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BACKGROUND The prevalence of type-2 diabetes mellitus (T2DM) has been increasing rapidly in developed and developing countries. Many new loci associated with T2DM have been uncovered by genetic and genome-wide association studies, eg. the transcription factor 7-like 2 (TCF7L2) gene variant rs7895340 has been associated with type-2 diabetes mellitus. The aim of this research was to find an association between polymorphisms of TCF7L2 gene variant rs7895340 with T2DM in ethnic Minangkabau. METHODS This was a case-control study using a consecutive sampling technique among ethnic Minangkabau patients who came for treatment to the Metabolic Endocrinology Polyclinic at Dr. M. Jamil Hospital. Analysis was performed on 132 subjects (66 T2DM patients and 66 controls). DNA was isolated from venous blood samples and amplified using the amplification refractory mutation system - polymerase chain reaction (ARMS-PCR) method to detect single nucleotide polymorphisms (SNPs) of TCF7L2 gene variant rs7895340. To determine the significance of association of SNPs of rs7895340 with T2DM in ethnic Minangkabau, data analysis was performed using chi-square test. RESULTS The ARMS - PCR for TCF7L2 gene variant rs7895340 found polymorphisms in 28 (42.42%) subjects with type 2 diabetes mellitus and in 15 (22.72%) non-diabetic subjects.There was a significant association between rs7895340 SNPs with the probability of T2DM in ethnic Minangkabau (OR=2.505, p=0.026). CONCLUSIONS Single nucleotide polymorphism rs7895340 of TCF7L2 gene can be used as genetic marker of type-2 diabetes mellitus in ethnic Minangkabau.

AbstrakDiabetes melitus (DM) merupakan suatu penyakit metabolik dengan karakteristik hiperglikemia akibat kelainan sekresi dan/atau gangguan kerja insulin. Sekitar 80% dari seluruh kasus DM merupakan DM tipe 2. Salah satu komplikasi DM adalah tuberkulosis (TB) paru. Tujuan penelitian ini adalah mengevaluasi insidensi TB paru pada pasien DM tipe 2 di Ruang Rawat Inap Penyakit Dalam RSUP Dr. M. Djamil Padang. Instrumen penelitian yang digunakan adalah data sekunder dari Instalasi Rekam Medis RSUP Dr. M. Djamil Padang berupa data pasien DM tipe 2 dan rekam medik pasien DM tipe 2 dengan TB paru yang dirawat di Ruang Rawat Inap Penyakit Dalam RSUP Dr. M. Djamil Padang sejak 1 Januari 2011 – 31 Desember 2011. Hasil penelitian ini menunjukkan bahwa terdapat 29 kasus TB paru dari 748 orang pasien DM tipe 2 (3,88%). Kasus DM tipe 2 dengan TB paru terbanyak ditemukan pada kelompok jenis kelamin laki-laki (58,62%), usia < 60 tahun (72,41%; mean: 54,66 ± 12,77 tahun), dan normoweight (51,72%). Sebagian besar pasien menunjukkan hasil pemeriksaan BTA sputum negatif (65,52%). Insidensi pada penelitian ini sedikit lebih rendah daripada hasil penelitian sebelumnya.Kata kunci: insidensi, tuberkulosis paru, diabetes melitus tipe 2 AbstractDiabetes mellitus (DM) is a group of metabolic disorders characterized by hyperglycemia resulting from defects in insulin secretion, insulin action or both. Type 2 DM makes up about 80% of all cases of DM. One of the DM’s complications is pulmonary tuberculosis (TB). The objective of this study was to evaluate the incidence of pulmonary TB among type 2 DM patients in Internal Medicine Ward RSUP Dr. M. Djamil Padang. The instruments used in this research were the secondary data derived from the Medical Record Departement RSUP Dr. M. Djamil Padang in the form of type 2 DM patients’ data and medical records of type 2 DM patients with pulmonary TB who were treated in Internal Medicine Ward RSUP Dr. M. Djamil Padang since January 1st 2011 – December 31st 2011. The results of this research showed that there were 29 cases of pulmonary TB among 748 type 2 DM patients (3,88%). Most of type 2 DM with pulmonary TB cases were found in males (58,62%), < 60 years old (72,41%; mean: 54,66 ± 12,77 years old), and normoweight group (51,72%). Most of these patients showed negative results on the sputum AFB examination (65,52%). The incidence in this research is lower than the previous studies.Keywords: incidence, pulmonary tuberculosis, type 2 diabetes mellitus

AbstrakDiabetes Melitus (DM) merupakan suatu kelompok penyakit metabolik dengan karakteristik hiperglikemia. DM tipe 2 adalah yang paling sering ditemukan. Komplikasi kronis DM tipe 2 yaitu mikrovaskular dan makrovaskular yang dapat menurunkan kualitas hidup penderita. Tujuan penelitian ini adalah untuk memperoleh gambaran tentang insidensi penderita DM tipe 2 dengan komplikasi kronis. Penelitian deskriptif ini dilakukan dengan mengambil data pada rekam medik penderita DM tipe 2 dengan komplikasi kronis yang dirawat inap di bagian Penyakit Dalam RS.Dr. M. Djamil, Padang Januari 2011-Desember 2012. Penelitian dilakukan dari Februari 2013-April 2013 di bagian rekam medik RS. Dr. M. Djamil Padang. Data didapatkan sebanyak 261 pasien, dari jumlah tersebut didapatkan 197 pasien memiliki komplikasi kronis DM tipe 2. Data dikategorikan berdasarkan jenis komplikasi makrovaskular dan mikrovaskular. Hasil penelitian menunjukkan bahwa penderita dengan komplikasi kronis makrovaskular (66,5%) dan mikrovaskular (81,7%). Terdapat perubahan insidensi dalam dua tahun yaitu dari tahun 2011 dengan 2012. Komplikasi kronis yang paling sering terjadi adalah nefropati diabetik (42,6%) pada perempuan Kata kunci: diabetes melitus, komplikasi mikrovaskular, komplikasi makrovaskularAbstractDiabetes mellitus is a group of metabolic diseases with characterized by hyperglycemia. Type 2 diabetes is the most common disease in the world. Chronic complications of type 2 diabetes are microvascular and macrovascular complications that can reduce the quality of life of patients. The objective of this study was to obtain a picture of the incidence of type 2 diabetic chronic complications. This descriptive study was conducted by taking medical record data of hospitalized type 2 diabetic patients with chronic complications inInternal Medicine Department Dr. M. Djamil hospital, Padang on January 2011-December 2012. The study was conducted from February 2013-April 2013 at the hospital medical record Dr. M. Djamil, Padang. This study was conducted on 261 patients, from that number 197 patients have chronic complications of type 2 diabetes mellitus. Data were categorized by type of macrovascular and microvascular complications.The results showed that patients with chronic complications of macrovascular (66,5%) and microvascular (81,7%). There is a change in incidence from 2011 to 2012. The most common of chronic complications is diabetic nephropathy (42.6%). Chronic microvascular and macrovascular complications are different incidence in two years, the most common is diabetic nephropathy which often occurs in women Keyword: diabetes mellitus, microvascular complication, macrovacular complication.

