Next- generation sequencing is an effective method for diagnosing patients with different forms of monogenic diabetes.

AIM: Monogenic diabetes (MD) represents 5-7% of antibody-negative diabetes cases and is a heterogeneous group of disorders. METHODS: We used targeted next-generation sequencing (NGS) on Illumina NextSeq 550 platform involving the SureSelect assay to perform genetic and clinical characteristics of a study group of 684 individuals, including 542 patients referred from 12 Polish Diabetes Centers with suspected MD diagnosed between December 2016 and December 2019 and their 142 family members (FM). RESULTS: In 198 probands (36.5%) and 66 FM (46.5%) heterozygous causative variants were confirmed in 11 different MD-related genes, including 31 novel mutations, with the highest number in the GCK gene (206/264), 22/264 in the HNF1A gene and 8/264 in the KCNJ11 gene. Of the 183 probands with MODY1-5 diabetes, 48.6% of them were diagnosed at the pre-diabetes stage and most of them (68.7%) were on diet only at the time of genetic diagnosis, while 31.3% were additionally treated with oral hypoglycaemic drugs and/or insulin. CONCLUSIONS: In summary, the results obtained confirm the efficacy of targeted NGS method in the molecular diagnosis of patients with suspected MD and broaden the spectrum of new causal variants, while updating our knowledge of the clinical features of patients defined as having MD.

Dermoscopic and trichoscopic features of primary cutaneous lymphomas - systematic review.

Dermoscopy and trichoscopy are non-invasive methods used as auxiliary tools in diagnostics of different dermatoses. To date, no systematic review concerning the utility of dermoscopy and trichoscopy in the diagnostics of primary cutaneous lymphomas has been published. The aim of this study was to summarize the current state of knowledge on this topic based on systematic search of PubMed database and related references published before 8th of August 2020. Besides dermoscopic features, type of dermoscope, polarization mode, magnification, number of cases and histopathological correlation were analysed. A total of 34 records were included into the final analysis, evaluating 141 patients diagnosed with primary cutaneous T-cell lymphomas and 70 patients with primary cutaneous B-cell lymphomas. Most of the analysed records evaluated dermoscopic features (n = 206); trichoscopy was analysed in only 5 cases. Structures most commonly observed in classical mycosis fungoides (n = 108) were fine short linear vessels/linear vessels, spermatozoa-like vessels and orange-yellow patchy areas. In folliculotropic mycosis fungoides (n = 12), most frequently observed were comedonal lesions/comedo openings/central keratotic plugs and white halo around hair follicles/perifollicular accentuation. Primary cutaneous marginal zone B-cell lymphoma (n = 42) and primary cutaneous follicle centre lymphoma (n = 20) most commonly presented with salmon-coloured background and fine short/linear irregular/serpentine vessels. For other PCL, with less than 10 cases reported in the analysed records, details have been provided in the article. Most observations analysed in this systematic review rely on findings from case reports/case series (with the level of evidence V) and lack a control group. A few studies provided information concerning technical aspects of dermoscopic/trichoscopic examination. The role of dermoscopy/trichoscopy in diagnostics of cutaneous lymphomas requires further studies, especially in entities where dermoscopic features have been described in only single or a few cases. However, it seems that this practical, accessory tool in future may provide additional clues during clinical assessment.

Methotrexate in the treatment of mycosis fungoides - a multicenter observational study in 79 patients.

OBJECTIVE: The first report concerning methotrexate (MTX) in the treatment of Mycosis fungoides (MF) was published in 1964 by Wright. The mechanism of MTX action in the treatment of primary cutaneous T-cell lymphoma (CTCL) has been not explained in detail yet (the anti-inflammatory, immunomodulating, immunosuppressive, and cytostatic actions have been under discussion). PATIENTS AND METHODS: This is a retrospective analysis of 79 MF patients in 4 dermatology clinical centers in Poland. Data are presented in terms of the duration, use of MTX, the effectiveness of treatment with MTX in terms of time required to achieve remission, the disease stage, route of administration, age at diagnosis and the dosage. Moreover, the occurrence of side effects depending on the route of administration and duration of therapy with MTX was analyzed. RESULTS: The analysis has revealed that 56 patients (70,9%) had achieved remission on the MTX. The remission began in the 1st month of therapy in 20% of patients, lasted 4 to 6 months in 50% of cases. At least 12 months' remission was confirmed in 25% of patients (2-year-long only in 10% and 3-year-long in 5% of patients). The time to remission was related to the stage of disease at diagnosis as well as to minimal and maximal dose of MTX. The total therapeutic dose of MTX was found important for the course of the disease: higher total dose had prolonged the remission. CONCLUSIONS: Despite the common use of MTX in MF patients, relatively few clinical studies have been published. The response of MF subjects to MTX seems to depend on the stage and, more importantly, the dose of MTX treatment. Methotrexate appears to be an effective treatment at every stage of MF; however, it is not devoided of side effects such as infections and elevated level of aminotransferases, which are most common.

