The role of GlcNAc-PI-de-N-acetylase gene by gene knockout through homologous recombination and its consequences on survival, growth and infectivity of Leishmania major in in vitro and in vivo conditions

At present, there are no efficacious vaccines or effective drugs against leishmaniasis; therefore new and innovative control methods are urgently required. One way to achieve this important goal is through using reverse genetic engineering to evaluate important enzymes, proteins and macromolecules. One of the most important enzymes for Glycosylphosphatidylinositol (GPI) biosynthetic pathways is GlcNAc-PI-deN-acetylase (GPI12). The molecular constructs were cloned in Escherichia coli strain Top 10 and confirmed by molecular methods and were transfected by electroporation into Leishmania major. We demonstrated that two alleles of the GPI12 gene in L. major were successfully removed and enabling the generation of a null mutant, which supports the idea that GPI12 is not an essential gene for the growth and survival of Leishmania and the homozygous knockouts of Leishmania are able to survive. We were able to produce a mutant parasite that caused no damaged to the host. Further investigations are essential to check the safety profile in laboratory animal

Experimental Effects of Injunctive Norms on Simulated Risky Driving Among Teenage Males

Objective: Teenage passengers affect teenage driving performance, possibly by social influence. To examine the effect of social norms on driving behavior, male teenagers were randomly assigned to drive in a simulator with a peer-aged confederate to whom participants were primed to attribute either risk-accepting or risk-averse social norms. It was hypothesized that teenage drivers would engage in more risky driving behavior in the presence of peer passengers than no passengers, and with a risk-accepting compared with a risk-averse passenger. Method: 66 male participants aged 16 to18 years holding a provisional driver license were randomized to drive with a risk-accepting or risk-averse passenger in a simulator. Failure to Stop at a red light and percent Time in Red (light) were measured as primary risk-relevant outcomes of interest at 18 intersections, while driving once alone and once with their assigned passenger. Results: The effect of passenger presence on risky driving was moderated by passenger type for Failed to Stop in a generalized linear mixed model (OR = 1.84, 95% CI [1.19, 2.86], p \u3c .001), and percent Time in Red in a mixed model (B = 7.71, 95% CI [1.54, 13.87], p \u3c .05). Conclusions: Exposure of teenage males to a risk-accepting confederate peer increased teenage males’ risky simulated driving behavior compared with exposure to a risk-averse confederate peer. These results indicate that variability in teenage risky driving could be partially explained by social norms

Screening for Depression, Sleep-Related Disturbances, and Anxiety in Patients with Adenocarcinoma of the Pancreas: A Preliminary Study

Purpose. Screening for depression, sleep-related disturbances, and anxiety in patients with diagnosed adenocarcinoma of the pancreas. Materials and Methods. Patients were evaluated at initial consultation and subsequent visits at the multidisciplinary pancreatic cancer clinic at our University Cancer Center. Cross-sectional and longitudinal psychosocial distress was assessed utilizing Personal Health Questionnaire 9 (PHQ9) to screen for depression and monitor symptoms, the Penn State Worry Questionnaire (PSWQ) for generalized anxiety, and the University of Michigan Sleep Questionnaire to monitor sleep symptoms. Results. Twenty-two patients diagnosed with pancreatic cancer participated during the 6-month pilot study with longitudinal followup for thirteen patients. In this study, mild-to-moderate depressive symptoms, anxiety, and potential sleep problems were common. The main finding of the study was 23% of the patients who were part of this pilot project screened positive for moderately severe major depressive symptoms, likely anxiety disorder or a potential sleep disorder during the study. One patient screened positive for moderately severe depressive symptoms in longitudinal followup. Conclusions. Depression, anxiety, and sleep problems are evident in patients with pancreatic cancer. Prospective, longitudinal studies, with larger groups of patients, are needed to determine if these comorbid symptoms impact outcome and clinical course

Comparison of ex vivo harvested and in vitro cultured materials from Echinococcus granulosus by measuring expression levels of five genes putatively involved in the development and maturation of adult worms

