Matrix Metalloproteinases and Heart Transplantation-A Pathophysiological and Clinical View.

Heart transplantation is undergoing a continuous development, with rates of success increasing substantially due to advances in immunosuppressive therapy and surgical techniques. The most worrying complication occurring after cardiac transplantation is graft rejection, a phenomenon that is much affected by matrix metalloproteinases (MMPs), with the role of these proteases in the cardiac remodeling process being well established in the literature. A detailed investigation of the association between MMPs and cardiac rejection is necessary for the future development of more targeted therapies in transplanted patients, and to discover prognostic serum and immunohistochemical markers that will lead to more organized therapeutic management in these patients. The aim of this review is therefore to highlight the main MMPs relevant to cardiovascular pathology, with particular emphasis on those involved in complications related to heart transplantation, including cardiac graft rejection.

Lower urinary tract symptoms are associated with clinically relevant depression, anxiety, and stress symptoms.

OBJECTIVES: To investigate the correlation between lower urinary tract symptoms (LUTS), erectile dysfunction (ED), and testosterone deficiency (TD) with depressive, stress, and anxiety symptoms. MATERIAL AND METHODS: From October 2019 to March 2020, 113 males were included. Inclusion criteria: age 40-75, no clinical suspicion of prostate cancer, no serious cardiovascular comorbidities. All patients completed a set of questionnaires: International Prostate Symptom Score (IPSS), International Index of Erectile Function (IIEF-5), and Depression Anxiety Stress Scales (DASS-21). RESULTS: Median age was 62 years (range 40-74), mean IPSS score was 10.94 (SD 7.75), mean IIEF-5 score 13.12 (SD 7.08), and mean DASS-21 score 11.35 (SD 8.24). According to DASS-21 subscales, 28 (24.8%) patients had depressive symptoms, 25 (22.1%) anxiety symptoms, and 25 (22.1%) stress symptoms. Depression was associated with LUTS (14.5 vs. 8 score, p = .002). Similarly, stress symptoms were associated with LUTS (IPSS 15 vs. 7 score, p = .0001) and with ED (IIEF-5 5 vs. 15 score, p = .01). Positive Spearman's rho correlations between LUTS and all three, depression, anxiety, and stress symptoms were found (p values <.001). CONCLUSIONS: LUTS is associated with depression, anxiety, and stress symptoms. Screening for these symptoms could help with individual counseling and management.

Duplex High-Resolution Melting Assay for the Simultaneous Genotyping of IL28B rs12979860 and PNPLA3 rs738409 Polymorphisms in Chronic Hepatitis C Patients.

Chronic hepatitis C (CHC) is a major burden for public health worldwide. Although newer direct-acting antivirals show good efficacy, their cost precludes their wide adoption in resource-limited regions. Thus, strategies are being developed to help identify patients with high susceptibility to response to classic PEG-interferon + ribavirin therapy. IL28B polymorphism rs12979860 C/T is an important predictor for an efficient response to interferon-based therapy. A genetic variant in adiponutrin (PNPLA3) gene, rs738409 C/G, is associated with steatosis, severity, and progression of liver fibrosis in CHC patients, and predicts treatment outcome in difficult-to-cure HCV-infected patients with advanced fibrosis. We developed a rapid and inexpensive assay based on duplex high-resolution melting (HRM) for the simultaneous genotyping of these two polymorphisms. The assay validation was performed on synthetic DNA templates and 132 clinical samples from CHC patients. When compared with allele-specific PCR and sequencing, our assay showed 100% (95% CI: 0.9724-1) accuracy, with 100% sensitivity and specificity. Our assay was robust against concentration and quality of DNA samples, melting curve normalization intervals, HRM analysis algorithm, and sequence variations near the targeted SNPs (single nucleotide polymorphism). This duplex assay should provide useful information for patient-oriented management and clinical decision-making in CHC.

Circulating RNA molecules as biomarkers in liver disease.

