Zoledronic acid boosts γδ T-cell activity in children receiving αβ+ T and CD19+ cell-depleted grafts from an HLA-haplo-identical donor.
We demonstrated that γδ T cells of patients given HLA-haploidentical HSCT after removal of αβ+ T cells and CD19+ B cells are endowed with the capacity of killing leukemia cells after ex vivo treatment with zoledronic acid (ZOL). Thus, we tested the hypothesis that infusion of ZOL in patients receiving this type of graft may enhance γδ T-cell cytotoxic activity against leukemia cells. ZOL was infused every 28 d in 43 patients; most were treated at least twice. γδ T cells before and after ZOL treatments were studied in 33 of these 43 patients, till at least 7 mo after HSCT by high-resolution mass spectrometry, flow-cytometry, and degranulation assay. An induction of Vδ2-cell differentiation, paralleled by increased cytotoxicity of both Vδ1 and Vδ2 cells against primary leukemia blasts was associated with ZOL treatment. Cytotoxic activity was further increased in Vδ2 cells, but not in Vδ1 lymphocytes in those patients given more than one treatment. Proteomic analysis of γδ T cells purified from patients showed upregulation of proteins involved in activation processes and immune response, paralleled by downregulation of proteins involved in proliferation. Moreover, a proteomic signature was identified for each ZOL treatment. Patients given three or more ZOL infusions had a better probability of survival in comparison to those given one or two treatments (86% vs. 54%, respectively, p = 0.008). Our data indicate that ZOL infusion in pediatric recipients of αβ T- and B-cell-depleted HLA-haploidentical HSCT promotes γδ T-cell differentiation and cytotoxicity and may influence the outcome of patients.


Hypertension awareness and psychological distress.
There is conflicting evidence regarding the association of hypertension with psychological distress, such as anxiety and depressive symptoms. The association may be because of a direct effect of the raised blood pressure, adverse effects of treatment, or the consequences of labeling. In a representative study of 33 105 adults (aged 51.7+/-12.1 years; 45.8% men), we measured levels of psychological distress using the 12-item General Health Questionnaire and collected blood pressure, data on history of hypertension diagnosis, and medication usage. Awareness of hypertension was confirmed through a physician diagnosis or the use of antihypertensive medication, and unaware hypertension was defined by elevated clinic blood pressure (systolic/diastolic > or =140/90 mm Hg) without previous treatment or diagnosis. In comparison with normotensive participants, an elevated risk of distress (General Health Questionnaire score > or =4) was observed in aware hypertensive participants (multivariable adjusted odds ratio: 1.57 [95% CI: 1.41 to 1.74]) but not in unaware hypertensives (odds ratio: 0.91 [95% CI: 0.78 to 1.07]). Antihypertensive medication and comorbidity were also associated with psychological distress, although this did not explain the greater risk of distress in aware hypertensives. We observed a weak curvilinear association between systolic blood pressure and distress, which suggested that distressed participants were more likely to have low or highly elevated blood pressure. These findings suggest that labeling individuals as hypertensive, rather than having elevated blood pressure, per se, may partially explain the greater levels of distress in patients treated for hypertension.


Estimating absolute methylation levels at single-CpG resolution from methylation enrichment and restriction enzyme sequencing methods.
Recent advancements in sequencing-based DNA methylation profiling methods provide an unprecedented opportunity to map complete DNA methylomes. These include whole-genome bisulfite sequencing (WGBS, MethylC-seq, or BS-seq), reduced-representation bisulfite sequencing (RRBS), and enrichment-based methods such as MeDIP-seq, MBD-seq, and MRE-seq. These methods yield largely comparable results but differ significantly in extent of genomic CpG coverage, resolution, quantitative accuracy, and cost, at least while using current algorithms to interrogate the data. None of these existing methods provides single-CpG resolution, comprehensive genome-wide coverage, and cost feasibility for a typical laboratory. We introduce methylCRF, a novel conditional random fields-based algorithm that integrates methylated DNA immunoprecipitation (MeDIP-seq) and methylation-sensitive restriction enzyme (MRE-seq) sequencing data to predict DNA methylation levels at single-CpG resolution. Our method is a combined computational and experimental strategy to produce DNA methylomes of all 28 million CpGs in the human genome for a fraction (<10%) of the cost of whole-genome bisulfite sequencing methods. methylCRF was benchmarked for accuracy against Infinium arrays, RRBS, WGBS sequencing, and locus-specific bisulfite sequencing performed on the same human embryonic stem cell line. methylCRF transformation of MeDIP-seq/MRE-seq was equivalent to a biological replicate of WGBS in quantification, coverage, and resolution. We used conventional bisulfite conversion, PCR, cloning, and sequencing to validate loci where our predictions do not agree with whole-genome bisulfite data, and in 11 out of 12 cases, methylCRF predictions of methylation level agree better with validated results than does whole-genome bisulfite sequencing. Therefore, methylCRF transformation of MeDIP-seq/MRE-seq data provides an accurate, inexpensive, and widely accessible strategy to create full DNA methylomes.


[Determination of carminomycin and its basic metabolite 13-dihydrocarminomycin by a highly efficient liquid chromatographic method with a fluorescent detector].
Carminomycin, an antitumor antibiotic, and 13-dihydrocarminomycin, its main metabolite, were determined in the blood plasma of patients with highly efficient liquid chromatography and a fluorescent detector (lambda ex=492 nm, lambda em=538 nm). The liquid chromatograph manufactured by Spectro-Physics, model SP-8000, with Particyl Column 10/25 was used for the determination. The composition of the mobile phase of acetonitrile--0.1 M H3PO4 was 93:7 by the volume. The mobility speed was 2 ml/min. The retention time of carminomycin, rubomycin (used as the national standard) and dihydrocarminomycin was 6.8, 8 and 10.2 minutes, respectively. All three substances showed symmetrical peaks with high resolution. The early phase kinetics of carminomycin after its intravenous injection to the patients in a dose of 24 mg (18 mg/m2) is satisfactorily described by the diexponential equation. The pharmacokinetic parameters of carminomycin corresponding to the open two-compartmental system are presented. Highly efficient liquid chromatography may be used for the pharmacokinetic description of biotransformation of carminomycin and some other antibiotics of the anthracycline group.


Fermentation of dietary fibre components in the rat intestinal tract.
1. The fermentative breakdown of dietary fibre from various sources in the intestinal tract was studied using rat balance experiments and gas-liquid chromatograhic analysis of dietary fibre monomers in feed and faces. 2. On a basal diet with 690 g maize starch/kg but no added fibre, small but detectable amounts of polymeric glucose, rhamnose, arabinose, xylose, galactose, mannose and uronic acids, i.e. sugars occurring in dietary fibre, were excreted in faeces. 3. Dietary fibre in wheat bran was rather resistant to fermentation; 63% was recovered in the faeces. Guar gum, on the other hand, was almost completely fermented, whereas 19 and 25% of the uronic acids in low and high methoxylated pectin respectively, were excreted in faeces. The various constituents of sugar-beet dietary fibre (approximately equal amounts of arabinose-based hemicellulose, pectin and non-starch glucan (cellulose)) showed quite variable availability for micro-organisms in that 6-12% of the arabinose, 17-25% of the uronic acids, and 52-58% of the cellulose were recovered in the faeces. 4. Faecal nitrogen excretion increased on addition of any one of the dietary fibre preparations studied, resulting in decreased true and apparent protein digestibility values. 5. The faecal dry weight increment was most pronounced when feeding bran and could then almost be accounted for by the remaining fibre and by protein. The less-prominent bulking effect of guar gum and pectins, that were much more extensively fermented, could be only partly explained by dietary fibre and protein.


Communication in refugee and migrant mental healthcare: A systematic rapid review on the needs, barriers and strategies of seekers and providers of mental health services.
Migrants and refugees may not access mental health services due to linguistic and cultural discordance between them and health and social care professionals (HSCPs). The aim of this review is to identify the communication needs and barriers experienced by third-country nationals (TCNs), their carers, and HSCPs, as well as the strategies they use and their preferences when accessing/providing mental health services and language barriers are present. We undertook a rapid systematic review of the literature (01/01/2011 - 09/03/2022) on seeking and/or providing mental health services in linguistically discordant settings. Quality appraisal was performed, data was extracted, and evidence was reviewed and synthesised qualitatively. 58/5,650 papers met the inclusion criteria. Both TCNs (and their carers) and HSCPs experience difficulties when seeking or providing mental health services and language barriers are present. TCNs and HSCPs prefer linguistically and culturally concordant provision of mental health services but professional interpreters are often required. However, their use is not always preferred, nor is it without problems. Language barriers impede TCNs' access to mental health services. Improving language support options and cultural competency in mental health services is crucial to ensure that individuals from diverse linguistic and cultural backgrounds can access and/or provide high-quality mental health services.


