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Synapse-Assembly Proteins Preserve Synaptic Vesicle Chaos Balance and Get a grip on Synaptic Vesicle Transfer within Caenorhabditis elegans.
The infection of triatomines with trypanosomes can be performed with different forms of the parasite, and the procedure is important not only for vector-parasite interaction studies but also for maintaining the infectivity of parasite strains, which guarantees more realistic biological and molecular investigations. Here, I describe how to infect the vector Rhodnius prolixus, a model species, with two different species of Trypanosoma.African trypanosomes are naturally transmitted by bloodsucking tsetse flies in sub-Saharan Africa and these transmission cycles can be reproduced in the laboratory if clean tsetse flies and suitable trypanosomes are available for experiments. Tsetse transmission gives access to more trypanosome developmental stages than are available from in vitro culture, albeit in very small numbers; for example, the sexual stages of Trypanosoma brucei have been isolated from infected tsetse salivary glands, but have not yet been reported from culture. Tsetse transmission also allows for the natural transition between different developmental stages to be studied.Both wild-type and genetically modified trypanosomes have been successfully fly transmitted, and it is possible to manipulate the trypanosome environment inside the fly to some extent, for example, the induction of expression of genes controlled by the Tet repressor by feeding flies with tetracycline.In the 1990s my laboratory discovered that Leishmania promastigotes can combine two environmental cues, typical to lysosomes, acidic pH (~5.5) and body temperature (37 °C) into a single signal that induced differentiation. Based on this concept, we modified EARLS-based medium 199 to become an amastigote-specific medium. Shifting promastigotes to this medium followed by incubation in a CO2 incubator induced differentiation. Axenic amastigotes reach maturation within 5 days, resembling the time it takes in vivo. This chapter provides a complete protocol we developed for L. donovani promastigote-to-amastigote differentiation. This protocol should be useful for both old-world and new-world species of Leishmania.Cultivation of pleomorphic Trypanosoma brucei strains was introduced in 1996 when matrix dependence of growth of natural isolates was recognized. Semisolid agarose or liquid methylcellulose are currently used and here we provide optimized protocols for these culture methods and for transfection of pleomorphic strains. Although more laborious than standard liquid culture, culture of native pleomorphic strains is important for a number of research questions including differentiation, virulence, tissue tropism, and regulated metabolism. Some subclones of pleomorphic strains have acquired matrix independence upon passage in culture but maintained a pleomorphic phenotype. It appears that matrix dependence and pleomorphism are not tightly linked traits, yet phenotypes have to be verified before choosing one of these subclones for given experiments. Based on direct comparisons, we give recommendations for pleomorphic strain selection and culture conditions that guarantee truly pleomorphic and differentiation competent Trypanosoma brucei.Monoxenous (one host) trypanosomatids from insects and other invertebrates can be introduced into axenic culture relatively easily and efficiently, allowing for their transfer from the field into the laboratory. Here we describe simple methods and alternative cultivation protocols, the wider application of which will allow substantial expansion of trypanosomatids available for research.PURPOSE The clinical outcome after in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) is diverse in infertility patients with polycystic ovary syndrome (PCOS). The aim of this study was to develop a nomogram based on an association of patients' characteristics to predict the live birth rate in PCOS patients. METHODS All women in a public university hospital who attempted to conceive by IVF/ICSI for PCOS infertility from January 2014 to October 2018 were included. The nomogram was built from a training cohort of 178 consecutive patients and tested on an independent validation cohort of 81 patients. PCOS was confirmed in all participants. RESULTS Three variates significantly associated with live birth rate of PCOS patients were BMI, total serum cholesterol (TC) and basal FSH. This predictive model built on the basis of BMI, TC, basal FSH, type of embryo transferred and age showed good calibration and discriminatory abilities, with an area under the curve (AUC) of 0.708 (95% CI 0.632-0.785) for the training cohort. The nomogram showed satisfactory goodness-of-fit and discrimination abilities in the independent validation cohort, with an AUC of 0.686 (95% CI 0.556-0.815). CONCLUSION Our simple evidence-based nomogram presents graphically risk factors and prognostic models for IVF/ICSI outcomes in patients with PCOS, which can offer useful guidance to clinicians and patients for individual adjuvant therapy.Bacteria inside biofilms are more persistent and resistant to stress conditions found in the production environment of food processing plants, thus representing a constant risk for product safety and quality. Therefore, the aim of this study was to characterize, using 16S rRNA sequencing, the bacterial communities from biofilms found in four food processing plants (P1, P2, P3, and P4). In total, 50 samples from these four processing plants were taken after cleaning and disinfection processes. Four phyla Proteobacteria, Firmicutes, Actinobacteria, and Bacteroides represented over 94% of the operational taxonomic units found across these four plants. A total of 102 families and 189 genera were