The Prevalence of Intestinal Parasitic Infections Among Kosovar and Serbian School-children in Kosovo.

The aim of this study was to estimate the prevalence and distribution of species of intestinal parasites in the Balkans on the example of Kosovar and Serbian residents from the region of Gnjilane ineastern Kosovo,where the epidemiology of intestinal parasitic infections remains unknown. Parasitological examination of stools pecimens was performed in 2017 and 2018. Stool samples were collected from asymptomatic school-children aged 6-17 years: 530 Kosovars from the municipality of Kaçanik and310 Serbs from the municipalities of Kamenica and Strpce. Each patient provided two stool samples collected every second day,fixed in SAF preservative and 70 % spiritus vini, transported to the Department of Epidemiology and Tropical Medicine at the Military Institute of Medicine in Poland,and tested by light microscopy using three diagnostic methods: directs mearin Lugol's solution,decantation in distilled water, and Fülleborn's flotation. A total of 101 Kosovar children (19.1 % of the study group)were found to be infected with intestinal parasites: nematodes (n=20), cestodes (n=2), trematodes (n=2), and protozoa (n=79). Only 13 Serbian children (4.2 %) were found to beinfected with nematodes (n=4),cestodes (n=3),and protozoa (n=6). Giardia intestinalis was themost prevalent intestinal parasite in both groups (14,9 % vs. 1.9 % children). The prevalence ofasymptomatic parasitic infections was significantly higher in Kosovars in comparison to the Serbsliving in the same region of eastern Kosovo. This fact shows that there may be significant differencesin the quality of health care and sanitation as well as feed hygiene between these two communities.

Liver Transplantation for Incurable Alveolar Echinococcosis: An Analysis of Patients Hospitalized in Department of Tropical and Parasitic Diseases in Gdynia.

INTRODUCTION: Alveolar echinococcosis is a parasitic disease caused by the larval stage of tapeworm Echinococcus multilocularis. It usually involves the liver, but can spread to other organs. The treatment of choice is a surgical resection supported by antiparasitic drugs. In the advanced stages of the disease a liver transplantation is the only option. AIM: This article presents the problems related to care of patients after liver transplantation for advanced alveolar echinococcosis. MATERIAL: Sixty-seven patients with alveolar echinococcosis were hospitalized in our clinic in the years 2000-2015. Liver transplantation has been a therapeutic option for 9 patients. We retrospectively analyzed data of qualification for the liver transplantation and the postoperative treatment. RESULTS: Follow-up time after liver transplantation ranged from 7 months to 155 months (average, 6.4 years). One patient, with a history of advanced disease (P4N1M0), died due to liver failure. One patient was lost to follow-up. After liver transplantation all patients were receiving albendazole treatment. Two patients did not follow the medical recommendations. In 1 patient, who decided to stop therapy after 1 year, the relapse of alveolar echinococcosis in the left lobe of the transplanted liver passing through the diaphragm to the pericardium was detected. In another case we suspected a relapse of alveolar echinococcosis in transplanted liver due to positive serological tests. CONCLUSION: The prognosis of patient after liver transplantation for alveolar echinococcosis is good. The main problem caused by immunosuppressive therapy is a recurrence of disease in the transplanted liver.

Monoglyceride lipase deficiency causes desensitization of intestinal cannabinoid receptor type 1 and increased colonic µ-opioid receptor sensitivity.

BACKGROUND AND PURPOSE: Monoglyceride lipase (MGL) degrades 2-arachidonoyl glycerol (2-AG), an endogenous agonist of cannabinoid receptors (CB1/2 ). Because the CB1 receptor is involved in the control of gut function, we investigated the effects of pharmacological inhibition and genetic deletion of MGL on intestinal motility. Furthermore, we determined whether defective 2-AG degradation affects µ-opioid receptor (µ receptor) signalling, a parallel pathway regulating gut motility. EXPERIMENTAL APPROACH: Gut motility was investigated by monitoring Evans Blue transit and colonic bead propulsion in response to MGL inhibition and CB1 receptor or µ receptor stimulation. Ileal contractility was investigated by electrical field stimulation. CB1 receptor expression in ileum and colon was assessed by immunohistochemical analyses. KEY RESULTS: Pharmacological inhibition of MGL slowed down whole gut transit in a CB1 receptor-dependent manner. Conversely, genetic deletion of MGL did not affect gut transit despite increased 2-AG levels. Notably, MGL deficiency caused complete insensitivity to CB1 receptor agonist-mediated inhibition of whole gut transit and ileal contractility suggesting local desensitization of CB1 receptors. Accordingly, immunohistochemical analyses of myenteric ganglia of MGL-deficient mice revealed that CB1 receptors were trapped in endocytic vesicles. Finally, MGL-deficient mice displayed accelerated colonic propulsion and were hypersensitive to µ receptor agonist-mediated inhibition of colonic motility. This phenotype was reproduced by chronic pharmacological inhibition of MGL. CONCLUSION AND IMPLICATIONS: Constantly elevated 2-AG levels induce severe desensitization of intestinal CB1 receptors and increased sensitivity to µ receptor-mediated inhibition of colonic motility. These changes should be considered when cannabinoid-based drugs are used in the therapy of gastrointestinal diseases.