Estimating the diversity, completeness, and cross-reactivity of the T cell repertoire.
In order to recognize and combat a diverse array of pathogens the immune system has a large repertoire of T cells having unique T cell receptors (TCRs) with only a few clones specific for any given antigen. We discuss how the number of different possible TCRs encoded in the genome (the potential repertoire) and the number of different TCRs present in an individual (the realized repertoire) can be measured. One puzzle is that the potential repertoire greatly exceeds the realized diversity of naïve T cells within any individual. We show that the existing hypotheses fail to explain why the immune system has the potential to generate far more diversity than is used in an individual, and propose an alternative hypothesis of "evolutionary sloppiness." Another immunological puzzle is why mice and humans have similar repertoires even though humans have over 1000-fold more T cells. We discuss how the idea of the "protecton," the smallest unit of protection, might explain this discrepancy and estimate the size of "protecton" based on available precursor frequencies data. We then consider T cell cross-reactivity - the ability of a T cell clone to respond to more than one epitope. We extend existing calculations to estimate the extent of expected cross-reactivity between the responses to different pathogens. Our results are consistent with two observations: a low probability of observing cross-reactivity between the immune responses to two randomly chosen pathogens; and the ensemble of memory cells being sufficiently diverse to generate cross-reactive responses to new pathogens.


Some challenges and opportunities for grazing dairy cows on temperate pastures.
Grazing plays an important role in milk production in most regions of the world. In this review, some challenges to the grazing cow are discussed together with opportunities for future improvement. We focus on daily feed intake, efficiency of pasture utilization, output of milk per head, environmental impact of grazing and the nutritional quality to humans of milk produced from dairy cows in contrasting production systems. Challenges are discussed in the context of a trend towards increased size of individual herds and include limited and variable levels of daily herbage consumption, lower levels of milk output per cow, excessive excretion of nitrogenous compounds and requirements for minimal periods of grazing regardless of production system. A major challenge is to engage more farmers in making appropriate adjustments to their grazing management. In relation to product quality, the main challenge is to demonstrate enhanced nutritional/processing benefits of milk from grazed cows. Opportunities include more accurate diet formulations, supplementation of grazed pasture to match macro- and micronutrient supply with animal requirement and plant breeding. The application of robotics and artificial intelligence to pasture management will assist in matching daily supply to animal requirement. Wider consumer recognition of the perceived enhanced nutritional value of milk from grazed cows, together with greater appreciation of the animal health, welfare and behavioural benefits of grazing should contribute to the future sustainability of demand for milk from dairy cows on pasture.


A charge coupled device camera with electron decelerator for intermediate voltage electron microscopy.
Electron microscopists are increasingly turning to intermediate voltage electron microscopes (IVEMs) operating at 300-400 kV for a wide range of studies. They are also increasingly taking advantage of slow-scan charge coupled device (CCD) cameras, which have become widely used on electron microscopes. Under some conditions, CCDs provide an improvement in data quality over photographic film, as well as the many advantages of direct digital readout. However, CCD performance is seriously degraded on IVEMs compared to the more conventional 100 kV microscopes. In order to increase the efficiency and quality of data recording on IVEMs, we have developed a CCD camera system in which the electrons are decelerated to below 100 kV before impacting the camera, resulting in greatly improved performance in both signal quality and resolution compared to other CCDs used in electron microscopy. These improvements will allow high-quality image and diffraction data to be collected directly with the CCD, enabling improvements in data collection for applications including high-resolution electron crystallography, single particle reconstruction of protein structures, tomographic studies of cell ultrastructure, and remote microscope operation. This approach will enable us to use even larger format CCD chips that are being developed with smaller pixels.


Positional Change Used to Manage Postextubation Respiratory Failure in a Child With Cerebral Palsy.
Dental treatment for patients with cerebral palsy (CP) is often performed under general anesthesia due to involuntary movements that can render dental treatment difficult. Since CP is often accompanied by spasticity, care must be taken when positioning patients during general anesthesia. We report the management of a 14-year-old girl with CP and epilepsy undergoing general anesthesia for dental treatment who experienced respiratory failure due to acute thoracoabdominal muscle hypertonia after extubation. She had a history of cardiac arrest due to respiratory failure caused by acute muscle hypertonia and successful resuscitation. General anesthesia was induced after careful positioning of the patient to prevent spastic muscle stretching, and the dental treatment was completed without complications. However, upon awakening after extubation, the patient developed respiratory failure due to acute muscle hypertonia. The patient was resedated and repositioned from a supine to a sitting position, and her symptoms improved. There was no recurrence of muscle hypertonia, and she recovered fully without complications. In this case, respiratory failure associated with acute muscle hypertonia was successfully managed by position change after initial treatment with positive-pressure ventilation and propofol. It is important to be prepared for the possibility of respiratory failure associated with acute muscle hypertonia and its countermeasures when providing general anesthesia for patients with CP.


Protocol for a feasibility study to inform the development of a multicentre randomised controlled trial of asthma-tailored pulmonary rehabilitation versus usual care for individuals with severe asthma.
Pulmonary rehabilitation with core components of exercise training and multiprofessional education is an integral part of the management of patients with chronic lung disease. International guidelines for individuals with asthma recommend exercise as exercise improves symptoms, indices of cardiopulmonary efficiency, health status and psychosocial outcome. However, there is little published evidence evaluating safety and acceptability of exercise training for individuals with severe asthma and there are concerns regarding exercise-induced asthma. We propose a feasibility study for a multicentre randomised controlled trial (RCT) of asthma-tailored pulmonary rehabilitation (asthma-tailored PR) versus usual care in individuals with severe asthma. The study will be conducted in three stages. Adults with severe asthma will be included if they have persistent symptoms despite being at step 4 or 5 of the British Thoracic Society guidelines. Stage 1: semistructured interviews will be used in a sample of 20-30 individuals with severe asthma to understand the experience and attitudes of this population towards exercise. Stage 2: eight focus groups of at least six healthcare professionals involved in the care of patients with severe asthma will be conducted to understand their attitudes towards exercise for this population. Stage 3: a small-scale RCT of the proposed multicentre RCT of asthma-tailored PR versus usual care for individuals with severe asthma will be conducted. The primary outcome measures will be recruitment, retention and adverse event rates. Semistructured interviews with participants of stage 3 will be used to identify further barriers or facilitators to participation in PR and the trial. Thematic analysis will be used for the interpretation of all interviews. The study results will inform the design of a larger multicentre RCT. The National Research Ethics Service Committee East Midland approved the study protocol. ISRCTN96143888.


Regulatory elements required for development of caenorhabditis elegans hermaphrodites are conserved in the tra-2 homologue of C. remanei, a male/female sister species.
The Caenorhabditis elegans hermaphrodite is essentially a female that produces sperm. In C. elegans, tra-2 promotes female fates and must be repressed to achieve hermaphrodite spermatogenesis. In an effort to learn how mating systems evolve, we have cloned tra-2 from C. remanei, the closest gonochoristic relative of C. elegans. We found its structure to be similar to that of Ce-tra-2 but its sequence to be divergent. RNA interference demonstrates that Cr-tra-2 promotes female fates. Two sites of tra-2 regulation are required for the onset of hermaphrodite spermatogenesis in C. elegans. One, the MX region of TRA-2, is as well conserved in C. remanei as it is in C. briggsae (another male/hermaphrodite species), suggesting that this control is not unique to hermaphrodites. Another, the DRE/TGE element of the tra-2 3' UTR, was not detected by sequence analysis. However, gel-shift assays demonstrate that a factor in C. remanei can bind specifically to the Cr-tra-2 3' UTR, suggesting that this translational control is also conserved. We propose that both controls are general and do not constitute a novel "switch" that enables sexual mosaicism in hermaphrodites. However, subtle quantitative or qualitative differences in their employment may underlie differences in mating system seen in Caenorhabditis.