Parts of the natural life cycle of Echinococcus granulosus can be retraced in vitro such as the development of protoscoleces into semiadult worms with three or more proglottids, or the redifferentiation of in vitro cultured protoscoleces into metacestode-like cystic structures. Most in vitro generated samples share-at the microscopical level-high similarities with those naturally grown, but developmental differences have also been documented, such as missing egg production in in vitro grown adults or unusual bladder/vesicle formation in protoscoleces cultured into the metacestode direction. The aim of the present study was to explore how far different in vitro generated stage-specific materials/structures match the natural situation on the transcriptome level, based on testing five exemplarily chosen different genes: the frizzled receptor eg-fz4 (posterior marker), the FGF receptor-like factor eg-fgfrl (anterior association), the cell differentiation protein eg-rcd1 (part of the CCR4-NOT complex, a key regulator of eukaryotic gene expression), the rapidly accelerated fibrosarcoma serin/threonin kinase eg-braf (part of the MAPK pathway involved, e.g., in EGF signaling) and the co-smad eg-smadD (downstream factor of TGFβ/BMP2/activin signaling). These genes-tested via qPCR-were selected such as to allow a discussion on their potential role in the development of E. granulosus into the adult stage. Thus, testing took place with three ex vivo isolated samples, namely (i) egg-containing adult worms, (ii) invaginated protoscoleces, and (iii) protoscolex-free germinal layer tissue. Respective data were compared (a) with in vitro generated metacestode-like microcysts developed from protoscolices, and (b) different development stages of protoscoleces in vitro cultured toward adult maturation. As a finding, only eg-smadD and partially eg-fz4 showed high expression similarities between ex vivo harvested and in vitro cultured E. granulosus, thus suggesting a putative role in adult maturation. Conclusively, the fact of using "only" five genes did not allow answering the question if ex vivo and in vitro materials are similar on the transcriptome level. Another experimental restriction arises from different growth conditions of the in vitro cultured materials, and comparing these to the ex vivo harvested ones. Future experiments may solve the problems by using fully standardized E. granulosus sample collection and fully standardized culture conditions

Observation of motorcycle helmet use rates in Michigan after partial repeal of the universal motorcycle helmet law

Motorcycle crashes result in a significant health burden, including many fatal injuries and serious non-fatal head injuries. Helmets are highly effective in preventing such trauma, and jurisdictions that require helmet use of all motorcyclists have higher rates of helmet use and lower rates of head injuries among motorcyclists. The current study examines helmet use and characteristics of helmeted operators and their riding conditions in Michigan, following a weakening of the state's universal motorcycle helmet use law in April 2012. Data on police-reported crashes occurring during 2012–14 and from a stratified roadside observational survey undertaken in Southeast Michigan during May-September 2014 were used to estimate statewide helmet use rates. Observed helmet use was more common among operators of sports motorcycles, on freeways, and in the morning, and least common among operators of cruisers, on minor arterials, and in the afternoon. The rate of helmet use across the state was estimated at 75%, adjusted for roadway type, motorcycle class, and time of day. Similarly, the helmet use rate found from examination of crash records was 73%. In the observation survey, 47% of operators wore jackets, 94% wore long pants, 54% wore boots, and 80% wore gloves. Protective clothing of jackets and gloves was most often worn by sport motorcycle operators and long pants and boots most often by riders of touring motorcycles. Findings highlight the much lower rate of helmet use in Michigan compared with states that have a universal helmet use law, although the rate is higher than observed in many states with partial helmet laws. Targeted interventions aimed at specific groups of motorcyclists and situations where helmet use rates are particularly low should be considered to increase helmet use

Differential Expression of Hox and Notch Genes in Larval and Adult Stages of Echinococcus granulosus

