Močové extracelulární vezikuly (uEVs) představují slibný nástroj pro neinvazivní diagnostiku a monitorování onemocnění ledvin. Tyto vezikuly, secernované buňkami ledvin, obsahují biomolekuly odrážející stav mateřských buněk. Výzkum se zaměřuje na využití uEVs jako biomarkerů pro chronická onemocnění ledvin, akutní poškození ledvin, diabetickou nefropatii a monitorování po transplantaci ledvin. Kromě diagnostického potenciálu jsou uEVs zkoumány pro terapeutické aplikace v regeneraci ledvinové tkáně. Přes výzvy v standardizaci izolačních a analytických metod uEVs, pokrok ve vývoji metod charakterizace uEVs podporuje jejich klinické využití.
Urinary extracellular vesicles (uEVs) represent a promising tool for non-invasive diagnosis and monitoring of kidney disease. These vesicles, secreted by kidney cells, contain biomolecules reflecting the status of the parent cells. Research focuses on using uEVs as biomarkers for chronic kidney disease, acute kidney injury, diabetic nephropathy and monitoring kidney transplantation. In addition to diagnostic potential, uEVs are being investigated for therapeutic applications in kidney tissue regeneration. Despite challenges in standardizing uEVs isolation and analysis methods, progress in the development of uEVs characterization methods supports their clinical use.
INTRODUCTION: The aim of study was to assess the impact of an enhanced recovery after surgery (ERAS) protocol and minimally invasive approaches on short-term outcomes in rectal surgery. PATIENTS AND METHODS: A consecutive series of patients that underwent open or minimally invasive rectal resections in a single institution between January 2015 and April 2020 were included in the study. An ERAS program was introduced in April 2016. The study cohort was divided into three groups: open surgery without ERAS, open surgery with ERAS, and minimally invasive surgery with ERAS. Outcome measures compared were recovery parameters, surgical stress parameters, 30-day morbidity and mortality, oncological radicality and length of hospital stay. RESULTS: A total of 202 patients were included: 43 in the open non-ERAS group, 92 in the open ERAS group and 67 in the minimally invasive ERAS group. All recovery parameters apart from postoperative nausea and vomiting were significantly improved in both ERAS groups. Surgical stress parameters, prolonged postoperative ileus, and hospital stay were significantly reduced in the minimally invasive ERAS group. The overall 30-day morbidity and mortality and oncological radicality did not significantly differ among the three groups. CONCLUSIONS: Minimally invasive approaches and enhanced recovery care in rectal surgery improve short-term outcomes. Their combination leads to an improvement in recovery parameters and a reduction of prolonged postoperative ileus and hospital stay.
- MeSH
- chirurgie trávicího traktu * metody MeSH
- délka pobytu MeSH
- ileus * etiologie MeSH
- laparoskopie * metody MeSH
- lidé MeSH
- miniinvazivní chirurgické výkony škodlivé účinky metody MeSH
- perioperační péče metody MeSH
- pooperační komplikace epidemiologie etiologie MeSH
- střeva MeSH
- urychlená pooperační rehabilitace * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The incidence of cutaneous malignant melanoma is increasing worldwide. While the treatment of initial stages of the disease is simple, the advanced disease frequently remains fatal despite novel therapeutic options . This requires identification of novel therapeutic targets in melanoma. Similarly to other types of tumours, the cancer microenvironment plays a prominent role and determines the biological properties of melanoma. Importantly, melanoma cell-produced exosomes represent an important tool of intercellular communication within this cancer ecosystem. We have focused on potential differences in the activity of exosomes produced by melanoma cells towards melanoma-associated fibroblasts and normal dermal fibroblasts. Cancer-associated fibroblasts were activated by the melanoma cell-produced exosomes significantly more than their normal counterparts, as assessed by increased transcription of genes for inflammation-supporting cytokines and chemokines, namely IL-6 or IL-8. We have observed that the response is dependent on the duration of the stimulus via exosomes and also on the quantity of exosomes. Our study demonstrates that melanoma-produced exosomes significantly stimulate the tumour-promoting proinflammatory activity of cancer-associated fibroblasts. This may represent a potential new target of oncologic therapy .
