Peripheral Blood CD8+ T-Lymphocyte Immune Response in Benign and Subpopulations of Breast Cancer Patients.

Peripheral blood CD8+ T lymphocytes play a crucial role in cell-mediated immunity and tumor-related immune responses in breast cancer. In this study, label-free quantification analysis and gene set enrichment analysis (GSEA) of CD8+ T lymphocytes in the peripheral blood of benign patients and patients with different breast cancer (BC) subtypes, i.e., luminal A, luminal B, and triple-negative breast cancer (TNBC), were performed using nano-UHPLC and Orbitrap mass spectrometry. Differential protein expression in CD8+ T lymphocytes revealed significant downregulation (log2 FC ≥ 0.38 or ≤-0.38, adj. p < 0.05), particularly in proteins involved in cytotoxicity, cytolysis, and proteolysis, such as granzymes (GZMs) and perforin 1 (PRF1). This downregulation was observed in the benign group (GZMH, GZMM, and PRF1) and luminal B (GZMA, GZMH) subtypes, whereas granzyme K (GZMK) was upregulated in TNBC in comparison to healthy controls. The RNA degradation pathway was significantly downregulated (p < 0.05, normalized enrichment score (NES) from -1.47 to -1.80) across all BC subtypes, suggesting a potential mechanism for regulating gene expression during T cell activation. Also, the Sm-like proteins (LSM2, LSM3, and LSM5) were significantly downregulated in the RNA degradation pathway. Proteomic analysis of CD8+ T lymphocytes in peripheral blood across different breast cancer subtypes provides a comprehensive view of the molecular mechanisms of the systemic immune response that can significantly contribute to advancements in the diagnosis, treatment, and prognosis of this disease.

B Cell Lymphocytes as a Potential Source of Breast Carcinoma Marker Candidates.

Despite advances in the genomic classification of breast cancer, current clinical tests and treatment decisions are commonly based on protein-level information. Nowadays breast cancer clinical treatment selection is based on the immunohistochemical (IHC) determination of four protein biomarkers: Estrogen Receptor 1 (ESR1), Progesterone Receptor (PGR), Human Epidermal Growth Factor Receptor 2 (HER2), and proliferation marker Ki-67. The prognostic correlation of tumor-infiltrating T cells has been widely studied in breast cancer, but tumor-infiltrating B cells have not received so much attention. We aimed to find a correlation between immunohistochemical results and a proteomic approach in measuring the expression of proteins isolated from B-cell lymphocytes in peripheral blood samples. Shotgun proteomic analysis was chosen for its key advantage over other proteomic methods, which is its comprehensive and untargeted approach to analyzing proteins. This approach facilitates better characterization of disease-associated changes at the protein level. We identified 18 proteins in B cell lymphocytes with a significant fold change of more than 2, which have promising potential to serve as breast cancer biomarkers in the future.

MMP9 and pancreatic cancer.

BACKGROUND: Pancreatic carcinoma is one of the most severe oncological diseases of the gastrointestinal tract. At the time of diagnosis, up to 28% of patients have metastatic liver damage, and only 5% of patients survive five years. Scientific research focuses on non-invasive markers that could help screen for the disease and identify patients more quickly. Potential biomarkers also include matrix metalloproteinases, which play a role in oncogenesis. MATERIAL AND METHODS: We prospectively followed 46 patients with pancreatic cancer and benign pancreatic diseases from September 2022 to March 2023. We determined the level of MMP9 in serum and tissue biopsied during surgeries. RESULT: As a result, MMP9 levels were elevated from the T2 stage. The correlation between disease stage and MMP9 level was not confirmed in lower stages, possibly due to the small group of patients. CONCLUSION: MMP9 seems suitable for detecting late stages of pancreatic cancer, possibly for secondary prevention. We could not confirm a correlation between MMP9 levels and the initial stages of the disease (Tab. 1, Fig. 3, Ref. 21). Text in PDF www.elis.sk Keywords: pancreatic cancer, MMP9, marker, non-invasive, screening.

Risk factors for post-ERCP complications.