Introduction and Objective: Hippobosca equina (Diptera: Hippoboscidae), is a widespread blood-feeding ectoparasite associated with the forest ecosystem. The insect is characterized by a wide host range and low host specificity, which increases the risk of feeding on animals that constitute a reservoir of transmissible pathogens, including Bartonella spp., Material and Methods: Hippobosca equina adults were collected from humans and companion animals within a continental mesotrophic oak-pine mixed forest in eastern Poland. DNA was isolated by the ammonia method, and isolates obtained from single individuals were tested by PCR method for the presence of 5 vector-borne pathogens. In case of the positive results, the amplicons were sequenced and examined by a BLAST search., Results: The PCR analysis of DNA isolates obtained from 100 H. equina specimens revealed the presence of the RNA polymerase beta-subunit gene ( rpoB ) of the genus Bartonella , in 1% of the studied insects, i.e. one H. equina female. The rpoB gene haplotype of Bartonella sp. reported in this study, was identical to a Bartonella sp. sequence obtained from deer keds in Lithuania, and very closely related to strains with zoonotic potential. None of the H. equina specimens studied was positive for the presence of B. burgdorferi s.l., Anaplasma phagocytophilum , Babesia spp., and Coxiella burnetii ., Conclusions: The study indicates the need to screen the occurrence of Bartonella spp., both in potential vectors and reservoirs of this pathogen in various habitats.

Dermacentor reticulatus (Acari: Ixodidae) is an important arthropod vector in medical and veterinary contexts. Its geographic range is divided into western and eastern populations separated by a " Dermacentor -free zone" in central Poland. Recent faunistic studies showed a new endemic locality of the species in Upper Silesia to the west of the Vistula River (central-southern Poland) and its co-occurrence with I. ricinus. The prevalence of five tick-borne pathogens (TBPs), e.g., B. burgdorferi s.l., Bartonella spp., Rickettsia spp., and Babesia spp., in the ticks was assessed with polymerase chain reaction (PCR) methods. The molecular studies revealed the presence of Rickettsia spp. in 23.8% of the D. reticulatus specimens. In turn, 94.1% of the I. ricinus adults were infected with B. burgdorferi s.l., 11.7 % with Babesia spp., and 5.8% with Rickettsia spp. Coinfections with two TBPs were noted in 17.6% of the I. ricinus . These findings highlight not only the risk of infestation by both tick species in an area previously considered Dermacentor -free, but also the high prevalence of TBPs in the study area. Increased focus on medical and veterinary services appears necessary to diagnose and prevent tick-borne diseases in this region.

Introduction and Objective: Dermacentor reticulatus is one of the tick species of the greatest epidemiological importance in Europe. To date, the Eastern European and Western European populations of this tick species have been separated by an area located in Poland where the species has never been found. In this study, newly discovered D. reticulatus localities in areas transformed by human activities in central-southern Poland are described., Material and Methods: The specimens of the ornate dog tick were identified among ticks collected from companion animals in 2010, 2012, 2013, and 2014. They were examined using PCR methods to detect Borrelia burgdorferi s.l., Rickettsia spp., Anaplasma phagocytophilum , Bartonella spp., Babesia spp., and Toxoplasma gondii . In the case of the positive results, the amplicons were sequenced and examined by a BLAST search., Results: In total, 6 specimens of D. reticulatus were collected (3 females and 3 males). As declared by the owners, animal hosts stayed in the same area throughout the study period and had never travelled outside their place of residence. As many as 3/6 (50%) of D. reticulatus adults removed from dogs were infected with Rickettsia raoultii ., Conclusions: The results expand the available data on the spread of the ornate dog tick and indicate that, since 2010, this tick species and Rickettsia raoultii transmitted by this tick species have probably been present in this area, which has a strongly transformed agricultural structure and and had previously been regarded as a D. reticulatus -free zone. The presence of the ornate dog tick in urban and suburban habitats in central-southern Poland poses new threats to the health of companion animals and humans associated with the transmission of pathogens by this species.

Introduction and Objective: The common tick Ixodes ricinus is one of Europe's most important vectors of tick-borne diseases. The increased risk of attacks by this tick suggests the need for identification of factors contributing to the transmission of tick-borne pathogens, and the routes of pathogen circulation in nature., Material and Methods: Polymerase chain reaction was used to investigate the prevalence of four pathogens, i.e. Borrelia burgdorferi s.l. (Bb), Anaplasma phagocytophilum (Ap), Babesia spp. (Bs), and Bartonella spp. (Ba) in I. ricinus females and males mating on dogs in south-central Poland., Results: The study revealed the presence of three pathogens: Bb, Ap, and Bs in 9.4%, 5.4%, and 5.4% of all I. ricinus adults in copula , respectively. Co-infection with two pathogens was detected in one tick specimen. Borrelia burgdorferi spirochetes were isolated in two females and two males in copula , but the sexual transfer of the spirochetes between these specimens could not be clearly confirmed., Conclusions: By increasing the feeding dynamics in females, the copulation of I. ricinus males with females attached to the host's skin may stimulate pathogen replication in tick tissues and migration from the gut to the salivary glands. Further investigations of the I. ricinus copulation on the host on female feeding and pathogen transmission may contribute to the elucidation of the eco-epidemiology of tick-borne diseases transmitted by this tick species.

