Low prevalence of male microchimerism in women with Mayer-Rokitansky-Küster-Hauser syndrome.

STUDY QUESTION: Is there an increased prevalence of male microchimerism in women with Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome, as evidence of fetal exposure to blood and anti-Müllerian hormone (AMH) from a (vanished) male co-twin resulting in regression of the Müllerian duct derivatives? SUMMARY ANSWER: Predominant absence of male microchimerism in adult women with MRKH syndrome does not support our hypothesis that intrauterine blood exchange with a (vanished) male co-twin is the pathophysiological mechanism. WHAT IS KNOWN ALREADY: The etiology of MRKH is unclear. Research on the phenotype analogous condition in cattle (freemartinism) has yielded the hypothesis that Müllerian duct development is inhibited by exposure to AMH in utero. In cattle, the male co-twin has been identified as the source for AMH, which is transferred via placental blood exchange. In human twins, a similar exchange of cellular material has been documented by detection of chimerism, but it is unknown whether this has clinical consequences. STUDY DESIGN, SIZE, DURATION: An observational case-control study was performed to compare the presence of male microchimerism in women with MRKH syndrome and control women. Through recruitment via the Dutch patients' association of women with MRKH (comprising 300 members who were informed by email or regular mail), we enrolled 96 patients between January 2017 and July 2017. The control group consisted of 100 women who reported never having been pregnant. PARTICIPANTS/MATERIALS, SETTING, METHODS: After written informed consent, peripheral blood samples were obtained by venipuncture, and genomic DNA was extracted. Male microchimerism was detected by Y-chromosome-specific real-time quantitative PCR, with use of DYS14 marker. Possible other sources for microchimerism, for example older brothers, were evaluated using questionnaire data. MAIN RESULTS AND THE ROLE OF CHANCE: The final analysis included 194 women: 95 women with MRKH syndrome with a mean age of 40.9 years and 99 control women with a mean age of 30.2 years. In total, 54 women (56.8%) were identified as having typical MRKH syndrome, and 41 women (43.2%) were identified as having atypical MRKH syndrome (when extra-genital malformations were present). The prevalence of male microchimerism was significantly higher in the control group than in the MRKH group (17.2% versus 5.3%, P = 0.009). After correcting for age, women in the control group were 5.8 times more likely to have male microchimerism (odds ratio 5.84 (CI 1.59-21.47), P = 0.008). The mean concentration of male microchimerism in the positive samples was 56.0 male genome equivalent per 1 000 000 cells. The prevalence of male microchimerism was similar in women with typical MRKH syndrome and atypical MRKH syndrome (5.6% versus 4.9%, P = 0.884). There were no differences between women with or without microchimerism in occurrence of alternative sources of XY cells, such as older brothers, previous blood transfusion, or history of sexual intercourse. LIMITATIONS, REASON FOR CAUTION: We are not able to draw definitive conclusions regarding the occurrence of AMH exchange during embryologic development in women with MRKH syndrome. Our subject population includes all adult women and therefore is reliant on long-term prevalence of microchimerism. Moreover, we have only tested blood, and, theoretically, the cells may have grafted anywhere in the body during development. It must also be considered that the exchange of AMH may occur without the transfusion of XY cells and therefore cannot be discovered by chimerism detection. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study to test the theory that freemartinism causes the MRKH syndrome in humans. The study aimed to test the presence of male microchimerism in women with MRKH syndrome as a reflection of early fetal exposure to blood and AMH from a male (vanished) co-twin. We found that male microchimerism was only present in 5.3% of the women with MRKH syndrome, a significantly lower percentage than in the control group (17.2%). Our results do not provide evidence for an increased male microchimerism in adult women with MRKH as a product of intrauterine blood exchange. However, the significant difference in favor of the control group is of interest to the ongoing discussion on microchimeric cell transfer and the possible sources of XY cells. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: Dutch trial register, NTR5961.

