Study of enteric pathogens among children in the tropics and effects of prolonged storage of stool samples.

The study was performed to compare real-time PCR after nucleic acid extraction directly from stool samples as well as from samples stored and transported on Whatman papers or flocked swabs at ambient temperature in the tropics. In addition, the possible suitability for a clear determination of likely aetiological relevance of PCR-based pathogen detections based on cycle threshold (Ct) values was assessed. From 632 Tanzanian children <5 years of age with and without gastrointestinal symptoms, 466 samples were subjected to nucleic acid extraction and real-time PCR for gastrointestinal viral, bacterial and protozoan pathogens. Equal or even higher frequencies of pathogen detections from Whatman papers or flocked swabs were achieved compared with nucleic acid extraction directly from stool samples. Comparison of the Ct values showed no significant difference according to the nucleic acid extraction strategy. Also, the Ct values did not allow a decision whether a detected pathogen was associated with gastrointestinal symptoms.

Keratin 6a mutations lead to impaired mitochondrial quality control.

BACKGROUND: Epidermal differentiation is a multilevel process in which keratinocytes need to lose their organelles, including their mitochondria, by autophagy. Disturbed autophagy leads to thickening of the epidermis as seen in pachyonychia congenita (PC), a rare skin disease caused by mutations in keratins 6, 16 and 17. OBJECTIVES: To ask if mitophagy, the selective degradation of mitochondria by autophagy, is disturbed in PC and, if so, at which stage. METHODS: Immortalized keratinocytes derived from patients with PC were used in fluorescence-based and biochemical assays to dissect the different steps of mitophagy. RESULTS: PC keratinocytes accumulated old mitochondria and displayed disturbed clearance of mitochondria after mitochondrial uncoupling. However, early mitophagy steps and autophagosome formation were not affected. We observed that autolysosomes accumulate in PC and are not sufficiently recycled. CONCLUSIONS: We propose an influence of keratins on autolysosomal degradation and recycling. What's already known about this topic? Terminal epidermal differentiation is a multistep process that includes the elimination of cellular components by autophagy. Autophagy-impaired keratinocytes have been shown to result in thickening of epidermal layers. Hyperkeratosis also occurs in pachyonychia congenita (PC), a rare skin disease caused by mutations in keratins 6, 16 and 17. What does this study add? Keratins contribute to mitochondrial quality control as well as maintenance of mitochondria-endoplasmic reticulum contact sites. Keratins influence autolysosomal maturation or reformation. What is the translational message? Overaged mitochondria and autolysosomes accumulate in PC. Mutations in keratin 6a lead to severely impaired mitophagy, which might contribute to PC pathogenesis.

Clinical and genetic spectrum of SCN2A-associated episodic ataxia.

BACKGROUND: Pathogenic variants in SCN2A are associated with various neurological disorders including epilepsy, autism spectrum disorder and intellectual disability. Few reports have recently described SCN2A-associated episodic ataxia (EA). Our study identifies its broader clinical and genetic spectrum, and describes pharmacological approaches. RESULTS: We report 21 patients with SCN2A-associated EA, of which 9 are unpublished cases. The large majority of patients present with epileptic seizures (18/21, 86%), often starting within the first three months of life (12/18, 67%). In contrast, onset of episodic ataxia ranged from 10 months to 14 years of age. The frequency of EA episodes ranged from brief, daily events up to 1-2 episodes per year each lasting several weeks. Potential triggers include minor head traumas and sleep deprivation. Cognitive outcome is favorable in most patients with normal or mildly impaired cognitive development in 17/21 patients (81%). No clear genotype-phenotype correlations were identified in this cohort. However, two mutational hotspots were identified, i.e. 7/21 patients (33%) harbor the identical pathogenic variant p.A263V, whereas 5/21 (24%) carry pathogenic variants that affect the S4 segment and its cytoplasmic loop within the domain IV. In addition, we identified six novel pathogenic variants in SCN2A. While acetazolamide was previously reported as beneficial in SCN2A-associated EA in one case, our data show a conflicting response in 8 additional patients treated with acetazolamide: three of them profited from acetazolamide treatment, while 5/8 did not. CONCLUSIONS: Our study describes the heterogeneous clinical spectrum of SCN2A-associated EA, identifies two mutational hotspots and shows positive effects of acetazolamide in about 50%.