MiRNAs could play an important role in tumorigenesis and progression. The oncoprotein MDM2 (murine double minute 2) was identified as a negative regulator of the tumour suppressor p53. This study aims to analyse the expression of the MDM2 target miRNA candidates (miR-3613-3p, miR-371b-5p and miR-3658) and the MDM2 gene in oral squamous cell carcinoma tumour and margin samples and their association with the selected socio-demographic and clinicopathological characteristics. The study group consisted of 50 patients. The miRNAs and MDM2 gene expression levels were assessed by qPCR. The expression analysis of the miRNAs showed the expression of only one of them, i.e., miR-3613-3p. We found no statistically significant differences in the miR-3613-3p expression in tumour samples compared to the margin samples. When analysing the effect of smoking on miR-3613-3p expression, we demonstrated a statistically significant difference between smokers and non-smokers. In addition, we showed an association between the miR-3613-3p expression level and some clinical parameters in tumour samples (T, N and G). Our study demonstrates that miR-3613-3p overexpression is involved in the tumour progression of OSCC. This indicates that miR-3613-3p possesses potential prognostic values.

Osteogenesis imperfecta ( OI ) is a group of connective tissue disorders leading to abnormal bone formation, mainly due to mutations in genes encoding collagen type I (Col I). Osteogenesis is regulated by a number of molecules, including microRNAs (miRNAs), indicating their potential as targets for OI therapy. The goal of this study was to identify and analyze the expression profiles of miRNAs involved in bone extracellular matrix (ECM) regulation in patients diagnosed with OI type I caused by mutations in COL1A1 or COL1A2 . Primary skin fibroblast cultures were used for DNA purification and sequence analysis, followed by analysis of miRNA expression. Sequencing analysis revealed mutations of the COL1A1 or COL1A2 genes in all OI patients, including four previously unreported. Amongst the 40 miRNAs analyzed, 9 were identified exclusively in OI cells and 26 in both OI patients and the controls. In the latter case, the expression of six miRNAs (hsa-miR-10b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, has-miR-204-5p, has-miR-216a-5p, and hsa-miR-449a) increased, while four (hsa-miR-129-5p, hsa-miR-199b-5p, hsa-miR-664a-5p, and hsa-miR-30a-5p) decreased significantly in OI cells in comparison to their expression in the control cells. The identified mutations and miRNA expression profiles shed light on the intricate processes governing bone formation and ECM regulation, paving the way for further research and potential therapeutic advancements in OI and other genetic diseases related to bone abnormality management.

Introduction and Objective: Little is known about interspecific contacts between ticks. Therefore, this study focused on the investigation of factors that may influence interspecific contacts between Ixodes ricinus and Dermacentor reticulatus ticks., Material and Methods: Ixodes ricinus males and D. reticulatus females involved in oral-anal contacts (group I) and questing specimens with no such behaviour (group II) collected in eastern Poland were examined using molecular techniques to detect Borrelia burgdorferi s.l. (Bb), Rickettsia spp. (Rs), Anaplasma phagocytophilum , Babesia microti , and Toxoplasma gondii ., Results: An extremely high infection rate of Bb and Rs was determined in I. ricinus males (in groups I: 100% and 46.15% and group II: 90% and 40%, respectively) and D. reticulatus females (in group I: 84.61% and 61.53% and in group II: 90% and 20%, respectively). The prevalence of other pathogens in these ticks was substantially lower. Co-infection with pathogens was detected in approximately 53% of ticks., Conclusions: The study suggests that tick-borne pathogens may have influenced the sexual behaviour of their vectors. The oral-anal contacts between I. ricinus and D. reticulatus ticks are probably stimulated by Bb and/or Rs. The presence of five pathogens and numerous co-infections in the analysed ticks indicates a risk of various human infectious diseases in the study region. Further studies are required to clarify the implications of oral-anal interspecific tick interactions.

Introduction and Objective: Progressive climate change plays a major role in the expansion of tick populations, as well as the transmission of tick-borne disease pathogens (TBPs) to humans and animals throughout the world. Zoonoses are a growing environmental problem of great importance for public health. In Poland, domestic dogs and cats are mainly infested by Ixodes ricinus, Ixodes crenulatus, Ixodes hexagonus, Ixodes rugicollis and Dermacentor reticulatus ticks from the Ixodidae family. Other tick species, such as I. apronophorus and Haemaphysalis concinna, recorded sporadically on domestic dogs or cats, may expand their range in the future and successfully infest pets. Individual cases of infestations in Poland with foreign tick species, such as Rhipicephalus sanguineus, are known and may soon be recorded more frequently., Objective: The aim of this review is to summarize the occurrence and identification of tick species and TBDs that are present or are likely to spread in Poland, which, given the medical and veterinary importance of ticks, may help in the development of public health strategies., Review Methods: A comprehensive literature review of publications and own research and data analysis was performed, obtained from reports and scientific descriptions on the epidemiology of tick-borne diseases., Brief Description of the State of Knowledge: The ecology of ticks and hosts in urban and suburban environments is critical for quantifying the parameters necessary for an initial risk assessment, and identifying public health strategies to control and prevent TBDs. In the future, these species may expand their range of occurrence and their host, becoming common members of the Polish tick fauna., Summary: Anaplasma, Babesia, Borrelia and Rickettsia spp. are the major TBPs in Poland, and their prevalence is usually higher in dogs than cats.