Liver disease is a major cause of morbidity and mortality worldwide. As in other fields of medicine, there is a stringent need for non-invasive markers to improve patient diagnostics, monitoring and prognostic ability in liver pathology. Cell-free circulating RNA molecules have been recently acknowledged as an important source of potential medical biomarkers. However, many aspects related to the biology of these molecules remain to be elucidated. In this review, we summarize current concepts related to the origin, transportation and possible functions of cell-free RNA. We outline current development of extracellular RNA-based biomarkers in the main forms of non-inherited liver disease: chronic viral hepatitis, hepatocellular carcinoma, non-alcoholic fatty liver, hepato-toxicity, and liver transplantation. Despite recent technological advances, the lack of standardization in the assessment of these markers makes their adoption into clinical practice difficult. We thus finally review the main factors influencing quantification of circulating RNA. These factors should be considered in the reporting and interpretation of current findings, as well as in the proper planning of future studies, to improve reliability and reproducibility of results.

Platelet-to-Albumin Ratio: The Prognostic Utility in the Prediction of 2-Month Postoperative Heart Transplant Complications.

BACKGROUND: The platelet-to-albumin ratio (PAR), leucocyte-to-albumin ratio (LAR), neutrophil percentage-to-albumin ratio (NPAR), and monocyte-to-albumin ratio (MAR) represent easily reproducible markers, which may predict the outcomes in various diseases. Early postoperative complications might appear after heart transplantation, such as infections, diabetes mellitus type 2 (DM2), acute graft rejection, and atrial fibrillation (AFib). OBJECTIVE: The aim of our study was to investigate the PAR, LAR, NPAR, and MAR values before and after heart transplantation, and the associations of the preoperative levels of these markers with the presence of postoperative complications in first two months after surgery. METHODS: Our retrospective research was directed from May 2014 to January 2021, with a total number of 38 patients being included. We used cut-off values for the ratios from previously published studies, as well as our own determination of these levels by using a receiver operating characteristic (ROC) curve. RESULTS: By ROC analysis, the optimal preoperative PAR cut-off value was 38.84 (AUC: 0.771, p = 0.0039), with 83.3% sensitivity, and 75.0% specificity. Applying a Chi square (χ2) test, PAR > 38.84 represented an independent risk factor for complications, regardless of cause, and postoperative infections. CONCLUSIONS: Preoperative PAR > 38.84 was a risk factor of developing complications of any cause, and postoperative infections in the first two months after heart transplantation.

Fascin-1 and its role as a serological marker in prostate cancer: a prospective case-control study.

AIM: This study aims to investigate any modification of serological FSCN1 in prostate cancer patients compared with patients without neoplasia. MATERIAL & METHODS: Clinical data and blood specimens from patients with and without prostate cancer were obtained. A quantitative sandwich ELISA method was used to determine serological values of FSCN1. RESULTS: Although serum values of FSCN1 were dissimilar in the two cohorts of patients (6.90 vs 7.33 ng/ml), the difference was not statistically significant (p = 0.20). Serum values of FSCN1 stratified for Gleason score groups were not significantly distinguishable (p = 0.65). A negative correlation (rho = -0.331; p = 0.009) was reported between FSCN1 and age. CONCLUSION: Further studies are required to evaluate a possible diagnostic role of FSCN1 in prostate cancer.

Fascin is secreted in male's serum: results of a pilot study.

AIM: Fascin is a 55 kDa globular protein with an important role in cell migration. Aim of study was to investigate serum fascin in healthy males. MATERIALS & METHODS: From 1 July 2016 to 31 December 2016, we collected serum from 46 males. Serum fascin level was performed using ELISA kit from USBiological (Salem, MA, USA). RESULTS: Median age was 64 years. Mean fascin serum level was 9.84 ng/ml, mean prostate-specific antigen (PSA) was 2.74 ng/ml and mean prostate volume was 37.64 cc. The 51-60 years group had a mean of 10.53 ng/ml, the 61-70 group a mean of 9.7 ng/ml and the 71-80 group had a mean of 9.41 ng/ml fascin serum level. CONCLUSION: Fascin serum level did not differ according to age in males.

GABRG2 C588T gene polymorphisms might be a predictive genetic marker of febrile seizures and generalized recurrent seizures: a case-control study in a Romanian pediatric population.