Investigation of the interactions between silver nanoparticles and Hela cells by scanning electrochemical microscopy.
The interactions between Hela cells and silver nanoparticles (AgNPs) have been studied by scanning electrochemical microscopy (SECM) with both IrCl(6)(2-/3-) and Fe(CN)(6)(3-/4-) as the dual mediators. IrCl(6)(2-), which can be produced in situ and react with AgNPs, is used as the mediator between the AgNPs on the cells and the SECM tip. Another redox couple, Fe(CN)(6)(3-/4-), which has a similar hydrophilicity to IrCl(6)(2-/3-), but cannot react with AgNPs, is also employed for the contrast experiments. The cell array is cultured successfully onto a Petri dish by microcontact printing (muCP) technique, which can provide a basic platform for studying of single cells. The approach curve and line scan are the two methods of SECM employed here to study the Hela cells. The former can provide the information about the interaction between Hela cells and AgNPs whereas the later gives the cell imaging. The permeability of cell membranes and morphology are two main factors which have effects on the feedback mode signals when K(3)Fe(CN)(6) is used as the mediator. The permeability of the cell membranes can be ignored after interaction with high concentration of AgNP solution and the height of the Hela cells is slightly decreased in this process. The kinetic rate constants (k(0)) between IrCl(6)(2-) and Ag on the Hela cell can be evaluated using K(3)IrCl(6) as the mediator, and they are increased with the higher concentrations of the AgNP solutions. The k(0) is changed about 10 times from 0.43 +/- 0.04 x 10(-4) to 1.25 +/- 0.07 x 10(-4) and to 3.93 +/- 1.9 x 10(-4) cm s(-1) corresponding to 0, 1 and 5 mM of AgNO(3) solution. The experimental results demonstrate that the AgNPs can be adsorbed on the cell surface and detected by SECM. Thus, the amount of AgNPs adsorbed on cell membranes and the permeability or morphology changes can be investigated simultaneously using this approach. The dual mediator system and cell array fabricated by muCP technique can provide better reproducibility because they can simplify experiments, and provide a platform for further single cell detection.


Hemodialysis in children: how, when and why.
End-stage renal diseases requiring chronic dialysis are rare in childhood and adolescence, but they are associated with high mortality and impaired quality of life (1, 2). The most common disease that causes chronic kidney disease (CKD) is primary glomerular disease (GD), followed by congenital abnormalities of the kidney and urinary tract, cystic, hereditary or congenital disorders and, more rarely, secondary GD. However, patients with secondary GD, urologic disorders, and metabolic diseases have greater mortality risk than patients with primary GD (3). Here, we focused on the different options of treatment available, and specifically we compared peritoneal dialysis and hemodialysis, showing pros and cons between them.


Marker-dependent recombination in T4 bacteriophage. II. The evaluation of mismatch repairabilities in crosses within indicator distances.
The contribution of mismatch repair to genetic recombination in T4 phage has been evaluated by three independent approaches: (1) testing for non-additivity of recombinant frequencies; (2) measurements of double exchange frequencies in three-factor crosses: (3) comparisons of recombination abilities of mutations occupying the same site. Quantitative agreement among the results of these approaches suggests that within distances much less than the mean length of hybrid regions, mismatch repair accounts perfectly for high negative interference as measured in three-factor crosses and as manifested by non-additivity in two-factor crosses. The mismatch repair mechanism readily recognizes only particular mismatches, the repair frequency being dependent on the base sequence in both strands of the mismatched region.


Evaluation of the 2015 ATA Guidelines in Patients With Distant Metastatic Differentiated Thyroid Cancer.
Current American Thyroid Association (ATA) Management Guidelines for the treatment of differentiated thyroid cancer (DTC) stratify patients to decide on additional radioiodine (RAI) therapy after surgery, and to predict recurring/persisting disease. However, studies evaluating the detection of distant metastases and how these guidelines perform in patients with distant metastases are scarce. To evaluate the 2015 ATA Guidelines in DTC patients with respect to 1) the detection of distant metastases, and 2) the accuracy of its Risk Stratification System in patients with distant metastases. We retrospectively included 83 DTC patients who were diagnosed with distant metastases around the time of initial therapy, and a control population of 472 patients (312 low-risk, 160 intermediate-risk) who did not have a routine indication for RAI therapy. We used the control group to assess the percentage of distant metastases that would have been missed if no RAI therapy was given. Two hundred forty-six patients had no routine indication for RAI therapy of which 4 (1.6%) had distant metastases. Furthermore, among the 83 patients with distant metastases, 14 patients (17%) had excellent response, while 55 (67%) had structural disease after a median follow-up of 62 months. None of the 14 patients that achieved an excellent response had a recurrence. In patients without a routine indication for RAI therapy according to the 2015 ATA Guidelines, distant metastases would initially have been missed in 1.6% of the patients. Furthermore, in patients with distant metastases upon diagnosis, the 2015 ATA Guidelines are an excellent predictor of both persistent disease and recurrence.


Characterization of fig operon mutants of Francisella novicida U112.
Francisella species secrete a polycarboxylate siderophore that resembles rhizoferrin to acquire ferric iron. Several of the Francisella siderophore synthesis genes are contained in a Fur-regulated operon (designated fig or fsl) comprised of at least seven ORFs including fur. Reverse transcriptase-PCR showed transcriptional linkage between figD and figE and between figE and figF. Mutations were constructed in four of these ORFs (figB, figC, figD, and figE) in Francisella novicida U112. All four of these new mutants and a F. novicida figA mutant grew at rates comparable to that of wild type under iron-replete conditions but growth of all five mutants was stunted in iron-limiting media. When ferric rhizoferrin was added to the iron-limited media, growth of the figA, figB, figC, and figD mutants was restored to levels similar to those obtained in iron-replete media. However, this exogenously added siderophore could not rescue the figE mutant. When Chrome Azurol S assays were used to measure siderophore production, the figA, figB, and figC mutants were markedly deficient in their ability to synthesize siderophore whereas the figD and figE mutants produced siderophore at levels equivalent to the wild-type parent strain.


Selecting subjects for participation in clinical research: one sphere of justice.
Recent guidelines from the US National Institutes of Health (NIH) mandate the inclusion of adequate numbers of women in clinical trials. Ought such standards to apply internationally? Walzer's theory of justice is brought to bear on the problem, the first use of the theory in research ethics, and it argues for broad application of the principle of adequate representation. A number of practical conclusions for research ethics committees (RECs) are outlined. Eligibility criteria in clinical trials ought to be justified by trial designers. Research ethics committees ought to question criteria that seem to exclude unnecessarily women from research participation. The issue of adequate representation should be construed broadly, so as to include consideration of the representation of the elderly, persons with HIV, mental illness and substance abuse disorders in clinical research.


Effects of four Bifidobacteria on obesity in high-fat diet induced rats.
To compare the effects of four Bifidobacteria strains (Bifidobacteria L66-5, L75-4, M13-4 and FS31-12, originated from normal human intestines) on weight gain, lipid metabolism, glucose metabolism in an obese murine model induced by high-fat diet. Forty-eight Sprague-Dawley rats were randomly divided into six groups. Control group received standard chow, model group received high-fat diet, and intervention groups received high-fat diet added with different Bifidobacteria strains isolated from healthy volunteers' fresh feces. All rats were executed at the 6th weekend. Body weight (BW), obese indexes, oral glucose tolerance test, serum and liver lipid and serum insulin (INS) were tested. Liver lipid deposition was classified pathologically. Compared with the model group, B. M13-4 improved BW gains (264.27 +/- 26.91 vs 212.55 +/- 18.54, P = 0.001) while B. L66-5 induced a decrease in BW (188.47 +/- 11.96 vs 212.55 +/- 18.54, P = 0.043). The rest two strains had no significant change in BW. All the four strains can reduce serum and liver triglyceride and significantly alleviate the lipid deposition in liver. All strains showed a trend of lowing serum and liver total cholesterol while B. L66-5 and B. FS31-12 did so more significantly. In addition, all the four strains showed no significant differences in serum INS and glucose level. The response of energy metabolism to administration of Bifidobacteria is strain dependent. Different strains of Bifidobacteria might drive different directions of fat distribution.