BACKGROUND: In the 21st century, endoscopic retrograde cholangiopancreatography (ERCP) has emerged as a diagnostic and therapeutic method for diseases of the pancreaticobiliary duct system. However, like any other diagnostic and therapeutic method, ERCP carries the risk of unwanted complications. MATERIAL AND METHODS: We retrospectively followed patients who underwent ERCP examinations from January 2013 to April 2023. We focused on early post-ERCP complications and their risk factors, prevention, treatment, and mortality. RESULTS: A total of 4,814 patients were recorded, of which 175 patients had early post-ERCP complications, including acute pancreatitis, acute cholangitis, perforation, and bleeding. We focused on the statistical significance of risk factors such as BMI, repeated ERCP, bleeding disorders, and repeated pancreatitis or cholangitis. CONCLUSION: Ensuring proper preparation and appropriate indication for ERCP examination can significantly mitigate the risk of post-ERCP complications. Additionally, early diagnosis and prompt treatment of any post-ERCP complications are essential strategies for reducing mortality associated with these conditions (Tab. 3, Fig. 3, Ref. 32). Text in PDF www.elis.sk Keywords: post-ERCP complication, risk factor, BMI, prevention.

Non-Coding RNAs in Human Cancer and Other Diseases: Overview of the Diagnostic Potential.

Non-coding RNAs (ncRNAs) are abundant single-stranded RNA molecules in human cells, involved in various cellular processes ranging from DNA replication and mRNA translation regulation to genome stability defense. MicroRNAs are multifunctional ncRNA molecules of 18-24 nt in length, involved in gene silencing through base-pair complementary binding to target mRNA transcripts. piwi-interacting RNAs are an animal-specific class of small ncRNAs sized 26-31 nt, responsible for the defense of genome stability via the epigenetic and post-transcriptional silencing of transposable elements. Long non-coding RNAs are ncRNA molecules defined as transcripts of more than 200 nucleotides, their function depending on localization, and varying from the regulation of cell differentiation and development to the regulation of telomere-specific heterochromatin modifications. The current review provides recent data on the several forms of small and long non-coding RNA's potential to act as diagnostic, prognostic or therapeutic target for various human diseases.

Imipridone enhances vascular relaxation via FOXO1 pathway.

Cardiovascular complications are a side effect of cancer therapy, potentially through reduced blood vessel function. ONC201 (TIC10) is currently used in phase 2 clinical trials to treat high-grade gliomas. TIC10 is a phosphatidylinositol 3-kinase (PI3K)/AKT/extracellular signal-regulated kinase (ERK) inhibitor that induces apoptosis via upregulation of TNF-related apoptosis-inducing ligand, which via stimulation of FOXO and death receptor could increase eNOS upregulation. This has the potential to improve vascular function through increased NO bioavailability. Our aim was to investigate the role of TIC10 on vascular function to determine if it would affect the risk of CVD. Excised abdominal aorta from White New Zealand male rabbits were cut into rings. Vessels were incubated with TIC10 and AS1842856 (FOXO1 inhibitor) followed by cumulative doses of acetylcholine (Ach) to assess vessel function. Vessels were then processed for immunohistochemistry. Incubation of blood vessels with TIC10 resulted in enhanced vasodilatory capacity. Combination treatment with the FOXO1 inhibitor and TIC10 resulted in reduced vascular function compared to control. Immunohistochemical analysis indicated a 3-fold increase in death receptor 5 (DR5) expression in the TIC10-treated blood vessels but the addition of the FOXO1 inhibitor downregulated DR5 expression. The expression of DR4 receptor was not significantly increased in the presence of TIC10; however, addition of the FOXO1 inhibitor downregulated expression. TIC10 has the capacity to improve the function of healthy vessels when stimulated with the vasodilator Ach. This highlights its therapeutic potential not only in cancer treatment without cardiovascular side effects, but also as a possible drug to treat established CVD.

Impact of drugs on venous thromboembolism risk in surgical patients.

