Trichinella spp. Infection in European Polecats (Mustela Putorius Linnaeus, 1758) from Romania.

The European polecat (Mustela putorius Linnaeus, 1758) is in decline in Romania, often living near human settlements, from mountains to lowlands. They feed on a wide variety of small animals, including rodents, such as mice or rats. The occurrence of this parasite in polecats from Romania was mentioned only once in 1991, but the parasite species was not confirmed by molecular biology. The study aimed to investigate the occurrence of Trichinella spp. in European polecats from Romania and to identify the parasite species by molecular tools. A total of 75 wild European polecats were examined by trichinoscopy and artificial digestion. A large number of animals were examined because of their wide distribution in Romanian territory and their presence near human settlements. For species determination, the positive muscle samples and the larvae recovered from artificial digestion were collected for DNA isolation and further processed by means of Multiplex PCR. Only two polecats from southern Romania tested positive for Trichinella spp. infection. During trichinoscopy examination, 48 (in a polecat from Giurgiu County) and 78 (in a polecat from Ialomița County) cysts were found in the tested (56 samples/animal) tissue samples. Artificial digestion revealed infection with 2466 larvae/100 g of muscle in the polecat from Ialomița and 254/100 g in the polecat from Giurgiu. The Multiplex PCR indicated the occurrence of Trichinella spiralis in the polecat from Giurgiu and a co-infection with T. spiralis and T. britovi in the polecat from Ialomița. The current study confirms through molecular biology, the occurrence of T. spiralis and T. britovi, as well as the occurrence of co-infection with these two Trichinella species in European polecats from Romania.

Risk of hepatitis B virus reactivation in hepatitis B virus + hepatitis C virus-co-infected patients with compensated liver cirrhosis treated with ombitasvir, paritaprevir/r + dasabuvir + ribavirin.

Hepatitis B virus may reactivate in patients with chronic hepatitis C treated with direct-acting antivirals. The aim of this study was to investigate the risk of hepatitis B virus (HBV) reactivation in HBV + hepatitis C virus (HCV)-co-infected patients with compensated liver cirrhosis treated with paritaprevir/ombitasvir/ritonavir, dasabuvir with ribavirin. We reviewed prospectively gathered data from a national cohort of 2070 hepatitis C virus patients with compensated liver cirrhosis who received reimbursed paritaprevir/ombitasvir/r, dasabuvir with ribavirin for 12 weeks from the Romanian National Health Agency during 2015-2016. Twenty-five patients in this cohort were HBs antigen positive (1.2%); 15 untreated with nucleotide analogues agreed to enter the study. These patients were followed up: ALT monthly, serology for HBV and DNA viral load at baseline, EOT and SVR at 12 weeks. Hepatitis B virus (HBV)-co-infected patients were all genotype 1b and 52% females, with a median age of 60 years (51 ÷ 74); 76% were pretreated with peginterferon + ribavirin; 72% were with severe necroinflammatory activity on FibroMax assessment; 40% presented comorbidities; and all were HBe antigen negative. Hepatitis C virus (HCV) SVR response rate was 100%. Hepatitis B virus (HBV)-DNA viral load was undetectable in 7/15 (47%) before therapy, and for the other 8 patients, it varied between below 20 and 867 IU/mL. Five patients (33%) presented virological reactivation (>2 log increase in HBV-DNA levels) during therapy. One patient presented with hepatitis associated with HBV reactivation, and two started anti-HBV therapy with entecavir. Hepatitis B virus (HBV) virological reactivation was present in 33% in our patients. Generally, HBV-DNA elevations were mild (<20 000 IU/mL); however, we report one case of hepatitis associated with HBV reactivation.

EMERGENCY SURGICAL TRACHEAL DECOMPRESSION IN A HUGE RETROSTERNAL GOITER.