Evaluation of kisspeptin levels in obese pregnancy as a biomarker for pre-eclampsia.
Circulating concentrations of the peptide kisspeptin have been proposed as a novel biomarker for early detection of pre-eclampsia. Our aims were to assess analytical and clinical performance characteristics of a commercial kisspeptin assay and to determine sensitivity and specificity of the test for pre-eclampsia. Prospective, longitudinal study in a United Kingdom tertiary referral Antenatal Metabolic Clinic. Severely obese (body mass index, BMI > 40 kg/m(2), n = 194) and lean (BMI < 25 kg/m(2), n = 78) pregnant women. A commercial kisspeptin ELISA (Phoenix Pharmaceuticals) was assessed for analytical sensitivity, specificity, precision, linearity, recovery and stability in maternal plasma samples at 16, 28 and 36 weeks gestation. Pre-eclampsia, defined using International Society for the Study of Hypertension in Pregnancy guidelines; blood pressure; delivery gestation; birthweight. Kisspeptin concentrations were lower in early pregnancy in obese women (P < 0.001), and in women who later developed pre-eclampsia (P < 0.05), compared with women with uncomplicated pregnancies. For 16-week plasma kisspeptin in prediction of pre-eclampsia, area under the receiver-operator characteristic curve was 0.80 (P < 0.01), positive and negative likelihood ratios were 3.0 and 0.2, and test sensitivity and specificity were 85.7 and 71.4%, respectively. In regression analyses, kisspeptin (16 weeks) associated positively with delivery gestation (P < 0.05) and birthweight (P < 0.0001), and negatively with 28- and 36-week blood pressure (P < 0.0001). Kisspeptin concentration in early pregnancy is a promising biomarker for pre-eclampsia and low birthweight but cannot be recommended, in isolation, for universal screening because of inadequate test sensitivity and specificity. Large-scale studies are required to assess its potential in a panel of biomarkers.


Pharmacochemical profile of iopromide.
Iopromide (Schering, Berlin) is a new nonionic, monomeric contrast medium containing three different substituents on the triiodinated benzene ring. Iopromide exhibits low osmolality and viscosity in aqueous solutions of high concentrations. It has been shown to have a remarkably low intravenous toxicity in mice and rats. Neural tolerance was found to be equal to or better than that of metrizamide when injected in rats intracisternally and intracerebrally, respectively. The effects of iopromide after selective peripheral and cerebral arterial injections in rats were demonstrated to be very moderate at high dosages. The interaction of iopromide with proteins and membranes was found to be considerably low due to its hydrophilicity. Excretion of iopromide is fast and predominantly by the renal route. On the basis of the preclinical profile iopromide is a very promising contrast agent, being most suitable for all angiographic indications, including digital subtraction angiography, urography, and computed tomography.


FCHO controls AP2's initiating role in endocytosis through a PtdIns(4,5)P2-dependent switch.
Clathrin-mediated endocytosis (CME) is the main mechanism by which mammalian cells control their cell surface proteome. Proper operation of the pivotal CME cargo adaptor AP2 requires membrane-localized Fer/Cip4 homology domain-only proteins (FCHO). Here, live-cell enhanced total internal reflection fluorescence-structured illumination microscopy shows that FCHO marks sites of clathrin-coated pit (CCP) initiation, which mature into uniform-sized CCPs comprising a central patch of AP2 and clathrin corralled by an FCHO/Epidermal growth factor potential receptor substrate number 15 (Eps15) ring. We dissect the network of interactions between the FCHO interdomain linker and AP2, which concentrates, orients, tethers, and partially destabilizes closed AP2 at the plasma membrane. AP2's subsequent membrane deposition drives its opening, which triggers FCHO displacement through steric competition with phosphatidylinositol 4,5-bisphosphate, clathrin, cargo, and CME accessory factors. FCHO can now relocate toward a CCP's outer edge to engage and activate further AP2s to drive CCP growth/maturation.


Perceptions of video-based appointments from the patient's home: a patient survey.
We examined patient interest in a telehealth model in which the patient supplies the hardware and Internet connectivity to meet with a healthcare provider from his or her home via video call (video appointment). We hoped to understand prospectively the desirability, feasibility, and viability from the patient perspective. A phone survey was conducted of a random sample of patients who had been seen in the outpatient setting at a single institution. The sample was stratified by proximity to the local institution with oversampling for patients living outside a 120-mile radius. Out of 500 total patients, 301 patients responded, and 263 met the inclusion criteria. Of those 263 respondents, 38% indicated "very likely" to accept an invitation to see their provider via video, 28.1% "somewhat likely," and 33.8% "not at all likely." Of respondents, 75% have broadband, although only 36% reported having a Web camera. The primary factors affecting willingness to participate in a video appointment include comfort in setting up a video call, age, and distance participants would have traveled for an in-clinic appointment. Patient survey data indicate that most patients are likely to be accepting of telehealth care to the home using video call and that most have the required technology. Nevertheless, there are still significant hurdles to effectively implement this adaptation of telehealth care as part of mainstream practice.


A Structural Study of Escherichia coli Cells Using an In Situ Liquid Chamber TEM Technology.
Studying cell microstructures and their behaviors under living conditions has been a challenging subject in microbiology. In this work, in situ liquid chamber TEM was used to study structures of Escherichia coli cells in aqueous solutions at a nanometer-scale resolution. Most of the cells remained intact under electron beam irradiation, and nanoscale structures were observed during the TEM imaging. The analysis revealed structures of pili surrounding the E. coli cells; the movements of the pili in the liquid were also observed during the in situ tests. This technology also allowed the observation of features of the nucleoid in the E. coli cells. Overall, in situ TEM can be applied as a valuable tool to study real-time microscopic structures and processes in microbial cells residing in native aqueous solutions.


Renal artery stenosis in kidney transplants.
Transplant renal artery stenosis (TRAS) is an increasingly recognized complication of renal transplantation, with a reported incidence of between 1% and 23%. The clinical features include refractory hypertension, new-onset hypertension, allograft dysfunction, and the presence of a bruit over the graft. In this report, we describe the investigation and treatment of one such patient and review the current diagnostic approaches and therapy in this setting.


Association between Behavioral Ambidexterity and Brain Health.
Appropriately handling and switching exploration of novel knowledge and exploitation of existing knowledge is a fundamental element of genuine innovation in society. Moreover, a mounting number of studies have suggested that such "ambidexterity" is associated not only with organizational performance but also with the human brain. Among these reports, however, there have not been any definitive MRI-based parameters that objectively and easily evaluate such ambidexterity. Therefore, an MRI-based index derived from gray matter volume, called the gray-matter brain healthcare quotient (GM-BHQ), was used to measure the association between ambidexterity and the entire human brain. For this purpose, 200 healthy adults were recruited as subjects to undergo structural T1-weighted imaging and to answer multiple psychological questionnaires. Ambidexterity was evaluated using two scales: the Curiosity and Exploration Inventory II and the Short Grit Scale, as exploration-exploitation indicators of curiosity and grit, respectively. Additionally, to enrich the understanding of these associations, three additional positive thinking scales were used-the General Self-Efficacy Scale, the Rosenberg Self-Esteem Scale, and the Life Orientation Test-to evaluate self-efficacy, self-esteem, and optimism, respectively. The authors discovered the GM-BHQ was weakly associated with curiosity, grit, and self-efficacy individually after controlling for age and sex. Furthermore, the GM-BHQ was directly associated with curiosity but indirectly associated with grit in the path model. However, no significant association was found between the GM-BHQ and the other outcome indicators (i.e., self-esteem and optimism). These results suggest that brain health is weakly associated with ambidexterity evaluated using psychological tests.


Construction and validation of a predictive model for the risk of bowel resection in adults with incarcerated groin hernia.
It is difficult to definitively determine the degree of ischemia in the bowel in which an incarcerated groin hernia is embedded. Failure to diagnose and intervene promptly and accurately increases the rate of bowel resection and patient mortality. The aim of this study is to investigate the risk factors for incarcerated inguinal hernia complicating bowel necrosis with resection and to establish a predictive model as a reference for clinical work. Patients with incarcerated groin hernia who were admitted to our hospital were retrospectively analyzed. They were divided into bowel resection and non-bowel resection groups based on whether bowel resection was performed in the surgical record and postoperative pathological results. Risk factors for the development of bowel resection in incarcerated groin hernia were analyzed by univariate analysis and multivariate logistic regression, respectively. The screened independent risk factors were used to establish a prediction model, and finally, the predictive ability and accuracy of the model were validated and the clinical benefit was analyzed. A total of 345 patients with incarcerated groin hernia were included, of whom 58 underwent bowel resection for bowel necrosis and 287 did not. Multifactorial logistic regression analysis identified bowel obstruction (OR, 7.285 [95% CI, 2.254-23.542], P = 0.001), peritonitis (OR, 16.786 [95% CI, 5.436-51.838], P = 0.000), duration of incarcerated groin hernia (OR, 1.009 [95% CI, 1. 001-1.018], P = 0.034), heart rate (OR, 1.109 [95% CI, 1.021-1.205], P = 0.014), and preoperative total protein (OR, 0.900 [95% CI, 0.836-0.969], P = 0.005) were independent risk factors for bowel resection in incarcerated groin hernia. The predictive value of the established prediction model was basically in agreement with the measured value with a consistency index of 0.938 (0.901-0.974) and had a good clinical benefit. Clinical screening and management of independent risk factors for bowel resection in patients with incarcerated groin hernia should be strengthened. The predictive model developed in this study has high diagnostic efficacy for bowel resection associated with incarcerated inguinal hernia, with the aim of reducing the incidence of bowel resection and unplanned secondary surgery.


