Health care access of thyroid disease patients in Serbia during the COVID-19 pandemic.

PURPOSE: The aim of this study was to assess the impact of repurposing health care facilities in response to COVID-19 on the access of patients with thyroid disease to health care. METHODS: This study consisted of a web-based survey. The survey was anonymous and consisted of forty questions. RESULTS: This survey included 206 respondents. 91.3% of the respondents had health insurance through the Republic Fund of Health Insurance, 9.7% had private or both health insurances, and 3.4% did not have any health insurance. A significant proportion of respondents (60.4%) had to switch from public to private health care to reach a physician and 73.8% had to switch from public to private laboratories. For the 91.9%, this was perceived as a financial burden. Before the pandemic, 83.1% of respondents reported regular follow-up by physicians, which decreased to 44.9% during the pandemic (p < 0.01). 76.3% of the respondents regarded that their thyroid disease was managed optimally before the pandemic, while this figure declined to only 48% during the pandemic (p < 0.01). CONCLUSIONS: The COVID-19 pandemic disrupted the medical care of thyroid patients in Serbia. For the patients treated in the public health care system, access to general practice was hindered, while access to specialist care was disrupted. It led to a switch from public to private health care, which was perceived as a financial burden for almost all the respondents. However, private health care proved to be an important safety net when the public system was overwhelmed.

Keratitis-ichthyosis-deafness Syndrome with Heterozygous p.D50N in the GJB2 Gene in Two Serbian Adult Patients.

Purpose: Keratitis-ichthyosis-deafness (KID) syndrome is a rare congenital ectodermal dysplastic syndrome presenting with keratitis, ichthyosis and sensorineural hearing loss. The most common causes of KID syndrome are heterozygous missense mutations in the GJB2 gene that codes for connexin 26. Case report: During the ophthalmological examination, two adult females complained of recent worsening of visual acuity in both eyes. Anamnesis revealed that their eyes were red and irritated from early childhood onwards. Both of them had thickening and keratinisation of eyelid margins, lash loss, diffuse opacification of cornea and conjunctiva caused by keratinisation of eye surface, superficial and deep corneal vascularisation and corneal oedema. Partial sensorineural hearing loss and difficulties in speech were also noted along with typical ichthyosiform erythroderma. Genetic testing of the GJB2 gene revealed a heterozygous p.D50N mutation in both patients.Patients were treated with a combined topical corticosteroid and artificial tears therapy, with steroid therapy being intensified during the last month. The therapy increased the visual acuity by decreasing corneal oedema and by forming a more regular air-tear interface during the six months follow up. Subsequently, the disease progressed despite the continuation of the therapy. Conclusion: This is the first report of Serbian patients with KID syndrome. Despite the administration of the combined topical corticosteroid and artificial tears therapy the disease is relentlessly progressive and therapeutic success of ophthalmological signs with local therapeutic modalities used so far had been disappointing.

[Deep lumbar back pain and neurological sensory deficits in a 56-year-old male patient]. / Tief lumbaler Rückenschmerz und sensible neurologische Ausfälle bei einem 56-jährigen Patienten.

Echinococcus infestations are rare in humans, infestation of bone occurs in less than 1% and a primary spinal infestation is extremely rare. This article reports on a clinical case of lumbar and sacral spinal infestation by Echinococcus multilocularis in a 56-year-old male Caucasian with neurological sensory deficits and deep lumbar back pain. Due to the suspicion of spondylodiscitis a computer tomography-guided biopsy was carried out without success, so that a sample was surgically obtained. The diagnosis of a spinal Echinococcus infestation could be made. A radical surgical débridement was carried out and anthelminthic treatment was started. This article describes this unusual case in detail and gives a brief summary of the current literature on this disease.

[Large multicystic space-occupying liver lesion in a 19-year-old woman]. / Große multizystische Leberraumforderung bei einer 19-jährigen Frau.