INTRODUCTION: Over the past decades, several definitions and classifications of cervico-mediastinal goiters have been proposed. We analyzed and discussed the clinical presentation, the diagnostic procedures and the surgical technique in relation to post-operative complications and long-term results in a case of a sixty-six years old obese, hypertensive female admitted in the Thoracic Surgery Department with respiratory distress (inspiratory dyspnea, stridor) progressively aggravating during the latest month. METHODS: Cervico-thoracic CT scan revealed the existence of a cervico-mediastinal huge goiter which developed mostly intrathoracic (2/ 3 of the goiter). It determined a tracheal compression, reducing its caliber by two thirds, and its displacement to the right side. The proposed surgical procedure was total thyroidectomy and it involved a bipolar approach (transcervical and transsternal) through a partial upper cervico-sternotomy. RESULTS: The complete removal of the goiter and the decompression of the trachea have been achieved. Postoperative results were very satisfactory, with the absence of the respiratory distress. The histological examination revealed a multinodular goiter with epithelium hyperplasia. CONCLUSION: The presence of a complicated cervico-mediastinal goiter with severe respiratory distress required a surgical excision as the main and immediate treatment option. The surgical procedure represented a milestone for both the anesthesiologist (difficult intubation, with a thin tracheal tube in the absence of the jet ventilation technology) and for the surgeon. The goiter's excision from the visceral mediastinum was very difficult because of its huge dimensions and close relations with trachea and great vessels (anterior) and esophagus, erector spinal muscles and the spine (posterior).

16(th) IHIW: immunogenetics of aging.

Ageing is a process characterised by progressive loss of function in multiple different organ systems, such as the nervous, endocrine and immune systems. Current data showing that ageing processes may be associated with alterations in the immune system suggest that some of the genetic determinants of senescence might be polymorphic genes that regulate immune responses. The 'Immunogenetics of Aging' programme was a component introduced in the 14th International HLA and Immunogenetics Workshop (IHIWS) and developed further within the 15th and 16th. The aim of this component was to determine the contribution of immune genes to successful ageing and an increased capacity to reach the extreme limits of lifespan. Within the 16th IHIWS, new populations were included, and the number of samples analysed was increased. Analysis was focused on innate immunity genes (KIR and MBL2) and their correlation with CMV serostatus. Collaborative studies suggested that both activating and inhibitory KIR and functionally relevant MBL2 haplotypes are important factors for control of CMV infection in the elderly and therefore for chronic low-grade inflammation. Results showed that these genes might be predictive biomarkers in ageing and longevity. Prevalence of MBL2 haplotypes determining absence of the protein (LYPB, LYQC and HYPD) was observed in elderly people with a higher CMV antibody titre. The high CMV titre was also associated with a decreased frequency of the activatory KIR2DS5 and A1B10 haplotypes in elderly. Due to the role of KIR and low or deficient MBL haplotypes in viral infections, these genetic markers could be considered as indicators of a need for CMV prophylaxis at younger age and therefore increased probability of longer lifespan.

'Immunogenetics of Aging': report on the activities of the 15th International HLA and Immunogenetics Working Group and 15th International HLA and Immunogenetics Workshop.

'Immunogenetics of Aging' is a component that was first included in the 14th International HLA and Immunogenetics Workshop (IHIWS) and developed further within the 15th Workshop. The aim of this component was to assess the impact of human leukocyte antigen (HLA) genes, cytokine genes, and some innate immunity genes such as killer-cell immunoglobulin-like receptors (KIRs) and mannose-binding lectin 2 (MBL2) in successful aging and their contribution to the better understanding of immune dysfunction in old age. Within the 15th IHIWS new populations were included in the analysis. Additional cytokine gene polymorphisms were assessed and innate immunity genes were analyzed for possible relevance in longevity. The results showed that longevity might be associated with anti-inflammatory cytokine gene profiles, decreased frequency of interleukin-10 (IL-10) and transforming growth factor-B1 haplotypes associated with a low level of gene expression, and increased frequency of haplotypes determining a high level of expression. Extended tumor necrosis factor-A and IL-12B genotypes were also likely relevant to longevity. Data also showed that innate immunity genes are associated with susceptibility to infections in the elderly and showed that these genes might be an important genetic marker in aging. Decreased frequencies of KIR2DS5 and A1B10 haplotypes, and an increased proportion of MBL2-deficient haplotypes were found in the group with higher cytomegalovirus-specific IgG antibody levels. Together, these studies emphasize the relevance of genes regulating immune functions in maintaining human longevity and stress the importance of further clarifying their impact on successful aging.