PURPOSE: This review focuses on the most common drugs administered to surgical patients during the perioperative period that affect the risk of venous thromboembolism (VTE). RESULTS: Among analgesics, the risk of VTE is increased in patients treated with diclofenac, ibuprofen, and rofecoxib, but not naproxen, while metamizole can confer a protective effect. The relationship between sedatives and VTE has not been sufficiently studied. Tricyclic antidepressants, low-potency serotonin reuptake inhibitors, and antipsychotics have been associated with increased risk of VTE. The use of diuretics in the perioperative period is poorly researched; however, hyponatremia is considered a risk factor. Other factors that may influence the risk of VTE include bridging anticoagulation, allogeneic transfusion, and hemostatic management before surgery. Pharmacotherapy for HIV or cancer may also increase VTE risk. CONCLUSION: Increased monitoring for VTE is therefore advisable in surgical patients and those receiving antipsychotics, antidepressants, diuretics, or analgesics.

Intrapancreatic accessory spleen. A case report and review of literature.

CONTEXT: Accessory spleen is a congenital focus of healthy splenic tissue that is separated from the main body of spleen. Although an accessory spleen usually appears as an isolated asymptomatic abnormality, it may have clinical significance in some situations. CASE REPORT: We report the case of 53-year-old woman with a 2-year history of upper abdominal discomfort after meals and weight loss. The pathologic lesion was diagnosed by the abdominal sonography and the magnetic resonance tomography in the pancreatic tail. The patient was operated with suspicion of a solid pseudopapillary neoplasm or a nonfunctioning islet cell tumor. Histopathological examination found an intrapancreatic accessory spleen, which is a congenital abnormality consisting of normal splenic tissue in ectopic sites. CONCLUSION: We present possibilities of differential diagnosis of this entity.

The Role of Surgery in Chronic Pancreatitis.

Chronic pancreatitis is an inflammatory disease characterized by the progressive replacement of the functional pancreatic parenchyma with fibrotic tissue. This leads to exocrine and endocrine insufficiency. A typical clinical feature is recurrent, severe upper abdominal pain, which negatively affects the patient's quality of life. Conservative treatment as the method of first choice does not prevent irreversible changes in the pancreatic tissue. While endoscopic drainage can have some benefits in the early stages of the disease, it is generally unsuccessful in the long term. Based on recent studies, surgical intervention appears to be the most suitable treatment option for improving the patient's quality of life. It currently includes a wide range of effective, proven drainage and resection procedures. Advances in surgical techniques and postsurgical intensive care have increased the frequency of pancreatic surgeries, while improvements in diagnostic methods have increased the number of patients who meet the indications for such surgery, including elderly and chronically ill patients. However, despite mortality rates decreasing, high morbidity rates remain a problem. Currently, in patients with an inflammatory mass in the head of the pancreas, pyloric and duodenal-preserving resection offers the best results. Different variants of these techniques appear to produce similar results. Various techniques have shown similar outcomes in terms of mortality, morbidity, pain relief, life expectancy and improved quality of life. The optimal timing of surgery has been addressed by several studies and most surgeons now favor early surgical intervention in order to prevent extensive destruction of pancreatic tissue.

Bouveret's Syndrome: A Rare Form of Gallstone Ileus Caused by Large Renal Cysts.

Bouveret's syndrome is a rare variant of gallstone ileus caused by gastric outlet obstruction that arises from gallstones impacted in the distal stomach or proximal duodenum after passing through a cholecystoduodenal or cholecystogastric fistula. Simple kidney cysts are one of the most common lesions found in the elderly. They are usually asymptomatic, but the cysts can put pressure on the surrounding organs if they grow to large dimensions.This case report highlights a rare case of Bouveret's syndrome due to the presence of a large gallstone in the pyloric region that was caused by the creation of a cholecystogastric fistula from pressure exerted by two giant cysts of the right kidney.

Discharge protocol in acute pancreatitis: an international survey and cohort analysis.

There are several overlapping clinical practice guidelines in acute pancreatitis (AP), however, none of them contains suggestions on patient discharge. The Hungarian Pancreatic Study Group (HPSG) has recently developed a laboratory data and symptom-based discharge protocol which needs to be validated. (1) A survey was conducted involving all members of the International Association of Pancreatology (IAP) to understand the characteristics of international discharge protocols. (2) We investigated the safety and effectiveness of the HPSG-discharge protocol. According to our international survey, 87.5% (49/56) of the centres had no discharge protocol. Patients discharged based on protocols have a significantly shorter median length of hospitalization (LOH) (7 (5;10) days vs. 8 (5;12) days) p < 0.001), and a lower rate of readmission due to recurrent AP episodes (p = 0.005). There was no difference in median discharge CRP level among the international cohorts (p = 0.586). HPSG-protocol resulted in the shortest LOH (6 (5;9) days) and highest median CRP (35.40 (13.78; 68.40) mg/l). Safety was confirmed by the low rate of readmittance (n = 35; 5%). Discharge protocol is necessary in AP. The discharge protocol used in this study is the first clinically proven protocol. Developing and testifying further protocols are needed to better standardize patients' care.