PCR-based rapid diagnostic tests as a strategy for preventing infections with sexually transmitted diseases-a 'diagnostics-as-prevention' modelling approach.

The potential was modelled of molecular rapid diagnostic tests (RDTs) that target agents of sexually transmitted diseases (STDs) to contribute to a reduction in the exposure risk for sexually active populations who prefer condom-free high-risk contacts. The biostatistical approach is exemplified by considering the use of a PCR-based RDT for Chlamydia trachomatis and Neisseria gonorrhoeae in a population of German men having sex with men. We modelled the exposure risk as risk of sexual contact with individuals infected by a STD in spite of multiple RDTs having been conducted to identify STDs that could lead to avoidance of an infectious sexual contact. The risk for a given specific disease is influenced by the diagnostic accuracy of all diagnostic tests conducted. The simulation showed a reduction in exposure risk through conducting the selected PCR-based RDT as a strategy to prevent infections with C. trachomatis and N. gonorrhoeae in comparison with unprotected sexual contact of >97% for C. trachomatis and ≥96% for N. gonorrhoeae. Reliable knowledge of the performance characteristics of the diagnostic tests applied is essential for a thorough risk assessment based on such diagnostic approaches. SIGNIFICANCE AND IMPACT OF THE STUDY: A low-threshold use of molecular rapid diagnostic tests in settings where risky sexual activity is undertaken with deliberate non-use of condoms is modelled. Such an approach might contribute to a considerable reduction in the exposure risk if positively tested individuals are excluded from unprotected sexual intercourse. Protective effects are influenced by the prevalence of the disease and the performance characteristics of the applied tests. Precise knowledge of the performance characteristics of the test assays employed are prerequisites of a reliable risk assessment. The modelling approach is of particular use for preventive health experts.

HIV prevention strategies and risk of infection: a model-based analysis.

Risk populations for HIV infections tend to neglect condom use, making alternative preventive approaches necessary. Accordingly, we modelled the risk of sexual HIV transmission for condom use vs. use of rapid diagnostic test (RDT) systems with subsequent exclusion of potential sexual partners with a correctly or falsely positive test from unprotected sex with and without the use of HIV pre-exposure prophylaxis (PrEP) in a bio-statistical approach. We combined a previously described model of transmission risk for HIV-exposed individuals with a newly suggested model of risk of HIV exposure for sexually active HIV-negative individuals. The model was adapted for several stages of infection and different strategies of HIV infection prevention.HIV prevention with RDTs can reduce the transmission risk by up to 97% compared with having sex without any prevention and up to 80% compared with condom use. Nevertheless, RDT-based prevention strategies demonstrate a lack of protection in several stages of infection; in particular, RNA-based RDT systems may fail under treatment. RDT-based pre-screening of potential sex partners prior to unprotected sexual contacts substantially reduces HIV transmission risk. Combination of different prevention strategies is advisable for high-risk groups.

Comparing a single-day swabbing regimen with an established 3-day protocol for MRSA decolonization control.