Background: The rice weevil ( Sitophilus oryzae ) originates from subtropical and tropical areas of Asia and Africa, but it also appears on other continents, mostly as a result of trade in rice. It may occur in grain fields as well as in storage facilities, and cause allergenic reactions. The aim of this study was to identify the potential antigens in all developmental stages of S. oryzae, which may cause an allergic response in humans., Methods: Sera of 30 patients were tested for the presence of IgE antibodies to antigens from three life stages of the rice weevil. To identify protein fractions containing potential allergens, proteins collected from larvae, pupae, and adults separated by sex of S. oryzae were fractionated by SDS-PAGE. Then, they were probed with anti-human, anti-IgE monoclonal antibodies, fractionated by SDS-PAGE and detected by Western blotting., Results: In total, 26 protein fractions of males and 22 fractions of other life stages of S. oryzae (larvae, pupae, and females) positively reacted with the examined sera., Conclusions: The conducted study showed that S. oryzae may be a source of many antigens which may cause the potential allergic reactions in humans., Competing Interests: The authors declare no conflicts of interest.

Chronic tonsillitis is a problem related to bacterial and viral infections. Ficolins play a key role in the defence against various pathogens. In the present study, we investigated the associations between the selected single nucleotide polymorphisms (SNPs) of the FCN2 gene and chronic tonsillitis in the Polish population. The study included 101 patients with chronic tonsillitis and 101 healthy individuals. The selected SNPs of FCN2 (rs3124953, rs17514136 and rs3124954) were genotyped using TaqMan SNP Genotyping Assays (Applied Biosystem, Foster City, CA, USA). The analysis of rs17514136 and rs3124953 showed no significant differences in genotype frequencies between the chronic tonsillitis patients and controls ( p > 0.01). The CT genotype of rs3124954 was significantly more frequent, while the CC genotype was less frequent in chronic tonsillitis patients ( p = 0.003 and p = 0.001, respectively). The frequency of the A/G/T haplotype (rs17514136/rs3124953/rs3124954) was significantly more common in chronic tonsillitis patients ( p = 0.0011). Moreover, the FCN2 CT genotype of rs3124954 was associated with a higher risk of chronic tonsillitis, while the CC genotype of rs3124954 decreased this risk. Our findings demonstrate that FCN2 rs3124954 may be associated with chronic tonsillitis in the Polish adult population.

Tick-borne microorganisms belong to important etiological agents of many infectious diseases affecting humans and animals. Among them, there are haemoprotozoans of the Babesia genus, which infect erythrocytes of a host and may cause many clinical symptoms. Canine babesiosis is an emerging tick-borne disease in Southern and Central Europe. In this study, we report two cases of symptomatic canine babesiosis caused by Babesia canis in domestic dogs from the Silesian Voivodeship, Poland, as well as the presence of Dermacentor reticulatus ticks detected on one of the Babesia-infected dogs (D. reticulatus-free zone). The molecular analysis confirmed the presence of Babesia canis in the dogs’ blood, and the sequencing analysis showed that the obtained sequence is 100% identical to the sequence of Babesia canis isolate 3469 (sequence ID: KX712122.1). Our findings should raise awareness of B. canis infection among dog owners and veterinarians in the region where B. canis was not previously reported in residential, non-traveling dogs, as well as ensuring that adequate diagnostic methods are available.

Oral squamous cell carcinoma (OSCC) is one of the most prevalent types of cancers worldwide. LTF arrests the G1 to S phase transition of the cell cycle. This study is the first that has aimed to determine the possible association between the LTF polymorphisms (rs2073495, rs1126478, rs34827868, rs1042073, rs4637321, rs2239692 and rs10865941), the mRNA LTF expression, the risk of OSCC and the influence on the TNM staging and histological grading. This study was composed of 176 Polish patients, including 88 subjects diagnosed with OSCC and 88 healthy individuals. QuantStudio Design and Analysis Software v1.5.1 was used for the single nucleotide polymorphism (SNP) analysis and mRNA LTF expression. The G/G genotype of rs2073495 and the G/G genotype of rs4637321 were linked, with an increased risk of OSCC. There were no significant influences between the TNM staging and the histological grading and the LTF genotype. We found no statistically significant dissimilarities in the expression level of LTF genes in the tumour and margin specimens. No association was found between the gene expression levels, the other parameters or LTF polymorphisms in the tumour and margin samples. In conclusion, rs2073495 and rs4637321 polymorphisms may affect the risk of OSCC. These results should be validated on larger and different cohorts to better comprehend the role of the LTF gene in OSCC.

Deer keds are hematophagous ectoparasites (Diptera: Hippoboscidae) that mainly parasitize Cervidae. These flies are particularly important for animal health due to the occurrence of numerous pathogenic microorganisms. They may also attack humans and their bites may cause allergenic symptoms. The aim of the study was to identify the molecular characteristics of Borrelia burgdorferi sensu lato and Bartonella spp. pathogens detected in Lipoptena spp. sampled both from the hosts and from the environment. For identification of Bartonella spp and B. burgdorferi s. l., the primers specific to the rpoB and flaB gene fragments were used, respectively. The overall prevalence of B. burgdorferi s.l. DNA in Lipoptena cervi was 14.04%, including 14.8% infection in the tested group of winged specimens. The overall prevalence of Bartonella spp. was 57.02%. The presence of these bacteria was detected in 53.5% of specimens of L. cervi and 75.7% of L. fortisetosa . The phylogenetic analysis showed five new haplotypes of the rpoB gene of Bartonella sp. isolated from L. cervi/Lipoptena fortisetosa. We also identified one new haplotype of B. afzelii and three haplotypes of B. burgdorferi isolated from winged specimens of L. cervi. This is the first study to detect the genetic material of B. burgdorferi s.l. in L. cervi in Poland and the first report on the identification of these bacteria in host-seeking specimens in the environment.

The public health problem of tick-borne diseases has attracted much attention in recent years due to an increasing incidence in humans and animals. The aim of this study was to compare the risk of exposure to ticks and tick-borne infections in dogs and cats in recreational and urbanized areas in the Lesser Poland and Silesian Provinces. For molecular testing for the presence of the selected pathogens, 207 I. ricinus females collected from 119 dogs and 50 cats, and 2 I. hexagonus females collected from 2 domestic dogs, were examined. Overall, A. phagocytophilum was found in 3.7% of the I. ricinus specimens, B. microti in 27.1%, and B. burgdorferi s.l. in 0.9%. In urban areas of both provinces, A. phagocytophilum was found in 4.8% of the I. ricinus specimens, B. microti in 41.6% and B. burgdorferi s.l. in 3.9%. Pathogens were detected B. microti in both studied I. hexagonus specimens. These findings may indicate the important role that these animals play in the circulation of these pathogens in nature., (© 2022. The Author(s).)