A 19-year-old woman presented with liver capsule pain and a liver lesion on sonography, which contained microvesicular cystic, necrotic and solid fibrotic formations typical for alveolar echinococcosis (AE). The diagnosis was confirmed by serology and histopathology. This parasitic infection which is endemic in Germany is feared because of its malignant growth. The increasing expansion of E. multilocularis in Europe will lead to a higher incidence of AE with an occurrence of cases outside classical endemic regions.

[The effects of cycloplegic eyedrops on corneal tomography]. / Effets des gouttes cycloplégiques sur la tomographie cornéenne.

PURPOSE: Whether cycloplegics affect standard keratorefractometric and tomographic measurements is unknown. The purpose of our study was to compare the effects of cycloplegics (cyclopentolate and atropine) on corneal shape and refractive power of the eye. METHODS: This study was performed on 84 eyes of 49 study participants. Patients were randomized into two groups: atropine 1% (32 eyes) and cyclopentolate 1% (52 eyes). Corneal tomography was performed with the Orbscan IIz. To evaluate the corneal shape, simulated keratometry values, anterior and posterior best-fit sphere, white-to-white and tangential and axial corneal power were performed for the anterior and posterior corneal surfaces before and during cycloplegia. Pupil diameter, anterior chamber depth, corneal thickness at the 3, 5 and 7mm optical zones, thinnest area of the cornea and corneal thickness at the visual axis were examined. Data were analyzed using an SPSS statistical package. RESULTS: The anterior and posterior BFS (in the atropine 1% group, anterior BFS was P=0.188; anterior BFS in the cyclopentolate group was P=0.227) and tangential and axial corneal power showed no change during cycloplegia in either group. SimK showed no statistical significance. The ACD was deeper when using atropine than cyclopentolate. Corneal thickness remained unchanged during cycloplegia in both groups. Pupil diameter was larger in light-colored irides in the cyclopentolate group than the atropine group. There was no change in W to W before (P=0.473) and during cycloplegia (P=0.287) in either group. CONCLUSIONS: Our results suggest that usage of atropine or cyclopentolate does not alter corneal shape.

Clinical scores for the prediction of esophageal varices in patients with liver cirrhosis.

INTRODUCTION: Esophageal bleeding is one of the most important and dramatic complications of liver cirrhosis in our everyday practice. Considering the costs of repeated upper endoscopy (UE) there is an increasing number of studies focusing on noninvasive para-meters for the assessment of esophageal varices (EV). PATIENTS AND METHODS: Retrospective study included 74 patients with alcoholic and viral liver cirrhosis treated at Clinic of Gastroenterology and Hepatology, Clinical Center of Serbia. The data were obtained from patients medical records including history, biochemical, ultrasonography and UE findings. RESULTS: The average value of the RLLD/INR for patients who showed evidence of EV during UE and in those who didn't was 10.46 ±â€ˆ3.09 and 12.24 ±â€ˆ3.43, respectively (p = 0.019, p < 0.05). Cutoff value (11.5) of RLLD/INR showed a sensitivity of 64.15% and specificity of 66.67% (1.92LR+, and 0.54 LR-, AUROC 0.639) for the detection of EV. The average value of PC/SBD for patients who showed evidence of EV during UE and in those who didn't was 619.79 ±â€ˆ492.96 and 1423.1 ±â€ˆ908.2, respectively (p = 0.0, p < 0.05). The average value of RLLD/SA was 5.5 ±â€ˆ0.17 and 4.57 ±â€ˆ0.17 (p = 0.015, p < 0.05) for patients who showed evidence of EV -during UE and in those who didn't, respectively. CONCLUSION: Noninvasive assessment of EV using scores based on ultrasonography and laboratory is simple, inexpensive, and could be a useful tool in limiting the number of repeated UE.

Gut-associated lymphoid tissue, gut microbes and susceptibility to experimental autoimmune encephalomyelitis.