Minimally invasive genomic and transcriptomic profiling of visceral cancers by next-generation sequencing of circulating exosomes.

BACKGROUND: The ability to perform comprehensive profiling of cancers at high resolution is essential for precision medicine. Liquid biopsies using shed exosomes provide high-quality nucleic acids to obtain molecular characterization, which may be especially useful for visceral cancers that are not amenable to routine biopsies. PATIENTS AND METHODS: We isolated shed exosomes in biofluids from three patients with pancreaticobiliary cancers (two pancreatic, one ampullary). We performed comprehensive profiling of exoDNA and exoRNA by whole genome, exome and transcriptome sequencing using the Illumina HiSeq 2500 sequencer. We assessed the feasibility of calling copy number events, detecting mutational signatures and identifying potentially actionable mutations in exoDNA sequencing data, as well as expressed point mutations and gene fusions in exoRNA sequencing data. RESULTS: Whole-exome sequencing resulted in 95%-99% of the target regions covered at a mean depth of 133-490×. Genome-wide copy number profiles, and high estimates of tumor fractions (ranging from 56% to 82%), suggest robust representation of the tumor DNA within the shed exosomal compartment. Multiple actionable mutations, including alterations in NOTCH1 and BRCA2, were found in patient exoDNA samples. Further, RNA sequencing of shed exosomes identified the presence of expressed fusion genes, representing an avenue for elucidation of tumor neoantigens. CONCLUSIONS: We have demonstrated high-resolution profiling of the genomic and transcriptomic landscapes of visceral cancers. A wide range of cancer-derived biomarkers could be detected within the nucleic acid cargo of shed exosomes, including copy number profiles, point mutations, insertions, deletions, gene fusions and mutational signatures. Liquid biopsies using shed exosomes has the potential to be used as a clinical tool for cancer diagnosis, therapeutic stratification and treatment monitoring, precluding the need for direct tumor sampling.

Advanced airway management is necessary in prehospital trauma patients.

BACKGROUND: Treatment of airway compromise in trauma patients is a priority. Basic airway management is provided by all emergency personnel, but the requirement for on-scene advanced airway management is controversial. We attempted to establish the demand for on-scene advanced airway interventions. Trauma patients managed with standard UK paramedic airway interventions were assessed to determine whether airway compromise had been effectively treated or whether more advanced airway management was required. METHODS: A prospective observational study was conducted to identify trauma patients requiring prehospital advanced airway management attended by a doctor-paramedic team. The team assessed and documented airway compromise on arrival, interventions performed before and after their arrival, and their impact on airway compromise. RESULTS: Four hundred and seventy-two patients required advanced airway intervention and received 925 airway interventions by ground-based paramedics. Two hundred and sixty-nine patients (57%) still had airway compromise on arrival of the enhanced care team; no oxygen had been administered to 52 patients (11%). There were 45 attempted intubations by ground paramedics with a 64% success rate and 11% unrecognized oesophageal intubation rate. Doctor-paramedic teams delivering prehospital anaesthesia achieved definitive airway management for all patients. CONCLUSIONS: A significant proportion of severely injured trauma patients required advanced airway interventions to effectively treat airway compromise. Standard ambulance service interventions were only effective for a proportion of patients, but might not have always been applied appropriately. Complications of advanced airway management occurred in both provider groups, but failed intubation and unrecognized oesophageal intubation were a particular problem in the paramedic intubation group.

Identification and functional characterization of three novel alleles for the serotonin transporter-linked polymorphic region.

