Male infertility is a multifactorial condition contributing to approximately 50% of all cases of couple infertility. In recent years, significant advances have been made in both diagnostics and treatment. This review summarizes key developments from 2019 to 2024 with direct relevance to routine clinical practice in Czech urology and andrology. Particular attention is paid to the updated semen analysis standards (World Health Organisation 6th edition, 2021), sperm DNA fragmentation testing, genetic evaluation (karyotyping, Y chromosome microdeletions, and exome sequencing), surgical management of varicocele, and sperm retrieval techniques for azoospermia, including microdissection testicular sperm extraction (micro-TESE). The article also discusses pharmacological options (gonadotropins, selective estrogen receptor modulators, antioxidants), the impact of lifestyle factors, and the importance of interdisciplinary collaboration with assisted reproduction centers. Future perspectives, including the role of preventive strategies in male reproductive health, are also addressed. The aim is to provide a comprehensive and clinically applicable overview of current recommendations and therapeutic approaches in andrology, with a focus on their implementation in the Czech urological setting.

• MeSH
• analýza spermatu metody   MeSH
• antioxidancia farmakologie   terapeutické užití   MeSH
• asistovaná reprodukce MeSH
• genetické testování metody   MeSH
• gonadotropiny terapeutické užití   MeSH
• lidé MeSH
• mužská infertilita * diagnóza   etiologie   terapie   MeSH
• odběr spermií MeSH
• selektivní modulátory estrogenních receptorů farmakologie   terapeutické užití   MeSH
• varikokéla chirurgie   MeSH
• životní styl MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• systematický přehled MeSH

V článku je popsán současný vývoj mikrochirurgických operačních technik mikrochirurgické operace varikokély a mikrochirurgického odběru zárodečné tkáně z varlat (microTESE). U operace varikokély přinesl vývoj peroperační mikrovaskulární Dopplerovskou diagnostiku, trojrozměrné zvětšení a bezesvorkovou metodu ligatury spermatických žil při prezervaci spermatických arterií. U microTESE je přínosem možnost předoperační triplexní mikrovaskulární Dopplerovské diagnostiky, hydrodisekce kanálků testis a vysoké zvětšení ("high power") kanálků in vivo s možností mikrometrie kanálků, selekce odběru kanálků s vyšší pravděpodobností přítomnosti spermií a vysoký stupeň ochrany ostatní tkáně testis. Postupy jsou demonstrovány ve fotodokumentaci.

The article deals with the recent developments in microsurgical techniques for microscopic varicocele surgery and microdissection testicular sperm extraction (microTESE). In varicocele surgery, recent developments have led to intraoperative microvascular Doppler diagnosis, three-dimensional magnification, and clipless ligation of the spermatic veins with preservation of the spermatic arteries. In microTESE, the benefits include preoperative triplex microvascular Doppler diagnosis, hydrodissection of testicular tubules and high magnification of the tubules in vivo with possible micrometry of the tubules, selection of tubule collection sites with a greater likelihood of sperm presence, and a high degree of protection of surrounding testicular tissue. The procedures are demonstrated in the photo documentation.

• Klíčová slova
• mikrovaskulární Doppler, micro TESE, hydrodisekce, optoelektronická mikrometrie kanálků testes,
• MeSH
• lidé MeSH
• ligace metody   MeSH
• mikrochirurgie * metody   MeSH
• varikokéla * chirurgie   MeSH
• zobrazování trojrozměrné MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• komentáře MeSH

The advancement in molecular techniques has been attributed to the quality and significance of cancer research. Pancreatic cancer (PC) is one of the rare cancers with aggressive behavior and a high mortality rate. The asymptomatic nature of the disease until its advanced stage has resulted in late diagnosis as well as poor prognosis. The heterogeneous character of PC has complicated cancer development and progression studies. The analysis of bulk tissues of the disease was insufficient to understand the disease, hence, the introduction of the single-cell separating technique aided researchers to decipher more about the specific cell population of tumors. This review gives an overview of the Laser Capture Microdissection (LCM) technique, one of the single-cell separation methods used in PC research.