Gut microbiota and gut-associated lymphoid tissue have been increasingly appreciated as important players in pathogenesis of various autoimmune diseases, including multiple sclerosis. Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis that can be induced with an injection of spinal cord homogenate emulsified in complete Freund's adjuvant in Dark Agouti (DA) rats, but not in Albino Oxford (AO) rats. In this study, mesenteric lymph nodes (MLN), Peyer's patches (PP) and gut microbiota were analysed in these two rat strains. There was higher proportion of CD4(+) T cells and regulatory T cells in non-immunised DA rats in comparison to AO rats. Also, DA rat MLN and PP cells were higher producers of pro-inflammatory cytokines interferon-γ and interleukin-17. Finally, microbial analyses showed that uncultivated species of Turicibacter and Atopostipes genus were exclusively present in AO rats, in faeces and intestinal tissue, respectively. Thus, it is clear that in comparison of an EAE-susceptible with an EAE-resistant strain of rats, various discrepancies at the level of gut associated lymphoid tissue, as well as at the level of gut microbiota can be observed. Future studies should determine if the differences have functional significance for EAE pathogenesis.

How to not miss alveolar echinococcosis in hepatic lesions suspicious for cholangiocellular carcinoma.

PURPOSE: Hepatic alveolar echinococcosis (AE) resembles intrahepatic cholangiocarcinoma (ICC) on radiological imaging. The purpose of this study was to identify criteria to discriminate AE from ICC with CT and MR Imaging. METHODS: One hundred and sixteen imaging studies of 94 patients (CT n = 65; MRI n = 51) diagnosed with AE (n = 55) or ICC (n = 39) were retrospectively reviewed by two blinded radiologists for lesion features including enhancement pattern and matrix composition. A consensus read was conducted in cases of disagreement. Uni- and multivariate logistic regression with bootstrapping were used for analysis. RESULTS: Using CT, no or septal enhancement and calcification yielded the highest values of sensitivity/specificity (90.9%/90.6% and 81.8%/96.9%) for AE. Using MRI, no or septal enhancement and cystic components achieved the highest sensitivity/specificity (90.9%/100.0% and 84.8%/66.7%) for AE. Multivariate logistic regression identified the following strong independent predictors for AE: for MRI, no or septal enhancement (odds ratio [OR] 322.4; p < 0.001); for CT, no or septal enhancement and calcification (OR 35.9 and 42.5; p < 0.001 and p < 0.01, respectively). No or septal enhancement and calcification demonstrated the highest interreader agreement (>90%). CONCLUSION: Enhancement characteristics and matrix calcifications offer the strongest discriminating potential between AE and ICC with a high sensitivity, specificity, and interreader agreement.

Pitfalls in diagnosis and treatment of alveolar echinococcosis: a sentinel case series.

BACKGROUND: Alveolar echinococcosis (AE) is a neglected zoonosis presenting with focal liver lesions (FLL) with a wide range of imaging patterns resembling benign as well as malignant FLLs. Complementary serology and histopathology may be misleading. OBJECTIVE: The objective of our study is to highlight pitfalls leading to wrong diagnoses and harmful interventions in patients with AE. DESIGN: This retrospective sentinel case series analyses diagnostic and treatment data of patients with confirmed AE. RESULTS: 80 patients treated between 1999 and 2014 were included in the study. In 26/80 patients treatment decisions were based on a wrong diagnosis. AE was mistaken for cystic echinococcosis (CE) in 12/26 patients followed by cholangiocellular carcinoma (CCA) in 5/26 patients; 61/80 patients had predominantly infiltrative liver lesions and 19/80 patients had a predominantly pseudocystic radiological presentation. Serology correctly differentiated between Echinococcus multilocularis and Echinococcus granulosus in 53/80 patients. Histopathology reports attributed the right Echinococcus species in 25/58 patients but failed to differentiate E. multilocularis from E. granulosus in 25/58 patients. Although contraindicated in AE 8/25 patients treated surgically had instillation of a protoscolicidal agent intraoperatively. One of the eight patients developed toxic cholangitis and liver failure and died 1 year after liver transplantation. CONCLUSIONS: Misclassification of AE leads to a critical delay in growth inhibiting benzimidazole treatment, surgical overtreatment and bares the risk of liver failure if protoscolicidal agents are instilled in AE pseudocysts.