First finding of Echinococcus multilocularis DNA in soil: preliminary survey in Varmia-Masuria Province, northeast Poland.

The aim of this study was to estimate the presence of Echinococcus multilocularis DNA in the soil in rural areas of Varmia-Masuria Province, Poland, which is the region with the highest recorded number of human alveolar echinococcosis (AE) cases in this country. In total, 62 soil samples were analyzed. Samples were collected in forests near the lairs of foxes, as well as from arable fields, kitchen gardens and farmyards. The parasite DNA was detected in 11.3% of the samples. This study is the first environmental survey of the presence of E. multilocularis DNA in soil. The results demonstrate that the soil can be a direct source of human infections. These results should motivate the appropriate institutions to introduce informative and educational campaigns about the risk of infection, as well as developing methods for protecting the population, especially those living in at-risk areas.

Endothelialization of electrospun polycaprolactone (PCL) small caliber vascular grafts spun from different polymer blends.

Small caliber vascular grafts represent a challenge to material scientists. In contrast to large caliber grafts, prostheses with diameter <6 mm, lead to increased hemodynamic disturbances and thrombogenic complications. Thus, endothelialization of small caliber grafts should create a compatible interface for hemodynamic processes. The purpose of our study was to compare different compositions of electrospun scaffolds with conventional ePTFE grafts with an inner diameter of 4 mm as well as different pre-coatings to create an optimized physiological interface for endothelialization. Polycaprolactone, polylactide, and polyethylenglycol (PCL/PLA and PCL/PLA/PEG) electrospun grafts and ePTFE grafts were pre-coated with blood, gelatine or fibronectin and seeded with endothelial cells from the human term placenta. Best results were obtained with fibronectin-coated PCL/PLA/PEG grafts. Here, the number of attached viable cells was 78-81% higher than on fibronectin pre-treated ePTFE grafts. Cells attached to PCL/PLA/PEG grafts appeared in physiological cobblestone morphology. Viability analysis showed a high cell viability of more than 98%. Fibronectin-coated PCL/PLA/PEG grafts may be a promising improvement to conventionally used ePTFE grafts.

The first detection of Toxoplasma gondii DNA in environmental fruits and vegetables samples.

Toxoplasma gondii infections are prevalent in humans and animals all over the world. The aim of the study was to estimate the occurrence of T. gondii oocysts in fruits and vegetables and determine the genotype of the parasites. A total number of 216 fruits and vegetables samples were taken from shops and home gardens located in the area of northern Poland. Oocysts were recovered with the flocculation method. Then, real-time polymerase chain reaction (PCR) targeting the B1 gene was used for specific T. gondii detection and quantification. Toxoplasma DNA was found in 21 samples. Genotyping at the SAG2 locus showed SAG2 type I and SAG2 type II. This is the first investigation describing T. gondii DNA identification in a large number of fruits and vegetables samples with rapid molecular detection methods. The results showed that fruits and vegetables contaminated with T. gondii may play a role in the prevalence of toxoplasmosis in Poland.

Detection of Toxoplasma gondii oocysts in environmental soil samples using molecular methods.

Infections caused by Toxoplasma gondii are prevalent in humans and animals throughout the world. So far, there is no sufficient information concerning T. gondii oocysts prevalence in the environment, especially in soil. Therefore, the aim of this study was to estimate occurrence of T. gondii oocysts in soil and determine the genotype of detected parasites. A total of 101 soil samples were taken from different sites (sand-pits, "farming ground", areas around rubbish dumps) located in the Tri-City (Poland). Oocysts were recovered using the flotation method. Then, PCR reactions targeting the B1 gene were performed for specific T. gondii detection. The positive samples were further confirmed by PCR amplification of a repetitive element (REP) sequence [GenBank accession number AF146527]. Toxoplasma DNA was found in 18 samples. Among them, seven samples were successfully genotyped at the SAG2 locus. They were classified as SAG2 type I (5 samples) and SAG2 type II (2 samples). This is one of the first investigations describing T. gondii oocyst detection in environmental soil samples with rapid molecular detection methods and genotyping. The results of our findings showed that soil contaminated with T. gondii oocysts may play a role in the epidemiology of human toxoplasmosis in Poland.

