The Impact of COVID-19 Third Dose Vaccination on the Magnitude of Antigen Specific T Cells in Kidney Transplant Patients.

Measuring T cell response can add information about antivirus immunity provided by antibody test results. The study evaluates the impact of a third mRNA COVID-19 vaccine dose on T cell response and antibody production in kidney transplant recipients (25 KTRs) versus healthy controls (26 Hc). Results show a significant rise in S-activated CD4+CD154+IFN?+TNF?+ double producer cells in both KTRs (p=0.025) and Hc (p=0.009) as well as increased spike antibody response in KTRs (p=0.00019) and Hc (p=3.10-8) third-month post-third dose. Moreover, the study revealed a drop in seronegative KTRs (non-responders) from 9/25 (36%) pre-third dose to 2/25 (7%) at 3 months post-third dose while 5/9 (56%) of non-responders post-second dose showed specific T cell responses. Notably, the third dose significantly improved seroconversion rates in both KTRs and Hc, although Hc individuals exhibited higher antibody levels. Key words: mRNA COVID-19 vaccine, T cells, SARS-CoV-2 antibodies, Kidney transplantation, mRNA vaccination.

Insights into IL-1 family cytokines in kidney allograft transplantation: IL-18BP and free IL-18 as emerging biomarkers.

Proinflammatory cytokines and their inhibitors are involved in the regulation of multiple immune reactions including response to transplanted organs. In this prospective study, we evaluated changes in serum concentrations of six IL-1 family cytokines (IL-1 alpha, IL-1 beta, IL-1RA, IL-18, IL-18BP, and IL-36 beta) in 138 kidney allograft recipients and 48 healthy donors. Samples were collected before transplantation and then after one week, three months and one year, additional sera were obtained at the day of biopsy positive for acute rejection. We have shown, that concentrations of proinflammatory members of the IL-1 family (IL-1ß, IL-18, IL-36 ß) and anti-inflammatory IL-18BP decreased immediately after the transplantation. The decline of serum IL-1RA and IL-1α was not observed in subjects with acute rejection. IL-18, including specifically its free form, is the only cytokine which increase serum concentrations in the period between one week and three months in both groups of patients without upregulation of its inhibitor, IL-18BP. Serum concentrations of calculated free IL-18 were upregulated in the acute rejection group at the time of acute rejection. We conclude that IL-1 family cytokines are involved mainly in early phases of the response to kidney allograft. Serum concentrations of free IL-18 and IL-18BP represent possible biomarkers of acute rejection, and targeting IL-18 might be of therapeutic value.

Changes in phenotypic patterns of blood monocytes after kidney transplantation and during acute rejection.

Peripheral blood monocytes, which serve as precursors for tissue macrophages and dendritic cells (DC), play a key role in the immune response to kidney allograft, reparation processes and homeostasis regulation. In this prospective study, we used multicolor flow cytometry to monitor the phenotypic patterns of peripheral monocytes in subjects with uncomplicated outcomes and those with acute rejection. We found a reciprocal increase in the proportion of "classical monocytes" (CD14+CD16-) along with a decline in pro-inflammatory "intermediary" (CD14+CD16+) and "non-classical" (CD14lowCD16+) monocytes in subjects with normal outcomes. In subjects with acute rejection, we observed no reduction in "intermediary" monocytes and no increase in "classical" monocytes. Patients with uncomplicated outcomes exhibited downregulated HLA-DR in all three monocyte subpopulations. However, non-classical monocytes were unaffected in subjects with acute rejection. Expression of CD47 was downregulated after transplantation, while patients with antibody-mediated rejection and donor-specific antibodies showed higher pre-transplant values. In monocytes isolated at the time of biopsy, CD47 expression was higher in individuals with acute rejection compared to patients with normal outcomes one year post-transplant. Expression of CD209 (DC-SIGN) and the proportion of CD163+CD206+ subpopulations were upregulated during the first week after kidney transplantation. CD209 was also upregulated in samples taken on the day of biopsy confirming acute rejection. Our data demonstrate that kidney allograft transplantation is associated with phenotypic changes in peripheral blood monocytes during acute rejection.

Concomitant thyroid disease in patients operated for primary hyperparathyroidism.