Exposure to house dust allergens, mainly from domestic mites, is an important cause of allergic reactions in sensitized asthmatic patients. A total of 63 dust samples were collected from 16 flats in Bytom (south Poland); in each flat a person (age 4-17 years) suffering from bronchial asthma lived with his/her family. Mite density was calculated as the number of specimens per g of dust. The results were compared with household features and the data were statistically analyzed. In total 566 mite specimens were isolated, including 526 members of the family Pyroglyphidae (93%). The dominant species were Dermatophagoides pteronyssinus (60% of the total count) and Dermatophagoides farinae (32%). Pyroglyphids were found in all mite positive samples (68%) of which 35% also contained non-pyroglyphids, including glycyphagids, cheyletids and gamasids. The results suggest associations between the density of some mite taxa (per g of dust) and the following indoor environmental factors: presence of pets, number of inhabitants, coal-stoves as a type of heating, cleaning frequency, higher relative humidity, presence of flowers and PVC windows. The severity of asthma seems to be associated with the numbers of D. farinae, total domestic mites and live mites per g of dust., (© 2021. The Author(s).)

Polymorphisms in the apolipoprotein E (apoE) gene are risk factors for chronic inflammatory diseases including atherosclerosis. The gene product apoE is synthesized in many cell types and has both lipid transport-dependent and lipid transport-independent functions. Previous studies have shown that apoE expression in myeloid cells protects against atherogenesis in hypercholesterolemic ApoE -/- mice. However, the mechanism of this protection is still unclear. Using human APOE gene replacement mice as models, this study showed that apoE2 and apoE4 expressed endogenously in myeloid cells enhanced the inflammatory response via mechanisms independent of plasma lipoprotein transport. The data revealed that apoE2-expressing myeloid cells contained higher intracellular cholesterol levels because of impaired efflux, causing increasing inflammasome activation and myelopoiesis. In contrast, intracellular cholesterol levels were not elevated in apoE4-expressing myeloid cells, and its proinflammatory property was found to be independent of inflammasome signaling and related to enhanced oxidative stress. When ApoE -/- mice were reconstituted with bone marrow from various human APOE gene replacement mice, effective reduction of atherosclerosis was observed with marrow cells obtained from APOE3 but not APOE2 and APOE4 gene replacement mice. Taken together, these results documented that apoE2 and apoE4 expression in myeloid cells promotes inflammation via distinct mechanisms and promotes atherosclerosis in a plasma lipoprotein transport-independent manner., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The sheep ked (Melophagus ovinus) hematophagous insect may act as a potential vector of vector-borne pathogens. The aim of this study was to detect the presence of Trypanosoma spp., Bartonella spp., Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato in sheep ked collected from sheep in Poland. In total, Trypanosoma spp. was detected in 58.91% of M. ovinus, whereas Bartonella spp. and B. burgdorferi s.l. were found in 86.82% and 1.55% of the studied insects, respectively. A. phagocytophilum was not detected in the studied material. In turn, co-infection by Trypanosoma spp. and Bartonella spp. was detected in 50.39%, while co-infection with Trypanosoma spp. and Bartonella spp. and B. burgdorferi s.l. was found in 1.55% of the studied insects. The conducted study showed for the first time the presence of B. burgdorferi s. l. in M. ovinus, as well as for the first time in Poland the presence of Trypanosoma spp. and Bartonella spp. The obtained results suggest that these insects may be a potential vector for these pathogens, but further-more detailed studies are required., (© 2021. The Author(s).)

Insects of the genus Lipoptena are parasitic arthropods with a broad host range. Due to the type of parasitism (hematophagy), their potential role as vectors of pathogens, i.e., Bartonella sp., Anaplasma phagocytophilum , Rickettsia spp., and Borrelia burgdorferi is considered. As the range of their occurrence has been changing dynamically in recent years and infestations of humans have increasingly been reported, these organisms are now the subject of numerous studies. Our research aimed to present the molecular characteristics of Bartonella sp. detected in Lipoptena fortisetosa parasitizing wild cervids in south-eastern Poland. Adults of Lipoptena spp. were collected from carcasses of roe deer and red deer between spring and autumn in 2013. The PCR method was used to detect Bartonella sp. in the insects. We report two new haplotypes of the rpo B gene of Bartonella sp. isolated from L. fortisetosa feeding on wild cervids in south-eastern Poland and the presence of this invasive ectoparasitic species in the studied area since 2013. Phylogenetic analyses of newly obtained Bartonella sp. haplotypes confirmed their unique position on the constructed tree and network topology. The rpo B gene sequences found belonging to lineage B support the view that this phylogenetic lineage represents a novel Bartonella species.

The European roe deer (Capreolus capreolus) is the most common deer species in Europe. The species can be a reservoir of some tick-borne diseases but it is primarily recognized for its contribution as an amplifier host. In Central Europe, two roe deer ecotypes are living in adjacent areas: field and forest. We investigated differences in tick load and species composition on these two ecotypes. We collected ticks from 160 (80 the forest ecotype and 80 the field ecotype) roe deer culled in Wielkopolska Region (West-Central Poland). The most common was Ixodes ricinus (n = 1610; 99%) followed by Ixodes hexagonus (n = 22; 1%). The dominant life stage of the ticks was female. Prevalence was higher for forest roe deer. Mean number of ticks found on the forest ecotype was almost fivefold higher than on the field ecotype (3.75 ± 0.83 vs. 0.77 ± 0.20 ticks). The mean probability of tick occurrence was threefold higher in the forest (0.915 ± 0.050) than the field ecotype (0.279 ± 0.125). The most infested body parts of roe deer from both ecotypes were the neck and the head.

