Pilot Study of the Occurrence of Somatic Mutations in Ciliary Signalling Pathways as a Contribution Factor to Autosomal Dominant Polycystic Kidney Development.

Autosomal-dominant polycystic kidney disease (ADPKD) is an inherited disease that results in multiple kidney cysts, and it is a common cause of end-stage renal disease. Recent studies have shown that disease progression can be slowed by simultaneous disruption of the primary cilium and polycystins. The exact genetic mechanism of this process is still unknown. The aim of the present study was to characterize the mutation profile of ciliary signalling pathways in the renal epithelial cells of ADPKD patients. In our study, we performed an analysis of 110 genes encoding the components of Sonic Hedgehog, Hippo, Notch, Wnt and planar cell polarity signalling (PCP) by targeted next-generation sequencing. We analysed 10 formalin-fixed, paraffinembedded (FFPE) tissue samples of patients with ADPKD. We identified a unique mutation profile in each of the analysed ADPKD samples, which was characterized by the presence of pathogenic variants in eight to 11 genes involved in different signalling pathways. Despite the significant genetic heterogeneity of ADPKD, we detected five genes whose genetic variants affected most ADPKD samples. The pathogenic variants in NCOR2 and LRP2 genes were present in all analysed samples of ADPKD. In addition, eight out of 10 samples showed a pathogenic variant in the MAML2 and FAT4 genes, and six out of 10 samples in the CELSR1 gene. In our study, we identified the signalling molecules that may contribute to the cystogenesis and may represent potential targets for the development of new ADPKD treatments.

Neuron-specific enolase in the intestinal wall in Crohn´s disease.

The authors described the localization of neuron-specific enolase in the intestinal wall in Crohn´s disease. We have used samples obtained by biopsy from the colon lining of five people affected by Crohn's disease for our examination. We have processed samples using the formol paraffin technique. From paraffin blocks, we have prepared histological sections approximately 5 µm thick. For immunohistochemic examinations, we have revitalised the sections by acquiring the heat-induced epitope. We detected NSE by monoclonal mouse antibodies against human neuron-specific enolase, clone BBS/NC/VI-H14 (DakoCytomation, Denmark) (Fig. 4, Ref. 7).

Laryngotracheal Amyloidosis: Amyloid Airway Center 23-Year Experience.

OBJECTIVES: Localized laryngotracheal amyloidosis (LA) is a rare disease that can impact phonation and respiration. Treatment options include observation, surgery, and radiation therapy (RT). Given the rare incidence of LA, evidence regarding optimal management and long-term outcomes is limited. STUDY DESIGN: Retrospective cross-sectional analysis. METHODS: All patients with LA presenting to an international amyloid center from 1999 to 2022 were analyzed. Patients were categorized by treatment modality: surgery, RT, or observation. Patient and disease factors including demographics, clinical presentation, and progression with need for additional treatment were evaluated. RESULTS: Seventy-one patients (27M:44F) with LA were treated with surgery (n = 40), RT (n = 11), and observation (n = 20). Gender distribution, age at diagnosis, and systemic workup did not differ significantly between treatment cohorts. A correlation was identified between LA location and treatment modality, with higher rate of subglottic/tracheal amyloid in RT patients vs. surgery and observation patients [(90% and 52% respectively), p < 0.005]. Surgery patients had a median of two surgeries for disease management (range: 1-32) and RT patients had median five surgeries prior to RT (range: 0-17). Six patients required tracheotomy: 3/40 surgery, 3/11 RT and 0/20 in observation cohort. Surgery and RT patients had a longer duration of follow-up (mean 6.7 and 11.7 years) compared with the observation cohort (5.7 years). CONCLUSION: Laryngotracheal amyloidosis is a rare disease with variable presentation. Selective surgery of involved subsites is the primary treatment, though multiple surgeries may be needed to optimize function. Observation is appropriate for those with minimal symptoms. For recalcitrant disease, and particularly subglottic/tracheal amyloid, radiotherapy can be beneficial. LEVEL OF EVIDENCE: 4 Laryngoscope, 134:1606-1613, 2024.