Autoimmunity and thyrotropin level in developing thyroid malignancy.

OBJECTIVE: Malignancies and autoimmune thyroid disease are still controversial, but recent studies prove that a long lasting thyroid disease may be linked with malignancy, e.g. papillary thyroid carcinoma in patients with Hashimoto thyroiditis. Having in mind that thyrotropin is a thyroid growth factor, the relationship between its serum values, as well as the levels of anti-peroxidase and anti-thyroglobulin antibodies and thyroid malignancy in patients with nodular thyroid goiter was examined. PATIENTS AND METHODS: Six-hundred-thirty-seven medical records, which included the thyroid fine-needle aspiration cytology were retrospectively evaluated. Patients were grouped regarding the levels of thyrotropin, anti-peroxidase and anti-thyroglobulin antibodies (in or out of the reference ranges) and compared with cytology findings for establishing their prognostic potential for malignancy. RESULTS: Elevated serum thyrotropin (≥ 4.5 mIU/L) was found in 27.3% of patients with thyroid malignancy compared with 10.8% with benign and 16.1% with unspecified cytology finding (p < 0.01). In the group of patients with malignant cytology findings 7.0% of them had elevated anti-peroxidase antibodies level, and 1.4% had anti-peroxidase antibodies level in reference range. In the group of patients with malignant cytology findings 4.2% of them had elevated anti-thyroglobulin antibodies level, and 1.4% had anti-thyroglobulin antibodies level in reference range. CONCLUSIONS: In patients with elevated serum thyrotropin concentration and/or chronic thyroiditis the occurrence of thyroid malignancy is increased.

Thyroid dysfunction and thyroid autoimmunity in euthyroid women in achieving fertility.

OBJECTIVE: Thyroid disease is the second most common endocrine condition in women of childbearing age. Thyroid hormones are involved in control of menstrual cycle and in achieving fertility affecting the actions of follicle-stimulating hormone and luteinizing hormone on steroid biosynthesis by specific triiodothyronine sites on oocytes; therefore, affect all aspects of reproduction. It remains controversial if pregnant women should be screened for thyroid dysfunction. Purpose of this review was to examine recent studies on the assessment of thyroid dysfunction in pregnancy, its treatment and newly perspective of thyroid autoimmunity in pregnant euthyroid women in achieving fertility. METHODS: An electronic search was conducted using the internet medical databases: Medline/PubMed, EMBASE, EBSCO, and the Cochrane library. RESULTS: Thyroid gland faces great challenge in pregnancy when many hormonal changes occur. Precondition for normal follicular development and ovulation is pulsate gonadothropin realizing hormone secretion. Thyroid dysfunction in pregnancy is classified as forms of hypothyroidism (positivity of thyroid autoantibody, isolated hypothyroidism, and subclinical or overt hypothyroidism), hyperthyroidism, and autoimmune disease, but also thyroid nodules and cancer, iodine insufficiency and postpartum thyroiditis. These conditions can cause adverse effects on mother and fetus including pregnancy loss, gestational hypertension, or pre-eclampsia, pre-term delivery, low birth weight, placental abruption and postpartum hemorrhage. There is an evidence that thyroid autoimmunity, in thyroid dysfunction adversely affects conception and pregnancy outcomes, but it is unclear what impact has isolated eumetabolic thyroid autoimmunity in achieving fertility, especially in women undergoing in vitro fertilization. Treatment of euthyroid pregnant women with positive thyroid peroxides antibodies is still controverse, but not few studies show that levothyroxine substitution is able to lower the chance of miscarriage and premature delivery. CONCLUSIONS: Further randomized trials are needed to expand our knowledge of physiologic changes in thyroid function during the pregnancy and to reveal mechanisms by which thyroid autoimmunity in euthyroid women affect fertility, especially the success of assisted reproductive technology in achieving the same and validity of levothyroxine administration in thyroid autoimmunity positive women.