Chondroprotection by urocortin involves blockade of the mechanosensitive ion channel Piezo1.

Osteoarthritis (OA) is characterised by progressive destruction of articular cartilage and chondrocyte cell death. Here, we show the expression of the endogenous peptide urocortin1 (Ucn1) and two receptor subtypes, CRF-R1 and CRF-R2, in primary human articular chondrocytes (AC) and demonstrate its role as an autocrine/paracrine pro-survival factor. This effect could only be removed using the CRF-R1 selective antagonist CP-154526, suggesting Ucn1 acts through CRF-R1 when promoting chondrocyte survival. This cell death was characterised by an increase in p53 expression, and cleavage of caspase 9 and 3. Antagonism of CRF-R1 with CP-154526 caused an accumulation of intracellular calcium (Ca2+) over time and cell death. These effects could be prevented with the non-selective cation channel blocker Gadolinium (Gd3+). Therefore, opening of a non-selective cation channel causes cell death and Ucn1 maintains this channel in a closed conformation. This channel was identified to be the mechanosensitive channel Piezo1. We go on to determine that this channel inhibition by Ucn1 is mediated initially by an increase in cyclic adenosine monophosphate (cAMP) and a subsequent inactivation of phospholipase A2 (PLA2), whose metabolites are known to modulate ion channels. Knowledge of these novel pathways may present opportunities for interventions that could abrogate the progression of OA.

Updated 24-year trend of Type 1 diabetes incidence in children in Poland reveals a sinusoidal pattern and sustained increase.

AIMS: To present the incidence trend for Type 1 diabetes in Polish children aged 0-14 years, updated using data collected during 2005-2012, and assess the reliability of the predictive model constructed previously using the 1989-2004 database. METHODS: Children aged < 15 years with newly diagnosed Type 1 diabetes are recorded prospectively (EURODIAB criteria) in several regional registers in Poland. Age- and gender-standardized incidence rates for Type 1 diabetes were calculated per 100 000 persons/year. Incidence rates were analysed in terms of the dependency on age, gender, geographical region and population density. Incidence rate trends over time were modelled using generalized linear models. RESULTS: The mean standardized incidence for 1989-2012 was 12.72 per 100 000 persons/year [95% confidence interval (CI), 11.35 to 14.21]. Over the 24-year observation period, the incidence increased from 5.36 to 22.74 per 100 000 persons/year. The lowest incidence rate was in children aged 0-4 years (8.35, 95% CI 7.27 to 9.57 per 100 000 persons/year). There was no difference between genders, or urban and rural regions. Incidence rates were higher in northern compared with southern Poland [14.04 (95% CI 12.59 to 15.63) vs. 11.94 (95% CI 10.62 to 13.39) per 100 000 persons/year]. The new data corrected the earlier predictive model by changing the estimates of some factors related to patient age, gender and their interactions with the remaining factors. The incidence rate shows periodic 5.33-year fluctuations. The periodicity component allows for a more accurate prediction of the incidence rate over time. CONCLUSIONS: This cohort study reveals a sustained increase in Type 1 diabetes incidence in Polish children aged 0-14 years with regular, sinusoidal fluctuations and a slight levelling off in past few years. It is of concern that are the highest increases in incidence are found in children aged 0-4 years.

Monthly use of a real-time continuous glucose monitoring system as an educational and motivational tool for poorly controlled type 1 diabetes adolescents.