Impact of different pancreatic microenvironments on improvement in hyperglycemia and insulin deficiency in diabetic rats after transplantation of allogeneic mesenchymal stromal cells.

BACKGROUND: Mesenchymal stromal cells (MSCs) in the pancreatic microenvironment can improve diabetes mellitus (DM). The aim of the present study was to determine whether different pancreatic microenvironments influence the improvement of hyperglycemia and insulin deficiency. METHODS: MSCs isolated from rat bone marrow were transplanted directly into different pancreatic microenvironments in male DM rats. DM was induced in the rats by streptozotocin injection. The rats were divided into 5 groups: normal control rats, DM control rats, and 3 experimental groups (DM rats plus MSCs injected into the head of the pancreas, the tail of the pancreas, or the whole pancreas). The body weight and blood glucose of the rats were monitored during the experiment after transplantation of the MSCs. Histopathologic and immunohistochemical analyses were used to detect the presence and number of islets and insulin production in the pancreatic tissue of the rats after MSC transplantation. RESULTS: At 28 days after MSC transplantation, we observed a statistically significant decrease in the blood glucose level and an increase in weight in DM rats compared with DM control rats (P < 0.0001 and P < 0.03, respectively). A comparison of each of the DM rat groups treated with MSCs showed no significant differences in the blood glucose levels or body weight. CONCLUSION: Our results suggest that transplantation of MSCs could improve DM in the pancreatic microenvironment in an animal model with streptozotocin-induced DM. The different pancreatic areas into which the MSCs were implanted had no significant influence on the improvement in hyperglycemia and insulin deficiency.

Classic and New Markers in Diagnostics and Classification of Breast Cancer.

Breast cancer remains the most frequently diagnosed form of female's cancer, and in recent years it has become the most common cause of cancer death in women worldwide. Like many other tumours, breast cancer is a histologically and biologically heterogeneous disease. In recent years, considerable progress has been made in diagnosis, subtyping, and complex treatment of breast cancer with the aim of providing best suited tumour-specific personalized therapy. Traditional methods for breast cancer diagnosis include mammography, MRI, biopsy and histological analysis of tumour tissue in order to determine classical markers such as estrogen and progesterone receptors (ER, PR), cytokeratins (CK5/6, CK14, C19), proliferation index (Ki67) and human epidermal growth factor type 2 receptor (HER2). In recent years, these methods have been supplemented by modern molecular methodologies such as next-generation sequencing, microRNA, in situ hybridization, and RT-qPCR to identify novel molecular biomarkers. MicroRNAs (miR-10b, miR-125b, miR145, miR-21, miR-155, mir-30, let-7, miR-25-3p), altered DNA methylation and mutations of specific genes (p16, BRCA1, RASSF1A, APC, GSTP1), circular RNA (hsa_circ_0072309, hsa_circRNA_0001785), circulating DNA and tumour cells, altered levels of specific proteins (apolipoprotein C-I), lipids, gene polymorphisms or nanoparticle enhanced imaging, all these are promising diagnostic and prognostic tools to disclose any specific features from the multifaceted nature of breast cancer to prepare best suited individualized therapy.

[Current alternatives in the surgical treatment of chronic pancreatitis--a review article]. / Súcasné moznosti chirurgickej liecby chronickej pankreatitídy--literárny prehl'ad.

Chronic pancreatitis is characterized as an inflammatory process affecting the pancreas that causes progressive destruction of the gland and fibrosis, with subsequent endocrine and exocrine insufficiency. The most common cause of chronic pancreatitis is alcohol use in combination with nicotine. Manifestations are persistent or recurrent painful attacks. The only parameter of successful treatment of chronic pancreatitis is a relieve from long-lasting pain and improvement of the quality of life. Surgical treatment options include drainage operations on the pancreas, pancreatic resection or a combination of both. With optimal surgical treatment performed and good patient's compliance, operations for chronic pancreatitis have low number of post-operative complications and relatively good long-term results. The continued consumption of alcohol and drugs bring about worse outcomes, sometimes even a complete failure of therapy. Chronic pancreatitis also has considerable socio-economic consequences. Due to the persisting pain and frequent hospitalization it can lead to long-term disability and early retirement predominantly in young patients.