Autotitrating CPAP as a Tool for CPAP Initiation for Children.
Few studies have assessed autotitrating positive airway pressure (autoPAP) for treatment of obstructive sleep apnea (OSA) in children. We aimed to review our use of autoPAP for initiation of continuous positive airway pressure (CPAP) therapy in children, and compare autoPAP-derived treatment pressures to CPAP treatment pressure determined by attended polysomnography (PSG). Retrospective review of children initiated on autoPAP from 2013 to 2015. Mean autoPAP pressure (AutoMean pressure) and average device pressure ≤ 90% of time (Auto90 pressure) were taken from downloaded data and compared to the recommended treatment pressure following titration PSG (PSG pressure). Fifty-two children started CPAP, of whom 26 (age ± standard deviation 11.9 ± 3.4 years) used autoPAP and had titration PSG. AutoPAP was used on average 84% of nights (standard deviation 20%) in the first month, with a mean ± standard deviation 6.3 ± 2.0 hours of use on nights used. The median (interquartile range) obstructive apnea-hypopnea index decreased from 16.6 (11, 35) events/h before treatment to 2.2 (0.4, 3.8) events/h on the titration PSG. Median (interquartile range) PSG pressure was 9.0 cm H2O (7.0, 10.0), AutoMean pressure was 6.3 cm H2O (5.3, 7.5), and Auto90 pressure was 8.1 cm H2O (7.1, 9.5). These were significantly different (P < .001), with the significant difference lying between AutoMean and the other two pressures. PSG pressure was greater than or equal to the AutoMean pressure in all cases, and greater than or equal to the Auto90 pressure in 20 out of 26 cases (77%). AutoPAP is a safe and effective means of initiating CPAP in children. AutoMean and Auto90 pressures are usually below treatment pressure determined by titration PSG.


Antiplatelet activity of J78 (2-Chloro-3-[2'-bromo, 4'-fluoro-phenyl]-amino-8-hydroxy-1,4-naphthoquinone), an antithrombotic agent, is mediated by thromboxane (TX) A2 receptor blockade with TXA2 synthase inhibition and suppression of cytosolic Ca2+ mobilization.
We previously reported that J78 (2-chloro-3-[2'-bromo, 4'-fluoro-phenyl]-amino-8-hydroxy-1,4-naphthoquinone), a newly synthesized 1,4-naphthoquinone derivative, exhibited a potent antithrombotic effect, which might be due to antiplatelet rather than anticoagulation activity. In the present study, possible anti-platelet mechanism of J78 was investigated. J78 concentration-dependently inhibited rabbit platelet aggregation induced by collagen (10 microg/ml), thrombin (0.05 U/ml), arachidonic acid (100 microM), and U46619 (9,11-dideoxy-9,11-methanoepoxy-prostaglandin F(2); 1 microM), a thromboxane (TX) A(2) mimic, with IC(50) values of 0.32 +/- 0.01, 0.44 +/- 0.02, 0.50 +/- 0.04, and 0.36 +/- 0.02 microM, respectively. J78 also produced a shift to the right of the concentration-response curve of U46619, indicating an antagonistic effect on the TXA(2) receptor. J78 concentration-dependently inhibited collagen-induced arachidonic acid liberation. In addition, J78 potently suppressed TXA(2) formation by platelets that were exposed to arachidonic acid in a concentration-dependent manner but had no effect on the production of PGD(2), indicating an inhibitory effect on TXA(2) synthase. This was supported by a TXA(2) synthase activity assay that J78 concentration-dependently inhibited TXB(2) formation converted from PGH(2). Furthermore, J78 was also able to inhibit the [Ca(2+)](i) mobilization induced by collagen or thrombin at such a concentration that completely inhibited platelet aggregation. Taken together, these results suggest that the antiplatelet activity of J78 may be mediated by TXA(2) receptor blockade with TXA(2) synthase inhibition and suppression of cytosolic Ca(2+) mobilization.


Using three-dimensional ultrasound in predicting complex gastroschisis: A longitudinal, prospective, multicenter cohort study.
To determine whether complex gastroschisis (ie, intestinal atresia, perforation, necrosis, or volvulus) can prenatally be distinguished from simple gastroschisis by fetal stomach volume and stomach-bladder distance, using three-dimensional (3D) ultrasound. This multicenter prospective cohort study was conducted in the Netherlands between 2010 and 2015. Of seven university medical centers, we included the four centers that performed longitudinal 3D ultrasound measurements at a regular basis. We calculated stomach volumes (n = 223) using Sonography-based Automated Volume Count. The shortest stomach-bladder distance (n = 241) was determined using multiplanar visualization of the volume datasets. We used linear mixed modelling to evaluate the effect of gestational age and type of gastroschisis (simple or complex) on fetal stomach volume and stomach-bladder distance. We included 79 affected fetuses. Sixty-six (84%) had been assessed with 3D ultrasound at least once; 64 of these 66 were liveborn, nine (14%) had complex gastroschisis. With advancing gestational age, stomach volume significantly increased, and stomach-bladder distance decreased (both P < .001). The developmental changes did not differ significantly between fetuses with simple and complex gastroschisis, neither for fetal stomach volume (P = .85), nor for stomach bladder distance (P = .78). Fetal stomach volume and stomach-bladder distance, measured during pregnancy using 3D ultrasonography, do not predict complex gastroschisis.


[Analysis of characteristics and trends of randomized controlled trials of gastric cancer between 2000 and 2019].
Objectives: To analyze the current development status of gastric cancer (GC) randomized controlled trials (RCT) between 2000 and 2019, and to review the basic characteristics of published RCT. Methods: ClinicalTrials.gov was searched for phase 3 or 4 RCT conducted between January 2000 and December 2019 with the keyword "gastric cancer", and the development trend of different types of RCT during different time periods was described. Basic features of registered RCT such as intervention, study area, single-center or multicenter, sample size, and funding were presented. PubMed and Scopus databases were searched to judge the publication status of studies completed until June 2016. The adequacy of the report was estimated by the Consolidated Standards of Reporting Trials (CONSORT) checklist. Design flaws were evaluated by Cochrane tool and/or whether a systematic literature review was cited. The data was analyzed by χ2 test or Fisher exact test. Results: There were 262 RCT including in the present study. The number of GC-RCT registered on ClinicalTrials.gov had been on the rise from 1 case in 2000 to 30 cases in 2015. The proportion of RCT associated with targeted therapy or immunotherapy increased from 0 during 2000-2004 to 37.1% (36/97) during 2015-2019. The RCT registered in Asia was 191 cases, while that in non-Asia region was 71 cases. The proportion of multi-center RCT from non-Asia was higher than that from Asia (70.4% (50/71) vs. 50.3% (96/191), χ²=8.527, P=0.003). The proportion of RCT published was 59.1% (81/137). Among the published RCT, 65 (80.2%) studies were reported adequately, but 63 (77.8%) studies had avoidable design limitations. Conclusions: Targeted therapy and immunotherapy have become research hotspots in the treatment of GC. At present, there are inadequate multicenter RCT in Asia, and the publication rate of RCT is low. A considerable number of published RCT are reported inadequately and have avoidable design flaws.