INTRODUCTION: This case-control study aimed to assess two single nucleotide polymorphisms of the gene encoding the GABRG2 protein - GABRG2 (3145 G>A) and GABRG2 rs 211037 Asn196Asn (C588T) - in a cohort of pediatric patients from Romania, and evaluate their possible impact on drug-resistant forms of generalized epilepsy and recurrent febrile seizures. MATERIAL AND METHODS: One hundred and fourteen children with idiopathic generalized epilepsy (group 1) or febrile seizures (group 2) were compared to 153 controls. Peripheral blood samples were assessed using polymerase chain reaction-restriction fragment length polymorphism analysis, with results interpreted based on the disappearance of a restriction site in the C allele (122 bp) compared to the T allele (100 bp + 22 bp). RESULTS: A significant association was found with the TT homozygous genotype and T allele for both febrile seizures and epilepsy for the C588T locus, while GABRG2 G>A 3145 showed no significant association with any type of seizure. The TT homozygous genotype of GABRG2 Asn196Asn polymorphism was more frequent in patients with a history of febrile seizures (p = 0.0001), without a significant association identified for GABRG2-G>A 3145. Composite analysis showed associations with epilepsy for CC-AG (p = 0.02) and CT-AG (p = 0.007) with the CC-AA combination as reference. CONCLUSIONS: C588T polymorphism of the GABRG2 gene might be a predictive genetic marker in triggering febrile convulsions. GABRG2 rs211037 TT homozygotes and T allele variants have an increased risk for developing febrile seizures. Recurrent crises and repeated episodes of seizures are more frequent in the GABRG2 Asn196Asn TT genotype polymorphism, with a 45 and 8 times higher risk of developing idiopathic generalized epilepsy and recurrent febrile seizures, respectively.

Comparison of 10-year overall survival between patients with G1 and G2 grade Ta bladder tumors.

To compare long-term overall survival (OS) in patients with G1 and G2 grade Ta bladder cancer after transurethral resection of bladder tumors (TURBTs). Secondary aim was to investigate clinical and pathologic prognostic factors for OS of Ta patients, except G3/high grade (HG).A total of 243 patients, retrospectively selected, with Ta nonmuscle invasive bladder cancer (NMIBC) underwent TURBT between January 2006 and December 2008 (median follow-up 109 months). Inclusion criteria were: Ta at first manifestation, G1 or G2 grade with no associated carcinoma in situ (CIS). Seventy-nine patients were excluded due to concomitant CIS (1), G3/HG tumors (47), and lost to follow-up (31). Ethical approval was obtained from the Ethical Committee of the Mures County Hospital. Statistical analysis was performed using STATA 11.0.Following inclusion criteria, 164 patients with primary G1 or G2 Ta tumors, were enrolled. Recurrence was observed in 26 (15.8%) and progression in 5 (3%) patients. Ten-year survival in G1 patients was 67.8% (CI 54.3-78.1) and in G2 patients 59% (CI 49-67.3) (P = .31). Univariable and multivariable logistic regression analysis underlined that advanced age at diagnosis (hazard ratio [HR] 1.10) and no Bacillus Calmette-Guerin (BCG) treatment (HR 0.24 and 0.29) were independent predictors for death at 10 years after diagnosis.Long-term analysis confirms that patients with well differentiated (G1) and moderately well differentiated (G2) Ta tumors have similar OS. A longer OS was even reported in those who underwent BCG adjuvant therapy.

Triplex High-Resolution Melting Assay for the Simultaneous Assessment of IFNL3 rs12979860, ABCB11 rs2287622, and RNF7 rs16851720 Genotypes in Chronic Hepatitis C Patients.