Low-level laser therapy for protection against skeletal muscle damage after ischemia-reperfusion injury in rat hindlimbs.

BACKGROUND AND OBJECTIVE: Despite numerous hypotheses regarding the action of laser light, the use of low-level laser therapy (LLLT) in ischemic reperfusion (I/R) injury is still being verified. The present study investigates the effects of low-level laser irradiation (LLLI) on I/R injury of the musculus gracilis in rats. MATERIALS AND METHODS: I/R injury of the musculus gracilis flap was induced in male adult Sprague-Dawley rats (n = 84). Rats were subdivided depending on treatment into four subgroups: (1) healthy group, (2) I/R injury without irradiation, (3) R group irradiated only during reperfusion after injury, and (4) IR group irradiated during ischemia and reperfusion injury. LLLT (AlGaInP; λ = 670 nm; 4 J/cm²; 40 mW/cm²) was applied to the injured muscle four times daily until euthanasia. RESULTS: Lactate dehydrogenase (LD) levels were significantly lower (P<0.05) in the irradiated groups during the first 12-120 hours, while the lower creatine kinase (CK) level reached statistical significance only at 24 hours in the irradiated group when compared to the control group. The number of polymorphonuclear leukocytes in the gracilis muscle was significantly lower in the treated group only on the second day (P<0.0001). The lowered percentage of necrosis in the muscle tissue was statistically significant after 6 and 10 days of treatment (P<0.0001), while lower atrophy and higher neovascularization were observed at 6-14 days of irradiation (P<0.05). There was no statistically significant difference between the group irradiated only during reperfusion and that irradiated during ischemia and reperfusion. CONCLUSION: LLLT confers a protective effect against early inflammatory tissue response, further atrophy, and necrosis of the muscle and it stimulates neovascularization after I/R injury.

[Have changes in the management of treatment of the patients with acute pancreatitis brought the expected result?]. / Priniesla zmena manazmentu u chorých s t'azkou akútnou pankreatitídou ocakávaný výsledok?

BACKGROUND: Mortality from severe acute pancreatitis (AP) has remarkably decreased (10-20%) during the last decades. However, many questions on the treatment of this disease remained opened. To extend the most recent professional literature which discuses the abovementioned problem, experience from the treatment of patients at the department where the authors work is presented. METHODS: Influenced by the new methods published in professional literature and under the circumstances at own workplace, the decision to change the management of the treatment of AP has been made. The management referred to the enteral nutrition, epidural analgesy, antibiotic prophylaxis, pushing the surgical operation to the later period in the case of infected necrosis. Comparing of two groups of patients--A (2003-2005 years) and B (2006-2008 years) the authors came to interesting results. RESULTS: Applying the new protocol we observed: increased percentage of patients with sterile necrosis from 46% to 58%, decreased number of surgical re-operations from 43% to 33% and decreased mortality from 53,8% to 18%. CONCLUSIONS: Applying changes in the management of treatment of the patients with the complicated form of acute pancreatitis, the authors achieved remarkable results. Nevertheless, these results have to be evaluated very cautiously, because the group of patients who have been treated by the novel approach has not been very large.

Response of ependymal progenitors to spinal cord injury or enhanced physical activity in adult rat.