Cytokine gene polymorphisms support diagnostic monitoring of Romanian multiple myeloma patients.

INTRODUCTION: Cytokines and their receptor genes are very polymorphic. SNPs in the promotor region of the gene may influence the rate of cytokine secretion and may affect the biological activity of the encoded cytokine. A number of cytokines and cytokine receptors have been directly linked to the development of human cancers. The aim of our study was to determine the cytokine gene polymorphism in Romanian multiple myeloma patients. MATERIAL AND METHODS: Cytokine genotyping was performed in 80 patients and 100 healthy blood donors using molecular biology methods (SSP-Invitrogen, USA). RESULTS: Analyzing each polymorphic site, there was an increased frequency of the following genotypes in patients compared to control group: Interleukin-1beta (IL-1ß) pos.+3962 TT, IL-12 pos.-1188 CC, gamma-Interferon (γ-IFN) pos.+874 AA, Transforming Growth Factor- beta1 (TGF- ß1) codon10 TT, IL-2 pos.-330 TG and pos.+166 TT, Interleukin-4Receptor alpha (IL-4Rα) pos.-33 TC, IL-10 pos.-1082 GG and pos.-592 CC, IL-6 pos.-174 GG. It should be noted that almost one third of multiple myeloma patients had IL-6 pos.-174 GG genotype and 62% IL-10 GCC haplotype. These identified haplotypes are high interleukins producer, and this fact was confirmed by serum IL-6 and IL-10 levels performed by ELISA and enhanced chemiluminiscence methods. CONCLUSION: These markers could be successfully used, together with other specific clinical and biological parameters, as reliable individualized prognostic factors in multiple myeloma patients.

Hand transplant--a challenge in immunological management of patients.

The concept of composite tissue allotransplantation (CTA) for restoration of congenital or acquired deformities is not new and the recent success of clinical composite tissue allotransplantation (CTA) attests to the fact that composite tissue allografts have tremendous potential in these life-enhancing reconstructions. A hand transplant, unlike a solid organ transplant, involves multiple tissues (skin, muscle, tendon, bone, cartilage, fat, nerves and blood vessels) and can be considered the 'gold standard' in CTA. In this regard, no other organ or tissue transplant matches the hand transplant in its immunogenicity as well as complexity. Development of assays that allow us to monitor the current state of an immune response (rejection/tolerance) is of great interest and requires an in-depth understanding of the complex and rare phenomenon of tolerance.

Conjugation to hyperbranched polyglycerols improves RGD-mediated inhibition of platelet function in vitro.

RGD (arginine-glycine-aspartic acid) is a known peptide sequence that binds platelet integrin GPIIbIIIa and disrupts platelet-fibrinogen binding and platelet cross-linking during thrombosis. RGD peptides are unsuitable for clinical applications due to their high 50% inhibitory concentration (IC50) and low in vivo residence times. We addressed these issues by conjugating RGD peptides to biocompatible macromolecular carriers: hyperbranched polyglycerols (HPG) via divinyl sulfone. The GPIIbIIIa binding activity of RGD was maintained after conjugation and the effectiveness of the HPG-RGD conjugate was dependent upon molecular weight and the number of RGD peptides attached to each HPG molecule. These polyvalent inhibitors of platelet aggregation decreased the IC50 of RGD in an inverse linear manner based on the number of RGD peptides per HPG. Since HPG-RGD conjugates do not cause platelet activation by degranulation and certain substitution ratios do not increase fibrinogen binding to resting platelets, HPG-RGD may serve as a model for a novel class of antithrombotics.

