanded pelvic MRI was performed, which found an isolated tumor derived from Sertoli Leydig cells. The patient was enrolled to unilaterally remove the right ovary by laparotomy. Histopathological examination and immunohistochemical staining confirmed the diagnosis of Sertoli Leydig cells tumor, and in pathological examination we found glandular mucosa cells of the colon. Owing to scientific reports on the stromal tumors of the ovary, we decided to perform genetic testing and verify the patient's karyotype. In the follow-up 90 days after the surgery, levels of testosterone and AFP were correct. In case of Sertoli Leydig cell tumors, especially in young women of childbearing potential, special attention should be paid to Anti-Mullerian hormone testing before surgery, as well as genetic diagnostics to exclude disorders of sex development.

Influence of TNF-α promoter variability on stage and grade in individuals with colorectal cancer.

Carcinogenesis is a multistep process in which inflammation plays an important role. Tumour necrosis factor a (TNF-α) is a cytokine that plays a major role in inflammation. Activity of the TNF cytokine family could influence progression of colorectal cancer (CRC). The aim of the study was to establish an association between TNF-α promoter variability and stage/grade in individuals with sporadic CRC. The study included 152 CRC patients and 107 healthy volunteers. Four single nucleotide polymorphisms (rs361525, rs1800629, rs1799724, and rs1799964) located at the promoter of TNFA gene were genotyped using commercially available TaqMan allelic discrimination assays by real-time PCR. CRC stage was described on the basis of preoperative imaging studies and postoperative histopathological report. The grade was described on the basis postoperative pathological examination of the specimen. In the case of rs361525, there was a statistically significant association with M-score (p = 0.0209). Rs361525 has significant association with tumour grade (p = 0.0260). We failed to demonstrate significant association between the other 3 SNPs and cancer grade. Rs361525 potentially could be under consideration when the survival rate and prognosis is assessed.

Neuroendocrine neoplasms and somatostatin receptor subtypes expression.

Neuroendocrine neoplasms (NENs) show wide spectrum of clinical course - from benign biological potential to recurrences and rapidly progressive disease. Somatostatin analogs that bind to somatostatin receptor are part of the therapy; detection and evaluation of activation of somatostatin receptor subtypes are part of the process of new therapy induction. When using RT-PCR method and immunohistochemistry, it is possible to detect more than two SSTR subtypes in majority or all neuroendo-crine neoplasms regardless tumor origin. Generally with some exceptions, from the viewpoint of tumor grade - apart the site of origin, there is a tendency to decrease the percentage of SSTRs expression; 100% (G1, 2)-85.7% (G3) for SSTR 1; 81.8% (G1, 2)-61.9% (G3) for SSTR 2; 54.5% (G1, 2)-52.4% (G3) for SSTR 3; 9% (G1, 2)-4.8% (G3) for SSTR 5. Different studies indi-cate significant differences in the expression of SSTR 1 and 2A and 2B between NEC G3 small cell type and non-small cell type. Further research on SSTRs expression in NEN could serve as base to development and improvement of somatostatin analogs' pharmacotherapy in patients with unsatisfactory course.

The number of regulatory Foxp3+ T-cells in different stages of malignant transformation of large intestinal polyps.

PURPOSE: The aim of this study was to determine the relationship between the number of regulatory T-cells (Tregs) at various stages of malignant transformation of large intestinal polyps. MATERIAL/METHODS: The study included tissue specimens from individuals subjected to complete colonoscopy with polypectomy and from patients who underwent surgical resection of colorectal tumors. This group included 27 individuals, among them 10 women (37%). Median age of the patients was 64 years (range 37-82 years). Surgical specimens included hyperplastic polyps (n=4), adenomatous polyps with low- (n=5) and high-grade dysplasia (n=8) and invasive colorectal cancers (n=10). Tregs were identified immunohistochemically. RESULTS: Mean number of Foxp3+ T-cells per 10 high-power fields (HPFs) increased in line with malignant transformation, from 12.5 for hyperplastic polyps, 29.4 and 36.5 for adenomatous polyps with low- and high-grade dysplasia, respectively, to 56.3 for invasive colorectal cancers (p=0.00). An increase in the mean number of CD4+ T-cells was also observed, from 45.75, 57.8, 84.125, to 110.6 per 10 HPFs, respectively, however this change did not prove to be statistically significant (p=0.13). Mean Foxp3+/CD4+ T-cell ratio increased in line with malignant transformation (from 0.27, 0.3, 0.43, to 0.5), although a statistically significant change of this parameter was only observed in the case of invasive colorectal cancers (p=0.01). CONCLUSIONS: An increase in the number of Tregs in the lymphocytic infiltrate of large intestinal polyps is interestingly already observed at early stages of carcinogenesis. Proportions of various T-cell subpopulations in the infiltrate vary considerably depending on the degree of dysplasia, especially in the case of invasive colorectal cancer.