- MeSH
- exozómy metabolismus MeSH
- fibroblasty metabolismus patologie MeSH
- lidé MeSH
- melanom experimentální metabolismus patologie MeSH
- nádorové buňky kultivované MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Amyloid A amyloidosis is a serious clinical condition resulting from the systemic deposition of amyloid A originating from serum amyloid A proteins with the kidneys being the most commonly and earliest affected organ. Previously described amyloid A amyloidosis is linked to increased production and deposition of serum amyloid A proteins secondary to inflammatory conditions arising from infectious, metabolic, or genetic causes. Here we describe a family with primary amyloid A amyloidosis due to a chr11:18287683 T>C (human genome version19) mutation in the SAA1 promoter linked to the amyloidogenic SAA1.1 haplotype. This condition leads to a doubling of the basal SAA1 promoter activity and sustained elevation of serum amyloid A levels that segregated in an autosomal dominant pattern in 12 genetically affected and in none of six genetically unaffected relatives, yielding a statistically significant logarithm of odds (LOD) score over 5. Affected individuals developed proteinuria, chronic kidney disease and systemic deposition of amyloid composed specifically of the SAA1.1 isoform. Tocilizumab (a monoclonal antibody against the interleukin-6 receptor) had a beneficial effect when prescribed early in the disease course. Idiopathic forms represent a significant and increasing proportion (15-20%) of all diagnosed cases of amyloid A amyloidosis. Thus, genetic screening of the SAA1 promoter should be pursued in individuals with amyloid A amyloidosis and no systemic inflammation, especially if there is a positive family history.
- MeSH
- amyloidóza * komplikace MeSH
- lidé MeSH
- mutace MeSH
- promotorové oblasti (genetika) MeSH
- sérový amyloid A genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) represents an autoimmunity disease characterized by high mortality. For successful treatment, the detailed knowledge of its complex pathogenesis and the set of biomarkers for differential diagnostics are desired. Analysis of molecular content of small urinary extracellular vesicles (uEV) offers the possibility to find markers in the form of microRNAs (miRNAs) and study the pathways involved in pathogenesis. We used next-generation sequencing in the first preliminary study to detect the miRNAs with altered expression in uEVs of patients with AAV in comparison with age-matched controls. We confirmed the results using single-target quantitative polymerase chain reaction tests on different sets of samples and found five miRNAs (miR-30a-5p, miR-31-3p, miR-99a-5p, miR-106b-5p, miR-182-5p) with highly elevated levels in uEVs of patients. We performed the comparison of their targets with the differentially expressed proteins in uEVs of patients included in the first phase. We realized that upregulated miRNAs and proteins in uEVs in AAV patients target different biological pathways. The only overlap was detected in pathways regulating the actin cytoskeleton assembly and thus potentially affecting the glomerular functions. The associations of upregulated miRNAs with pathways that were neglected as components of complex AAV pathogenesis, e.g., the epidermal growth factor receptor signaling pathway, were found.