INTRODUCTION: Parathyroid and thyroid diseases are ones of the most common endocrine diseases, but simultaneous surgical treatment of both endocrine systems is still under discussion. METHODS: We retrospectively evaluated 1,574 patients operated for primary hyperparathyroidism at the 3rd Department of Surgery, 1st Faculty Medicine, Charles University and University Hospital Motol in Prague with the thyroid and parathyroid ultrasound reports available. The patients were divided into two groups - with and without thyroid surgery. RESULTS: Thyroid surgery was performed in 34% of patients with primary hyperparathyroidism. Group 2, where thyroid surgery was performed, showed a higher proportion of reported abnormal thyroid sonographic findings (74%), a higher proportion of bilateral throat exploration (69%) and a longer hospital stay (3.3 days). CONCLUSION: A patient indicated for surgery for parathyroid disease should also be indicated for possible thyroid surgery.

Thyroid tumors at the limit of operability.

Comprehensive information about current thyroid carcinoma treatment options depending on its histology and extent of the disease, focusing on locally advanced findings at the limit of operability. Treatment of such a heterogeneous group requires interdisciplinary cooperation. We provide 6 unique case reports including imaging scans, description of the therapy and description of development of the condition.

Up-regulation of CD163 expression in subpopulations of blood monocytes after kidney allograft transplantation.

M2 macrophages expressing CD163 are known to suppress immune responses but have been also found in biopsies of patients with chronic kidney allograft injury associated with interstitial fibrosis. The aim of our study was to evaluate the expression of CD163 in blood monocytes, precursors of tissue macrophages, in kidney allograft recipients with uncomplicated outcome (n=94) compared with those developing acute rejection (n=44). Blood samples were collected before the transplantation and at 1 week, 1 month and 1 year. The expression of CD163 increased during the first week after the transplantation not only in classical (CD14+CD16-) but also in intermediate (CD14+CD16+) and nonclassical (CD14lowCD16+) monocytes in all patients regardless of their rejection status. In patients developing acute rejection, higher pre-transplant expression of CD163 on blood monocytes was found. In vitro experiments confirmed strong induction of membrane CD163 on monocytes together with CD206 (an alternative marker of M2 macrophages) in response to IL-10. We assume from our data that dramatic upregulation of CD163 by peripheral blood monocytes may have a pathophysiological role in early phases after kidney allograft transplantation and high pre-transplant expression of CD163 on blood monocytes might be involved in events leading to acute rejection.

Retrosternal goiter.

INTRODUCTION: Retrosternal goiter is an enlarged thyroid mass of which more than 50% is located in the mediastinum. Indications for surgery of retrosternal goiter include ineffective pharmacological treatment, mechanical syndrome, suspicion of malignancy and thyrotoxicosis. Computed tomography is the gold standard in the diagnosis of retrosternal goiter. The surgery can be performed from a cervical incision, using sternotomy or thoracotomy. METHODS: The aim of our study was to evaluate our own group of retrosternal goiters. In 2011-2019, 1739 thyroid surgeries were performed at the 3rd Department of Surgery, University Hospital Motol, Prague. We retrospectively followed the age, gender, retrosternal spreading, postoperative complications (especially hypocalcemia, bleeding and dysphonia) and the definitive histological finding. RESULTS: Retrosternal goiters were presented in 202 (19.2%) of the total of 1739 thyroid surgery patients. Sternotomy was performed in 31 patients. Women (61%) predominated over men (39%). The mean operating time was 125 minutes. We did not detect any serious postoperative bleeding necessitating surgical revision. Temporary hypocalcemia was observed in 10 patients (32%) based on laboratory testing. Unilateral iatrogenic vocal cord paresis was observed in 6 patients (19.4%) and was permanent in 3 patients (9.3%). One patient passed away soon after the operation, nevertheless the death was related to a simultaneously planned cardiothoracic procedure. CONCLUSION: The retrosternal goiter can be removed using the cervical approach in most cases. Total thyroidectomy using sternotomy is associated with higher morbidity and mortality, higher blood loss and longer hospital stay; nevertheless, it does not increase the risk of long-term postoperative complications. Procedures requiring sternotomy or thoracotomy should be done at centers experienced in these types of procedures.