• MeSH
• duktální karcinom slinivky břišní * patologie   MeSH
• laserová záchytná mikrodisekce MeSH
• lidé MeSH
• nádory slinivky břišní * patologie   MeSH
• pankreas patologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Přestože se spektrum biomarkerů užívaných v onkologii neustále rozšiřuje, zůstává stanovení morfologie nádorů rozhodujícím kritériem diagnostiky. U karcinomů prsu bylo nedávno zjištěno, že stupeň diferenciace je provázen změnami exprese miRNA. Není však nic známo o tom, zda popsané změny souvisí se změnou morfologie a zda mají individuální nebo obecný charakter. Cílem navrhovaného projektu je použít zatím unikátní přístup a srovnat expresní profil miRNA z různých částí téhož nádoru s rozdílnou morfologickou charakteristikou i nádorů se stejnou morfologií od různých pacientek a zároveň zjistit, zda je možné tyto specifické miRNA detekovat v krvi. Pro řešení navrhujeme použít mikrodisekční metody izolace buněk z parafínových bloků TNBC a mikročipovou analýzu miRNA. Tkáňové a sérové hladiny vybraných miRNA budou sledovány pomocí qPCR v rozšířeném validačním souboru a jejich funkce bude studována pomocí experimentů na buněčných liniích. Projekt by mohl přispět k odhalení nových principů řídících morfogenezi v nádorech a rovněž k identifikaci nových diagnostických a prognostických znaků.; Although the spectrum of cancer biomarkers is constantly expanding, determining the morphology is a decisive criterion for diagnosis. Recent research in breast cancer has shown that the degree of differentiation is accompanied by changes in miRNA expression. However, nothing is known about relationship between the described changes and tumor morphology or whether these changes have an individual or general character. We suggest to compare the miRNA expression profiles from different parts of the same tumor which have different morphology as well as tumors with the same morphology from different patients and analyse whether specific miRNAs are released into blood. Microdissection of cells from paraffin blocks of TNBC and miRNA microarray analysis will be used. Tissue and serum levels of selected miRNAs will be monitored by qPCR in an extended validation set and their functions will be studied by experiments on cell lines. The project could contribute to the discovery of new principles governing morphogenesis in tumors as well as identify novel diagnostic and prognostic markers.

• MeSH
• časná detekce nádoru MeSH
• exprese genu MeSH
• lidé MeSH
• mikro RNA MeSH
• mikročipová analýza MeSH
• mikrodisekce MeSH
• morfogeneze MeSH
• nádorové biomarkery analýza   MeSH
• prognóza MeSH
• triple-negativní karcinom prsu diagnóza   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• onkologie
• genetika, lékařská genetika
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Molecular assessment of renal allografts has already been suggested in antibody-mediated rejection (ABMR), but little is known about the gene transcript patterns in particular renal compartments. We used laser capture microdissection coupled with quantitative RT-PCR to distinguish the transcript patterns in the glomeruli and tubulointerstitium of kidney allografts in sensitized retransplant recipients at high risk of ABMR. The expressions of 13 genes were quantified in biopsies with acute active ABMR, chronic active ABMR, acute tubular necrosis (ATN), and normal findings. The transcripts were either compartment specific (TGFB1 in the glomeruli and HAVCR1 and IGHG1 in the tubulointerstitium), ABMR specific (GNLY), or follow-up specific (CXCL10 and CX3CR1). The transcriptional profiles of early acute ABMR shared similarities with ATN. The transcripts of CXCL10 and TGFB1 increased in the glomeruli in both acute ABMR and chronic active ABMR. Chronic active ABMR was associated with the upregulation of most genes (SH2D1B, CX3CR1, IGHG1, MS4A1, C5, CD46, and TGFB1) in the tubulointerstitium. In this study, we show distinct gene expression patterns in specific renal compartments reflecting cellular infiltration observed by conventional histology. In comparison with active ABMR, chronic active ABMR is associated with increased transcripts of tubulointerstitial origin.

• MeSH
• biopsie MeSH
• chronická nemoc MeSH
• dospělí MeSH
• glomerulus imunologie   metabolismus   patologie   MeSH
• HLA antigeny imunologie   MeSH
• isoprotilátky imunologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• laserová záchytná mikrodisekce MeSH
• ledviny imunologie   metabolismus   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• rejekce štěpu genetika   imunologie   metabolismus   patologie   MeSH
• senioři MeSH
• stanovení celkové genové exprese MeSH
• studie případů a kontrol MeSH
• transkriptom * MeSH
• transplantace ledvin škodlivé účinky   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

• Klíčová slova
• Hematologie,
• MeSH
• cytologické techniky MeSH
• hematologické testy MeSH
• hematopoéza MeSH
• histologické techniky MeSH
• kostní dřeň MeSH
• krev MeSH

• NLK Obory
• hematologie a transfuzní lékařství

The platform for precise proteomic profiling of targeted cell populations from heterogeneous tissue sections is developed. We demonstrate a seamless and systematic integration of LCM with an automated cap-IA for the handling of a very small-sized dissected tissues section from the kidney, liver and pancreatic Langerhans islet of rats. Our analysis reveals that the lowest LCM section area ≥ 0.125 mm2 with 10 µm thickness can be optimized for the detection of proteins through LCM-cap-IA integration. We detect signals ranging from a highly-abundant protein, β-actin, to a low-abundance protein, LC-3AB, using 0.125 mm2 LCM section from rat kidney, but, so far, a relatively large section is required for good quality of results. This integration is applicable for a highly-sensitive and accurate assessment of microdissected tissue sections to decipher hidden proteomic information of pure targeted cells. To validate this integration, PCK2 protein expression is studied within Langerhans islets of normal and diabetic rats. Our results show significant overexpression of PCK2 in Langerhans islets of rats with long-term diabetes.