OBJECTIVES: Success of methicillin-resistant Staphylococcus aureus (MRSA) decolonization procedures is usually verified by control swabs of the colonized body region. This prospective controlled study compared a single-day regimen with a well-established 3-day scheme for noninferiority and adherence to the testing scheme. METHODS: Two sampling schemes for screening MRSA patients of a single study cohort at a German tertiary-care hospital 2 days after decolonization were compared regarding their ability to identify MRSA colonization in throat or nose. In each patient, three nose and three throat swabs were taken at 3- to 4-hour intervals during screening day 1, and in the same patients once daily on days 1, 2 and 3. Swabs were analysed using chromogenic agar and broth enrichment. The study aimed to investigate whether the single-day swabbing scheme is not inferior to the 3-day scheme with a 15% noninferiority margin. RESULTS: One hundred sixty patients were included, comprising 105 and 101 patients with results on all three swabs for decolonization screening of the nose and throat, respectively. Noninferiority of the single-day swabbing scheme was confirmed for both pharyngeal and nasal swabs, with 91.8% and 89% agreement, respectively. The absolute difference of positivity rates between the swabbing regimens was 0.025 (-0.082, 0.131) for the nose and 0.006 (-0.102, 0.114) (95% confidence interval) for the pharynx as calculated with McNemar's test for matched or paired data. Compliance with the single-day scheme was better, with 12% lacking second-day swabs and 27% lacking third-day swabs from the nostrils. CONCLUSIONS: The better adherence to the single-day screening scheme with noninferiority suggests its implementation as the new gold standard.

Vertical force distribution in the paws of sound Labrador retrievers during walking.

In contrast to gait analysis in humans, where pedobarography is an integral part of biomechanical studies, veterinary researchers have rarely investigated vertical force distribution (VFD) in the paws of dogs. The aim of this study was to investigate the VFD of peak of vertical force (PFz), vertical impulse (IFz) and time of occurrence of PFz during stance phase (TPFz) in 20 sound, adult Labrador retrievers walking normally on a pressure plate. A technique was used that divided the canine paw prints into quadrants. A general linear model was introduced to investigate the effects of forelimbs/hindlimbs, body side, and medial/lateral and cranial/caudal quadrants on VFD as they related to the total force (sum of all PFz/IFz values). For PFz and IFz, there were significantly greater effects on VFD in the lateral quadrants compared to the medial quadrants, respectively (6.49 ± 2.56% vs. 6.01 ± 2.60% and 6.62 ± 3.06% vs. 5.88 ± 3.21%; P < 0.001), in the forelimbs compared to the hindlimbs (8.02 ± 2.13% vs. 4.48 ± 1.61% and 8.02 ± 2.83% vs. 4.48 ± 2.36%; P < 0.001), and in the cranial quadrants compared to the caudal quadrants (7.87 ± 2.09% vs. 4.63 ± 1.93% and 8.57 ± 2.17% vs. 3.88 ± 1.98%; P < 0.001). The cranial/caudal ratio was higher in the hindlimbs than in the forelimbs (PFz: 2.10 ± 0.45 vs. 1.65 ± 0.32; P = 0.001; and IFz: 3.35 ± 0.80 vs. 2.04 ± 0.46; P < 0.001). The TPFz was reached earlier in the hindlimbs than in the forelimbs (46.86 ± 19.16% vs. 54.08 ± 19.62%; P < 0.001) and in the caudal quadrant than in the cranial quadrant (32.57 ± 5.77% vs. 68.37 ± 10.01%; P < 0.001). These data from sound Labrador retrievers could be used as a basis for future research investigating orthopedically- and/or neurologically-impaired animals.

Mutations in the sodium channel gene SCN2A cause neonatal epilepsy with late-onset episodic ataxia.

Mutations in SCN2A cause epilepsy syndromes of variable severity including neonatal-infantile seizures. In one case, we previously described additional childhood-onset episodic ataxia. Here, we corroborate and detail the latter phenotype in three further cases. We describe the clinical characteristics, identify the causative SCN2A mutations and determine their functional consequences using whole-cell patch-clamping in mammalian cells. In total, four probands presented with neonatal-onset seizures remitting after five to 13 months. In early childhood, they started to experience repeated episodes of ataxia, accompanied in part by headache or back pain lasting minutes to several hours. In two of the new cases, we detected the novel mutation p.Arg1882Gly. While this mutation occurred de novo in both patients, one of them carries an additional known variant on the same SCN2A allele, inherited from the unaffected father (p.Gly1522Ala). Whereas p.Arg1882Gly alone shifted the activation curve by -4 mV, the combination of both variants did not affect activation, but caused a depolarizing shift of voltage-dependent inactivation, and a significant increase in Na(+) current density and protein production. p.Gly1522Ala alone did not change channel gating. The third new proband carries the same de novo SCN2A gain-of-function mutation as our first published case (p.Ala263Val). Our findings broaden the clinical spectrum observed with SCN2A gain-of-function mutations, showing that fairly different biophysical mechanisms can cause a convergent clinical phenotype of neonatal seizures and later onset episodic ataxia.