Measurement of intact parathormone during operation for primary hyperparathyroidism.

In this prospective study, the role of the intact parathormone (iPHT) levels for the verification of pathologic parathyroid tissue removal during parathyroidectomy, was analyzed in 441 patients diagnosed with primary hyperparathyroidism. The level of intact parathormone was obtained before the initial incision (baseline level) and 10 minutes after the pathologic parathyroid gland removal (control value). In 80 % of cases, the decrease of intact parathormone was more than 50 % of the baseline level. The comparison of preoperative and postoperative intact parathormone levels can also be used as marker of parathyroid hyperplasia or persistent hyperparathyroidism. This method is necessary mainly for performance of focused, miniinvasive approaches as well as in reoperations. This method is of significant benefit in cases of negative preoperative examination methods. The determination of intact parathormone level increases the success of parathyroidectomy (Ref. 26).

[Videothoracoscopic excision of mediastinal parathyroid adenoma in primary hyperparathyroidism]. / Videothorakoskopické odstranení mediastinálne ulozeného adenomu prístítného telíska pri primární hyperparathyreóze.

INTRODUCTION: Primary hyperparathyroidism is a disease caused by elevated secretion of parathyroid hormone from pathological parathyroid glands. After the diagnosis, the success of its surgical solution depends predominantly on surgical management and experience of the surgeons. A special group is formed by ectopic localizations of pathologically enlarged parathyroid glands in the mediastinum, which require a modified surgical approach. When the adenoma is deep in the mediastinum, sternotomy or thoracotomy is indicated; alternatively, a minimally invasive approach can be used - videothoracoscopic thymectomy. CASE REPORT: We present a case of a patient with normocalcemic recurrent primary hyperparathyroidism. This patient underwent a minimally invasive video-assisted thymectomy after scintigraphic confirmation of parathyroid adenoma in the mediastinum. CONCLUSION: The removal of parathyroid adenoma in the mediastinum using the videothoracoscopic method is safe. Compared to sternotomy, this method improves the postoperative period, reduces the length of stay and provides more comfort to patients. We recommend considering the videothoracoscopic method in cases where the pathologically enlarged parathyroid gland is localized in the inferior and anterior mediastinum. KEY WORDS: primary hyperparathyroidism adenoma of glandula parathyroidea - mediastinum miniinvasive.

[Ganglioneuroma, a rare cause of soft neck tissues tumor in adult age]. / Ganglioneurom, raritní prícina nádoru mekkých tkání krku v dospelosti.

We present the case of a 26-year-old female patient with coincidental ultrasound finding of a neck tumour located below the left thyroid lobe. The tumour slightly increased in size over a year. Fine needle aspiration indicated the neuroectodermal origin of the tumour and surgical removal was performed. Although the tumour mass was carefully and successfully removed, post-operational Horners syndrome appeared. The tumour tissue was diagnosed as ganglioneuroma by histopathology.

[Ectopic parathyroid glands. Imaging methods and surgical access]. / Ektopická parathyreoidea, zobrazovací metody a chirurgické prístupy.

We discuss the benefits of imaging methods in localizing ectopic parathyroid glands in patients with primary hyperparathyroidism. The ectopic localizations are discussed within the context of the orthotopic norm. In the sample of 123 patients, a 23% rate of ectopic parathyroid glands was detected. Three selected case studies are presented, supporting the benefit of SPECT/CT imaging in terms of surgical access strategy selection.

Response to competition of bulbous geophyte Allium oleraceum differing in ploidy level.