A promoter polymorphism in the serotonin transporter gene (5-HTTLPR) has been reported to confer relative risk for phenotypes (depression/anxiety) and endophenotypes (amygdala reactivity). In this report, we identify and characterize three rare 5-HTTLPR alleles not previously described in the human literature. The three novel alleles were identified while genotyping 5-HTTLPR in a family-based attention deficit hyperactivity disorder clinical population. Two of the novel alleles are longer than the common 16-repeat long (L) allele (17 and 18 repeats) and the third is significantly smaller than the 14-repeat short (S) allele (11 repeats). The sequence and genetic architecture of each novel allele is described in detail. We report a significant decrease in the expression between the XL17 (17r) allele and the L(A) (16r) allele. The XS11 (11r) allele showed similar expression with the S (14r) allele. A 1.8-fold increase in expression was observed with the L(A)(16r) allele compared with the L(G) (16r) allele, which replicates results from earlier 5-HTTLPR expression experiments. In addition, transcription factor binding site (TFBS) analysis was performed using MatInspector (Genomatix) that showed the presence or absence of different putative TFBSs between the novel alleles and the common L (16r) and S (14r) alleles. The identification of rare variants and elucidation of their functional impact could potentially lead to understanding the contribution that the rare variant may have on the inheritance/susceptibility of multifactorial common diseases.

Berberine and evodiamine influence serotonin transporter (5-HTT) expression via the 5-HTT-linked polymorphic region.

The aim of this study was to investigate the effect of berberine and evodiamine on serotonin transporter (5-HTT) expression and then test how allelic variations previously identified in the promoter region could modulate that effect in the serotonergic neuronal cell line RN46A. Both berberine and evodiamine, alone and in combination, increased 5-HTT mRNA and protein expression significantly across the various alleles. When tested against the S, XS(11), L(G), L(A), XL(17), and XL(18) alleles, respectively, 100 µM berberine increased 5-HTT promoter activities by 67%, 128.7%, 106.9%, 100.4%, 26.2% and 82%, 2 µM evodiamine increased 5-HTT promoter activities by 216.7%, 81.6%, 305.6%, 181.5%, 175.3% and 102.2%. Berberine and evodiamine increased 5-HTT promoter activity differently depending on the genetic variation of the 5-HTTLPR polymorphism. This study has provided a convincing example of how herbal compounds influence the expression of one of the most intensively studied psychiatric candidate genes, the serotonin transporter.

Pre-hospital use of ketamine in paediatric trauma.

OBJECTIVES: To describe the use of ketamine in children by a pre-hospital physician-based service. METHODS: A five and a half year retrospective database review of all patients aged <16 years who were attended by London's Helicopter Emergency Medical Service and given ketamine. RESULTS: One hundred and sixty-four children met the inclusion criteria. The median age was 10 years (range 0-15 years). One hundred and four (63%) had a Glasgow Coma Scale (GCS) of 15 and 153 (93%) had a GCS>8 before administration of ketamine. Patients received from 2 to 150 mg ketamine IV (mean=1.0 mg/kg) and 112 (68%) received concomitant midazolam (0.5-18 mg, mean=0.1 mg/kg). One hundred and forty-one (86%) received ketamine intravenously and 23 (14%) intramuscularly. Only 12 patients (7%) were trapped. The most common mechanisms of injury in those who received ketamine were road traffic collisions, burns and falls. CONCLUSION: The safe delivery of adequate analgesia and appropriate sedation is a priority in paediatric pre-hospital care. Ketamine was predominantly used in awake non-trapped patients with blunt trauma for procedural sedation and analgesia. Detailed database searches did not demonstrate loss of airway, oxygen desaturation or clinically significant emergence reactions after ketamine administration. This study failed to demonstrate any major side effects of the drug and reassured us that the safety profile of the drug in this environment is likely to be satisfactory. The use of ketamine in trapped children was rare.

Medical response times to major incidents: potential benefits of a regional air ambulance mutual aid scheme.

In the event of major incidents, neighbouring air ambulances can be used to assist. To assess the potential benefit of this cooperation, three fictitious major incidents were described to emergency service dispatch desks to assess the availability and response times for neighbouring air ambulances. A medical infrastructure at each site could be in place in a shorter time when the mutual aid scheme was used. This short study demonstrates the increased availability of doctors and flight paramedics that can be achieved by cooperation schemes. The costs of such schemes are minimal where air ambulances already exist. Ambulance services can use this type of scheme rapidly to place a comprehensive medical infrastructure for major incidents.