Activity, but not mRNA expression of gelatinases correlates with susceptibility to experimental autoimmune encephalomyelitis.

Experimental autoimmune encephalomyelitis (EAE) is a model of multiple sclerosis (MS), inflammatory, demyelinating and neurodegenerative disease of the central nervous system (CNS). Clinically manifested EAE can be induced in Dark Agouti (DA) rats, but not in Albino Oxford (AO) rats by immunization with spinal cord homogenate (SCH) and complete Freund's adjuvant (CFA). Matrix metalloproteinases (MMP) play important roles in various steps of MS and EAE pathogenesis. Expression of gelatinases MMP2 and MMP9, their activator MMP14 and their inhibitor tissue inhibitor of MMP (TIMP)1 in the CNS of AO and DA rats immunized with SCH+CFA was determined. Expression of mRNA for MMP2, MMP9 and MMP14 was higher and expression of TIMP1 mRNA was lower in AO rats. However, gelatinase activity in spinal cords was higher in samples obtained from DA rats. Further, while there was no strain difference in MMP2 and MMP9 mRNA expression in lymph nodes of the immunized rats, gelatinase activity was higher in DA rats. This activity was reduced by antiinflammatory cytokines interleukin (IL)-10 and IL-4. Interestingly, gelatinase activity was detected in the nuclei of cells within the CNS, but not of those in lymph nodes. Our results imply that posttranscriptional regulation of MMP2 and MMP9 expression and/or function determines low gelatinase activity within the CNS and in immune cells of EAE-resistant AO rats.

A novel human glycoprotein ACA is an upstream regulator of human hematopoiesis.

A central issue in stem cell biology is a better understanding of the molecular mechanisms that regulate self-renewal of human hematopoietic stem cells (HSCs). Control of the specific function of HSCs like self-renewal and differentiation might be regulated by a common set of critical genes. However, the regulation among these genes is yet to be elucidated. Here, we show that activation by a novel human GPI-linked glycoprotein ACA at the surface of human peripheral blood progenitor cells induces via PI3K/Akt/mTor/PTEN upregulation of WNT, Notch1, Bmi-1 and HoxB4 genes thus, promoting self-renewal and generation of primitive HSCs. ACA-generated self-renewing cells retained their lympho-myeloid repopulating potential in NOD/SCID mouse xeno-transplantation model with long term functional capacity. We conclude that ACA is an essential regulator of the genes involved in maintaining hematopoiesis and its use in clinical praxis could overcome many of the barriers present so far in transplantation medicine.

Activation by ACA induces pluripotency in human blood progenitor cells.

Reprogramming of human somatic cells by transcription factors to pluripotent state holds great promise for regenerative medicine. However, low efficiencies of current reprogramming methods, immunogenicity and lack of understanding regarding the molecular mechanisms responsible for their generation, limits their utilization and raises questions regarding safety for therapeutic application. Here we report that ACA signaling via PI3K/Akt/mTor induces sustained de-differentiation of human blood progenitor cells leading to generation of ACA pluripotent stem cells. Blood-derived pluripotent stem cells differentiate in vitro into cell types of all three germ layers, exhibiting neuronal, liver, or endothelial characteristics. Our results reveal insight into the molecular events regulating cellular reprogramming and also indicate that pluripotency might be controlled in vivo through binding of a natural ligand(s) to ACA receptor enabling reprogramming through defined pathway(s) and providing a safe and efficient method for generation of pluripotent stem cells which could be a breakthrough in human therapeutics.

A human iPSC model of Ligase IV deficiency reveals an important role for NHEJ-mediated-DSB repair in the survival and genomic stability of induced pluripotent stem cells and emerging haematopoietic progenitors.