Experimental studies that explore the possible causes of ploidy distributions and niche differentiation are rare. Increased competitive ability may be advantageous for survival in dense vegetation and may strongly affect local and regional abundances of cytotypes and potentially contribute to invasion success. We compared survival, growth and reproduction of plants originating from bulbils of three cytotypes (2n = 4x, 5x, 6x) of Allium oleraceum growing with and without a competitor (Arrhenatherum elatius). There was a strong negative effect of competition but no effect of ploidy or ploidy × competition on survivorship, height and total dry mass of A. oleraceum, i.e. no support for different competitive abilities of the ploidy levels. However, slightly different responses of populations to competition treatments within all cytotypes suggest differentiation within cytotypes. Under competition, plant survivorship was low, surviving plants were small, had low dry mass and produced neither sexual nor asexual propagules. Without competition, plant survivorship was high, and cytotypes differed in three traits after 2 year's growth: dry mass of flowers, number of flowers and ratio of the dry mass of sexual to asexual propagules all decreased with increasing ploidy level. We additionally tested tetra- and pentaploids as to whether plants originating from different types of propagule (bulbils, seeds) differ in survivorship, growth and reproduction when growing with and without a competitor. Plants originating from bulbils had higher survivorship, were more robust, flowered earlier and produced more propagules when compared to plants originating from seeds and grown without competition. Under competition, differences in performance between plants originating from seeds and bulbils mostly disappeared, with higher survivorship only for plants originating from bulbils.

[Repeated surgery for parathyroid carcinoma]. / Opakované operace pro karcinom prístítného telíska.

INTRODUCTION: Carcinoma of the parathyroid glands is a rare cause of primary hyperparathyroidism with an incidence of 1%. MATERIAL AND METHODS: This article presents an up-to-date review of the literature illustrated by three clinical cases in the form of case reports. RESULTS: Parathyroid carcinoma is usually not detected before the first operation. Symptoms of carcinoma of the parathyroid glands are similar to those of benign adenoma. Patients with parathyroid carcinoma usually have a higher level of calcium in serum and a higher level of parathormone. Imaging methods such as neck ultrasound and 99mTc sestamibi scan can help localize pathologically enlarged glands, but they are not capable of distinguishing malignant disease. Fine needle aspiration is not recommended due to the possible associated risk of tumour seeding along the needle track. The radical excision of the tumour together with the ipsilateral thyroid gland removal remains the standard of treatment. Local recurrence is frequent. CONCLUSION: Parathyroid carcinoma is a very rare disease and should be managed surgically in a specialized centre.

Adaptation of the heart to hypertension is associated with maladaptive gap junction connexin-43 remodeling.

We hypothesized that hypertension-related myocardial remodeling characterized by hypertrophy and fibrosis might be accompanied by cell-to-cell gap junction alterations that may account for increased arrhythmogenesis. Intercellular junctions and expression of gap junction protein connexin-43 were analyzed in rat heart tissues from both spontaneous (SHR) and L-NAME model of hypertension. Isolated heart preparation was used to examine susceptibility of the heart to lethal ventricular fibrillation induced by low potassium perfusion. Ultrastructure observation revealed enhanced neoformation of side-to-side type while internalization of end-to-end type (intercalated disc-related) of gap junctions prevailed in the myocardium of rats suffering from either spontaneous or L-NAME-induced hypertension. In parallel, immunolabeling showed increased number of connexin-43 positive gap junctions in lateral cell membrane surfaces, particularly in SHR. Besides, focal loss of immunopositive signal was observed more frequently in hearts of rats treated with L-NAME. There was a significantly higher incidence of hypokalemia-induced ventricular fibrillation in hypertensive compared to normotensive rat hearts. We conclude that adaptation of the heart to hypertension-induced mechanical overload results in maladaptive gap junction remodeling that consequently promotes development of fatal arrhythmias.

Arachidonic acid and long-chain n-3 polyunsaturated fatty acid contents in meat of selected poultry and fish species in relation to dietary fat sources.