Evaluation of protein expression and DNA methylation profiles detected by pyrosequencing in invasive breast cancer.

Breast carcinoma is the most common cancer with high mortality caused by metastatic disease. New molecular biomarkers predicting the tumour's metastatic potential would therefore improve metastasis prevention and personalised care. The aim of the study was to investigate the relationship between DNA methylation levels in invasivity and metastasising associated genes with aberrant protein expression and also to evaluate whether a similar DNA methylation level is present in the tumour and circulating cell-free DNA for utilising plasma DNA methylation as prognostic biomarker. By using pyrosequencing, we analysed DNA methylation levels of 11 genes, namely APC, ADAM23, CXCL12, ESR1, PGR B, CDH1, RASSF1A, SYK, TIMP3, BRMS1 and SOCS1 in tumour, plasma and peripheral blood cells from 34 patients with primary breast cancer, as well as plasma and peripheral blood cells from 50 healthy controls. Simultaneously, the expression of related proteins in paraffin-embedded tumour samples was evaluated by immunohistochemistry. Statistical analysis was performed by SPSS statistics 15.0 software. Tumour DNA hypermethylation was found in most commonly methylated RASSF1A (71.9%), APC (55.9%), ADAM23 (38%) and CXCL12 (34.4%) genes with methylation levels up to 86, 86, 53 and 64 %, respectively. In tumours, significantly higher methylation levels were found in nine genes, compared with the patients´ peripheral blood cell DNA. Furthermore, in patients methylation levels in peripheral blood cell DNA were significantly higher than in controls in CXCL12, ESR1 and TIMP3 genes, but the values did not exceed 15%. On the other hand, no correlations were observed in patients between DNA methylation in tumours and cell-free plasma DNA. Moreover, in patients and controls nearly identical values of cumulative DNA methylation (43.6 % ± 20.1 vs. 43.7 % ± 15.0) were observed in plasma samples. A variable spectrum from high to none expressions presented in tumour tissues in all of the proteins evaluated, however in APC and CXCL12 genes a visible decreasing trend of mean DNA methylation level with increasing expression of the corresponding protein was observed. The DNA methylation profiles manifested in our group of breast carcinomas are cancer specific, but they are not the only cause that affects the silencing of evaluated genes and the decrease of relevant protein products. The clinical utility of DNA methylation testing in peripheral blood cell DNA for cancer diagnosis and therapy need to be further investigated.

Morphology of intestinal microcirculation in colitis ulcerosa and Crohn's disease.

AIM OF THE WORK: To describe the state of microcirculation in the intestinal wall in ulcerative colitis and Crohn's disease in own material. MATERIALS AND METHODS: For morphological examination, we obtained tissue samples from both diseases during bioptic bowel examination from 11 patients. We fixed the samples with 10% buffered formalin, de-hydrated and covered with paraffin. From paraffin blocks, we made histological sections about 5µm thick with a microtome. They were always stained with haematoxylin and eosin. For immunohistochemistry sections, we either did or did not revitalise the sections according to the manufacturer's recommendations for specific to-reagents. For immunohistochemic examinations, we revitalised the sections by acquiring the heat-induced epitope in DakoCytomation Target Retrieval Solution, Code No S 1700, or in mmol.l-1 citrate buffer, pH 6.0. We did not dry the sections during such procedure. We used antibodies as follows: Monoclonal mouse antibodies against human von Willebrand factor, Clone F8/86 (DakoCytomation, Denmark), Monoclonal mouse antibodies against smooth muscle actine, Clone HHF35 (DakoCytomation, Denmark). We used detection system Dako EnVisionTM + Dual Link System-HPR (Dako, Denmark) or Dako Liquid DAB + Substrate Chromogen System (Dako, Denmark) to display areas of specific connection of antibodies. Antibodies against von Willebrand factor react in our samples with the endothelium of vessels and with precursors in the lining of peaks of villi. We have also seen a diffuse positive reaction. Lymphatic vessels do not display monoclonal antibodies (Fig. 4, Ref. 8).