Effects of recombinant LH treatment on folliculogenesis and responsiveness to FSH stimulation.

BACKGROUND: The role of LH in sensitizing antral follicles to FSH is unclear. LH is required for normal hormone production and normal oocyte and embryo development, but follicular responses to LH may depend upon the stage of development. Potential roles at the early follicular phase were explored in a clinical setting by employing a sequential approach to stimulation by recombinant human (r-h) LH followed by r-hFSH in women who were profoundly down-regulated by depo GnRH agonist. METHODS: We employed a multi-centre, prospective, randomized approach. Women (n = 146) were treated in a long course high-dose GnRH agonist (Decapeptyl, 4.2 mg s.c.) protocol and were randomized to receive r-hLH (Luveris, 300 IU/day) for a fixed 7 days, or no r-hLH treatment. This was followed by a standard r-hFSH stimulation regime (Gonal-F, 150 IU/day). Ultrasound and hormone assessments of responses were measured at the start of r-hLH treatment, on FSH stimulation Days 0 and 8 and at the time of HCG administration. RESULTS: The LH treatment was associated with increased small antral follicles prior to FSH stimulation (P = 0.007), and an increased yield of normally fertilized (2 PN) embryos (P = 0.03). There was no influence of the r-hLH pretreatment upon hormone profiles or ultrasound assessments during the FSH phase. Anti-mullerian hormone increased in both groups during the week prior to FSH stimulation (P = 0.002). CONCLUSIONS: This sequential approach to the use of r-hLH in standard IVF showed a possible modest clinical benefit. The results support other recent work exploring up-regulated androgen drive upon follicular metabolism indicating that clinical benefit may be obtainable after further practical explorations of the concept.

Routine use of r-hFSH follitropin alfa filled-by-mass for follicular development for IVF: a large multicentre observational study in the UK.

The technical advance of recombinant biotechnology to manufacture FSH has resulted in a biochemically pure FSH preparation with high batch-to-batch consistency. The protein content of recombinant human FSH (r-hFSH) can be reliably quantified in mass units by size exclusion high performance liquid chromatography (SE-HPLC) enabling r-hFSH follitropin alfa (Gonal-F) to be filled and released in the vial on the basis of mass with dose variability of only +/-2% [filled-by-mass (FbM)]. Observational studies have merit in addressing the effectiveness of a new product in routine clinical practice. This non-interventional study assessed r-hFSH follitropin alfa FbM for the multifollicular stimulation of patients undergoing assisted reproduction. A total of 1427 patients were recruited in 21 centres in the UK and the mean age was 34.3 years (+/-4.48, range 18-48). Of those who started treatment, 1388 (97.3%) received human chorionic gonadotrophin and 1330 (93.2%) underwent oocyte retrieval. A total of 1213 (85%) patients had embryo transfer and in the majority of recorded cases, most embryos replaced were graded as 1 or 2 (74.6%). The ongoing clinical pregnancy rate was 29.2% per cycle and 34.4% per embryo transfer. The routine use of r-hFSH follitropin alfa FbM in normal clinical practice in IVF has been demonstrated to be effective, safe and well received by patients.

Leukemia inhibitory factor in human reproduction.

OBJECTIVE: To describe the clinical findings, expressions, interactions, and clinical implications of leukemia inhibitory factor (LIF) in human reproduction. DESIGN: Review of published articles. SETTING: Clinical development unit of biotechnology company. INTERVENTION(S): None. RESULT(S): In the endometrium, LIF is expressed in a menstrual cycle-dependent manner, with the highest level occurring at the time of implantation. LIF is also detected in uterine flushing, and its level is significantly lower in women with unexplained infertility. Likewise, endometrial explants derived from women with unexplained infertility showed reduced levels of LIF secretion. Binding of LIF to LIF receptor and gp130 activates signal transduction pathways. LIF receptor is expressed in endometrium, oocytes, and blastocysts. Cytotrophoblasts cultured in the presence of LIF differentiate toward an anchoring extravillous phenotype. CONCLUSION(S): On the basis of reports gathered from animal and human studies, LIF appears to play an important role in implantation and in the establishment of pregnancy.