DNA double strand breaks (DSBs) are the most common form of DNA damage and are repaired by non-homologous-end-joining (NHEJ) or homologous recombination (HR). Several protein components function in NHEJ, and of these, DNA Ligase IV is essential for performing the final 'end-joining' step. Mutations in DNA Ligase IV result in LIG4 syndrome, which is characterised by growth defects, microcephaly, reduced number of blood cells, increased predisposition to leukaemia and variable degrees of immunodeficiency. In this manuscript, we report the creation of a human induced pluripotent stem cell (iPSC) model of LIG4 deficiency, which accurately replicates the DSB repair phenotype of LIG4 patients. Our findings demonstrate that impairment of NHEJ-mediated-DSB repair in human iPSC results in accumulation of DSBs and enhanced apoptosis, thus providing new insights into likely mechanisms used by pluripotent stem cells to maintain their genomic integrity. Defects in NHEJ-mediated-DSB repair also led to a significant decrease in reprogramming efficiency of human cells and accumulation of chromosomal abnormalities, suggesting a key role for NHEJ in somatic cell reprogramming and providing insights for future cell based therapies for applications of LIG4-iPSCs. Although haematopoietic specification of LIG4-iPSC is not affected per se, the emerging haematopoietic progenitors show a high accumulation of DSBs and enhanced apoptosis, resulting in reduced numbers of mature haematopoietic cells. Together our findings provide new insights into the role of NHEJ-mediated-DSB repair in the survival and differentiation of progenitor cells, which likely underlies the developmental abnormalities observed in many DNA damage disorders. In addition, our findings are important for understanding how genomic instability arises in pluripotent stem cells and for defining appropriate culture conditions that restrict DNA damage and result in ex vivo expansion of stem cells with intact genomes.

A 7-year prospective study of routine histopathological evaluation of the lacrimal sac wall incisional biopsy specimens obtained during external dacryocystorhinostomy in adults and a review of the literature.

The most common lacrimal sac pathology is chronic inflammation with or without occlusive fibrosis. However, a substantial number of lacrimal sac-specific pathologies were reported throughout the literature which may mimic chronic inflammation and be misdiagnosed. From a tertiary ophthalmic care centre in Serbia, in a single ophthalmic pathology laboratory, during a 7-year period (January 2004 to October 2010), a 599 consecutive lacrimal sac wall biopsy samples routinely obtained during external dacryocystorhinostomy in adult patients with clinically presumed primary acquired lacrimal drainage system obstruction were analysed. Although non-specific lacrimal sac pathology was present in the vast majority of cases (578 biopsy specimens; 96.49%), this report also reveals a relatively substantial number (21 biopsy specimens; 3.51%) of clinically non-suspected or intraoperatively unexpected primary lacrimal sac-specific pathology--among them, six lesions with malignant biological behaviour were identified: one microinvasive squamous cell carcinoma and five malignant lymhoproliferative lesions. Usefulness of routine lacrimal sac wall biopsy during surgery for primary acquired lacrimal drainage system obstruction is undoubtful and commensurate with the constant need for better understanding of the pathological processes that involve lacrimal drainage system.

Explicit review of risperidone long-acting injection prescribing practice.