PURPOSE: Experience with the use of real-time continuous glucose monitoring systems (RT-CGMS) in teenagers with type 1 diabetes mellitus (T1DM) is limited. We aimed to assess the possibility of glycaemic control improvement and to characterize the group of adolescents, who may gain long-term benefits from the use of the RT-CGMS. METHODS: Forty T1DM patients, aged 14.6 ± 2.1 years, with diabetes duration 7.4 ± 3.6 years and initial HbA1c 9.3 ± 1.5% were recruited. The analysis was based on one-month glucose sensors use, combined with the thorough family support. Patients were analysed in groups according to baseline HbA1c: below and above 7.5%, and 10.0%. Comparison between patients with or without improvement in HbA1c after 3-month follow-up was also performed. Patients' satisfaction based on the questionnaire was assessed. RESULTS: HbA1c level in entire study group decreased after three months, from 9.3 ± 1.0% to 8.8 ± 1.6% (P<0.001). In the group with HbA1c improvement, reduction was the highest: 9.0 ± 1.3% vs. 8.0 ± 1.2% (P<0.001). Only the group with initial HbA1c>10% did not achieve significant improvement: 11.2 ± 0.5% vs. 10.9 ± 1.1 (P=0.06). In satisfaction questionnaire the lowest scores (negative opinion) were reported by group of patients with initial HbA1c above 10%, while the highest scores (positive opinion) were found in the group with improvement of HbA1c after 3 month follow-up. CONCLUSION: Short-term use of CGMS RT, united with satisfaction questionnaire, performed in poorly controlled teenagers with T1DM, can be useful in defining the group of young patients, who can benefit from long-term CGMS RT use in metabolic control improvement.

Cardiovascular risk in nonobese hypertensive adolescents: a study based on plasma biomarkers and ultrasonographic assessment of early atherosclerosis.

The objective of this study was to investigate the vascular status, left-ventricular mass and biomarkers of endothelial activation in hypertensive (HT) adolescents, with particular attention to comparing nonobese with obese patients. Seventy-nine newly diagnosed HT adolescents aged 15.1±2.1 years (divided into 34 nonobese and 45 obese) were compared with 35 healthy volunteers. Intima-media thickness (IMT), flow-mediated dilation (FMD) and left-ventricular mass index (LVMi) were determined using ultrasound. Adhesion molecules and inflammatory interleukins (ILs), together with lipids and insulin resistance (HOMA), were also studied. HT obese adolescents had higher triglycerides, HOMA, and elevated levels of interleukin-6, tumor necrosis factor-α, soluble intercellular adhesion molecule-1 and soluble E-selectin compared with controls and nonobese HT patients. FMD was lower in HT groups (8.5±4.5% in nonobese, P=0.004; 8.1±4.9%, P=0.01 in obese vs 12.5±4.9%; in control), and IMT was higher (0.52±0.06 mm, P<0.001 in nonobese; 0.54±0.05 mm, P<0.001 in obese vs 0.42±0.05 mm in control). Higher LVMi was found in both HT groups, with the highest value in the nonobese group being 37.8±5.3 g m(-2.7) vs 28.4±5.3 g m(-2.7) in controls (P=0.003). In conclusion, nonobese HT adolescents had the same early cardiovascular deteriorations assessed ultrasonographically as their obese HT peers, although metabolic alterations and endothelial activation measured as plasma biomarkers were more pronounced in obese individuals. The potential mechanisms of early atherosclerosis in nonobese HT adolescents need further evaluation in prospective studies because these factors may differ considerably from those found in young obese individuals with HT.

Disturbances in some gene expression in T regulatory cells separated from children with metabolic syndrome.

The metabolic syndrome (MS) is defined as a cluster of risk factors, including abdominal obesity, dyslipidaemia, glucose intolerance and hypertension, which increase the risk for coronary heart disease. The immunological aspects of obesity and MS, including the role of T regulatory cells, have been intensively investigated. The aim of this study was to determine whether there is any disturbance in T regulatory cells number and/or function in children with MS. The percentages of T regulatory cells in the peripheral blood of children fulfilling the International Diabetes Federation criteria of the disease (n = 47) were assessed. Treg cells were also separated for further analysis of multiple genes important in their function with the use of real-time RT-PCR. We did not observe any difference in Treg percentages between study and control group but there was lower expression of some molecules including transforming growth factor-beta and interleukin-12 family members in Treg cells separated from children with MS compared to the healthy subjects. Our study is the first to report significant disturbances in some gene expression in T regulatory cells separated from children with MS. The results should be useful for further research in this field, including immunotherapeutic interventions.