Protein tyrosine nitration in mouse peritoneal macrophages activated in vitro and in vivo: evidence against an essential role of peroxynitrite.

Tyrosine nitration is considered a key reaction of peroxynitrite-triggered tissue injury in inflammatory diseases. We investigated the potential involvement of peroxynitrite in protein tyrosine nitration in isolated murine peritoneal macrophages activated either in vitro with interferon-gamma/lipopolysaccharide or in vivo by priming mice with Corynebacterium parvum (10 mgxkg-1). Both protocols led to release of NO and accumulation of nitrite accompanied by formation of protein-bound 3-nitrotyrosine. Oxidation of dihydrorhodamine 123, a measure of peroxynitrite release, remained close to basal levels upon in vitro activation of the macrophages but was increased approximately twofold in vivo. Tyrosine nitration in macrophages activated in vitro was inhibited by catalase and the time course of nitration correlated with nitrite accumulation, whereas superoxide (O2*-) and H2O2 release occurred at much earlier times. To address the contribution of O2*- and peroxynitrite to in vivo nitration, a O2*- scavenger (MnTBAP; 1 mgxkg-1) was given to C. parvum-primed mice. MnTBAP led to almost complete inhibition of C. parvum-triggered O2*- and peroxynitrite release, whereas nitrite accumulation and formation of 3-nitrotyrosine were less affected ( approximately 50% of controls). These results argue against an essential role of peroxynitrite in protein tyrosine nitration in vivo.

Sperm banking and assisted reproduction treatment for couples following cancer treatment of the male partner.

In recent years, the survival of young males suffering from cancer has been improved. Development of new techniques such as IVF and intracytoplasmic sperm injection enables even low quality spermatozoa to be used successfully. It is possible therefore to preserve fertility potential of cancer patients before embarking on adjuvant chemotherapy and radiotherapy. Recognizing the importance of protecting the fertility potential of these young males, we present our recommendations for sperm cryopreservation based on the 11 year experience of Bourn Hall and the British Joint Council for Clinical Oncology consultation report. This paper discusses the options available for patients who recover from cancer to become fathers. In many cases patients are concerned about possible abnormalities and teratogenic risks to their future children who have been conceived naturally or by fertility treatment. The data available in the literature may reassure the medical community that there is no such increased risk. However, due to the relatively small number of children born after such treatment, a long-term follow-up is required. There is an ongoing debate regarding the justification for the programme due to the small number of patients who make use of their banked spermatozoa. The authors believe in the importance of protecting the fertility potential of cancer patients, enabling them to father their genetic children in the future while fighting their illness.

Protein tyrosine nitration in cytokine-activated murine macrophages. Involvement of a peroxidase/nitrite pathway rather than peroxynitrite.

Peroxynitrite, formed in a rapid reaction of nitric oxide (NO) and superoxide anion radical (O(2)), is thought to mediate protein tyrosine nitration in various inflammatory and infectious diseases. However, a recent in vitro study indicated that peroxynitrite exhibits poor nitrating efficiency at biologically relevant steady-state concentrations (Pfeiffer, S., Schmidt, K., and Mayer, B. (2000) J. Biol. Chem. 275, 6346-6352). To investigate the molecular mechanism of protein tyrosine nitration in intact cells, murine RAW 264.7 macrophages were activated with immunological stimuli, causing inducible NO synthase expression (interferon-gamma in combination with either lipopolysaccharide or zymosan A), followed by the determination of protein-bound 3-nitrotyrosine levels and release of potential triggers of nitration (NO, O(2)*, H(2)O(2), peroxynitrite, and nitrite). Levels of 3-nitrotyrosine started to increase at 16-18 h and exhibited a maximum at 20-24 h post-stimulation. Formation of O(2) was maximal at 1-5 h and decreased to base line 5 h after stimulation. Release of NO peaked at approximately 6 and approximately 9 h after stimulation with interferon-gamma/lipopolysaccharide and interferon-gamma/zymosan A, respectively, followed by a rapid decline to base line within the next 4 h. NO formation resulted in accumulation of nitrite, which leveled off at about 50 microm 15 h post-stimulation. Significant release of peroxynitrite was detectable only upon treatment of cytokine-activated cells with phorbol 12-myristate-13-acetate, which led to a 2.2-fold increase in dihydrorhodamine oxidation without significantly increasing the levels of 3-nitrotyrosine. Tyrosine nitration was inhibited by azide and catalase and mimicked by incubation of unstimulated cells with nitrite. Together with the striking discrepancy in the time course of NO/O(2) release versus 3-nitrotyrosine formation, these results suggest that protein tyrosine nitration in activated macrophages is caused by a nitrite-dependent peroxidase reaction rather than peroxynitrite.