[Epidemiology of Ebola virus disease and of other highly contagious, life-threatening diseases with low incidence in Germany]. / Epidemiologie des Ebolafiebers und anderer, in Deutschland seltener hochkontagiöser, lebensbedrohlicher Erkrankungen.

Apart from sporadic exported cases, the occurrence of Ebola, Marburg and Lassa virus diseases is limited to the African continent. Crimean-Congo Hemorrhagic Fever occurs in Southeastern Europe but, so far, not in Germany. Other hemorrhagic fever disease-viruses occur in distinct regions in South America. Pulmonary plague is the bacterial infectious disease with the most contagious and lethal course and it is endemic to Madagascar and East Africa, but also occurs in other countries (e.g. India, USA). Monkey pox epidemics have occurred in remote areas of the Congo Basin. Such outbreaks could potentially become more common with the discontinuation of the cross-protective smallpox vaccination. The Severe Acute Respiratory Syndrome (SARS) that emerged in 2002/2003 is another pathogen with significant epidemic potential. Typical for these diseases is a natural circulation between reservoir animals in remote areas. Sporadic transmission to humans can occur through contact with an infected animal. Subsequent human-to-human transmission can lead to epidemics, such as the current outbreak of Ebola virus disease in West Africa.

Surveillance and Outbreak Response Management System (SORMAS) to support the control of the Ebola virus disease outbreak in West Africa.

In the context of controlling the current outbreak of Ebola virus disease (EVD), the World Health Organization claimed that 'critical determinant of epidemic size appears to be the speed of implementation of rigorous control measures', i.e. immediate follow-up of contact persons during 21 days after exposure, isolation and treatment of cases, decontamination, and safe burials. We developed the Surveillance and Outbreak Response Management System (SORMAS) to improve efficiency and timeliness of these measures. We used the Design Thinking methodology to systematically analyse experiences from field workers and the Ebola Emergency Operations Centre (EOC) after successful control of the EVD outbreak in Nigeria. We developed a process model with seven personas representing the procedures of EVD outbreak control. The SORMAS system architecture combines latest In-Memory Database (IMDB) technology via SAP HANA (in-memory, relational database management system), enabling interactive data analyses, and established SAP cloud tools, such as SAP Afaria (a mobile device management software). The user interface consists of specific front-ends for smartphones and tablet devices, which are independent from physical configurations. SORMAS allows real-time, bidirectional information exchange between field workers and the EOC, ensures supervision of contact follow-up, automated status reports, and GPS tracking. SORMAS may become a platform for outbreak management and improved routine surveillance of any infectious disease. Furthermore, the SORMAS process model may serve as framework for EVD outbreak modeling.

Emotion regulation: quantitative meta-analysis of functional activation and deactivation.

Emotion regulation is hypothesized to be a multifaceted process by which individuals willfully modulate the intensity and direction of emotional response via prefrontally mediated inhibition of subcortical response-related regions of the brain. Here we employ activation likelihood estimation (ALE) meta-analysis of functional magnetic resonance imaging studies to (1) reveal a consistent network of structures active during emotion regulation, (2) identify the target regions inactivated by the willful regulation process, and (3) investigate the consistency of activated structures associated with downregulation and upregulation. Results reveal signal change in bilateral amygdala/parahippocampal gyrus that decreased in downregulated states and increased in upregulated states, while cortical regions including superior frontal gyrus, cingulate, and premotor areas exhibited enhanced activity across all regulation conditions. These results provide consistent evidence for the role of amygdala activity in experienced emotional intensity, where intentional dampening and exaggeration are clearly expressed. However, the execution of emotional upregulation and downregulation may involve distinct subsets of frontocortical structures.