Foxp3+ lymphocyte count in Barrett's esophagus tissue is higher than in in amed esophageal tissue.

INTRODUCTION: The number of Foxp3+ lymphocytes is increased in patients with esophageal carcinoma. Little is known about Foxp3+ cells count in Barret's esophagus, which is a precancerous state for esophageal cancer. A i m: To count the number of Foxp3+ lymphocytes in tissue samples from patients with Barrett's and compare it with samples from individuals with esophagitis and esophageal cancer. MATERIALS AND METHODS: 43 patients were enrolled to the study: 14 with esophageal carcinoma, 15 with Barrett's esophagus and 14 with non-metaplastic esophagitis. Every patient undergone gastroscopy during which a tissue sample was taken. Foxp3+ lymphocytes and CD4+ lymphocytes were detected by using immunohistochemistry. RESULTS: Mean density of Foxp3+ cells in patients with esophagitis was 7.37/10HPF (range: 1-9), 18.5/10HPF (range: 5-29) and 26.8/10HPF (range: 16-40) in patients with dysplastic and non-dysplastic BE, respectively and 47.92/10HPF in individuals with esophageal a carcinoma. These intergroup differences turned out to be statistically significant (p = 0.000; Fig. 3). Patients, either with dysplasia or without, presented with significantly higher Foxp3+ cell counts than the subjects with esophagitis (p = 0.0003 and p = 0.0006, respectively). Also the number of Foxp3+ lymphocytes in esophageal adenocarcinoma specimens turned out to be significantly higher than in esophagitis (p = 0.0001), non-dysplastic and dysplastic BE tissue (p = 0.016 and p = 0.00047, respectively). CONCLUSIONS: Barrett's metaplasia, either with dysplasia or without, is associated with an evident increase in the number of Foxp3 lymphocytes within the esophagogastric junction mucosa. Restoration of lymphocyte balance in esophageal tissue might prevent malignant transformation of Barrett's metaplasia.

[Risk and prophylaxis of venous thromboembolism in the material of 2nd Chair of Surgery JUMC]. / Ryzyko i profilaktyka zylnej choroby zakrzepowo-zatorowej w materiale II Katedry Chirurgii UJCM.

INTRODUCTION: Venous thromboembolism (VTE) is one of the most dangerous complication that could occur in patiens treated in surgical ward. Threat is increased due to presence of additional, apart from surgical procedures, risk factors. It is assesed, that approximately 50 000 patients are at risk of VTE yearly in Poland, 20 000 of them develop pulmonary thromboembolism during hospitalisation, which is fatal in 10% of cases. AIM: Aim of the study was to asses presence of VTE risk factors in the group of patients treated in the 2nd Chair of Surgery Jagiellonian University Medical College and to analyse applied prophylaxis. MATERIAL AND METHODS: The study enrolled 170 patients, 89 (52%) females and 81 (48%) males. Average age of the studied group was 57.9. Medical history of all patients was taken with an emphasis on presence of VTE risk factors. It had a form of questionnaire and enclosed questions. Apart from the anamnesis, physical examination was performed. Type of applied prophylaxis was assesed on the basis of medical records. RESULTS: Each patient was charged with 4 risk factors on average. The most common risk factors were: age, surgical procedure lasting longer than 45 minutes, ischaemic heart disease and hipertension. Clinical probability of deep venous thrombosis was assesed according to Wells scale; low: 59% of studied group, intermediate: 41%, high: 0%. Applied prophylaxis included pharmacotherapy, physiotherapy, and mechanical methods (compression). CONCLUSIONS: The most common risk factor that could be changed was duration of the surgery lasting longer than 45 minutes. Pharmacological prophylaxis should be more often complemented by mechanical methods and physiotherapy, especially in group burdened with high risk of VTE.