- MeSH
- ANCA-asociované vaskulitidy * genetika MeSH
- biologické markery MeSH
- extracelulární vezikuly * genetika MeSH
- ledviny MeSH
- lidé MeSH
- mikro RNA * genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
ANCA-associated vasculitis (AAV) is a rare, but potentially severe autoimmune disease, even nowadays displaying increased mortality and morbidity. Finding early biomarkers of activity and prognosis is thus very important. Small extracellular vesicles (EVs) isolated from urine can be considered as a non-invasive source of biomarkers. We evaluated several protocols for urinary EV isolation. To eliminate contaminating non-vesicular proteins due to AAV associated proteinuria we used proteinase K treatment. We investigated the differences in proteomes of small EVs of patients with AAV compared to healthy controls by label-free LC-MS/MS. In parallel, we performed an analogous proteomic analysis of urine samples from identical patients. The study results showed significant differences and similarities in both EV and urine proteome, the latter one being highly affected by proteinuria. Using bioinformatics tools we explored differentially changed proteins and their related pathways with a focus on the pathophysiology of AAV. Our findings indicate significant regulation of Golgi enzymes, such as MAN1A1, which can be involved in T cell activation by N-glycans glycosylation and may thus play a key role in pathogenesis and diagnosis of AAV. SIGNIFICANCE: The present study explores for the first time the changes in proteomes of small extracellular vesicles and urine of patients with renal ANCA-associated vasculitis compared to healthy controls by label-free LC-MS/MS. Isolation of vesicles from proteinuric urine samples has been modified to minimize contamination by plasma proteins and to reduce co-isolation of extraluminal proteins. Differentially changed proteins and their related pathways with a role in the pathophysiology of AAV were described and discussed. The results could be helpful for the research of potential biomarkers in renal vasculitis associated with ANCA.
Erythropoietin (EPO) downregulates hepcidin expression to increase the availability of iron; the downregulation of hepcidin is mediated by erythroferrone (ERFE) secreted by erythroblasts. Erythroblasts also express transferrin receptor 2 (TFR2); however, the possible role of TFR2 in hepcidin downregulation is unclear. The purpose of the study was to correlate liver expression of hepcidin with the expression of ERFE and TFR2 in murine bone marrow and spleen at 4, 16, 24, 48, 72 and 96 h following administration of a single dose of EPO. Splenic Fam132b expression increased 4 h after EPO injection; liver hepcidin mRNA was decreased at 16 h. In the spleen, expression of TFR2 and transferrin receptor (TFR1) proteins increased by an order of magnitude at 48 and 72 h after EPO treatment. The EPO-induced increase in splenic TFR2 and TFR1 was associated with an increase in the number of Tfr2- and Tfr1-expressing erythroblasts. Plasma exosomes prepared from EPO-treated mice displayed increased amount of TFR1 protein; however, no exosomal TFR2 was detected. Overall, the results confirm the importance of ERFE in stress erythropoiesis, support the role of TFR2 in erythroid cell development, and highlight possible differences in the removal of TFR2 and TFR1 from erythroid cell membranes.
- MeSH
- cytokiny genetika metabolismus MeSH
- erythropoetin farmakologie MeSH
- erytroblasty účinky léků metabolismus MeSH
- exozómy metabolismus MeSH
- hepcidiny genetika metabolismus MeSH
- játra metabolismus MeSH
- kultivované buňky MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- receptory transferinu genetika metabolismus MeSH
- slezina metabolismus MeSH
- svalové proteiny genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Matriptase-2, a serine protease expressed in hepatocytes, is a negative regulator of hepcidin expression. The purpose of the study was to investigate the interaction of matriptase-2 with hemojuvelin protein in vivo. Mice lacking the matriptase-2 proteolytic activity (mask mice) display decreased content of hemojuvelin protein. Vice versa, the absence of hemojuvelin results in decreased liver content of matriptase-2, indicating that the two proteins interact. To further characterize the role of matriptase-2, we investigated iron metabolism in mask mice fed experimental diets. Administration of iron-enriched diet increased liver iron stores as well as hepcidin expression. Treatment of iron-overloaded mask mice with erythropoietin increased hemoglobin and hematocrit, indicating that the response to erythropoietin is intact in mask mice. Feeding of an iron-deficient diet to mask mice significantly increased spleen weight as well as the splenic content of erythroferrone and transferrin receptor proteins, indicating stress erythropoiesis. Liver hepcidin expression was decreased; expression of Id1 was not changed. Overall, the results suggest a complex interaction between matriptase-2 and hemojuvelin, and demonstrate that hepcidin can to some extent be regulated even in the absence of matriptase-2 proteolytic activity.