Difficulties in species identification within the genus Haemophilus - A pilot study addressing a significant problem for routine diagnostics.

Diagnostic misidentifications of commensalic Haemophilus haemolyticus as pathogenic Haemophilus influenzae are frequent. This pilot study evaluates whether isolations of H. haemolyticus are frequent enough in Germany to cause a relevant diagnostic problem, considering the fact that even H. influenzae is a mere colonizer in about 30% of isolations. In microbiological laboratories of two hospitals located in Northern and Southern Germany, the distribution of Haemophilus spp. was analyzed during a six-month-period. Site of infection, sex, and age of the patients was taken into consideration. A total of 77 Haemophilus spp. isolates was acquired and discriminated on species level, comprising: 48 H. influenzae, 25 Haemophilus parainfluenzae, 3 H. haemolyticus, and 1 Haemophilus parahaemolyticus. The proportion of H. haemolyticus was calculated to range between 1.2% and 16.2 % within the 95% confidence limits. Commensalic Haemophilus spp. were isolated from oropharynx-associated sites only. H. influenzae, in contrast, was detected in clinically relevant materials like lower respiratory materials and conjunctiva swabs. Altogether, there was a low proportion of clinical H. haemolyticus isolates. Accordingly, the problem of unnecessary antibiotic therapies due to misidentifications of H. haemolyticus as H. influenzae is quantitatively negligible compared with the risk of confusing H. influenzae colonizations with infections.

Comparison of Obstetric Efficacy and Safety of the Kiwi OmniCup with Conventional Vacuum Extraction.

Purpose: The aim of the present study was to compare the safety and efficacy of the Kiwi OmniCup system with conventional vacuum delivery. Methods: A retrospective study of operative vaginal deliveries was done for 4682 births. The procedures included 217 operative vaginal deliveries (4.6 %), 79 of which were done using conventional vacuum extraction (37 %) and 138 using the Kiwi system (63 %). Results: Use of the Kiwi system was associated with a significant reduction in episiotomies (61 vs. 76 % in the control group; p < 0.05). The rates of successful completion of birth were comparable for the two systems (94 % with the Kiwi system and 99 % with conventional vacuum delivery). Cup detachment occurred significantly more often in the Kiwi group (p < 0.005), requiring a change to a different method of birth significantly more often. It was necessary to change the procedure significantly more often from the mid-pelvis (p < 0.05). The incidence of maternal and foetal injuries was similar for the two systems. Conclusions: With regard to obstetric efficacy and safety and foetal and maternal injuries, the Kiwi system is an acceptable alternative to the conventional vacuum cup. The advantages of the Kiwi system are its significantly lower episiotomy rate and its ease and rapidity of use.

Is rapid hepatitis C virus testing from corpses a screening option for index persons who have died after mass-casualty incidents in high-prevalence settings in the field?

INTRODUCTION: We tested a commercially available rapid hepatitis C virus (HCV) test assay for its potential use for analyses of corpses as a screening option for index persons who have died after mass-casualty incidents in high-prevalence settings in the field. MATERIALS AND METHODS: 50 blood samples were drawn from 16 recently deceased confirmed HCV-positive patients whose corpses were stored at 4°C in the mortuary and were analysed at admission and up to 48 h post mortem by rapid serological testing using the ImmunoFlow HCV test (Core Diagnostics, Birmingham, UK) in comparison with automated serological assays and PCR. Samples from 50 HCV-negative corpses were also analysed. RESULTS: The blood of only four of the 16 HCV-positive corpses reacted clearly with the ImmunoFlow HCV test, while in five cases the result was only weakly reactive and three cases showed very weak reactivity. Four of the infected corpses showed initially negative results, three of which became very weakly reactive 48 h post mortem. 49 out of 50 samples (98%) from HCV-negative corpses tested negative. DISCUSSION: The rapid test system we investigated showed insufficient sensitivity regarding the identification of HCV positivity. Automated serological testing or PCR should be preferred if it is realistically available in the deployed military setting.