[Virological sustained response to former young intravenous drug abusers with chronic hepatitis C treated by pegylated interferon-α plus ribavirin]. / Trvalá virologická odpoved kombinovanej imunomodulacnej liecby pegylovaným interferónom-α a ribavirínom u bývalých intravenóznych drogovo závislých jedincov s chronickou vírusovou hepatitídou C.

INTRODUCTION: The most risk group with high rate of infection of viral hepatitis C represents the former drug abusers (IDUs). OBJECTIVES: The objectives of the clinical study were to assess the sustained virological response (SVR) in the group of former IDUs with chronic hepatitis C (CHC) treated with pegylated interferon-α and ribavirin. PATIENTS AND METHODS: Of 293 previously untreated (naive) former IDUs with CHC who have started treatment, we assessed 239 (69 women, 170 men) with an average age of 27 years who completed treatment and received a follow-up examination after 24 weeks. Prior to therapy abstinence to drug abuse 6 and more months has been required. The CHC has been diagnostics by the standard virological diagnostic tests, increased activity of aminotranferases (ALT) and histology of liver biopsy. Patients were treated with combined immunomodulatory therapy in the standard duration and standard regimen. SVR was considered if qualitative test HCV RNA was negative 24 weeks after the completion of the treatment. RESULTS: End-of-treatment virological response was in 224/94 % patients. Overall SVR rate was 95 %, 227 subjects had negative HCV RNA and only 12 patients no response. During treatment there was a statistically significant decrease in ALT. Treatment was accompanied by the expected side effects. CONCLUSION: Young age, short duration of HCV infection, high prevalence of genotype 3 and low grade of liver fibrosis in majority of patients and good adherence of patients to treatment were crucial predictive factors resulting in high SVR.

Diverse nanocomposites as a potential dressing for diabetic wound healing.

Wound healing is a programmed process of continuous events which is impaired in the case of diabetic patients. This impaired process of healing in diabetics leads to amputation, longer hospitalisation, immobilisation, low self-esteem, and mortality in some patients. This problem has paved the way for several innovative strategies like the use of nanotechnology for the treatment of wounds in diabetic patients. The use of biomaterials, nanomaterials have advanced approaches in tissue engineering by designing multi-functional nanocomposite scaffolds. Stimuli-responsive scaffolds that interact with the wound microenvironment and controlled release of bioactive molecules have helped in overcoming barriers in healing. The use of different types of nanocomposite scaffolds for faster healing of diabetic wounds is constantly being studied. Nanocomposites have helped in addressing specific issues with respect to healing and improving angiogenesis. Method: A literature search was followed to retrieve the articles on strategies for wound healing in diabetes across several databases like PubMed, EMBASE, Scopus and Cochrane database. The search was performed in May 2022 by two researchers independently. They keywords used were "diabetic wounds, nanotechnology, nanocomposites, nanoparticles, chronic diabetic wounds, diabetic foot ulcer, hydrogel". Exclusion criteria included insulin resistance, burn wound, dressing material.

CT evaluated sarcopenia signals: Shorter survival for small cell lung cancer patients.