Ixodes ricinus (Acari: Ixodida) is the main vector in Europe of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti. Wolinski National Park (WNP) is situated by the Baltic Sea and is frequently visited by tourists. The aim of the study was to determine the potential risk of exposure to tick borne infection with B. burgdorferi s.l., A. phagocytophilum and B. microti on the areas of WNP. In total, 394 I. ricinus were tested. The pathogens in ticks were detected by PCR, nested PCR, RFLP and sequencing. Altogether, pathogens were detected in 12.69% of the studied ticks. B. burgdorferi s.l., was shown in 0.25% of the studied I. ricinus, while A. phagocytophilum and B. microti were detected in 1.01% and 10.65% of studied ticks, respectively. Co-infection by A. phagocytophilum and B. microti was shown in only one I. ricinus nymph. Analysis of B. burgdorferi s.l., genospecies showed that 0.25% of the studied ticks were infected with Borrelia garinii. The obtained results show the potentially high human risk of exposure to tick-borne infection with B. microti, and the low potential risk of infection with B. garinii and A. phagocytophilum on the studied areas of WNP.

Background: Schizophrenia is a mental disease that affects approximately 1% of the population. Despite over 100 years of research, its pathomechanism has still not been clarified. Cognitive deficits, which are one of the symptomatic dimensions of schizophrenia, usually appear a few years before the first psychotic episode. Therefore, this is why they are probably the clinical manifestation of the primary pathomechanism of schizophrenia. It is also supposed that N-methyl-D-aspartate receptor (NMDA-R) insufficiency in the prefrontal cortex is responsible for cognitive deficits in schizophrenia. The study aimed to examine whether four selected single nucleotide variants in GRIN1, GRIN2A and GRIN2B encoding NMDA-R subunits, of which two have not been tested before, are linked with the selected clinical phenotype of cognitive dysfunction in schizophrenia., Methods: The study included the targeted group of 117 patients diagnosed with schizophrenia, all with cognitive deficits and in symptomatic remission. DNA fragments including the studied polymorphisms of the NMDA receptors subunit genes were amplified by polymerase chain reaction and subjected to sequencing., Results: The study did not confirm the presence of any of the four selected single nucleotide variants in GRIN1, GRIN2A and GRIN2B subunits of NMDA-R., Conclusions: The finding indicates that selected single nucleotide variants in GRIN2A and GRIN2B encoding subunits of the NMDA receptor are not associated with the presence of cognitive deficits in schizophrenia.

Background: Schizophrenia is a common mental illness whose pathogenesis is still unknown. The vulnerability and stress model in schizophrenia assume that susceptibility to the disease is mainly associated with genes. Of the five symptomatic dimensions of schizophrenia, cognitive impairment appears to be most associated with the pathogenesis of schizophrenia. The aim of the study was to explore whether selected nucleotide variants in GRIN1, GRIN2A, and GRIN2B encoding subunits of the N-methyl-D-aspartate receptor (NMDA-R) receptor occur in a selected group of patients with treatment resistant schizophrenia with cognitive impairment., Methods: The study included 45 patients diagnosed with super refractory schizophrenia, all with cognitive deficits and chronically psychotic. DNA fragments including the studied polymorphisms of the NMDA receptors subunit genes were amplified by polymerase chain reaction and subjected to sequencing., Results: The study did not confirm the presence of any of the four selected single-nucleotide variants in GRIN1, GRIN2A, and GRIN2B subunits of NMDA-R in the study group., Conclusion: Results of the study indicated that the selected single-nucleotide variants are not associated both with resistance to clozapine and the presence of cognitive deficits in schizophrenia. It is possible, however, that a more extensive sequencing along with analyzing the expression of these genes may reveal different single-nucleotide variants than those assumed in the study.

Purpose: Fleas are insects with a high medical and veterinary importance. They may participate in spreading of many pathogenic agents, but still there is limited information about their possible reservoir or vector role for protozoans. The main aim of this study was an attempt of detection zoonotic pathogens, such as Babesia microti and Toxoplasma gondii in fleas Ctenocephalides felis felis and Ctenocephalides canis., Methods: In 2013-2017, 155 fleas were captured from domestic dogs and cats in veterinary clinics, animal shelters and pet grooming salons in Upper Silesia Region in Poland. Then, the DNA was extracted from each Ctenocephalides flea by using the ammonia method. Samples were screened for the presence of B. microti and T. gondii using PCR and nested PCR methods., Results: B. microti was reported in 6.6% of C. felis felis and 9.1% of C. canis, whereas the prevalence of coinfection with B. microti and T. gondii was 1.9% in cat fleas and 2.3% in dog fleas., Conclusion: This study shows the first cases of B. microti occurrence and B. microti and T. gondii coinfection in Ctenocephalides fleas. The estimation of prevalence of examined protozoans may be useful considering the possibility of infection among companion animals, as well as during presentation of the potential risk of infection in humans. In order to clarify the role of C. felis felis and C. canis in transmission of B. microti and T. gondii, the another studies with in vitro cultures and laboratory animals are needed.

The mountainous and foothill areas, in which the city of Zakopane, the capital of Tatra County, is located are characterized by continuous weather changes, lower air temperature, persistent snow cover, and poorer vegetation than in the lowlands. Ixodes ricinus and Ixodes hexagonus are vectors of tick-borne diseases and play an important role in the persistence of tick-borne diseases. The aim of the study was to determine the risk of exposure of domestic cats and dogs to the attacks of Ixodid ticks, to tick-borne infections with Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Babesia microti and Toxoplasma gondii in the city of Zakopane and the surrounding area. In 2017-2018 ticks were collected from a total of 10 domestic cats and 88 domestic dogs. Selected pathogens of tick-borne diseases were detected by PCR and nested PCR. The study material contained 119 I. ricinus and 36 I. hexagonus. The molecular examinations showed the presence of A. phagocytophilum in 3.8%, B. microti in 24.5% and T. gondii in 4.5% of the all ticks. In addition, in the study area, there is a high potential risk of tick-borne infection by B. microti, and a low potential risk of exposure to A. phagocytophilum and T. gondii infection.