Good manufacturing practices production of natural killer cells for immunotherapy: a six-year single-institution experience.
Natural killer (NK) cells, a subset of lymphocytes and part of the innate immune system, play a crucial role in defense against cancer and viral infection. Herein is a report on the experience of clinical-scale, good manufacturing practices (GMPs) production of NK cells to treat advanced cancer. Two types of NK cell enrichments were performed on nonmobilized peripheral blood mononuclear cell apheresis collections with a cell selection system (CliniMACS, Miltenyi): CD3 cell depletion to enrich for NK cells and CD3 cell depletion followed by CD56 cell selection to obtain a more pure NK cell product. After overnight incubation with interleukin-2 (IL-2), cells were washed, resuspended in 5 percent human serum albumin, and then released for infusion. A total of 70 NK cell therapy products have been manufactured for patient infusion since 2000. For the CD3 cell-depleted NK cell products, the mean purity, recovery, and viability were 38, 79, and 86 percent, respectively. For the CD3 cell-depleted/CD56 cell-enriched NK cell products, the mean purity, recovery, and viability were 90, 19, and 85 percent, respectively. Gram stain, sterility, and endotoxin testing were all within acceptable limits for established lot release. Compared to the resting processed cells, IL-2 activation significantly increased the function of cells in cytotoxicity assays. Clinical-scale production of NK cells is efficient and can be performed under GMPs. The purified NK cell product results in high NK cell purity with minimal contamination by T cells, monocytes, and B cells, but it requires more time for processing and results in a lower NK cell recovery when compared to NK cell enrichment with CD3 cell depletion alone. Additional laboratory studies and results from clinical trials will identify the best source and type of NK cell product.


Drosophila blood cells and their role in immune responses.
Drosophila melanogaster has been extensively used to study the humoral arm of innate immunity because of the developmental and functional parallels with mammalian innate immunity. However, the fly cellular response to infection is far less understood. Investigative work on Drosophila haemocytes, the immunosurveillance cells of the insect, has revealed that they fulfil roles similar to mammalian monocytes and macrophages. They respond to wound signals and orchestrate the coagulation response. In addition, they phagocytose and encapsulate invading pathogens, and clear up apoptotic bodies controlling inflammation. This review briefly describes the Drosophila haematopoietic system and discusses what is currently known about the contribution of haemocytes to the immune response upon infection and wounding, during all stages of development.


Virtual Microscopy in Undergraduate Pathology Education: An early transformative experience in clinical reasoning.
Whole-slide imaging and virtual microscopy (VM) have revolutionised teaching, diagnosis and research in histopathology. This study aimed to establish the feasibility of achieving early integration of clinical reasoning with undergraduate pathology teaching on a VM platform and to determine its student-centricity through student feedback. This study was conducted at the Mohammed Bin Rashid University of Medicine and Health Sciences, Dubai, United Arab Emirates, between August and December 2017. A total of 38 VM-centred clinical cases were introduced to 49 students in an integrated undergraduate medical curriculum. The cases were aligned to curricular objectives, reinforced the pathologic basis of disease with critical thinking and were delivered across 15 interactive small-group sessions. A simulated cross-disciplinary integration and judicious choice of pertinent diagnostic investigations were linked to principles of management. Feedback was obtained through a mixed-methods approach. User-friendliness, gradual learning curve of VM and annotation-capacity were scored as 4-5 (on a Likert scale of 1-5) by 91.84%, 87.76% and 83.67% of the participants, respectively. Most students agreed that the content matched the stage of learning (81.63%), theme of the week (91.84%) and development of a strong clinical foundation (77.55%). Integration (85.71%) and clinico-pathological correlation (83.67%) were the strengths of this educational effort. High student attendance (~100%) and improved assessment scores on critical thinking (80%) were observed. Software lacunae included frequent logouts and lack of note-taking tools. Easy access was a significant student-centric advantage. A VM-centred approach with a clinico-pathological correlation has been successfully introduced to inculcate integrated learning. Using the pathologic basis of disease as a fulcrum and critical reasoning as an anchor, a digitally-enabled generation of medical students have embraced this educational tool for tutor-guided, student-centred learning.


Bilateral motor cortex functional differences in left-handed approaching-avoiding behavior.
Automatic action tendencies occur at behavioral and neurophysiological levels during task performance with the dominant right hand, with shorter reaction times (RTs) and higher excitability of the contralateral primary motor cortex (M1) during automatic vs. regulated behavior. However, effects associated with the non-dominant left-hand in approaching-avoiding behavior remain unclear. Here, we used transcranial magnetic stimulation during the performance by 18 participants of an approaching-avoiding task using the non-dominant left hand. Single-pulse transcranial magnetic stimulation was applied over left or right M1 at 150 and 300 ms after the onset of an emotional stimulus. RTs and motor-evoked potentials (MEPs) were recorded. Significant automatic action tendencies were observed at the behavioral level. Higher MEP amplitudes were detected 150 ms after stimulus onset from the right hand (non-task hand, corresponding to left M1) during regulated behavior compared with during automatic behavior. However, no significant modulation was found for MEP amplitudes from the left hand (task hand, corresponding to right M1). These findings suggested that left M1 may play a principal role in the early phase of mediating left-handed movement toward an emotional stimulus.


Egeria densa organic extracts: an eco-friendly approach to suppress Microcystis aeruginosa growth through allelopathy.
Macrophytes play a significant role in shaping plankton communities by shading, controlling water turbulence, and nutrient availability, while also producing allelochemicals causing varying effects on different organisms. Many researchers have shown that when live macrophytes are present, they inhibit cyanobacteria. However, their widespread use is often limited due to numerous concerns, including invasive characteristics. This study focused on the applicability of Egeria densa extracts to suppress Microcystis aeruginosa. We employed pure water and dimethyl sulfoxide, to obtain compounds from E. densa. The results revealed that E. densa aqueous extracts stimulated M. aeruginosa growth, whereas organic extracts exhibited suppression. Specifically, at low concentrations of organics extracts (0.5 and 1 g/L), after day 4, the growth inhibition was confirmed by significantly higher (p < 0.05) stress levels shown in cells treated with low concentrations. The highest inhibition rate of 32% was observed at 0.5 g/L. However, high concentrations of organic extracts (3 and 6 g/L), showed increased growth compared with control. These results suggest that high concentrations of organic extracts from E. densa potentially suppress allelochemical-induced M. aeruginosa inhibition due to high nutrient availability. In comparison with an aqueous solvent, the use of organic solvent seems to be more effective in efficiently extracting allelochemicals from E. densa.


Emergence of diplopia and oscillopsia due to Heimann-Bielschowsky phenomenon after cataract surgery.
The Heimann-Bielschowsky phenomenon (HBP) refers to coarse vertical oscillation of the eye with impaired vision. The ocular movements are strictly monocular, occurring only in the eye with amblyopia. The vertical oscillation is of equal velocity in both vertical directions, or may sometimes be greater in the downward than upward direction. HBP develops several years after loss of vision. It can be differentiated from dissociated nystagmus in spasmus nutans, congenital nystagmus and internuclear ophthalmoplegia based on the strict unilaterality, vertical direction and low frequency. Previously, only a few reports described the development of oscillopsia due to HBP after cataract surgery, which resolved spontaneously or responded to gabapentin. However, visual impairments due to diplopia or oscillopsia from HBP after cataract surgery have received little attention. We report a man who developed persistent vertical diplopia and oscillopsia due to HBP after a cataract operation, which markedly impaired his vision.


B cells in pancreatic cancer stroma.
Pancreatic cancer is a disease with high unmet clinical need. Pancreatic cancer is also characterised by an intense fibrotic stroma, which harbours many immune cells. Studies in both human and animal models have demonstrated that the immune system plays a crucial role in modulating tumour onset and progression. In human pancreatic ductal adenocarcinoma, high B-cell infiltration correlates with better patient survival. Hence, B cells have received recent interest in pancreatic cancer as potential therapeutic targets. However, the data on the role of B cells in murine models is unclear as it is dependent on the pancreatic cancer model used to study. Nevertheless, it appears that B cells do organise along with other immune cells such as a network of follicular dendritic cells (DCs), surrounded by T cells and DCs to form tertiary lymphoid structures (TLS). TLS are increasingly recognised as sites for antigen presentation, T-cell activation, B-cell maturation and differentiation in plasma cells. In this review we dissect the role of B cells and provide directions for future studies to harness the role of B cells in treatment of human pancreatic cancer.