This investigation aimed to evaluate the differential expression of HoxB7 and notch genes in different developmental stages of Echinococcus granulosus sensu stricto. The expression of HoxB7 gene was observed at all developmental stages. Nevertheless, significant fold differences in the expression level was documented in the juvenile worm with 3 or more proglottids, the germinal layer from infected sheep, and the adult worm from an experimentally infected dog. The notch gene was expressed at all developmental stages of E. granulosus; however, the fold difference was significantly increased at the microcysts in monophasic culture medium and the germinal layer of infected sheep in comparison with other stages. The findings demonstrated that the 2 aforementioned genes evaluated in the present study were differentially expressed at different developmental stages of the parasite and may contribute to some important biological processes of E. granulosus

The impact of Michigan's partial repeal of the universal motorcycle helmet law on helmet use, fatalities, and head injuries

Objectives. To evaluate the impact of the partial repeal of Michigan's universal motorcycle helmet law on helmet use, fatalities, and head injuries. Methods. We compared helmet use rates and motorcycle crash fatality risk for the 12 months before and after the April 13, 2012, repeal with a statewide police-reported crash data set. We linked police-reported crashes to injured riders in a statewide trauma registry.We compared head injury before and after the repeal. Regression examined the effect of helmet use on fatality and head injury risk. Results. Helmet use decreased in crash (93.2% vs 70.8%; P < .001) and trauma data (91.1% vs 66.2%; P < .001) after the repeal. Although fatalities did not change overall (3.3% vs 3.2%; P = .87), head injuries (43.4% vs 49.6%; P < .05) and neurosurgical intervention increased (3.7% vs 6.5%; P < .05). Male gender (adjusted odds ratio [AOR] = 1.65), helmet nonuse (AOR = 1.84), alcohol intoxication (AOR = 11.31), intersection crashes (AOR = 1.62), and crashes at higher speed limits (AOR = 1.04) increased fatality risk. Helmet nonuse (AOR = 2.31) and alcohol intoxication (AOR = 2.81) increased odds of head injury. Conclusions. Michigan's helmet law repeal resulted in a 24% to27% helmet use decline among riders in crashes and a 14% increase in head injury

Peer passenger norms and pressure: Experimental effects on simulated driving among teenage males

Objective Serious crashes are more likely when teenage drivers have teenage passengers. One likely source of this increased risk is social influences on driving performance. This driving simulator study experimentally tested the effects of peer influence (i.e., risk-accepting compared to risk-averse peer norms reinforced by pressure) on the driving risk behavior (i.e., risky driving behavior and inattention to hazards) of male teenagers. It was hypothesized that peer presence would result in greater driving risk behavior (i.e., increased driving risk and reduced latent hazard anticipation), and that the effect would be greater when the peer was risk-accepting. Methods Fifty-three 16- and 17-year-old male participants holding a provisional U.S., State of Michigan driver license were randomized to either a risk-accepting or risk-averse condition. Each participant operated a driving simulator while alone and separately with a confederate peer passenger. The simulator world included scenarios designed to elicit variation in driving risk behavior with a teen passenger present in the vehicle. Results Significant interactions of passenger presence (passenger present vs. alone) by risk condition (risk-accepting vs. risk-averse) were observed for variables measuring: failure to stop at yellow light intersections (Incident Rate Ratio (IRR)\ua0=\ua02.16; 95% confidence interval [95CI]\ua0=\ua01.06, 4.43); higher probability of overtaking (IRR\ua0=\ua010.17; 95CI\ua0=\ua01.43, 73.35); shorter left turn latency (IRR\ua0=\ua00.43; 95CI\ua0=\ua00.31, 0.60); and, failure to stop at an intersection with an occluded stop sign (IRR\ua0=\ua07.90; 95CI\ua0=\ua02.06, 30.35). In all cases, greater risky driving by participants was more likely with a risk-accepting passenger versus a risk-averse passenger present and a risk-accepting passenger present versus driving alone. Conclusions Exposure of male teenagers to a risk-accepting confederate peer passenger who applied peer influence increased simulated risky driving behavior compared with exposure to a risk-averse confederate peer passenger or driving alone. These results are consistent with the contention that variability in teenage risky driving is in part explained by social influences