14th International HLA and Immunogenetics Workshop: report on the immunogenetics of aging.

The 'Immunogenetics of Aging' is a newly included component within the 14th International HLA and Immunogenetics Workshop. The aim of this component was to determine the contribution of human leukocyte antigen (HLA), cytokine genes and other major histocompatibility complex-encoded loci to successful aging and to determine an increased capacity to reach the extreme limits of life span. Two main data sets from four European populations were included in this study: unrelated healthy elderly individuals and ethnically matched young controls, and families with longevity members. Analysis was focused on HLA class I and II and cytokine gene polymorphisms. Preliminary results showed increased frequencies of DRB1*11- and DRB*16-associated haplotypes that were found to be protective for autoimmune diseases in some populations. Additionally, in families with longevity members, alleles and haplotypes positively associated with autoimmunity were not observed. Analysis of cytokine gene polymorphisms showed prevalence of anti-inflammatory profiles in healthy elderly individuals. Inheritance of extended haplotypes in families with longevity members allowed the identification of immunogenetic profiles that could be predictive for longevity. These preliminary studies indicate the relevance of genes regulating immune functions in human longevity and the importance of clarifying further their impact in successful aging.

Pycnogenol efficacy in the treatment of systemic lupus erythematosus patients.

A pilot study was performed to evaluate the efficacy of Pycnogenol treatment in systemic lupus erythematosus (SLE) patients. Eleven SLE patients were treated with first line medication according to disease activity and in addition, six of them received Pycnogenol and five a placebo. The SLE disease activity index (SLEDAI), serum anti-dsDNA antibodies, fibrinogen, C-reactive protein levels, erythrocyte sedimentation rate, production of reactive oxygen species (ROS) by neutrophils, spontaneous apoptosis and p56(lck) specific activity in peripheral blood lymphocytes were evaluated. Pycnogenol treatment determined a significant reduction of ROS production, apoptosis, p56(lck) specific activity and erythrocyte sedimentation rate. In addition, the decrease of SLEDAI was significant in the Pycnogenol treated group compared with the placebo group (p = 0.018). The results obtained suggest that Pycnogenol could be useful for second line therapy to reduce the inflammatory feature of SLE.

"Trial moves rapidly on, when the judge has determined the sentence beforehand" (W. Scott: Ivanhoe, 36) or pitfalls in serosurvey of anti-hepatitis C antibody in children.

Hepatitis C is and will be a major public health concern. Confirmed infections were reported from all Romanian counties but important differences between regions raise several explanations. Differences may reflect the different levels of testing, the performances of laboratories in confirming initially reactive samples or the risk factors higher prevalence. We have suggested that the prevalence of anti HCV infections can be a surrogate marker for the quality of parenteral medical or paramedical interventions. Present report identified additional problems in the surveillance of HCV infection in children. We screened 1787 samples from children hosted in orphanages (children under three years old) or in preschool children institutions (between 3-7 years old). We detected 31 repeatedly reactive samples with two EIA screening kits but confirmed only 8 in WB anti HCV. Four confirmed samples come from children under four months old suggesting maternally transmitted antibodies. In highly endemic area, many infants have maternally derived antibodies and the wane of reactivity comes with age above 12 months. Therefore, the prevalence of anti HCV antibody in infants reflects the prevalence in adult population. Confirmatory tests are mandatory for the serosurvey in children. More frequent than adults samples, children EIA reactive samples give indeterminate or negative Western Blot profiles. Only the viral load evaluation can confirm those samples as false positive or, on the contrary, samples at the beginning of seroconversion.

[Goodpasture's syndrome--a case report]. / Sindromul Goodpasture--prezentare de caz.

A female subject case aged 64 years is presented. The history shows several hospital admissions for lung diffuse infiltrative processes, accompanied with hemoptoic sputa. These infiltrations had a regressive evolution either spontaneously or under treatment. The diagnosis of Goodpasture syndrome was fixed post-mortem by alterations evidenced in the kidneys.