Assessment of ovarian reserve - is there a role for ovarian biopsy?

The pool of primordial follicles in the ovary or 'ovarian reserve' is a major factor in the human fertility potential. The ageing ovary is characterized by reduction of the number of primordial follicles and this loss accelerates in the late 30's and precedes the menopause by 10-12 years. Woman's age alone or with a combination of biochemical markers, dynamic tests and ultrasound measurements fail to predict this loss accurately. In this manuscript, a novel approach of histopathological examination of ovarian biopsy for the evaluation of infertility, especially in unexplained infertility and in women in the later part of reproductive life, is discussed.

How do patients choose private in vitro fertilization treatment? A customer survey in a tertiary fertility center in the United Kingdom.

OBJECTIVE: To assess the relevant factors affecting patients' decision when choosing a clinic for private, self-funded IVF treatment. DESIGN: Prospective anonymous closed questionnaires. SETTING: Single tertiary-care private IVF center. PATIENT(S): New patients attending primary consultation in a period of 7 months from September 1999 to March 2000. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Primary knowledge about the clinic, source of referral, knowledge of, availability of information pack, and familiarity with the clinic's success rates and treatment's price. The last question assessed the relative importance of each factor in the decision-making process according to the SERVQUAL model, modified version. All items were measured as perceptions on a 5-point Likert scale. 1 = not important, 3 = neutral, and 5 = very important. RESULTS(S): One hundred seventy-five patients completed the questionnaire. They were well informed and investigated the market before making their final choice. Patients collect information from many sources, including new channels such as the Internet. Up to one third of patients do not consult their doctor before treatment. By far the most relevant factor in decision making is the success rate (i.e., delivery rates) of the clinic, followed by recommendation by general practitioner (GP) or consultant (respectively, Likert scale score, 4.6; 95% confidence interval [CI], 4.5-4.7; and Likert scale score, 4.0; 95% CI, 3.9-4.2; P<.01). Other factors, such as cost of the treatment, friends and relatives' opinions, and distance from home were not relevant. CONCLUSION(S): Units that wish to thrive and increase their market share should deliver high-quality service and concentrate their efforts on excelling in performance.

XX males without SRY gene and with infertility.

The case of a 28 year old male with normal male phenotype, in whom repeated seminal analysis showed complete azoospermia, is presented. Peripheral blood culture for chromosome studies revealed 46 chromosomes with XX constitution. Polymerase chain reaction (PCR) analysis of genomic DNA failed to detect the presence of the sex-determining region of the Y chromosome (SRY). A literature review of all SRY-negative XX males with normal male phenotype showed that this case is the sixth reported case but the first to be diagnosed during the investigations of infertility. The frequency, aetiology and diagnosis of this rare syndrome are also reviewed.

Effect of coenzyme Q(10) and alpha-tocopherol content of mitochondria on the production of superoxide anion radicals.

The effects of coenzyme Q(10) (CoQ(10)) and alpha-tocopherol on the rate of mitochondrial superoxide anion radical (O2(./-)) generation were examined in skeletal muscle, liver, and kidney of 24-month-old mice. Mice were orally administered alpha-tocopherol (200 mg.kg(-1).day(-1)) alone, CoQ(10) (123 mg.kg(-1).day(-1)) alone, or the two together for 13 wk. Administration of alpha-tocopherol resulted in an approximately sevenfold elevation of mitochondrial alpha-tocopherol content. Intake of CoQ(10) alone caused an approximately fivefold increase in CoQ content (CoQ(9) and/or CoQ(10)) and alpha-tocopherol of mitochondria. The rate of (O2(./-)) generation by submitochondrial particles (SMPs) was inversely related to their alpha-tocopherol content but unrelated to CoQ content. Experimental in vitro augmentation of SMPs with varying amounts of alpha-tocopherol caused an up to approximately 50% decrease in the rate of O2(./-) generation. Similar in vitro augmentations of SMPs with CoQ(10) had previously been found to have no effect on the rate of O2(./-) generation The CoQ(10)-induced elevation of alpha-tocopherol in the present study was inferred to be due to a 'sparing/regeneration' by CoQ. Results indicate the involvement of alpha-tocopherol in the elimination of mitochondrially generated O2(./-)