Predicting loneliness with polygenic scores of social, psychological and psychiatric traits.

Loneliness is a heritable trait that accompanies multiple disorders. The association between loneliness and mental health indices may partly be due to inherited biological factors. We constructed polygenic scores for 27 traits related to behavior, cognition and mental health and tested their prediction for self-reported loneliness in a population-based sample of 8798 Dutch individuals. Polygenic scores for major depressive disorder (MDD), schizophrenia and bipolar disorder were significantly associated with loneliness. Of the Big Five personality dimensions, polygenic scores for neuroticism and conscientiousness also significantly predicted loneliness, as did the polygenic scores for subjective well-being, tiredness and self-rated health. When including all polygenic scores simultaneously into one model, only 2 major depression polygenic scores remained as significant predictors of loneliness. When controlling only for these 2 MDD polygenic scores, only neuroticism and schizophrenia remain significant. The total variation explained by all polygenic scores collectively was 1.7%. The association between the propensity to feel lonely and the susceptibility to psychiatric disorders thus pointed to a shared genetic etiology. The predictive power of polygenic scores will increase as the power of the genome-wide association studies on which they are based increases and may lead to clinically useful polygenic scores that can inform on the genetic predisposition to loneliness and mental health.

Military perspective on the civilian response to the London bombings July 2005.

With the break up of the Warsaw Pact and changing global relations, current military deployments are becoming smaller and more expeditionary (e.g. Afghanistan, East Timor and Sierra Leone). During the Cold War, the use of weapons of mass effect was highly likely to have been seen on the battlefield. Ironically, the proliferation of CBRN agents and the knowledge of their application, as well as the manufacture of improvised explosive devices, have lead to the targeting of civilian populations by extremist groups. One of the benefits of military clinicians embedded in NHS hospital trusts, as well as a strong reservist cadre, is a greater understanding of the implications and management of asymmetric attacks against the U.K. The experience and skills of military clinicians may be of benefit to NHS trusts while this type of threat exists. Military clinicians are also likely to benefit from the experience that they get in certain NHS posts that provide skills that are readily transferable to military medicine. The events of 7th July highlighted the dynamic use of deployable medical resources and a rapid return to normal service provision. This type of 'Health Resilience' can only be achieved with a combination of effective emergency planning, on scene clinical risk management and clinical leadership.

Pre-hospital pelvic girdle injury: Improving diagnostic accuracy in a physician-led trauma service.

BACKGROUND: Examination of missed injuries in our physician-led pre-hospital trauma service indicated that the significant injuries missed were often pelvic fractures. We therefore conducted a study whose aim was to evaluate the pre-hospital diagnostic accuracy of pelvic girdle injuries, and how this would be affected by implementing the pelvic injury treatment guidelines recently published by the Faculty of Pre-Hospital Care. STUDY DESIGN: All blunt trauma patients attended in a 5-month period were included in the study. The presence or absence of pelvic girdle injury on computed tomography (CT) or, if unavailable, pelvic X-ray was used as a primary outcome measure. A retrospective database and case note review was conducted to identify patients who had pelvic binder applied in the study period. For the purposes of the study, pelvic ring and acetabular fractures were grouped together as patients with suspected pelvic girdle injury that should be fitted with a pelvic binder in the pre-hospital setting. The sensitivity and specificity, relating to the presence of pelvic girdle injury in patients with pelvic binders, was calculated in order to determine pre-hospital diagnostic accuracy. RESULTS: 785 patients were attended during the study period. 170 met the study inclusion criteria. 26 (15.3%) sustained a pelvic girdle injury. 45 (26.5%) had a pelvic binder applied. There were eight missed fractures (31%), of which the majority (six) sustained less severe injuries that were managed non-operatively. Two patients required operative fixation. Radiological images and/or reports were available on 169 (99.4%) patients. As a test of the presence of pelvic fracture, pelvic binder application had a sensitivity of 0.69 (95% CI 0.50-0.85) and a specificity of 0.81 (95% CI 0.74-0.87). CONCLUSIONS: Even with a careful clinical assessment and a low threshold for binder application, this study highlights the problems of distracting injury when trying to diagnose and manage pelvic fractures. By implementing the pelvic treatment guidelines published by the Faculty of Pre-hospital Care, the missed injury rate could be reduced from 31% to 8%.

Association of actin with the platelet membrane.

Human platelet membrane-actin associations were studied by means of differential extraction of purified membranes and low-shear viscometry of membrane-F-actin mixtures. As indicated by resistance to extraction with 0.6 M potassium iodide, a significant amount of platelet actin appears to be tightly associated with the membrane. When tested by falling-ball viscometry, both whole and KI-extracted membranes increased the low-shear viscosity of preformed rabbit skeletal muscle F-actin at physiologically reasonable pH and ionic conditions. This membrane-associated actin gelation activity was dependent upon low free calcium concentration (10(-8)-10(-7) M). The results are consistent with specific associations between actin and platelet membranes and may be relevant to membrane-cytoskeletal interactions believed to occur in the intact cell.

London bombings July 2005: the immediate pre-hospital medical response.

On July 7th 2005 a series of terrorist bombs exploded in London. The transport system was targeted and at least 54 passengers were killed and around 700 injured. This paper describes the immediate pre-hospital medical response to the four scenes. From the perspective of the London Helicopter Emergency Medical Service the deployment, difficulties on scene and the initial lessons learned are discussed.

Retinal blood flow during hyperglycemia. A laser Doppler velocimetry study.

The effect of different rates of glucose infusion on the retinal circulation was studied in Gottingen breed minipigs. Seven minipigs were made hyperglycemic rapidly with an intravenous bolus injection of 50% dextrose, after which a slow dextrose infusion maintained hyperglycemia for 60 minutes. Seven minipigs were more gradually made hyperglycemic over 60 minutes with a slow intravenous infusion of 50% dextrose, and a further seven had a control infusion of urea of equal volume and osmolality over 60 minutes. Retinal blood flow (RBF) was determined from the maximum (centerline) velocity of the blood (Vmax) (determined by bidirectional laser doppler velocimetry) and the vessel diameter (D) (determined from monochromatic fundus photographs). Measurements were made in a single temporal retinal vein of each animal at baseline, during, and after each of the infusions. Plasma glucose rose from 6.1 +/- 0.5-25.3 +/- 1.5 mM (mean +/- standard error) during the bolus infusion and from 6.4 +/- 0.7-22.0 +/- 0.7 mM during the slow infusion. The bolus and the slow glucose infusions both produced large increases in RBF (63% and 62%, respectively) which were mainly attributable to increases in Vmax. The urea infusion had no significant effect on RBF, Vmax, or D. The ocular perfusion pressure rose slowly and was significantly elevated after 60 minutes of slow glucose infusion but not after the urea infusion.

The state of actin polymerization in tetracaine-treated platelets.

Although there is considerable evidence that platelet activation is associated with polymerization of actin, it is not known whether there is some pre-existing F-actin even before activation. We have examined the state of actin polymerization in nonactivated platelets by deoxyribonuclease assay of G-actin and total actin. To fully suppress activation, platelets were prepared in the presence of tetracaine. The G-actin/total actin ratio in tetracaine-treated platelets prepared by three different methods is significantly less than one (average of all results = 0.61). We conclude that about 39% of total actin in nonactivated tetracaine-treated platelets may be present as F-actin.

Allelic variability in D21S11, but not in APP or APOE, is associated with cognitive decline in Down syndrome.

Genetic variation in the APOE gene and variation in chromosome 21 genotypes, including the APP locus, may influence age-associated cognitive decline in adults with Down syndrome. Molecular genetic and longitudinal neuropsychological analysis was performed for 41 unrelated Caucasian individuals (mean age 48.1 +/- 1.1 years (s.c.m.)) with free trisomy 21. Allele frequencies and genotype distributions were compared among subgroups with or without evidence of cognitive decline. Genetic variability at APOE and APP was not significantly associated with evidence of cognitive decline. However, aged individuals with Down syndrome, without evidence of cognitive decline, demonstrated unusual allelic variability at D21S11. These findings are discussed in the context of current hypotheses of Alzheimer-type dementia in Down syndrome and in the general population.

Berberine increases expression of GATA-2 and GATA-3 during inhibition of adipocyte differentiation.

It is known that a number of transcription factors are key regulators in the complex process of adipocyte differentiation including peroxisome proliferator activated receptor gamma (PPARgamma) and the CCAAT enhancer binding protein alpha (C/EBPalpha). Studies have demonstrated that in pre-adipocyte 3T3-L1 cells constitutive expression of the DNA binding proteins GATA-2 and GATA-3 results in protein/protein interactions with C/EBPalpha resulting in down regulation of PPARgamma and subsequent suppressed adipocyte differentiation with cells trapped at the pre-adipocyte stage. Thus it appears that GATA-2 and GATA-3 are of critical importance in regulating adipocyte differentiation through molecular interactions with PPARgamma and C/EBPalpha. Recent reports suggest that berberine, an isoquinoline derivative alkaloid isolated from many medicinal herbs prevents differentiation of 3T3-L1 cells via a down regulation of PPARgamma and C/EBPalpha expression. The aim of this study was to determine the effect of berberine on GATA-2 and 3 gene and protein expression levels during differentiation of 3T3-L1 cells. MTT (Methylthiazolyldiphenyl-tetrazolium bromide) was used to detect the cytotoxic effects of berberine on the viability of 3T3-L1 cells during proliferation and differentiation. Differentiation of 3T3-L1 cells was monitored by Oil Red O staining and RT-PCR of PPARgamma and C/EBPalpha and the expression of GATA-2 and 3 was determined by RT-PCR and Western Blot. Results show that following treatment with 8microM berberine the mRNA and protein expression levels of GATA-2 and 3 were elevated and accompanied by inhibited adipocyte differentiation. These results may lead to the use of berberine to target the induction of specific genes such as GATA-2 and GATA-3 which affect adipocyte differentiation.

Is the prevalence of deliberate penetrating trauma increasing in London? Experiences of an urban pre-hospital trauma service.

BACKGROUND: Recent media interest in stabbings and shootings has lead to the general assumption that injury and death secondary to deliberate penetrating trauma are rising. The aim of this study was to establish the prevalence of deliberate penetrating trauma within a London-based physician-led pre-hospital trauma service, and evaluate whether the perceived increase reported by the media translates into a real increase in penetrating trauma caseload. METHOD: A retrospective review of a physician-led pre-hospital care trauma database was conducted to identify all patients who sustained stabbing or shooting injuries over a 16-year period. Patients who died in the pre-hospital phase and paediatric patients were included. Other local and national datasets were examined to determine whether similar trends were observed. RESULTS: 1564 penetrating trauma victims were identified, including 92 children. 1358 patients (86.8%) sustained stab wounds; 206 patients were shot (13.2%). Penetrating injury accounted for 9.9% of the overall trauma caseload during the study period. The annual increase in patients sustaining stabbing injuries was 23.2%. Gun shot wounds increased by 11.0% per year. CONCLUSION: The study demonstrates a significant annual rise in the number of cases of deliberate penetrating trauma managed by a UK physician-led pre-hospital trauma service.