Ependymal cells (EC) in the spinal cord central canal (CC) are believed to be responsible for the postnatal neurogenesis following pathological or stimulatory conditions. In this study, we have analyzed the proliferation of the CC ependymal progenitors in adult rats processed to compression SCI or enhanced physical activity. To label dividing cells, a single daily injection of Bromo-deoxyuridine (BrdU) was administered over a 14-day-survival period. Systematic quantification of BrdU-positive ependymal progenitors was performed by using stereological principles of systematic, random sampling, and optical Dissector software. The number of proliferating BrdU-labeled EC increased gradually with the time of survival after both paradigms, spinal cord injury, or increased physical activity. In the spinal cord injury group, we have found 4.9-fold (4 days), 7.1-fold (7 days), 4.9-fold (10 days), and 5.6-fold (14 days) increase of proliferating EC in the rostro-caudal regions, 4 mm away from the epicenter. In the second group subjected to enhanced physical activity by running wheel, we have observed 2.1-2.6 fold increase of dividing EC in the thoracic spinal cord segments at 4 and 7 days, but no significant progression at 10-14 days. Nestin was rapidly induced in the ependymal cells of the CC by 2-4 days and expression decreased by 7-14 days post-injury. Double immunohistochemistry showed that dividing cells adjacent to CC expressed astrocytic (GFAP, S100beta) or nestin markers at 14 days. These data demonstrate that SCI or enhanced physical activity in adult rats induces an endogenous ependymal cell response leading to increased proliferation and differentiation primarily into macroglia or cells with nestin phenotype.

Conservative management in three cases of prenatally recognized splenic cyst using 2D, 3D, multi-slice and Doppler ultrasonography.

The aetiology, differential diagnosis and management strategies of the foetal spleen affected with a cystic lesion are discussed. In the current literature, there are very few reports that relate to antenatally diagnosed splenic cyst. Our study presents 3 case reports that were first suspected due to anisoechogenic structures detected during routine ultrasonographic examination at the 27th, 31st and 34th weeks of gestation. All 3 cases were further characterized by the lack of pathological power Doppler findings inside and around the lesions, and were morphologically refined by prenatal 3D ultrasound imaging. All findings were reconfirmed postnatally. No complications such as cyst expansion, subcapsular bleeding or acute abdomen have developed, and all 3 cystic lesions have regressed spontaneously after birth.

Influence of a polymer membrane on the electrochemical determination of insulin in nanomodified screen printed carbon electrodes.

Diabetes mellitus can be considered one of the most widespread diseases globally. Hence, the diabetes research is currently focused on developing an effective, low-cost sensor having high stability and suitable analytical characteristics. Screen printed carbon electrodes (SPCEs) embody ideal candidates for insulin determination due to the small area of the working electrode eliminating the solution volume required for the given purpose. Modification of SPCEs by using nanoparticles resulted in an increase of the working electrode surface area and formation of a higher number of active species. The aim of this paper is to examine the impact of a chitosan membrane on the electrochemical determination of insulin on NiO nanoparticles (NiONPs) and multi-walled nanotube (MWCNTs) modified SPCE (NiONPs/MWCNTs/SPCE). This study is primarily conceived to compare the analytical characteristics and stability of NiONPs/chitosan-MWCNTs/SPCE and NiONPs/MWCNTs/SPCE. An electrode modified with chitosan displays a wider linear range, one of 0.25 µM - 5 µM (R2 0.997); a lower limit of detection, 94 nM; a high sensitivity (0.021 µA/µM) and better stability than that of an electrode without chitosan. According to these characteristics, the polymer is considered a necessary compound of the electrochemical insulin sensor, improving the sensor's analytical characteristics.

Prenatal diagnosis of annular pancreas: reliability of the double bubble sign with periduodenal hyperechogenic band.

OBJECTIVE: To evaluate the power of prenatal 2-D ultrasound examination in the 2nd trimester as a method of choice for accurate diagnosis of annular pancreas. METHODS: Co-incidence of the double bubble sign (often accompanying gastroduodenal dilatation) together with a hyperechogenic band around the duodenum (corresponding with the tissue of annular pancreas) was used as a diagnostic criterion. Findings from postnatal surgery served for verification. RESULTS: From 7,897 screened pregnancies, annular pancreas was proven in the cases where both signs were present, but never without the hyperechogenic band (N(1) = 3, N(2) = 3, p < or = 0.05). Sensitivity and specificity were 100%. CONCLUSIONS: More multicentric studies are required to test this approach. The following diagnostic strategy is reasonable at the present time: when the double bubble sign is discovered, always suspect annular pancreas and look for the second sign: hyperechogenic bands around the duodenum. Also look for known associated anomalies, and vice versa, if any of associated anomalies are noted, also search specifically for the signs of annular pancreas.