Growth factors secreted by the GH3 rat pituitary tumor cell line. Study of their autocrine mitogenic activity in cultured GH3 cells, A431 cells and human fibroblasts.

The pathogenesis of pituitary tumors is still controversial. Little is known about the effects of growth factors on pituitary growth, despite most of them having well-documented mitogenic actions in other tissues. We have investigated the secretion of growth factors by the rat pituitary tumor cell line, GH3 and also have studied their mitogenic effects in other two non-pituitary human cell lines, A431 and fibroblasts. Size-exclusion chromatography of acidic extracts of GH3 cells yielded two peaks of mitogenic activity when GH3 cells were used as potential targets. One peak of growth-promoting activity (> 5 KDa) stimulated [3H] thymidine incorporation into GH3 cells (201% above control). Another peak (2-3 KDa) also stimulated [3H] thymidine incorporation into GH3 cells (162% above control). The first peak of autocrine action represented 60% and the second one, 40% of the total peaks mitogenic activity. GH3 pooled fractions A, B, C, D, corresponding to the GH3 peaks of autocrine growth-stimulating activity significantly enhanced [3H] thymidine incorporation into A431 cells and human fibroblasts. In A431 cells, the first peak of mitogenic activity represented 46.7%, and the second one, 53.2% from the total peaks mitogenic activity. In fibroblasts, the GH3 pooled fractions produced just one peak of growth-stimulating activity, which increased [3H] thymidine incorporation (175% above control). Our data provide the indications that (1) cultured rat pituitary tumor cells GH3 secrete two autocrine growth factors which may have a role on their development and maintenance; (2) these factors, which have yet to be characterized, are potent mitogens for malignant non-pituitary cells such as human squamous carcinoma cell line, A431, as well as for human fibroblast cell line.

[Elastic osteosynthesis in intracapsular fracture of the femoral neck]. / L'ostéosynthèse élastique dans les fractures transcervicales du col du fémur.

The authors present a method of treatment for transcervical fractures of the femoral neck using implants with mechanical characteristics--curved shape, smooth surfaces and elastic resistance to deformity--which respect the biomechanical properties of the normal bone. These implants maintain a close contact between the bony fragments during convalescence and preserve the blood supply of the bone. In this way, bone healing is encouraged and the development of ischaemic necrosis of the femoral head is prevented. Between 1983 and 1985, 126 fractures, of the femoral neck, 120 recent and 6 old, were treated by internal fixation with two short elastic nails. One hundred and seven patients were followed up; fracture healing occurred in 94 cases (87.8 per cent) and 13 developed non-union. There were no necroses of the femoral head.

[Experimentation of apiarian preparations for the direct and the indirect capping of the dental pulp]. / Experimentarea unor preparate apicole în coafajul indirect si direct al pulpei dentare.

With a view to extend the range of biologically active preparations for the direct and the indirect capping of the dental pulp the authors have used a paste made from an alcoholic solution of propolis and zincoxyde. The study was carried on in 150 teeth with indirect capping of deep cavities, and 50 teeth with direct capping. The evolution of the cappings was followed clinically, radiologically and morphologically. The results obtained showed that the paste with propolis exerts effects similar to those of zinc eugenate. The morphologic study of the indirect capping showed that secondary dentin developed shortly after the application of the paste, and that it was followed by the development of pulpolites and the sclerous transformation of the pulp. In teeth with direct capping a protective film developed at the opening of the dental chamber. With time the pulpal wound undergoes cicatrization by a process of fibrosis and there is a trend to remineralization. No areas of pulpal degenerescence were found the rest of the pulpal tissue, and this suggests that the paste is more histophilic than the pastes based on calcium hydroxide, with which an area of necrosis occurred at the opening of the chamber, and calcium and fibrous degenerescence occurred in the coronal pulp.