Sarcopenia is an independent risk factor for morbidity and mortality in patients suffering from small cell lung cancer (SCLC), however, a universal indicator of sarcopenia usable in clinical practice is still missing. A novel indicator for describing the severity of cancer could be helpful in tailoring the anti-tumor therapy. The aim of this study was to evaluate the computed tomography (CT) scans of total muscle area and radiation attenuation in patients suffering from small cell lung cancer. We used staging CT scans performed at the time of diagnosis to measure total muscle area (TMA) and average psoas density (PD) at level of the 3rd lumbar vertebra. TMA and PD were statistically evaluated in association with overall survival and disease staging. We used Mann-Whitney test and Spearman´s correlation coefficient for statistical testing and p-value under 0.05 was considered statistically significant. Retrospectively we examined 47 patients suffering from SCLC (mean age 65.05+/-7.3 years, BMI 23.97+/-4.4 kg/m2, BSA 1.77+/-0.2 m2, 30-day mortality was 4.3 % with 10 months median survival). As sarcopenia was pointed TMA under 55 and 39 cm2/m2 for men and women respectively. The sarcopenic patients had significantly shorter median survival (7 vs. 11 months, p=0.05). We observed a significant relationship between survival and performance status (Spearman´s correlation, R=-0.39, p=0.05). The patients were divided into two groups according to the extensive (ED, n=34) or limited (LD, n=13) form of the disease. We observed significant difference in PD (42.49+/-6.1 vs. 47.67+/-4.5 HU, p=0.006) between ED vs. LD groups.

Renal angiomyoadenomatous tumor: morphologic, immunohistochemical, and molecular genetic study of a distinct entity.

We present a series of a distinct tumorous entity named renal angiomyoadenomatous tumor (RAT). Five cases were retrieved from the consultation files of the authors. Histologic and immunohistochemical features were evaluated. Sequencing analysis of coding region of the VHL gene was carried out in all cases. The tumors were composed of admixture of an epithelial clear cell component and prominent leiomyomatous stroma. Epithelial cells formed adenomatous tubular formations endowed with blister-like apical snouts. All tubular/glandular structures were lined by a fine capillary network. The epithelial component was positive for epithelial membrane antigen, CK7, CK20, AE1-AE3, CAM5.2, and vimentin in all cases. In all analyzed samples, no mutation of the VHL gene was found. RAT is a distinct morphologic entity, being different morphologically, immunohistochemically, and genetically from all renal tumors including conventional clear cell carcinoma and mixed epithelial and stromal tumor of kidney.

High virologic sustained response for former young intravenous drug users with chronic hepatitis C treated by pegylated interferon-alpha plus ribavirin.

AIMS: The aim of this clinical study was to assess virological response at end-of -treatment (ETR), sustained virological (SVR) and biochemical response in former drug users with chronic hepatitis C treated with PEG-IFN-alpha and R. PATIENTS: Ninety two former drug users (21 F, 71 M) average age 27 years (18 to 41 years) and previously not treated with IFN-alpha and R (naive patients, pts) were evaluated for their virological and biochemical response. Standard treatment regimen of either 24 or 48 weeks was applied in patients with genotype 3 or genotype 1, respectively. SVR was considered if viral tests (HCV RNA) were negative 24 weeks after the end of treatment. RESULTS: Overall SVR was attained in 87 (95%) of 92 treated patients, and therapy failed in 5 pts with genotype 1. In genotype 1 patients ETR and SVR were 81% and 86%, respectively (p < 0.001). In genotype 3 patients ETR and SVR were 98% and 100%, respectively (p < 0.001). ALT levels decreased significantly after 12 weeks of therapy (ALT 1.61 vs 0.64 micro/kat/l, p < 0.001) and were at normal levels during follow-up. CONCLUSIONS: Crucial predictive factors resulting in high SVR were the younger age in combination with low stage of liver fibrosis, relatively short duration of viral infection, high proportion of genotype 3 and excellent adherence of patients to treatment regimen than previously not treated with IFN-alpha and R (naive patients). High proportion of SVR in former drug users has been achieved in patients with genotype 3 (100%) and genotype 1 (86%). The most decisive prognostic factor which favors high therapeutic efficacy appears to be young age and early onset of anti-HCV treatment (Tab. 3, Fig. 1, Ref. 33). Full Text (Free, PDF) www.bmj.sk.

[The 2007 World Health Organisation classification of tumours of the central nervous system, comparison with 2000 classification]. / WHO klasifikácia tumorov centrálneho nervového systému 2007: porovnanie s klasifikáciou z roku 2000.