- MeSH
- deficit železa MeSH
- dietní železo farmakologie MeSH
- erythropoetin farmakologie MeSH
- GPI-vázané proteiny biosyntéza nedostatek genetika fyziologie MeSH
- hepcidiny biosyntéza genetika MeSH
- inhibitor diferenciace 1 biosyntéza genetika MeSH
- játra metabolismus MeSH
- kostní morfogenetický protein 6 biosyntéza genetika MeSH
- membránové proteiny nedostatek genetika fyziologie MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši MeSH
- orgánová specificita MeSH
- přetížení železem metabolismus MeSH
- promotorové oblasti (genetika) genetika MeSH
- protein hemochromatózy biosyntéza nedostatek genetika fyziologie MeSH
- proteinové domény MeSH
- regulace genové exprese účinky léků MeSH
- rekombinantní proteiny metabolismus MeSH
- serinové endopeptidasy nedostatek genetika fyziologie MeSH
- slezina metabolismus MeSH
- železo MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Hemicorporectomy is the amputation of the lower body - pelvis and lower limbs. It requires transection of the spine and dural sac at the level of aortic bifurcation and inferior lower vein, and permanent urinary and stool derivation. Performance indications are tumour trauma and terminal pelvic osteomyelitis. So far about 60 cases have been published; only 11 operations were performed for terminal osteomyelitis. We have successfully performed hemicorporectomy in a patient with chronic sepsis from terminal pelvic osteomyelitis after exhausting all other treatment options. The experience gained and the important moments of the procedure are given in the case report.
Hemicorporectomy or translumbar amputation is an extensive surgical procedure consisting in removing the lower portion of the body. Thakur et al. found a total of 71 hemicorporectomies described in literature before 2017. In the form of a case study we present the case of our patient with terminal pelvic osteomyelitis, in whom hemicorporectomy was subsequently performed, namely from the spine surgery perspective. The man, 19 years old, was exposed to high-voltage electricity and fell down from a height of 4 meters. He suffered an instable comminuted fracture of T10 (AO A3.3.) with paraplegia (Frankel A) and multiple third-degree burns affecting 25% of his total body surface area. Subsequently, the patient underwent a total of 16 surgical procedures performed by medical experts in various specialties (orthopaedic surgery, general surgery, plastic surgery, urology, vascular surgery), but in spite of that the extensive pelvic osteomyelitis has not been successfully managed. At first, urine and stool diversion were performed. After 3 weeks, i.e. 18 months after the injury, the removal of the lower portion of the body was scheduled. The hemicorporectomy was divided into 4 stages. The surgery started by posterior transecting the spine at L4-L5 segment with nerve root and dural sac ligation and treating the bleeding venous plexus in the spinal canal. After turning the patient to the supine position, the second stage of the operation followed, consisting in transecting large vessels and harvesting a musculocutaneous flap from the right thigh. During the third stage of the surgery the separation of the L4-L5 motion segment was completed by the transaction of the anterior longitudinal ligament and m. psoas major, subsequently followed by the amputation of the lower portion of the body. During the last stage of the surgery, the wound was closed by musculocutaneous flap from the fight thigh with preserved a. femoralis. The patient was discharged to home in a generally good condition 127 days after the amputation of the lower portion of the body. Now, 1 year after the surgery, the patient enjoys good physical as well as mental health. Hemicorporectomy is an extensive surgical technique, which can despite multiple complications be offered to patients with otherwise unmanageable condition. Terminal pelvic osteomyelitis is currently the most frequent diagnostic indication and the resulting condition makes possible a long-term survival of the patient in a satisfactory condition. The spinal surgeon is an irreplaceable member of the multidisciplinary team performing the surgical procedure, the primary treatment of the spinal column considerably limits blood losses. Key words: hemicorporectomy, en bloc sacrectomy, terminal pelvic osteomyelitis, sacral tumors.
- MeSH
- amputace MeSH
- dospělí MeSH
- fraktury páteře komplikace MeSH
- lidé MeSH
- mladý dospělý MeSH
- ortopedické výkony * MeSH
- osteomyelitida * chirurgie MeSH
- pánev MeSH
- zákroky plastické chirurgie * MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