• Publikační typ
• časopisecké články MeSH

Triple negative breast cancers (TNBC) are a morphologically and genetically heterogeneous group of breast cancers with uncertain prediction of biological behavior and response to therapy. Epithelial to mesenchymal transition (EMT) is a dynamic process characterized by loss of typical epithelial phenotype and acquisition of mesenchymal characteristics. Aberrant activation of EMT can aggravate the prognosis of patients with cancer, however, the mechanisms of EMT and role of microRNAs (miRNAs) in EMT activation is still unclear. The aim of our study was to analyze miRNA expression within areas of TNBCs with cellular morphology that may be related to the EMT process and discuss possible associations. Out of all 3953 re-examined breast cancers, 460 breast cancers were diagnosed as TNBC (11.64%). With regard to complete tumor morphology preservation, the tissue samples obtained from core-cut biopsies and influenced by previous neoadjuvant therapy were excluded. We assembled a set of selected 25 cases to determine miRNA expression levels in relation to present focal spindle cell and apocrine cell morphology within individual TNBCs. We used descriptive (histological typing and morphology), morphometric, molecular (microdissection of tumor and non-tumor morphologies, RNA isolation and purification, microchip analysis) and bioinformatic analysis (including pathway analysis). The results were verified by quantitative real-time PCR (RT-qPCR) on an extended set of 70 TNBCs. The majority of TNBCs were represented by high-grade invasive carcinomas of no special type (NST) with medullary features characterized by well-circumscribed tumors with central necrosis or fibrosis and frequent tendency to spindle-cell and/or apocrine cell transformation. Apocrine and spindle cell transformation showed a specific miRNA expression profile in comparison to other tumor parts, in situ carcinoma or non-tumor structures, particularly down-regulated expression of hsa-miRNA-143-3p and hsa-miRNA-205-5p and up-regulated expression of hsa-miR-22-3p, hsa-miRNA-185-5p, and hsa-miR-4443. Apocrine cell tumor morphology further revealed decreased expression of hsa-miR-145-5p and increased expression of additional 14 miRNAs (e.g. hsa-miR-182-5p, hsa-miR-3135b and hsa-miR-4417). Pathway analysis for target genes of these miRNAs revealed several shared biological processes (i.e. Wnt signaling, ErbB signaling, MAPK signaling, endocytosis and axon guidance), which may in part contribute to the EMT and tumor progression. We provide the first miRNA expression profiling of specific tissue morphologies in TNBC. Our results demonstrate a specific miRNA expression profile of apocrine and spindle cell morphology which can exhibit a certain similarity with the EMT process and may also be relevant for prognosis and therapy resistance of TNBC.

• MeSH
• apokrinní žlázy * mikrobiologie   patologie   MeSH
• dospělí MeSH
• epitelo-mezenchymální tranzice * genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikro RNA * genetika   MeSH
• regulace genové exprese u nádorů genetika   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• signální dráha Wnt genetika   MeSH
• stanovení celkové genové exprese MeSH
• triple-negativní karcinom prsu * genetika   patologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH

• MeSH
• cytologické techniky MeSH
• hematologické testy MeSH
• hematopoéza MeSH
• histologické techniky MeSH
• kostní dřeň MeSH
• krev MeSH

• Konspekt
• Patologie. Klinická medicína
• Učební osnovy. Vyučovací předměty. Učebnice
• NLK Obory
• hematologie a transfuzní lékařství
• NLK Publikační typ
• učebnice vysokých škol

Members of neuropeptide B/W signaling system have been predominantly detected and mapped within the CNS. In the rat, this system includes neuropeptide B (NPB), neuropeptide W (NPW) and their specific receptor NPBWR1. This signaling system has a wide spectrum of functions including a role in modulation of inflammatory pain and neuroendocrine functions. Expression of NPB, NPW and NPBWR1 in separate heart compartments, dorsal root ganglia (DRG) and stellate ganglia was proven by RT-qPCR, Western blot (WB) and immunofluorescence. Presence of mRNA for all tested genes was detected within all heart compartments and ganglia. The presence of proteins preproNPB, preproNPW and NPBWR1 was confirmed in all the chambers of heart by WB. Expression of preproNPW and preproNPB was proven in cardiac ganglionic cells obtained by laser capture microdissection. In immunofluorescence analysis, NPB immunoreactivity was detected in nerve fibers, some nerve cell bodies and smooth muscle within heart and both ganglia. NPW immunoreactivity was present in the nerve cell bodies and nerve fibers of heart ganglia. Weak nonhomogenous staining of cardiomyocytes was present within heart ventricles. NPBWR1 immunoreactivity was detected on cardiomyocytes and some nerve fibers. We confirmed the presence of NPB/W signaling system in heart, DRG and stellate ganglia by proteomic and genomic analyses.

• MeSH
• exprese genu MeSH
• fluorescenční protilátková technika MeSH
• ganglion stellatum metabolismus   MeSH
• myokard metabolismus   MeSH
• neuropeptidy genetika   imunologie   metabolismus   MeSH
• potkani Zucker MeSH
• receptory neuropeptidů genetika   imunologie   metabolismus   MeSH
• receptory spřažené s G-proteiny genetika   imunologie   metabolismus   MeSH
• reprodukovatelnost výsledků MeSH
• signální transdukce MeSH
• spinální ganglia metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH