A report on the impact of rapid prenatal exome sequencing on the clinical management of 52 ongoing pregnancies: a retrospective review.

OBJECTIVE: Studies have shown that prenatal exome sequencing (PES) improves diagnostic yield in cases of fetal structural malformation. We have retrospectively analysed PES cases from two of the largest fetal medicine centres in the UK to determine the impact of results on management of a pregnancy. DESIGN: A retrospective review of clinical case notes. SETTING: Two tertiary fetal medicine centres. POPULATION: Pregnancies with fetal structural abnormalities referred to clinical genetics via a multidisciplinary team. METHODS: We retrospectively reviewed the notes of all patients who had undergone PES. DNA samples were obtained via chorionic villus sampling or amniocentesis. Variants were filtered using patient-specific panels and interpreted using American College of Medical Genetics guidelines. RESULTS: A molecular diagnosis was made in 42% (18/43) ongoing pregnancies; of this group, there was a significant management implication in 44% (8/18). A positive result contributed to the decision to terminate a pregnancy in 16% (7/43) of cases. A negative result had a significant impact on management in two cases by affirming the decision to continue pregnancy. CONCLUSIONS: We demonstrate that the results of PES can inform pregnancy management. Challenges include variant interpretation with limited phenotype information. These results emphasise the importance of the MDT and collecting phenotype and variant data. As this testing is soon to be widely available, we should look to move beyond diagnostic yield as a measure of the value of PES. TWEETABLE ABSTRACT: Prenatal exome sequencing can aid decision-making in pregnancy management; review ahead of routine implementation in NHS.

Human fear conditioning: From neuroscience to the clinic.

Both clinicians and neuroscientists have been long interested in the topic of fear conditioning, with recent advances in neuroscience, in particular, igniting a shared interest in further translation between these domains. Here, we review some historical aspects of this relationship and the progress that has been made in translating the neuroscientific study of fear conditioning to the conceptualization and treatment of mental disorders, especially anxiety-related disorders. We also address some conceptual and methodological challenges faced by this research, and offer some suggestions to support future progress in the field.

Temperature dependence of ice-on-rock friction at realistic glacier conditions.

Using a new biaxial friction apparatus, we conducted experiments of ice-on-rock friction in order to better understand basal sliding of glaciers and ice streams. A series of velocity-stepping and slide-hold-slide tests were conducted to measure friction and healing at temperatures between -20°C and melting. Experimental conditions in this study are comparable to subglacial temperatures, sliding rates and effective pressures of Antarctic ice streams and other glaciers, with load-point velocities ranging from 0.5 to 100 µm s-1 and normal stress σn = 100 kPa. In this range of conditions, temperature dependences of both steady-state friction and frictional healing are considerable. The friction increases linearly with decreasing temperature (temperature weakening) from µ = 0.52 at -20°C to µ = 0.02 at melting. Frictional healing increases and velocity dependence shifts from velocity-strengthening to velocity-weakening behaviour with decreasing temperature. Our results indicate that the strength and stability of glaciers and ice streams may change considerably over the range of temperatures typically found at the ice-bed interface.This article is part of the themed issue 'Microdynamics of ice'.

An apparatus to measure frictional, anelastic, and viscous behavior in ice at temperate and planetary conditions.

In this paper, we describe a cryogenic, servo-controlled biaxial friction apparatus designed to measure the deformational behaviors of ice. The apparatus is specifically designed to accurately achieve and measure the low differential stresses applicable to deforming ice on earth and on icy satellites. We can apply loads in the range ∼2-1800 kPa and velocities up to 4 mm/s, with resolution of 39 Pa and 0.7 µm, respectively. Precise temperature control, measurement, and insulation allow testing at constant temperature (from -2 to -30 °C) for prolonged periods of time. The apparatus is tested with various plastics as well as with polycrystalline ice samples and the results are consistent with previously published values. Critical components of the instrument are described along with examples of data collection schemes and preliminary results. The flexibility of the design allows for both glaciological and planetary applications over a range of deformational behaviors including friction, anelastic, and viscous.

Sexuality issues in gynaecological oncology patients: post treatment symptoms and therapeutic options.

BACKGROUND/AIM: According to recent studies up to 80% of patients would like to receive more information about how cancer treatments can affect their sexual functioning. Moreover, 75 % of them would not feel comfortable being the first to bring up the subject. Our Gynaecological Advice Clinic was established in 2006 at the Christie Hospital and offers support to cancer patients who face sexuality issues. A previous evaluation established that the service sees approximately, 200 patients per year. The aims of this study are to evaluate the service by collecting data relating to levels of attendance, type and amount of clinical activity and to explore further patients' experiences and management. METHODS: This is a retrospective study which was carried out in 2012-2013. Different models are used to evaluate our patients including history taking integrated therapy model, consultations to understand the normal anatomy and physiology with the use of diagrams and photographs, psycho-education and the international classification "Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition" (DSMV). The treatment options advised include medications such as hormone replacement treatment, testosterone, antidepressant, local oestrogen, tibolone, aqueous cream, lubrication to introitus, diprobase, dermal cream and advice for massage to areas of discomfort. Moreover, the use of vaginal dilators, the role of pelvic floor exercises, vulval care and self examination are explained. The team works closely with the psycho-oncology department. The most frequently discussed topics that were covered during the consultation are analysed. RESULTS: 41 outpatient clinics were held between 2012 and 2013. 194 patients attended those clinics during the study period. Single and not group therapy was offered to all the participants. 216 patients were offered appointments while 194 patients actually attended (90%). Patients' age ranged from 24 to 91 years with a mean age of 59 years. 45% had endometrial and 32% cervical cancer, 18% vulval cancer, 4 % ovarian cancer. 74% attended on their own while 18% with their partners and 8% with another family member or friend. 98% of the patients had confidence/trust in the healthcare professional. 95% felt involved with treatment decisions. 82% reported completely understanding information about side-effects. 16% were unaware of who to contact once they had completed treatment. 8% were attending for on-going help with psycho-sexual concerns following completion of treatment. CONCLUSION: The clinic was positively evaluated by patients, they had a high level of trust in the nurses running the service and they found it beneficial. Consultations were generally lasting 20-30 min and most patients reported this was an appropriate length. It once again highlights that although the service was set-up to address psycho-sexual concerns, in reality it dealt with a variety of other patient concerns. Due to the success of the clinic, the service has expanded and we are now supporting patients from other tumour groups such as colorectal, urology and plastic surgery.

Complexity of the Culex pipiens complex in California.

In California, the Culex pipiens complex consists of Culex pipiens, Cx. quinquefasciatus, their hybrids and Culex pipiens form molestus. Using 15 microsatellite markers and a variety of statistical analyses of within- and among-population variation, there is widespread introgression throughout the Central Valley with mostly quinquefasciatus genotypes in the south and pipiens in the north. Those specimens in the Sacramento County area consisted primarily of pipiens-quinquefasciatus and pipiens-molestus hybrids. Populations in Coachella Valley and Los Angeles, CA and Benton, WA were Cx. quinquefasciatus and Cx. pipiens, respectively. Studies are underway to relate these genotypes to phenotypes of autogeny, diapause and vector competence for West Nile Virus.

Mobile Right Atrial Thrombi in a Patient with the Hemoglobin SC Disease.

The formation of Intracardiac thrombi is rare in the absence of structural heart disease or atrial fibrillation. We describe a case of spontaneous right atrial thrombus formation that occurred in a patient with a hypercoagulable condition who had been sub optimally anticoagulated.

Genetic analysis of lung function in inbred mice suggests vitamin D receptor as a candidate gene.

Vitamin D receptor (VDR) polymorphisms are associated with an increased asthma incidence in human populations; however, observations in Vdr knockout mice are unclear. The aim of our study was to determine the influence of the genetic variation in Vdr among inbred strains on lung resistance (i.e., dynamic and airway resistance). In an intercross between the strains C57BL/6J (B6) and KK/HlJ (KK), we identified that a significant QTL for dynamic resistance on Chr X was interacting with a QTL on Chr 15. The Chr 15 QTL peak was located in close proximity to the Vdr locus. We further examined if phenotypes of several inbred strains with varying Vdr genotypes differed. Strains with a B6-like genotype on the Vdr locus had significantly lower airway resistance than strains with a KK-like genotype. Vdr knockout mice were examined for dynamic resistance and showed significantly higher resistance than mice with one (i.e., heterozygous) or both copies (i.e., wild-type) of the Vdr. In comparison to B6, the strain A/J is more resistant but carries the same genotype at the Vdr locus. Dietary vitamin D manipulation in the strain A/J did not rescue the high airway resistance phenotype. Finally, we observed that serum vitamin D does not correlate significantly with lung resistance parameters in a survey of 18 strains. Conclusively, Vdr contributes to the phenotypic variation of lung resistance in inbred mice but other molecules in the Vdr pathway and extended network [i.e., Chr X gene(s)] may contribute as well.

Native fluorescence spectroscopic evaluation of chemotherapeutic effects on malignant cells using nonnegative matrix factorization analysis.

The native fluorescence spectra of retinoic acid (RA)-treated and untreated human breast cancerous cells excited with the selective wavelengths of 300 nm and 340 nm were measured and analyzed using a blind source separation method namely Nonnegative Matrix Factorization (NMF). The results show that the fluorophores of human malignant breast cells change their compositions when they are treated with RA. The reduced contribution from tryptophan, NADH and flavin to the fluorescence of the treated breast cancerous cells was observed in comparison with that of the untreated cells. The results indicate that the decrease of adenosine triphosphate (ATP) in the RA-treated cells. The possible clinical applications of this native fluorescence study are discussed.

Central venous sinus [added] thrombosis misdiagnosed as eclampsia in an emergency department.

Central venous sinus thrombosis can present in the emergency department with a diverse range of symptoms and signs, including a non-neurological scenario, and it thus poses a diagnostic challenge. It is amenable to treatment, so a patient history and awareness of the condition prompts early diagnosis and instigation of treatment which may reduce morbidity and mortality.

[Circulation of West Nile virus (Flaviviridae, Flavivirus) and some other arboviruses in the ecosystems of Volga delta, Volga-Akhtuba flood-lands and adjoining arid regions (2000-2002)]. / Osobennosti tsirkuliatsii virusa Zapadnogo Nila (Flaviviridae, Flavivirus) i nekotorykh drugikh arbovirusov v ékosistemakh del'ty Volgi, Volgo-Akhtubinskoi poimy i sopredel'nykh aridnykh landshaftakh (2000-2002 gg.).

Comprehensive virological, serological as well as genetic studies of the ecology of West Nile Virus (WNV) as well as of some other arboviruses were undertaken in different ecosystems in the territories of the Astrakhan Region and of the Kalmyk Republic. The main carriers (mosquitoes, ticks, birds and mammals) were defined as involved in the circulation of viruses within the natural and anthropogenic biocenosis. Phylogenetic examinations of isolated strains and samples, which were positive in RT-PCR, showed an absolute predominance of genotype I virus that was most closely related to American and Israeli strains. At the same time, epidemic strains had up to 6% of nucleotide differences versus the historic strains isolated in the same region 20-30 years ago. Besides, the circulation of genotype IV was discovered; it was characterized by a lower pathogenicity, which, possibly, ensures the shaping of a pronounced immune interlayer bearing no epidemic consequences. An analysis of the study results on the WNV ecology denotes the epicenter of the endemic territory located in the middle part of the Volga delta.

West Nile virus and other zoonotic viruses in Russia: examples of emerging-reemerging situations.

Studies of the interactions of vertebrates, viruses and arthropod vectors of these viruses were monitored in terms of different ecological groups of viruses transmitted by mosquitoes and ticks in Northern Eurasia in an area encompassing more than 15 million km2. About 90 viruses were isolated, including 24 new to science. Newly recognized infections of vertebrates, including humans, were described. Many unusual epidemic situations were analysed. Permanent efforts were established to prevent bioterrorist activities and their consequences. Extensive epidemic outbreaks of West Nile fever (WNF; i.e., fever caused by West Nile virus) and Crimean-Congo hemorrhagic fever (CCHF) with unusual high mortality appeared in the last four years in southern Russia. We determined infection rates in humans, domestic and wild animals, mosquitoes and ticks from natural and synanthropic biocenoses [Editorial note: "synanthropic" means, roughly, all species living with (c.f. lice, fleas) or near people, such as in houses (c.f. house mice), parks (c.f. Rattus spp.), and the like, rather like "peridomestic", but not strictly so; "biocenosis" is the biome, the "totality of living populations in a particular habitat, which itself is only a part of the ecosystem".]. CCHF virus strains were phylogenetically similar to strains isolated in this area 35 years ago but different from Central-South-Asian and African strains. Before the outset of the current emergence of epidemic WNF, three genetic variants of this virus had been isolated in USSR, two African and one Indian. Phylogenetic analysis of complete genome sequences of epidemic strains demonstrated considerable similarity to strains from USA and Israel and differences from strains isolated in the same USSR areas 20-30 years before. In addition to strains of genotype 1, we isolated strains of second and third lineages and a strain of a fourth genetic variant. Nucleotide differences of these strains from all three genotypes was about 30%. The emerging WNF situation in Russia for the last 4 years probably has been the result of not only natural and social factors, but also to introduction of more virulent strains or by evolution of the virus.

[Analysis of new variants of West Nile fever virus]. / Analiz novykh variantov virusa likhoradki Zapadnogo Nila.

The complete nucleotide sequences for 6 strains of the West Nile fever virus were determined. For the first time the complete nucleotide sequences of the Indian isolate and Krsn190 strain, that is the most far phylogenetically from all isolates known at present time were established. The scheme for separation of virus variants into 4 groups and criteria for determination the group to which the isolate belongs are suggested.

West Nile virus isolates from mosquitoes in New York and New Jersey, 1999.

An outbreak of encephalitis due to West Nile (WN) virus occurred in New York City and the surrounding areas during 1999. Mosquitoes were collected as part of a comprehensive surveillance program implemented to monitor the outbreak. More than 32,000 mosquitoes representing 24 species were tested, and 15 WN virus isolates were obtained. Molecular techniques were used to identify the species represented in the WN virus-positive mosquito pools. Most isolates were from pools containing Culex pipiens mosquitoes, but several pools contained two or more Culex species.

West Nile virus in overwintering Culex mosquitoes, New York City, 2000.

After the 1999 West Nile (WN) encephalitis outbreak in New York, 2,300 overwintering adult mosquitoes were tested for WN virus by cell culture and reverse transcriptase-polymerase chain reaction. WN viral RNA and live virus were found in pools of Culex mosquitoes. Persistence in overwintering Cx. pipiens may be important in the maintenance of WN virus in the northeastern United States.

West nile virus surveillance in Romania: 1997-2000.

In response to the 1996 West Nile (WN) fever epidemic that occurred in Bucharest and southeastern Romania, a surveillance program was established. The surveillance system detected 39 clinical human WN fever cases during the period 1997-2000: 14 cases in 1997, 5 cases in 1998, 7 cases in 1999, and 13 cases in 2000. Thirty-eight of the 39 case-patients lived in the greater Danube Valley of southern Romania, and 1 case-patient resided in the district of Vaslui, located on the Moldavian plateau. The estimated annual case incidence rate for the surveillance area during the period 1997-2000 was 0.95 cases per million residents. Thirty-four cases were serologically confirmed, and 5 cases were classified as probable. Twenty-four case-patients presented with clinical symptoms of meningitis (62%), 12 with meningoencephalitis (31%), 1 with encephalitis (3%), and 2 with febrile exanthema (5%). Five of the 39 cases were fatal (13%). Fourteen case-patients resided in rural areas, and 25 in urban and suburban areas, including 7 case-patients who resided in Bucharest. The ages of case-patients ranged from 8 to 76 years with a median age of 45 years. Twenty-four case-patients were males and 15 were females. Dates of onset of illness occurred from May 24 through September 25, with 82% of onset dates occurring in August and September. Limited entomological surveillance failed to detect WN virus. Retrospective sampling of domestic fowl in the vicinity of case-patient residences during the years 1997-2000 demonstrated seroprevalence rates of 7.8%-29%. Limited wild bird surveillance demonstrated seroprevalence rates of 5%-8%. The surveillance data suggest that WN virus persists focally for several years in poorly understood transmission cycles after sporadic introductions or that WN virus is introduced into Romania at relatively high rates, and persists seasonally in small foci.

Epidemic West Nile encephalitis in Romania: waiting for history to repeat itself.

Seroprevalence data suggest that West Nile virus activity in southern Romania dates to the 1960s or earlier. In the summer of 1996, southeastern Romania and especially Bucharest experienced an unprecedented epidemic of West Nile encephalitis/meningitis, with at least 393 hospitalized cases and 17 deaths. Contributing factors included a susceptible avian population and urban/suburban infrastructural conditions that favored the production of large numbers of Culex pipiens pipiens. The epidemic ended spontaneously in early autumn. Results of serosurveys conducted as the epidemic waned pointed to the recent, novel introduction of West Nile virus to Bucharest. During 1997-2000, 39 scattered human cases of clinical West Nile virus infection (mean, 10 per year; range, 5-14 per year)--including 5 (13%) fatal cases--were diagnosed serologically throughout the region, but epidemic disease did not recur. Results of limited ecologic surveillance efforts during 1997-2000 suggested the existence of numerous focal areas of enzootic West Nile virus activity within the region. The authors explore the possible factors that led to the 1996 epidemic, review the ecologic and human data gathered during the postepidemic period of 1997-2000, summarize the public health lessons offered by the epidemic and its aftermath, and speculate on the future of epidemic West Nile virus activity in southeastern Romania.

Structural origins of the interfacial activation in Thermomyces (Humicola) lanuginosa lipase.

The already known X-ray structures of lipases provide little evidence about initial, discrete structural steps occurring in the first phases of their activation in the presence of lipids (process referred to as interfacial activation). To address this problem, five new Thermomyces (formerly Humicola) lanuginosa lipase (TlL) crystal structures have been solved and compared with four previously reported structures of this enzyme. The bias coming from different crystallization media has been minimized by the growth of all crystals under the same crystallization conditions, in the presence of detergent/lipid analogues, with low or high ionic strength as the only main variable. Resulting structures and their characteristic features allowed the identification of three structurally distinct species of this enzyme: low activity form (LA), activated form (A), and fully Active (FA) form. The isomerization of the Cys268-Cys22 disulfide, synchronized with the formation of a new, short alpha(0) helix and flipping of the Arg84 (Arginine switch) located in the lid's proximal hinge, have been postulated as the key, structural factors of the initial transitions between LA and A forms. The experimental results were supplemented by theoretical calculations. The magnitude of the activation barrier between LA (ground state) and A (end state) forms of TlL (10.6 kcal/mol) is comparable to the enthalpic barriers typical for ring flips and disulfide isomerizations at ambient temperatures. This suggests that the sequence of the structural changes, as exemplified in various TlL crystal structures, mirror those that may occur during interfacial activation.