MANAGEMENT OF ENDOCRINE DISEASE: Management of Cushing's syndrome during pregnancy: solved and unsolved questions.
With fewer than 200 reported cases, Cushing's syndrome (CS) in pregnancy remains a diagnostic and therapeutic challenge. In normal pregnancies, misleading signs may be observed such as striae or hypokalemia, while plasma cortisol and urinary free cortisol may rise up to 2- to 3-fold. While the dexamethasone suppression test is difficult to use, reference values for salivary cortisol appear valid. Apart from gestational hypertension, differential diagnosis includes pheochromocytoma and primary aldosteronism. The predominant cause is adrenal adenoma (sometimes without decreased ACTH), rather than Cushing's disease. There are considerable imaging pitfalls in Cushing's disease. Aberrant receptors may, in rare cases, lead to increased cortisol production during pregnancy in response to HCG, LHRH, glucagon, vasopressin or after a meal. Adrenocortical carcinoma (ACC) is rare and has poor prognosis. Active CS during pregnancy is associated with a high rate of maternal complications: hypertension or preeclampsia, diabetes, fractures; more rarely, cardiac failure, psychiatric disorders, infection and maternal death. Increased fetal morbidity includes prematurity, intrauterine growth retardation and less prevalently stillbirth, spontaneous abortion, intrauterine death and hypoadrenalism. Therapy is also challenging. Milder cases can be managed conservatively by controlling comorbidities. Pituitary or adrenal surgery should ideally be performed during the second trimester and patients should then be treated for adrenal insufficiency. Experience with anticortisolic drugs is limited. Metyrapone was found to allow control of hypercortisolism, with a risk of worsening hypertension. Cabergoline may be an alternative option. The use of other drugs is not advised because of potential teratogenicity and/or lack of information. Non-hormonal (mechanical) contraception is recommended until sustained biological remission is obtained.


Aluminium in Brain Tissue in Multiple Sclerosis.
Multiple sclerosis (MS) is a devastating and debilitating neurodegenerative disease of unknown cause. A consensus suggests the involvement of both genetic and environmental factors of which the latter may involve human exposure to aluminium. There are no data on the content and distribution of aluminium in human brain tissue in MS. The aluminium content of brain tissue from 14 donors with a diagnosis of MS was determined by transversely heated graphite furnace atomic absorption spectrometry. The location of aluminium in the brain tissue of two donors was investigated by aluminium-specific fluorescence microscopy. The aluminium content of brain tissue in MS was universally high with many tissues bearing concentrations in excess of 10 μg/g dry wt. (10 ppm) and some exceeding 50 ppm. There were no statistically significant relationships between brain lobes, donor age or donor gender. Aluminium-specific fluorescence successfully identified aluminium in brain tissue in both intracellular and extracellular locations. The association of aluminium with corpora amylacea suggests a role for aluminium in neurodegeneration in MS.


Effect of hyperphenylalaninaemia on lipid synthesis from ketone bodies by rat brain.
The effect of hyperphenylalaninaemia on the metabolism of ketone bodies in vivo and in vitro by developing rat brain was investigated. The incorporation in vivo of [14C]acetoacetate into cerebral lipids was decreased by both chronic (for 3 days) and acute (for 6h) hyperphenylalaninaemia induced by injecting phenylalanine into 1-week-old rats. In studies in vitro it was observed that the incorporation of the radioactivity from [14C]acetoacetate and 3-hydroxy[14C]butyrate into cerebral lipids was inhibited by phenyl-pyruvate, but not by phenylalanine. Phenylpyruvate also inhibited the incorporation of 3H from 3H2O into lipids by brain slices metabolizing either 3-hydroxybutyrate or acetoacetate in the presence of glucose. These findings suggest that the decrease in the incorporation in vivo of [14C]acetoacetate into cerebral lipids in hyperphenylalaninaemic rats is most likely caused by phenylpyruvate and not by phenylalanine. Phenylpyruvate as well as phenylalanine had no inhibitory effects on ketone-body-catabolizing enzymes, namely 3-hydroxybutyrate dehydrogenase, 3-oxo acid CoA-transferase and acetoacetyl-CoA thiolase, in rat brain. Phenylpyruvate but not phenylalanine inhibited the activity of the 2-oxoglutarate dehydrogenase complex from rat and human brain. These findings suggest that the metabolism of ketone bodies is impaired in brains of untreated phenylketonuric patients, and in turn may contribute to the diminution of mental development and function associated with phenylketonuria.


Diagnosis of hepatic allograft rejection: role of liver biopsy.
The role of the liver biopsy for establishing a diagnosis of liver allograft rejection was investigated by examining under code 151 liver biopsies from 32 recipients and correlating the results with the clinical diagnosis determined by chart review. One-hundred-and-four biopsies were obtained to evaluate the cause of liver dysfunction and 47 were performed on a weekly protocol basis independent of the clinical status. A diagnosis of "definite histologic rejection" was made if a biopsy contained portal triaditis, bile duct injury, and endothelialitis, while biopsies were said to be suspicious for rejection if portal triaditis and bile duct injury were evident. A diagnosis of rejection using clinical and biochemical findings and response to therapy was made in association with 37 biopsies, of which 16 had definite histologic rejection and 13 were suspicious for rejection. Of the 114 biopsies performed in patients without clinical rejection, 71 had histology suspicious for rejection. Thus, biopsy was sensitive for the detection of rejection (78%), but was not very specific (33%). The positive predictive value was low (28%), while the negative predictive value was high (83%). These results indicate that liver biopsy is useful to evaluate graft dysfunction, to exclude rejection, to confirm clinically suspected rejection, and to diagnose other problems that can affect allograft function. Protocol biopsies appear to have little value.


Prediction of body condition in Jersey dairy cattle from 3D-images using machine learning techniques.
The body condition of dairy cows is a crucial health and welfare indicator that is widely acknowledged. Dairy herds with a well-management body condition tend to have more fertile and functional cows. Therefore, routine recording of high-quality body condition phenotypes is required. Automated prediction of body condition from 3D-images can be a cost-effective approach to current manual recording by technicians. Using 3D-images, we aimed to build a reliable prediction model of body condition for Jersey cows. The dataset consisted of 808 individual Jersey cows with 2,253 phenotypes from three herds in Denmark. Body condition was scored on a 1 to 9 scale and transformed into a 1 to 5 scale with 0.5-unit differences. The cows' back images were recorded using a 3D camera (Microsoft Xbox One Kinect v2). We used contour and back height features from 3D-images as predictors, together with class predictors (evaluator, herd, evaluation round, parity, lactation week). The performance of machine learning algorithms was assessed using H2O AutoML algorithm (h2o.ai). Based on outputs from AutoML, DeepLearning (DL; multi-layer feedforward artificial neural network) and Gradient Boosting Machine (GBM) algorithms were implemented for classification and regression tasks and compared on prediction accuracy. In addition, we compared the Partial Least Square (PLS) method for regression. The training and validation data were divided either through a random 7:3 split for 10 replicates or by allocating two herds for training and one herd for validation. The accuracy of classification models showed the DL algorithm performed better than the GBM algorithm. The DL model achieved a mean accuracy of 48.1% on the exact phenotype and 93.5% accuracy with a 0.5-unit deviation. The performances of PLS and DL regression methods were comparable, with mean coefficient of determination of 0.67 and 0.66, respectively. When we used data from two herds for training and the third herd as validation, we observed a slightly decreased prediction accuracy compared to the 7:3 split of the dataset. The accuracies for DL and PLS in the herd validation scenario were >38% on the exact phenotype and >87% accuracy with 0.5-unit deviation. This study demonstrates the feasibility of a reliable body condition prediction model in Jersey cows using 3D-images. The approach developed can be used for reliable and frequent prediction of cows' body condition to improve dairy farm management and genetic evaluations.


Assessment of neonatal jaundice in low birth weight infants comparing transcutaneous, capillary and arterial bilirubin levels.
Total serum bilirubin level was assessed in a group of jaundiced low birth weight infants using three different methods. Transcutaneous bilirubinometry was compared with conventional capillary and arterial methods to investigate the over-or underestimation of neonatal jaundice. Sampling site did not influence bilirubin levels. Capillary and arterial results showed a linear correlation (r = 0.9) suggesting no influence of environmental light on peripheral bilirubin isomerization. Similar results were obtained comparing both serum levels with transcutaneous values (r = 0.7). We conclude that treatment decisions may be made on the basis of one of the three mentioned methods in healthy low birth weight infants with neonatal jaundice.