Chronic hepatitis C (CHC) is a leading cause of liver disease. Despite the improved efficacy of new antivirals, their high costs preclude their adoption in resource-limited settings, where CHC prevalence is highest. We developed a triplex high-resolution melting assay for the simultaneous assessment of three genetic polymorphisms related to the response to treatment and development of advanced fibrosis in CHC: IFNL3 rs12979860, ABCB11 rs2287622, and RNF7 rs16851720. We validated the assay in clinical samples from 130 CHC patients treated with classic therapy. The assay showed excellent reproducibility and 100% accuracy, sensitivity, and specificity against the gold standard Sanger sequencing. When added to routine examination data, genotype information significantly improved their performance for prediction of advanced liver fibrosis and sustained virological response (P = 0.041 and P = 0.011, respectively). Correspondingly, the full models had area under the receiver operating characteristic curve values of 0.842 (95% CI, 0.773-0.911) and 0.921 (95% CI, 0.870-0.972) and integrated discrimination improvements of 7.5% (95% CI, 2.5%-12.5%; P = 0.003) and 11.5% (95% CI, 5.8%-17.2%; P < 0.001), respectively. This is the first report on a diagnostic test for simultaneous genotyping of IFNL3, ABCB11, and RNF7 in CHC patients. Reliable and inexpensive, the assay should provide useful information for the clinical management of CHC, like identification of patients at risk of rapid disease progression or with high chances of response to classic therapy.

Effectiveness of fine-needle aspiration cytology in the diagnosis of lateral cervical nonthyroid tumors.

Given that the clinical and radiological examinations of lateral cervical masses are not always sufficient for deciding on appropriate management, the cytological examination of the material obtained by fine-needle aspiration might be an efficient tool in the preoperative investigation of these lesions.In this prospective cross-sectional study we evaluated the efficacy and diagnostic accuracy of fine-needle aspiration cytology in the assessment of lateral cervical nonthyroid tumors, by comparing its results with those of histopathology.A total of 58 patients with lateral cervical masses were included. Preoperative cytological results were compared with the histopathologic examination of surgical specimens.Both cytology and histology indicated that malignant tumors outnumbered benign lesions (62% vs 38%), with 88.9% of malignancies presenting in patients aged >50 years, but cytology was less effective at differentiating between benign and nontumor lesions. Cytology had 76.5% specificity and 78.1% sensitivity for identifying malignant lateral cervical lesions, and there was a concordance between the two diagnostic tests (McNemar test, P = 0.17, κ = 0.50, P <0.001).Fine-needle aspiration cytology is a simple, quick, and effective procedure that can aid in the preoperative evaluation of lateral cervical masses by differentiating benign tumors and inflammatory processes from malignancies and thus help in determining a subsequent therapeutic strategy.

The value of histopathological diagnosis in the elderly patients with granulomatous dermatoses. Case series.

Granulomatous inflammations are a particular type of chronic septic or aseptic inflammation, in which infectious or non-infectious agents are difficult to eliminate by the immune system. These are type IV hypersensitivity reactions mediated by pre-sensitized T-lymphocytes cells CD4+ and CD8+ lymphocytes. Disorders included in this category are: tuberculosis, leprosy, syphilis, sarcoidosis, type I diabetes, multiple sclerosis, Crohn's disease and rheumatoid arthritis. At cutaneous level, this pattern of granulomatous reaction is characterized by a chronic inflammation with formation of granulomas consisting of a variable number of histiocytes, multinucleated giant cells and lymphocytes. Granulomatous dermatoses should be differentiated from other primary or secondary lesions affecting the skin such as inflammation or tumors. Often granulomatous dermatoses can be confused with other skin disorders, both clinically and histological. Histopathology examination can add important information and clarify the diagnosis. This paper presents a series of three clinical cases of granulomatous skin occurring in the elderly patients confirmed at histology examination. Clinical and histology criteria were analyzed, along with specific differential diagnosis, based on data from the literature.

Study regarding the microscopic aspects of pilo-sebaceous units after antiandrogen treatment in hirsute women.