Introduction: Ticks (Acari: Ixodida) are vectors and/or reservoirs of many pathogens, i.e. Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti . These pathogens are ethiological agents of such diseases as Lyme borreliosis, human granulocytic anaplasmosis and human babesiosis., Objective: The aim of the study was to evaluate the role of the Ixodes ricinu s in the transmission of Borrelia burgdorferi sensu lato, Borrelia afzelii, Borrelia garinii, Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum and Babesia microti in Opolskie Province in Poland., Material and Methods: DNA from 222 ticks was isolated by the ammonia method. The pair of primers specific to the flagelline gene was used to detect of B. burgdorferi s. l. To detect of genospecies of this spirochete, three pairs of internal primers were used. In turn, two pairs of primers specific to the 16S rDNA gene and the 18S rRNA were used, respectively, for the detection of A. phagocytophilum and B. microti. Borrelia burgdorferi s. l., A. phagocytophilum , and B. microti were detected in 4.5%, 2.7% and 5.4% of examined ticks, respectively., Results and Conclusions: Of the ten ticks infected with B. burgdorferi s. l., B. afzelii was found in seven, undefinied genospecies in two, and mixed infection with B. afzelii and B. burgdorferi s. s. in one. The study demonstrated the potential risk of exposure of humans and animals to infections of B. burgdorferi s. l., A. phagocytophilum and B. microti in the examined area of Poland.

Companion animals can be infested by various species of parasitic insects. Cat flea Ctenocephalides felis (C. felis felis) (Bouché, 1835) and dog flea Ctenocephalides canis (Curtis, 1826) belong to multihost external parasites of mammals, which most frequently occur on domestic cats Felis catus Linnaeus and dogs Canis familiaris Linnaeus. The main aim of this study was to investigate the presence of pathogens, such as Anaplasma phagocytophilum (syn. Ehrlichia phagocytophila) and Rickettsia spp., in adult C. felis and C. canis fleas. Flea sampling has been realised from January 2013 to April 2017 in veterinary clinics, animal shelters and pet grooming salons. Fleas were collected from domestic cats and dogs, directly from the pet skin or hair. Then, the DNA was isolated from a single flea by using the alkaline hydrolysis and samples were screened for the presence of pathogens using PCR method. Anaplasma phagocytophilum has occurred in 29% of examined C. felis and 16% of C. canis individuals. In turn, the prevalence of Rickettsia spp. in cat fleas population was only 3%, and the dog fleas 7%. The present study showed the presence of pathogenic agents in cat and dog fleas, which indicates the potential role of these insects in circulation of A. phagocytophilum and Rickettsia spp. in the natural habitat. Furthermore, exposition to these flea species, whose hosts are domestic cats and dogs, can pose a potential risk of infection for humans.

Essentials Tissue factor (TF) isoforms are expressed in pancreatic neuroendocrine tumors (pNET). TF knockdown inhibits proliferation of human pNET cells in vitro. mTOR kinase inhibitor sapanisertib/MLN0128 suppresses TF expression in human pNET cells. Sapanisertib suppresses TF expression and activity and reduces the growth of pNET tumors in vivo. SUMMARY: Background Full-length tissue factor (flTF) and alternatively spliced TF (asTF) contribute to growth and spread of pancreatic ductal adenocarcinoma. It is unknown, however, if flTF and/or asTF contribute to the pathobiology of pancreatic neuroendocrine tumors (pNETs). Objective To assess TF expression in pNETs and the effects of mTOR complex 1/2 (mTORC1/2) inhibition on pNET growth. Methods Human pNET specimens were immunostained for TF. Human pNET cell lines QGP1 and BON were evaluated for TF expression and responsiveness to mTOR inhibition. shRNA were used to knock down TF in BON. TF cofactor activity was assessed using a two-step FXa generation assay. TF promoter activity was assessed using transient transfection of human TF promoter-driven reporter constructs into cells. Mice bearing orthotopic BON tumors were treated with the mTORC1/2 ATP site competitive inhibitor sapanisertib/MLN0128 (3 mg kg -1 , oral gavage) for 34 days. Results Immunostaining of pNET tissue revealed flTF and asTF expression. BON and QGP1 expressed both TF isoforms, with BON exhibiting higher levels. shRNA directed against TF suppressed BON proliferation in vitro. Treatment of BON with sapanisertib inhibited mTOR signaling and suppressed TF levels. BON tumors grown in mice treated with sapanisertib had significantly less TF protein and cofactor activity, and were smaller compared with tumors grown in control mice. Conclusions TF isoforms are expressed in pNETs. Sapanisertib suppresses TF mRNA and protein expression as well as TF cofactor activity in vitro and in vivo. Thus, further studies are warranted to evaluate the clinical utility of TF-suppressing mTORC1/2 inhibitor sapanisertib in pNET management., (© 2018 International Society on Thrombosis and Haemostasis.)

Mite infestation of stored products is a serious threat to food safety and public health. These stored product mites are not only serious pests of stored food but also cause allergies in humans. Thirty serum samples from patients living in suburban areas of Upper Silesia (South Poland) were tested for sensitization to two species of storage mites: Lepidoglyphus destructor [LD] and Tyrophagus putrescentiae [TP]. Patient antibodies against particular antigens were identified using anti-human anti-IgE monoclonal antibodies. Fifteen protein fractions from LD gave positive reactions with IgE antibodies and 18 from TP. Seven of the 30 samples showed positive reactions to a protein fraction measuring about 29 kDa from LD and six reacted with a fraction measuring about 25 kDa from TP. These findings may imply the existence of many protein fractions with allergenic properties besides the characterized allergens in the two tested species.

The study was performed in the Trzciel Forest Districts which is located in the west part of Poland. The Scots pine is the main tree species creating forest landscape there. Dermacentor reticulatus, usually found in wet, boggy and damp habitat, in this case was discovered in fresh mixed coniferous forest and fresh coniferous forest. In Central Europe the Dermacentor reticulatus is after Ixodes ricinus the second most important vector for tick-borne diseases in Europe. The ticks were collected by flagging from lower vegetation during the autumn peak of their activity. All Dermacentor reticulatus were checked for presence of tickborne pathogens using PCR and nested PCR. In total 125 Dermacentor reticulatus ticks were collected. Among the pathogens examined, only Babesia microti has been found in 4% of them whereas Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum have not been found. In this research the presence of Babesia. microti in Dermacentor reticulatus has been proved. It should be stressed that it is a new focus for the Dermacentor reticulatus tick in Western Poland.