identified. Two genera, Pseudomonas spp. and Acinetobacter spp., were the most frequently found (93.47%) across the four plants. In P1, Pseudomonas spp. and Lactobacillus spp. were the dominant genera, whereas Lactobacillus spp. and Streptococcus spp. were identified in P2. On the other hand, biofilms found in P3 and P4 mainly consisted of Pseudomonas spp. and Acinetobacter spp. Our results indicate that different bacterial genera of interest to the food industry due to their ability to form biofilm and affect food quality can coexist inside biofilms, and as such, persist in production environments, representing a constant risk for manufactured foods. In addition, the core microbiota identified across processing plants evaluated was probably influenced by type of food produced and cleaning and disinfection processes performed in each one of these.The primary endpoint of this study is to evaluate the feasibility and safety of the laparoscopic approach in selected types of PeriHilar Cholangiocarcinoma (PHC). Secondary endpoint is to evaluate the potential advantages of laparoscopic approach over the open counterpart. From 2018, an MILS program for PHC was undertaken in selected patients 16 patients constituted the study group (out of 261 operated between 2004 and 2019) and was compared with a group of patients operated by open technique (control group) in the previous period through a propensity score matching with a 12 ratio. Intraoperative and postoperative outcomes were evaluated and compared, focusing on blood loss, length of surgery, conversion to open approach, and complications. Laparoscopic resections resulted in statistically significant longer procedures (360 vs 275 min, p = 0.048). Conversion rate was 18.8%, being oncological concerns the most frequent reason for conversion (3/3 cases). A lower blood loss (380 vs 470, p = 0.048) and minor intraoperative blood transfusions (12.5% vs 21.9%, p = 0.032) were recorded in the study group. A number of retrieved nodes and rate of R0 resections were similar between the two groups. Patients in the MILS group had shorter length of stay (median 10) compared with open group (median 14), p = 0.048. The laparoscopic approach in PHC, so far maintained in an exploratory phase with the biliary-enteric anastomosis performed through the service incision, demonstrates adequate feasibility and safety standards when conducted in carefully selected patients and in centers with expertise.This study investigated the occurrence of malignancy in a cohort of patients with amyotrophic lateral sclerosis (ALS). Forty-three consecutive ALS patients (mean age 61.3 years, 31 men/12 women) admitted between 2015 and 2019 were enrolled. Clinical, electrophysiologic and outcome features that differentiate cancer-associated ALS from classical ALS were investigated. ALS was associated with cancer in 5 (11.6%) patients. Two patients were diagnosed with non-small cell lung, one with endometrium, one with prostate and one with laryngeal carcinoma. GSK1059615 One another patient was diagnosed with monoclonal gammopathy of unknown significance, but she did not develop cancer within 12-month follow-up duration. Two patients had a previous diagnosis of cancer before the diagnosis of ALS. In the remaining 3 patients, cancer was diagnosed at the same time or after the onset of ALS. There was no significant difference with respect to mean age of onset, sex, disease duration, clinical and electrophysiological features and outcome between patients with and without malignancy. None of the patients with cancer survived more than two years after the onset of ALS except one who was diagnosed with flail-leg syndrome before the diagnosis of non-small cell lung carcinoma. ALS is not accepted as a classical paraneoplastic neurological syndrome. Although the association of ALS and cancer seems coincidental, a careful investigation for an underlying cancer is important in patients with ALS. In our patient group with cancer-associated ALS, no distinguishing features were identified.The aim of this study was to investigate (1) whether a sample of highly educated individuals with dyslexia living under optimal personal, educational, cultural, and socioeconomic conditions continues to display core deficits in reading and writing skills during adulthood (extending prior results in Dutch, English, Hebrew, and Spanish to the Portuguese writing system); (2) whether these individuals can compensate for the effects of persistent core deficits when reading complex academic texts; (3) which cognitive resources, such as reading strategies, are used as compensatory mechanisms; and (4) whether quality of life is affected in these individuals. These questions were examined in a sample of 28 adults with dyslexia (DG) and 28 control participants (CG) paired by sex, age, education, and occupation, with a mean of 15 years of formal education. Participants completed measures of phonological awareness; decoding of syllables, words, and pseudowords; writing; reading comprehension (inferential and literal questions, recall, and sensitivity to the rhetorical structure of the target text); and quality of life. Results showed that (1) core deficits associated with dyslexia persisted into adulthood participants with dyslexia performed worse than control subjects at all levels of phonological awareness, reading (except word reading accuracy), and spelling; (2) the groups did not differ on any measures of reading comprehension, suggesting a compensation of core deficits; (3) three compensatory mechanisms were identified slower reading, use of text structure, and verbal ability; (4) participants with dyslexia required more family support and professional help throughout their educational careers, and had more depressive symptoms than control subjects.
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