AIM: To analyze the morphological data of pilo-sebaceous units in hirsute women before and 12 months after the antiandrogen treatment with Cyproterone acetate (CPA) 100 mg÷day. MATERIALS AND METHODS: Fourteen female patients with idiopathic hirsutism that followed an antiandrogen treatment with CPA were biopsied from the androgen-dependent area of the chin before and 12 months after the treatment. Routine sections were stained with Hematoxylin-Eosin, Masson, Van Gieson, Sirius red and picric-indigocarmine, while additional sections were immunostained for S100 protein and vimentin. Electron microscopy was performed in two cases with Langerhans cell hyperplasia. RESULTS: On biopsies-stained sections, an increased number of hair follicles, the deeper part of the epithelial sheath of the hair follicle with epithelial buds, hyperplasia of sebaceous glands, and no inflammatory infiltrate were noticed. Langerhans cells identified with S100 protein and vimentin were normal in terms of numbers and distribution. After the administration of the treatment, atrophy of the pilo-sebaceous units was visible in nine (64.2%) cases, while inflammatory infiltrate and cells included in the vacuoles of the basal layer of the epidermis became apparent. In six of the cases treated with antiandrogens, a marked hyperplasia of Langerhans cells was noticed. In conclusion, the benefit of antiandrogen treatments is supported by atrophy of the hair follicle and the sebaceous glands. The activation of Langerhans cells associated with inflammatory infiltrate in the dermis and hair follicles could be considered as a local consequence of the involution process of hair follicles after the administration of the treatment.

Correlations between anthropometric and serologic elements of metabolic syndrome and histopathologic features of nonalcoholic fatty liver disease.

AIM OF THE STUDY: Studying the correlation between elements of metabolic syndrome and histological changes of the liver in nonalcoholic fatty liver disease. PATIENTS AND METHODS: Thirty-nine patients with nonalcoholic fatty liver disease were included in our study. Inclusion criteria were: presence of liver steatosis on ultrasound in patients with waist circumference over 94 cm in men and over 80 cm in women and with serologic elements of metabolic syndrome. Exclusion criteria were: chronic viral hepatitis, autoimmune hepatitis, Wilson disease, hemochromatosis, regular alcohol consumption. Body mass index, waist circumference, fasting plasma glucose, serum triglyceride and cholesterol levels and serum ALT were determined. On liver biopsy specimens, performed in each patient, the NASH score, representing the sum of fibrosis, steatosis, lobular inflammation and ballooning, was calculated. RESULTS: Necroinflammation was mild in 15 patients, medium in 19 patients and severe in five patients. Mild fibrosis was present in four cases, medium in 14 cases, severe in six, and two patients were diagnosed with cirrhosis. We found statistically significant correlation between waist circumference and the grade of histological activity, the presence of diabetes and both fibrosis grade and histological activity, and the serum ALT and histological activity. CONCLUSIONS: Noninvasive assessment of the severity of hepatic histological changes in nonalcoholic fatty liver disease could be made by anthropometric parameters or by serologic components of metabolic syndrome, but it is not an accurate method to identify patients with high-risk for disease progression. These noninvasive parameters cannot replace liver biopsy.

New ways of bronchial stump closure after lung resection: experimental study.

INTRODUCTION: Lung surgery has long been associated with a high rate of mortality and morbidity. Bronchial stump fistula is one of the leading causes of high morbidity and high mortality rate. The objectives of our study are to asses the effect of new ways of closure the bronchial stump after lung resections. MATERIALS AND METHODS: For our study, we used eighteen adult rabbits. We performed left trans-thoracic inferior lobectomy to each animal because left lung functional capacities are smaller than right lung functional capacities. The bronchial stump was closed in three different ways: simple running suture of 5.0 monofilament polypropylene (Sweet procedure), running suture of 5.0 monofilament polypropylene (horizontal "U" shape model) and Vycril patch, running suture of 5.0 monofilament polypropylene (horizontal "U" shape model) and bovine pericardium patch. All surviving rabbits were sacrificed in 30 day postoperatively. RESULTS: In our study, we did not notice any deaths among our experimental animals and we did not observe any bronchial stump fistula. In the group of rabbits in which the bronchial stump was closed using a patch of heterologous bovine pericardium, the histological exams revealed a cvasinormal bronchial wall with a normal bronchial epithelium and basal membrane integrity and no inflammatory lympho-plasmocytary infiltrate cell, absence of macrophages at this level. CONCLUSIONS: Using the heterologous bovine pericardium patch to reinforce the bronchial stump could be beneficial because of the faster healing and scarring process. To our knowledge, no studies have previously evaluated this procedure of bronchial stump closure. Our findings could therefore, serve as an impetus for further research in this area.