In 2008-2011 ticks were collected from southern Poland. Out of 6336 individuals collected and identified as Ixodes ricinus, 768 (2 larvae, 84 nymphs, 417 females, 265 males) were included in molecular study. The aim of this study was to investigate the prevalence and types of genospecies of Borrelia burgdorferi sensu lato in ticks. The polymerase chain reaction (PCR) was applied to detect the presence of pathogens in ticks. Subsequently the amplified DNA was digested with TasI enzyme. The infection rate was 15% (116) of examined ticks. PCR-RFLP analysis allowed distinguishing three genospecies of B. burgdorferi s.l.: B. burgdorferi sensu stricto, B. afzelii, and B. garinii. RFLP analyses of 116 positive samples revealed 96 (83%) monoinfections and 13 (11%) coinfections, whereas unidentified genospecies were present in 7 (6%) of positive samples. In the case of monoinfections, B. burgdorferi s.s. was the predominant species of pathogen in infected ticks - 61.4%. Other genospecies: B. garinii and B. afzelii were detected in 22.9% and 15.6% of the samples, respectively. To sum up, 15 % of ticks were infected by B. burgdorferi s.l which increases the risk of human infections in the recreational areas of southern Poland. Furthermore, there is a need to increase public awareness and implement more preventive measures concerning Lyme disease.

The article presents the results of the first study on seasonal activity of ticks and prevalence of tick-borne encephalitis virus (TBEV) in nymphs from the Silesian Province (Southern Poland). Previous studies on the prevalence of TBEV in ticks in Poland have been conducted mostly in northern and eastern regions, but none in the Silesian Province itself. The aims of this study were to analyse the seasonal variation in tick populations and compare TBEV prevalence in nymphs from different geographical locations in the Silesia. A total of 5160 questing Ixodes ricinus ticks were collected by the flagging method from 23 localities in southern Poland in 2010. Micro-climatic parameters (air temperature and humidity) were measured in order to estimate their influence on tick population. The highest tick activity was recorded in spring and was positively correlated with relative air humidity (RH). TBEV in the Silesian Province was analysed in 1750 nymphs and an overall prevalence was 0.11% (2 pools out of 175 analysed). The results of this study show that TBEV pool prevalence in nymphs is low in accordance with the low number of TBE cases reported within the region., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Introduction: Ticks (Acari: Ixodida) are vectors of many pathogens i. a: Babesia microti [Bm] and Toxoplasma gondii [Tg]. In Poland, Ixodes ricinus [Ir] is the main vector of both pathogens. This tick species and pathogens transmitted by them are a significant threat to human and animal health., Objectives of the Study: Detection of the protozoans Bm and Tg in ticks collected in the Tarnogórski district area., Material and Methods: The ticks were collected from vegetation and pets in the spring period of their activity on the territory of the Tarnogórski district. The parasites were preserved in 70% ethanol. DNA was isolated by ammonia method. Bm was detected by nested-PCR using specific primers for the 18S rRNA sequence. To detect the Tg a commercial kit was used. The PCR products were separated on 2% ethidium bromide stained agarose gels and visualised under UV light., Results: It was showed that all collected ticks belonged to the species Ir. Bm was detected in 50.87% and Tg in 64.91% of all examined ticks. Co-existence of these both protozoans in 36.84% of total examined ticks was noted., Conclusions: The study showed a high risk of exposure to Bm and Tg in the studied area. Ticks Ir play an important role in the transmission of Bm in this region. Demonstrating a high percentage of ticks collected from animals infected with Tg may indicate their important role in the transmission of this pathogen, but it requires a further studies.

Niepołomice Forest is located about 20 kilometers east of Cracow (Malopolska province, southern Poland). Its natural and touristic values, as well as wide range of hosts occurring within indicate this to be an area of high risk of exposure to Ixodes ricinus and tick-borne diseases it transfers. I. ricinus is a common species in Poland and Europe. Its seasonal activity begins in Poland in the early spring, and ends with late autumn. A total number of 129 specimens of I. ricinus was collected by flagging in Niepołomice Forest. DNA was isolated by ammonia method from 30 randomly-selected individuals. PCR was used to detect tick-borne pathogens with primers specific for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Babesia sp. Molecular studies confirmed the presence of all three pathogens in I. ricinus. A. phagocytophilum was found in 76.7%, Babesia sp., 60%, B. burgdorferi s. l., in 3.3% of studied ticks. A. phagocytophilum co-infection with Babesia sp., was found in 46.7% of the specimens. A co-infection of all three tested pathogens was recorded in one case (3.3%). In Poland the problem of tick-borne diseases is a growing issue, therefore people residing in southern Polish touristic areas should be informed about the prevention and protection against ticks.

Mitochondrial encephalomyopathies are complex disorders with wide range of clinical manifestations. Particularly time-consuming is the identification of mutations in mitochondrial DNA. A group of 20 children with clinical manifestations of mitochondrial encephalomyopathies was selected for molecular studies. The aims were (a) to identify mutations in mtDNA isolated from muscle and (b) to verify detected mutations in DNA isolated from blood, in order to assess the utility of a Surveyor nuclease assay kit for patient screening. The most common changes found were polymorphisms, including a few missense mutations altering the amino acid sequence of mitochondrial proteins. In two boys with MELAS (i.e., mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes), a mutation A→G3243 was detected in the tRNALeu gene of mtDNA isolated from muscle and blood. In one boy, the carrier status of his mother was confirmed, based on molecular analysis of DNA isolated from blood. A method using Surveyor nuclease allows systematic screening for small mutations in mtDNA, using as its source blood of the patients and asymptomatic carriers. The method still requires confirmation studying a larger group. In some patients, the use of this method should precede and might limit indications for traumatic muscle and skin biopsy., (Copyright © 2010 Elsevier Inc. All rights reserved.)