The association of early atherosclerosis and retinopathy in adolescents with type 1 diabetes: preliminary report.

Recent studies have shown a close correlation between advanced diabetic retinopathy and the late stages of atherosclerosis. The purpose of this study was to analyse the association between diabetic retinopathy and early atherosclerotic changes in adolescents with type 1 diabetes. We studied 28 adolescents with type 1 diabetes. Eight patients with nonproliferative retinopathy were compared with the remaining 20 patients, and with 11 healthy controls. The function of endothelium was assessed by measuring flow-mediated dilatation (FMD), the intima-media thickness (IMT) of the common carotid arteries and adhesion molecules (sICAM-1, sVCAM-1, sE-selectin). In the group with retinopathy FMD equalled 7.8+/-4.1% vs. 12.1+/-5.1% in the control group (p=0.04), and in the group without retinopathy, 7.6+/-5.5% (p=0.04 compared to controls). Higher IMT was found in all patients with diabetes in comparison with healthy controls: 0.49+/-0.06 mm vs. 0.42+/-0.03 mm (p=0.001). Patients with retinopathy had a significantly higher value of IMT in comparison not only with controls but also with patients without complications: 0.56+/-0.06 mm vs. 0.47+/-0.03 mm (p=0.0001). Adhesion molecule levels were not changed in patients with retinopathy. Higher IMT was found in adolescents with diabetic retinopathy in comparison with patients without complications, which may suggest that macrovascular changes are more advanced in these patients than in their diabetic peers without retinopathy.

[Epidemiologic aspect of lyme borreliosis among the inhabitants of Podlasie Province]. / Wybrane aspekty epidemiologiczne boreliozy z Lyme wsród mieszkanców województwa podlaskiego.

UNLABELLED: The aim of this study was to evaluate Lyme borreliosis morbidity among the inhabitants of Podlasie Province in the years of 1996-2000. MATERIAL AND METHODS: The new cases of Lyme borreliosis reported in the years of 1996-2000 in Podlasie Province were analyzed. The analysis included the data from Mz-56 and Mz-57 forms and the information from epidemiologic investigations sent to Regional Sanitary and Epidemiologic Station in Bialystok. In 1996-2000, screening examinations were carried out in the same group of 358 forestry workers including 44 (12.29%) females and 314 (87.71%) males aged from 21 to 64 (x = 41.2) years. Immunoenzymatic test of ELISA Borrelia recombinant IgM and IgG by Biomedica firm (Austria) were used to detect B. burgdorferi antibodies. RESULTS: In 1996-2000, 4933 of borreliosis cases were registered in Poland including 1377 (27.91%) in Podlasie Province. The morbidity rate in Podlasie province ranged from 15.05% in 1996 year to 21.29% in 2000 year of the whole country morbidity. At the same time, the incidence rate in Podlasie Province ranged from 9.09 in 1996 to 32.2 in 2000 year and was 6.72-fold higher than the incidence rate in the whole country. In Eastern and Central region of the province, 80.54% of cases were registered whereas, 14.09% by the Lakeside of Augustów and Suwalki only 5.37% in the western region of the province. It was proved that the morbidity increased in proportion to the age of patients (41.39% of patients were at the age of 30-49) and it decreased only above the age of 60. Thus, Lyme borreliosis affects mainly people at working age. The increase in B. burgdorferi antibody detectability was noticed in the population of forestry workers; it was detected in 38.55% in 1995 and in 2000 in 43.56% of the examined. Interestingly, 81 people infected with B.b in 1995-2000 included 74 men and only 7 women, supporting the statement that B.b infection affects mainly working professionals and the risk increases with the practice. CONCLUSIONS: Our studies indicate that Lyme borreliosis is a serious health problem among the inhabitants of Podlasie Province which is an endemic area for Ixodes ricinus ticks infected with B. burgdorferi spirochete.

The incidence of malignancy in heart transplant recipients.