Current data in clinicopathological characteristics of primary hepatic tumors.

AIM: The present study aimed to investigate the status of primary liver tumors diagnosed in the Emergency Hospital of Mures County, Romania. MATERIALS AND METHODS: We performed a retrospective study based on data collected from histopathological bulletins stored in the archives of the Pathology Laboratories of Mures County. We selected those patients' data that were diagnosed with liver tumors during January 2004-August 2011. The acquired data were collected and processed statistically using the GraphPad InStat Demo 3 statistical software. RESULTS: We identified 748 liver tumors out of which 264 were primary liver tumors (35.29%), whereof 108 were malignant primary liver tumors and 156 benign liver tumors. The male:female ratio in case of malignant tumors was 1.7:1 and for benign tumors it was 1:2.62. In case of malignant tumors, the mean age of male patients was 63.98±11.67 as for female patients it was 60.14±11.65 years. In case of benign tumors, the mean age of male patients was 59.29±9.58 as for female patients it was 55.55±12.86. 81.48% of the malignant tumors were hepatocellular carcinoma (HCC), followed by cholangiocarcinoma (14.81%). 40% of HCC presented associated cirrhosis, in the other 60% signs of chronic alcoholism being observed. CONCLUSIONS: Hepatocellular carcinoma remains the first on the list of malignant primary liver tumors in Romanian patients, probably due to a high rate of alcohol consumption.

Multiple organ histopathological changes in broiler chickens fed on genetically modified organism.

Diet can influence the structural characteristics of internal organs. An experiment involving 130 meat broilers was conducted during 42 days (life term for a meat broiler) to study the effect of feed with protein from genetically modified soy. The 1-day-old birds were randomly allocated to five study groups, fed with soy, sunflower, wheat, fish flour, PC starter. In the diet of each group, an amount of protein from soy was replaced with genetically modified soy (I - 0%, II - 25%, III - 50%, IV - 75%, V - 100% protein from genetically modified soy). The level of protein in soy, either modified, or non-modified, was the same. Organs and carcass weights were measured at about 42 days of age of the birds and histopathology exams were performed during May-June 2009. No statistically significant differences were observed in mortality, growth performance variables or carcass and organ yields between broilers consuming diets produced with genetically modified soybean fractions and those consuming diets produced with near-isoline control soybean fractions. Inflammatory and degenerative liver lesions, muscle hypertrophy, hemorrhagic necrosis of bursa, kidney focal tubular necrosis, necrosis and superficial ulceration of bowel and pancreatic dystrophies were found in tissues from broilers fed on protein from genetically modified soy. Different types of lesions found in our study might be due to other causes (parasites, viral) superimposed but their presence exclusively in groups fed with modified soy raises some serious questions about the consequences of use of this type of feed.

Cardiac allograft vasculopathy in adult human recipients: a case series of seven patients and review of literature.

BACKGROUND: Chronic rejection (CR) also called cardiac allograft vasculopathy (CAV) is, besides infections and malignant tumors, the leading cause of death during the late period of post-heart transplant. PATIENTS AND METHODS: In this paper, we present a series of seven cases with chronic post-transplant cardiac rejection in the light of our experience related to histopathological aspects, difficulties in diagnosing and survival time. RESULTS: Our study comprises patients whose ages ranked at the time of transplant between 33 and 58 years, with a mean age of 47.71 years, the ratio between men and women being 6:1. Chronic rejection - cardiac allograft vasculopathy occurred in all seven patients comprised in this study, the earliest in the second year post-transplant (three patients), followed by the third year (one patient), the seventh and eighth year (one patient) and the latest survival period being over 11 years (one patient). Four out of the seven patients with chronic rejection events were preceded by episodes of acute cellular rejection. The most convincing indirect evidence of chronic rejection in endomyocardial biopsies were: ischemic events of the myocardiocytes and impaired microvascular network because of perivascular and interstitial fibrosis. CONCLUSIONS: Our study shows that the most important morphological factors correlated with the manifestation of chronic rejection were the episodes of acute cellular rejection or perivascular and interstitial fibrosis, these injuries also being indirect signs mainly detectable at the level of endomyocardial biopsies.