An eight month randomized controlled exercise intervention alters resting state synchrony in overweight children.

Children with low aerobic fitness have altered brain function compared to higher-fit children. This study examined the effect of an 8-month exercise intervention on resting state synchrony. Twenty-two sedentary, overweight (body mass index ≥85th percentile) children 8-11 years old were randomly assigned to one of two after-school programs: aerobic exercise (n=13) or sedentary attention control (n=9). Before and after the 8-month programs, all subjects participated in resting state functional magnetic resonance imaging scans. Independent components analysis identified several networks, with four chosen for between-group analysis: salience, default mode, cognitive control, and motor networks. The default mode, cognitive control, and motor networks showed more spatial refinement over time in the exercise group compared to controls. The motor network showed increased synchrony in the exercise group with the right medial frontal gyrus compared to controls. Exercise behavior may enhance brain development in children.

Compliance with antimalarial chemoprophylaxis in German soldiers: a 6-year survey.

PURPOSE: Since 1992, German soldiers have been deployed in areas where malaria is endemic. Antimalarial chemoprophylaxis (CP) is directed according to the assessed risk and is provided free of charge. Compliance is crucial if its effect is to be reliable. This study analysed compliance with directed CP in German soldiers as well as its determinants. METHODS: Between 2003 and 2009, standardized questionnaire-based interviews were performed with 2,149 out of approximately 100,000 German soldiers who were deployed during this period in areas where malaria is endemic. The questionnaires dealt with information that the soldiers had received about malaria prior to their missions, with their adherence to mosquito-protective and antimalarial chemoprophylactic procedures, and their estimations of their individual level of exposure. RESULTS: About 1,308 out of 2,149 interviewed soldiers had been ordered to take CP, allowing for an assessment of the outcome parameter "CP-compliance". About 76.9 % out of 1,308 soldiers to whom regular CP was directed took it regularly. The exposure variables "age", "satisfaction with malaria counselling", "perceived threat due to insects or mosquitoes" and "use of insect repellents" were positively associated with compliance with directed antimalarial CP. CONCLUSIONS: The study confirms the findings of the French and US armies that even free-of-charge access to antimalarial medication will not lead to 100 % acceptance. The compliance problem is aggravated by the generally low age of deployed soldiers. Adequate counselling is crucial to increase adherence to antimalarial CP.

Technical complications during removal of locking screws from locking compression plates: a prospective multicenter study.

PURPOSE: To assess the risk for technical complications in patients undergoing removal of locking compression plates (LCP) with head locking screws. METHODS: A total of 205 patients who were scheduled for implant removal surgery after a healed fracture of the femur, tibia, humerus, distal radius, or clavicle in nine Austrian clinics were prospectively included in the study, all of whom had previously undergone fracture fixation by plates, with titanium implants used in 98 % of the patients. Intraoperative technical complications and the methods used to solve them were documented by the surgeon. RESULTS: During the course of this study, a total of 1,462 locking screws were removed from 204 LCPs. While 95 % of these screws could be removed without difficulties, technical complications were reported for 41 patients with 78 screws which could not be removed with standard screwdrivers and required the use of additional instruments. The estimated risk for the occurrence of at least one technical complication during implant removal surgery was 20.1 %. The most frequently observed complications were screws that could not be loosened because they were jammed in the LCP, screws with a damaged recess in which the screwdriver turned freely, as well as a combination of both events. The majority of these screws could be removed with the use of a conical extraction screw or by drilling off the screw head. In one patient, an intraoperative refracture of the humerus occurred during plate removal. Even though there is a rate of 20 % for technical complications when removing the implants, only a few patients experience a clinical impact. CONCLUSIONS: Titanium LCPs are prone to technical complications during implant removal, but the majority of the issues can be solved using special techniques.