OBJECTIVES AND METHODS: 219 heart transplant recipients with survival over 3 months were retro- and prospectively analysed for the incidence of primary neoplasms. Patients received immunosuppressive drugs (cyclosporine A, azathioprine, steroids) with a 4-5 days induction course of Rabbit Anti-Thymocyte Immunoglobulin (RATG) or monoclonal antibodies induction /OKT3/ in some cases. Anti-rejection treatment consisted of pulse doses of methyloprednisolon or RATG. RESULTS: 9 cases of malignancy (4.1%) with one case of pre-malignant liver condition (dysplasia gigantocellulare, 0.45%) were found (8M; 1F; age: 45-67 y.o., x57.7). Symptoms of neoplasms occurred 7-79 months (x31.4) postoperatively. Skin carcinomas: planoepitheliale, spinocellulare, soft tissue neoplasms/mesenchymal sarcoma, larynx Ca planoepitheliale, lung: adenocarcinoma and Ca microcellulare, kidney Ca clarocellulare and post transplant non-Hodgkin lymphoma were diagnosed. Chemo- and radiotherapy, surgery and reduction of immunosuppression did not change the outcome of malignancy in 6 pts.; (regression-1 pt was., remission-2 pts). Patients died 7-86 months after Htx (x41), 4-25 mos. (x12.5) after suffering from first symptoms and 0-10 months (x4.9) after pathology-based diagnosis of neoplasm. CONCLUSIONS: Heart transplant recipients have an increased risk of carcinogenesis. The incidence of malignancies in the studied group is similar or even lower than in other reports.

Respiratory syncytial virus induces selective production of the chemokine RANTES by upper airway epithelial cells.

The presence of histamine and eosinophil cationic protein in nasopharyngeal secretions of infants with respiratory syncytial virus (RSV)-induced bronchiolitis implies the activation of basophil and eosinophil leukocytes, but the specific mechanism of their recruitment has not been elucidated. Chemokines are potent and selective leukocyte chemotactic molecules that are also expressed by airway epithelial cells. Therefore, the pattern of chemokines produced in response to RSV infection was investigated in primary cultures of human nose- and adenoid-derived epithelial cells. Interleukin-8, growth-related peptide-alpha, and monocyte chemotactic protein-1 were constitutively released by uninfected epithelial cells and were not further enhanced by infection with RSV. RANTES (regulated upon activation, normal T cell-expressed and -secreted), which was present in negligible concentrations in uninfected cultures, was strongly induced by RSV infection, in a dose- and time-dependent manner. Through the release of RANTES, epithelial cells may control the selective concentration and activation of basophils and eosinophils in RSV-infected airway mucosa.

The fine structure of the spinal cord in human embryos and early fetuses.

The developing human spinal cord was studied by electron microscopy in 7 embryos (18--28 mm (C.-R.) and 5 fetuses (35--45 mm). It was found that, during the last week of the embryonic period, the spinal cord undergoes distinct maturation, as evidenced by cytoplasmic organelles, glial differentiation, and the formation of synaptic junctions. The ventricular (ependymal) cells are ciliated; densely grouped cells with scanty processes occupy the early intermediate (mantle) zone; and glioblasts are present in the marginal zone. Degenerating cells are evident in the spinal cord. The first axondendritic synapses appear in the marginal layer of the basal plate in the cervical part of the spinal cord in a human embryo of 28 mm, i.e., during the embryonic period proper. In the fetal period, synapses are found also in the alar plate, and axosomatic synapses are present. The formation of synapses and the general maturation of the spinal cord seem to proceed craniocaudally. The appearance of synapses is in keeping with the finding of reflex movements in the human during the late embryonic and early fetal periods.

Types of neural cells in the spinal ganglia of human embryos and early fetuses.

Spinal ganglial of human embryos and fetuses ranging in C.-R. length from 15 to 74 mm and in age from 6 1/2 to 11 postovulatory weeks were studied by light and electron microscopy. A sequence of events in differentiation and maturation enabled five types of cells to be distinguished: 1. apolar, undifferentiated neuroblasts are the main cells at 6 1/2 to 7 1/2 weeks; 2. early bipolar neuroblasts (strictly speaking, types 2 to 5 are immature neurons) predominate at the end of the embryonic period proper (8 postovulatory weeks); 3. intermediate bipolar neuroblasts are characteristic of the early fetal period; 4. late bipolar neuroblasts, in which two proceses arise separately from one pole of the cell, appear at about 10 postovulatory weeks; 5. unipolar neuroblasts are found within another week and, by that time, cells of types 1 and 2 are no longer present.