WHO Classification of Tumours of the Central Nervous System from the 2007 is distinguished from the previous 2000 classification by a few conceptual modifications, changes in the terminology and seven newly codified tumour entities. The text shows a short comparison of both classifications emphasising the most important changes from the surgical neuropathology point of view. The newly codified entities are: angiocentric glioma, pilomyxoid astrocytoma, papillary glioneuronal tumor, rosette-forming glioneuronal tumor of the 4th ventricle, papillary tumour of the pineal region, spindle cell oncocytoma and pituicytoma. Mostly, they are rare tumours already known from the literature. Based on new knowledge from the molecular pathology the paragraphs about tumour genetics were markedly changed. The complexity and diversity of tumours of the nervous system is enormous, and, not surprisingly, some problematic questions of classification and grading remain unresolved.

[Collagenofibrotic glomerulopathy--rare glomerulonephritis]. / Kolagenofibrotická glomerulopatia--raritná glomerulonefritída.

Glomerulopathies with fibrillary deposits form a heterogeneous group of renal diseases that can be identified only by means of electron microscopy. A case of a rare type of such a nephropathy, the collagenofibrotic glomerulopathy with focus on differential diagnostics is presented and current knowledge relating to this renal disease is reviewed.

[Treatment of chemically dependent patients: a program based on the Minnesota Model]. / Traitement des malades dépendants: un programme basé sur le modèle Minnesota.

An example of the Minnesota Model adapted to a Swiss private clinic (Clinic La Métairie) is presented with the aim of showing its place in the network facilities for the treatment of addiction. The prospective study based on the Addiction Severity Index questionnaire allows to describe the characteristics of hundred patients treated during the years 2002-2003 and follow their evolution one year after the treatment. This evolution is favourable in all investigated areas; 38% of patients remained totally abstinent during one year and even the outcome of 15% of patients in relapse has improved as well.

Sialic acid expression in normal and diseased human kidney.

Three sialic acid-specific lectins, Tritrichomonas mobilensis (TML) (without linkage preference), Maackia amurensis leukoagglutinin (MAL) (alpha2,3) and Sambucus nigra agglutinin (alpha2,6 linkage-specific), were used for detection of sialylated glycoconjugates in normal and diseased human kidneys. Normal kidneys demonstrated strong podocyte positivity of alpha2,3 linked sialic acid and weaker sialic acid expression on capillary endothelium, which was alpha2,6 linked. Renal biopsies (45) representing a variety of diseases, e.g. minimal change disease, membranous, membranoproliferative glomerulonephritis, tubulointerstitial nephrosis or diabetic glomerulopathy showed increased sialic acid expression in glomerular capillaries, Bowman's capsule epithelium and on podocytes. In several different kidney diseases the glomerular endothelium expressed also alpha2,3 linked sialic acid along with increased TML-positivity of epithelial cells related to alpha2,6 linked sialic acid. No difference in sialic acid linkage expression was observed in the tubules, which expressed putative alpha2,6 linked sialic acid on the luminal surface of cells distal to the descending limb of Henle's loop. The study did not show changes characteristic of specific diseases. Rather, increased sialic acid expression on glomerular endothelium and podocytes was linked to a variety of pathological changes. It is assumed that changes in sialysation of glycoconjugates in the glomeruli represent nonspecific changes and do not reflect fundamental pathogenetic features of renal diseases.

Tubular basement membrane thickening in diabetes mellitus.

Authors measured the thickness of the basement membranes of the proximal and distal tubuli in the renal cortex in 20 patients with verified diabetes mellitus. The test group consisted of 20 patients with small abnormalities of glomeruli (or in the range of variable norm), of the same sex and comparable age. Measurements were performed in locations without tubulo-interstitial changes. The average thickness of TBM in patients with diabetes mellitus was 1540 +/- 616 nm, and in the test group 360 +/- 86 nm, which is a statistically significant difference (p < 0.000000). In spite of the small number of cases and possible error in the manual measurement of electron microscopic pictures, a significant thickening of TBM in diabetes mellitus has been found. We consider this to be of diagnostic value.