Arachidonic acid (AA) content, long-chain n-3 polyunsaturated fatty acid (PUFA) equivalent [LCE; calculated as 0.15 x linolenic acid (LA) + eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA)], and PUFA n-6/PUFA n-3 ratio were determined in meat [breast meat (BM), thigh meat (TM), and fillets (F), respectively] within four sets of chickens, five sets of turkeys, one set of common carp, and four sets of rainbow trout, fed either commercial diet or diets with manipulated PUFA n-3 and PUFA n-6 contents. AA content was within the range of 20 mg/100 g (F of rainbow trout fed the diet with linseed oil, LO) to 138 mg/100 g (TM of chickens fed restrictively the diet based on maize to the age of 90 days). AA content in BM of turkeys fed the diet with LO or fish oil (FO) did not differ (P > 0.05) from that of rainbow trout F. LCE was in the range of 16 mg/100 g (BM of turkeys fed a commercial feed mixture) to 681 mg/100 g (F of rainbow trout fed a commercial feed mixture). With regard to BM, only turkeys fed the diet with LO deposited more (P < 0.01) LCE (71 mg/100 g) as compared to all other poultry sets except turkeys fed the diet with FO (123 mg/100 g). Apart from all fish samples, also both BM and TM of turkeys fed the diet with either LO or FO met the recommended value of the PUFA n-6/PUFA n-3 ratio (<4). AA content in the tissue increased significantly (P < 0.001) with increasing dietary LA in both all chicken tissues and all turkey tissues, which is contrary to the suggested strong metabolic regulation of the AA formation. When all tissues within all animal species were taken as a one set, both AA percentage and EPA + DHA percentage in the tissue (Y, %) decreased (P < 0.001) with increasing fat content in the tissue (X, %), according to the equation Y = 4.7 - 0.54X (R (2) = 0.41) and Y = 6.0 - 0.33X (R (2) = 0.35), respectively. AA content in chicken BM, chicken TM, and turkey BM, respectively, decreased linearly (P < 0.01) with increasing live weight reached at the slaughter age.

Determination of optimal conditions for detection of acute myeloid leukemia t(8;21) and t(15;17) translocations using RT-PCR.

Prognosis of patients with acute myeloid leukemia (AML) is not satisfactory. Long time remission can be achieved only in 25-40% children with AML. The aim of the study is to improve diagnosis of AML in cases characterized by specific chromosomal translocations t(8;21) and t(15;17). These translocations can be efficiently detected using reverse transcriptase-polymerase chain reaction (RT-PCR). To determine optimal conditions for detection of the t(8;21) and t(15;17) translocations in human cells, we performed a detail study of experimental conditions for RT-PCR on human cell lines NB-4 and KASUMI-1. For detection of t(8;21) using Pfx polymerase, the optimal PCR conditions included magnesium ion concentration in the range of 0.6 to 1 mM, 0.4 microM primer concentration, 5 degrees C annealing temperature and 1 microl of template cDNA. For detection of t(15;17) using the Pfx polymerase, the optimal conditions included 1 mM magnesium ion concentration, 0.6 microM primer concentration, 63 degrees C-66 degrees C annealing temperature and 2 microl of template cDNA. The results proved to be valid for clinical diagnostics of these chromosomal aberrations in blood and/or bone marrow samples of AML patients.

Cytotoxic effects of colicins E1 and E3 on v-myb-transformed chicken monoblasts.

Colicins show a considerable cytostatic activity, which is much less known and understood than their killing activity targeting bacteria of the Enterobacteriaceae family. In this communication, the cytotoxic effects of colicins E1 and E3 on v-myb-transformed chicken monoblasts BM2 are presented. We detected clear reduction of the viable cell number induced by colicins E1 and E3, occurring without apparent changes in cell cycle profiles. The level of inhibition was proportional to the colicin concentration within the limits of 0.5-1.25 microg/ml. This result documents that colicins produced by Enterobacteriaceae exert their cytotoxic effects on leukemic cells.

Conserved guanine-guanine stacking in tetraplex and duplex DNA.

Using a series of suitably chosen oligonucleotides, we demonstrate that the DNA duplex of d(CCCCGGGG) provides an almost identical CD spectrum as the parallel-stranded tetraplex of d(GGGG). The CD spectra are very sensitive to base stacking in DNA so that the above observation indicates that guanine-guanine stacking is essentially the same within the duplex of d(CCCCGGGG) and the tetraplex of d(GGGG). A very similar CD spectrum is also provided by the A-form of d(CCCCGGGG) induced by trifluoroethanol. These results reveal that guanine-guanine stacking is a structural invariant conserved in various nucleic acid conformers. The structural invariance is likely to cohere with evolution of the genetic molecules and be important for fundamental functions, e.g. initiation of transcription.

[Tension-free surgery of femoral hernias using the Lichtenstein method]. / "Tension free" plastika femorálních kýl Lichtensteinovou metodou.

Based on 12 operations, the author report on the advantages of "plug tension-free" plastic operations. The femoral canal is along its entire course sealed by a cylinder of rolled up polyester net. This operation reduces the postoperative discomfort and danger of the development of a relapse.