WHAT IS KNOWN AND OBJECTIVE: Long-acting injectable (LAI) antipsychotics are recommended for those people with a preference for this form of treatment and those who experience negative outcomes due to non-adherence with oral medication. LAI antipsychotics have been associated with improved outcomes and lower treatment discontinuation rates when compared with oral formulations. Risperidone long-acting injection (RLAI) treatment is effective and well-tolerated in clinical trials. The aim of this study was to review RLAI prescribing practice and compare prescribing to best practice recommendations (including indication, initiation, dose and co-prescribing) for adults receiving care from five clinical practice settings of New Zealand. METHODS: Patients starting publicly funded RLAI between 1 October 2005 and 31 October 2006 in five mental health services were included in the study. Data were retrospectively collected for 443 patients 1 year pre- and post-RLAI initiation at seven cross-sectional time-points (12, 6 and 3 months before; initiation; and 3, 6 and 12 months after). Patient characteristics (gender, age, ethnicity), DSM-IV-TR diagnosis, duration of mental illness, mental health act utilization, treatment setting and antipsychotic treatment (reasons for starting RLAI) were obtained from patient records. RESULTS AND DISCUSSION: The patients were mostly male (64,3%), of European background (42.9%) with a medium age of 34. In line with treatment recommendations, most had a diagnosis of schizophrenia or related psychoses, a history of medication adherence problems and previously been prescribed oral risperidone (72%). Treatment initiation also reflected recommended guidance; most were started on 25 mg/2 weeks (81.9%) and had treatment crossover (93.3%) until RLAI stabilized. For 58.3% of the group who continued for ≥ 12 months, mean fortnightly doses increased from 36.2 mg (3 months) to 41.3 mg (12 months); within the licensed range of 25-50 mg/2 weeks. Areas differing from recommended practice included high rates of antipsychotic co-prescribing at three cross-sectional time-points and ongoing at 12 months (12.3%). Patients prescribed higher RLAI starting doses were more likely to be prescribed higher doses 12 months later. WHAT IS NEW AND CONCLUSION: To our knowledge this is the largest multi-site explicit review of RLAI use in real world clinical practice. The review found that clinicians were using RLAI in clinical practice predominantly in accordance with best practice recommendations. However, high rates of antipsychotic co-prescribing with RLAI were identified which differ from practice reported in other small reviews of RLAI use and local studies of antipsychotic prescribing. We have demonstrated that clinical audit of practice is a powerful tool to identify areas of potentially poor practice, such as ongoing high rates of antipsychotic co-prescription cross-sectionally and 12 months after RLAI initiation and that this is an area of practice requiring further evaluation. Feedback to clinicians and stakeholders followed by re-audit of practice is needed in order to complete the audit cycle.

Mechanisms of modulation of experimental autoimmune encephalomyelitis by chronic Trichinella spiralis infection in Dark Agouti rats.

Trichinella spiralis is a helminth that provokes Th2 and anti-inflammatory type responses in an infected host. Our previous studies using Dark Agouti (DA) rats indicated that T. spiralis infection reduced experimental autoimmune encephalomyelitis (EAE) severity in rats. The aim of this study was to analyse the mechanisms underlying EAE suppression driven by T. spiralis infection. Reduced clinical and histological manifestations of the disease were accompanied by increased IL-4 and IL-10 production and decreased IFN-gamma and IL-17 production in draining lymph node cells. This indicates that T. spiralis infection successfully maintains a Th2 cytokine bias regardless of EAE induction. High IL-10 signifies parasite-induced anti-inflammatory and/or regulatory cell responses. Transfer of splenic T cell-enriched population of cells from T. spiralis-infected rats into EAE immunized rats caused amelioration of EAE and in some cases protection from disease development. This population of cells contained higher proportion of CD4(+) CD25(+) Foxp3(+) regulatory cells and produced high level of IL-10 when compared with uninfected rats.

Effects of different oocyte activation procedures on development and gene expression of porcine pre-implantation embryos.

Among the factors that affect the efficiency of somatic cell nuclear transfer (SCNT) in pigs, the activation protocol is the most variable among the current SCNT procedures. The aim of this study is focused on defining an efficient activation treatment of porcine oocytes. In Experiment 1, we studied the effects of nine different oocyte activation procedures (including chemical- and electrical-based treatments) on parthenogenetic embryo development. In Experiment 2, we studied the effect of the more efficient activation procedures on the gene expression profile of Oct4 and Igf2r in parthenogenetic blastocysts. In conclusion, ionomycin as a first calcium stimulus is not able to activate porcine oocytes efficiently in comparison with electric procedures. Electrical treatments with 6-DMAP significantly increased the level of Oct4 expression, whereas the single and double pulse treatments alone maintained the same profile as the IVF group.