Our experience with surgical treatment of the tumours of peripheral nerves in extremities and brachial plexus.

OUTLINE: The paper presents the results of 33 microsurgical removal of 33 tumours of brachial plexus and of peripheral nerves in extremities performed on 31 patients during the period of 11 years, from 1990 to 2001. PATIENT GROUP AND METHODS: The patient group included 26 patients treated for removal of 28 tumours of peripheral nerves in extremities and 5 patients treated for removal of 5 tumours of brachial plexus. Three of these brachial plexus tumours showed intraspinal spread. 32 tumours of neural sheath included 20 schwannomas and 12 neurofibromas. 6 of these 12 neurofibromas were observed in 4 patients suffering from Reklinghausen disease (VRD). One patient had a tumour of non-neural elements--lipoma. We did not encounter any malign tumours of neural sheaths or other neural sheath-derived tumours (benign or metastatic malign ones) that would compress or invade the nerves or plexus. The results were evaluated according to Donner classification of motor and sensory functions. RESULTS: Motor functions improved or remained unchanged in 18 (90%) patients with schwannoma. 5 patients (83.3%) with pain in the distribution of the affected nerve reported complete or partial improvement of the symptoms. Regarding neurofibroma patients, 3 exhibited either an improvement or unchanged motor functions, and all of them reported partial or complete retreat of pain symptoms. 4 patients with VRD were subjected to the removal of 6 tumours of peripheral nerves in extremities; all of them exhibited improved or stabilised motor functions and partial or complete retreat of pain symptoms. The outcome of treatments was generally better in smaller tumours of neural sheaths with more distal localisation. CONCLUSION: Patients should be treated in the early stage of tumour development when the size remains smaller and neurological deficits are absent.

[Colonoscopy monitoring intervals after polypectomy of adenomas of the large intestine in patients younger than 40 years of age]. / Intervaly kolonoskopických kontrol po polypektómii adenómov hrubého creva u pacientov mladsích ako 40 rokov.

A group of patients younger than 40 years of age, who had been subdued to endoscopic large intestine adenoma polypectomy were compared with the rest of the patients regarding the adenoma occurrence according to sex, localisation, histologic character, frequency of recurrence, and carcinoma formation in the large intestine in the site of previous polypectomy. The compared groups of patients differed merely in time of recurrence. In patients over 40 years of age the adenomas reoccurred most frequently in the first and fifth years following the primary polypectomy. In patients after primary polypectomy regular colonoscopic controls were suggested, namely after the first year and subsequently after every two years. This scheme is appropriate also for patients under the age of 40 years. Therefore the recommended intervals of colonoscopic controls at this age category do not require to be subdued to alteration. (Tab. 14, Ref. 5.)

[Angiocardiography of cardiopathy of unknown origin]. / Angiokardiografia kardiopatií nejasného pôvodu.

Authors described their experiences with angiocardiography, (selective coronarography and left ventriculography), in 41 patients aged from 32 to 59 years with the preliminary diagnosis of cardiopathy of unknown origin. The patients with diagnosis of cardiopathy of unknown origin were those, who cannot be assigned to any of the known and acknowledge diagnostic categories, not even by using complex clinical, laboratory and noninvasive cardiological diagnostic procedures. Angiocardiography enabled to produce new and unexpected data of diagnostic value three patients with heart disease (all of the three mentioned patients had significant changes of the coronary supply, i.e. the reduction in lumen diameter of the coronary vessel being of more than 50% with local or total wall motion abnormalities). The other thirty-eight patients had normal findings of the coronary supply. One patient had a normal coronarographic findings, normal left ventricular wall motion and also the subsequent histological examination from the endomyocardial bioptic sample of the heart tissue, did not confirm pathological abnormalities. Selective coronarography and left ventriculography is a useful and safe clinical diagnostic procedure in exactly defined groups of patients.