Rett syndrome-causing mutations compromise MeCP2-mediated liquid-liquid phase separation of chromatin.
Rett syndrome (RTT), a severe postnatal neurodevelopmental disorder, is caused by mutations in the X-linked gene encoding methyl-CpG-binding protein 2 (MeCP2). MeCP2 is a chromatin organizer regulating gene expression. RTT-causing mutations have been shown to affect this function. However, the mechanism by which MeCP2 organizes chromatin is unclear. In this study, we found that MeCP2 can induce compaction and liquid-liquid phase separation of nucleosomal arrays in vitro, and DNA methylation further enhances formation of chromatin condensates by MeCP2. Interestingly, RTT-causing mutations compromise MeCP2-mediated chromatin phase separation, while benign variants have little effect on this process. Moreover, MeCP2 competes with linker histone H1 to form mutually exclusive chromatin condensates in vitro and distinct heterochromatin foci in vivo. RTT-causing mutations reduce or even abolish the ability of MeCP2 to compete with histone H1 and to form chromatin condensates. Together, our results identify a novel mechanism by which phase separation underlies MeCP2-mediated heterochromatin formation and reveal the potential link between this process and the pathology of RTT.


Structural and Genomic Insights Into Pyrazinamide Resistance in Mycobacterium tuberculosis Underlie Differences Between Ancient and Modern Lineages.
Resistance to drugs used to treat tuberculosis disease (TB) continues to remain a public health burden, with missense point mutations in the underlying Mycobacterium tuberculosis bacteria described for nearly all anti-TB drugs. The post-genomics era along with advances in computational and structural biology provide opportunities to understand the interrelationships between the genetic basis and the structural consequences of M. tuberculosis mutations linked to drug resistance. Pyrazinamide (PZA) is a crucial first line antibiotic currently used in TB treatment regimens. The mutational promiscuity exhibited by the pncA gene (target for PZA) necessitates computational approaches to investigate the genetic and structural basis for PZA resistance development. We analysed 424 missense point mutations linked to PZA resistance derived from ∼35K M. tuberculosis clinical isolates sourced globally, which comprised the four main M. tuberculosis lineages (Lineage 1-4). Mutations were annotated to reflect their association with PZA resistance. Genomic measures (minor allele frequency and odds ratio), structural features (surface area, residue depth and hydrophobicity) and biophysical effects (change in stability and ligand affinity) of point mutations on pncA protein stability and ligand affinity were assessed. Missense point mutations within pncA were distributed throughout the gene, with the majority (>80%) of mutations with a destabilising effect on protomer stability and on ligand affinity. Active site residues involved in PZA binding were associated with multiple point mutations highlighting mutational diversity due to selection pressures at these functionally important sites. There were weak associations between genomic measures and biophysical effect of mutations. However, mutations associated with PZA resistance showed statistically significant differences between structural features (surface area and residue depth), but not hydrophobicity score for mutational sites. Most interestingly M. tuberculosis lineage 1 (ancient lineage) exhibited a distinct protein stability profile for mutations associated with PZA resistance, compared to modern lineages.


Pent-up demand for surgery in the Manaus metropolitan region: A population-based cross-sectional study.
Waiting lines in healthcare reflect an imbalance between the availability and the demand for medical services. This study aimed to analyze the prevalence and factors associated with the pent-up demand for surgical procedures in the Manaus metropolitan region.We performed a population-based cross-sectional study in 2015. Pent-up demand was based on self-report by the participants; those who reported waiting were contacted by phone to clarify the nature and reasons for the experienced delay.We interviewed 4001 adults in their households. The pent-up demand for surgical procedures in the Manaus metropolitan region was 14% (95% confidence interval: 13-15%). The surgical specialties with the highest demand included orthopedics (18.1%), gynecology (17.0%), ophthalmology (12.4%), neurosurgery (10.8%), and general surgery (10.2%). The main reason for not accessing services was their lack of availability in the public health system, leading some patients to pay for procedures in private facilities. The populations most affected by pent-up demand included elderly individuals, women, and housewives.Pent-up demand for surgical procedures is a common problem in the Manaus metropolitan region and thus requires coordinated actions to optimize access to and capacity of the healthcare system.


AGL1-AGL6, an Arabidopsis gene family with similarity to floral homeotic and transcription factor genes.
The predicted products of floral homeotic genes, AGAMOUS (AG) from Arabidopsis thaliana and DEFICIENS A (DEF A) from Antirrhinum majus, have been shown previously to share strong sequence similarity with transcription factors from humans (SRF) and yeast (MCM1). The conserved sequence between these proteins is localized within a domain known to be necessary for the DNA binding and for the dimerization of SRF. We have isolated six new genes from A. thaliana, AGL1-AGL6, which also have this conserved sequence motif. On the basis of the sequence comparison between the AG and AGL genes, they can be assigned to two subfamilies of a large gene family. RNA dot blot analysis indicates that five of these genes (AGL1, AGL2, AGL4, AGL5, and AGL6) are preferentially expressed in flowers. In addition, in situ RNA hybridization experiments with AGL1 and AGL2 show that their mRNAs are detected in some floral organs but not in others. Our results suggest that these genes may act to control many steps of Arabidopsis floral morphogenesis. In contrast, the AGL3 gene is expressed in vegetative tissues as well as in flowers, suggesting that it functions in a broader range of tissues. We discuss possible roles of this gene family during the evolution of flowers.


Wild-type IDH1 maintains NSCLC stemness and chemoresistance through activation of the serine biosynthetic pathway.
Tumor-initiating cells (TICs) reprogram their metabolic features to meet their bioenergetic, biosynthetic, and redox demands. Our previous study established a role for wild-type isocitrate dehydrogenase 1 (IDH1WT) as a potential diagnostic and prognostic biomarker for non-small cell lung cancer (NSCLC), but how IDH1WT modulates NSCLC progression remains elusive. Here, we report that IDH1WT activates serine biosynthesis by enhancing the expression of phosphoglycerate dehydrogenase (PHGDH) and phosphoserine aminotransferase 1 (PSAT1), the first and second enzymes of de novo serine synthetic pathway. Augmented serine synthesis leads to GSH/ROS imbalance and supports pyrimidine biosynthesis, maintaining tumor initiation capacity and enhancing gemcitabine chemoresistance. Mechanistically, we identify that IDH1WT interacts with and stabilizes PHGDH and fragile X-related protein-1 (FXR1) by impeding their association with the E3 ubiquitin ligase parkin by coimmunoprecipitation assay and proximity ligation assay. Subsequently, stabilized FXR1 supports PSAT1 mRNA stability and translation, as determined by actinomycin D chase experiment and in vitro translation assay. Disrupting IDH1WT-PHGDH and IDH1WT-FXR1 interactions synergistically reduces NSCLC stemness and sensitizes NSCLC cells to gemcitabine and serine/glycine-depleted diet therapy in lung cancer xenograft models. Collectively, our findings offer insights into the role of IDH1WT in serine metabolism, highlighting IDH1WT as a potential therapeutic target for eradicating TICs and overcoming gemcitabine chemoresistance in NSCLC.


[Pneumatosis intestinalis in a patient of myasthenia gravis treated with high-dose corticosteroid].
We report a patient who developed persistent and severe diarrhea in the course of high-dose corticosteroid therapy for myasthenia gravis (MG). The patient, a 37-year-old woman, developed diplopia, ptosis, and muscle weakness with fatigability. She was admitted to our hospital and was diagnosed as having MG. Five months after thymectomy and oral corticosteroid administration, she developed abdominal pain and severe diarrhea. Abdominal X-ray, showed extensive pericolic gas accumulation surrounding the ascending and transverse colon. She was diagnosed as having pneumatosis intestinalis (PI). Hyperbaric therapy was not effective. Symptoms began to improve gradually when the dose of prednisolone was reduced. PI is a rare condition characterized by gas-filled cysts or linear gas in the bowel wall caused by a variety of disorders and drugs such as necrotizing colitis, obstructive pulmonary diseases, and immunosuppressants. In the present case, PI appeared to be induced by corticosteroid therapy. To our knowledge, MG complicated by PI is unusual, but this rare case highlights the importance of taking it into consideration during steroid therapy for MG.


RNA in blood is altered prior to hemorrhagic transformation in ischemic stroke.
Hemorrhagic transformation (HT) is a major complication of ischemic stroke that worsens outcomes and increases mortality. Disruption of the blood-brain barrier is a central feature of HT pathogenesis, and leukocytes may contribute to this process. We sought to determine whether ischemic strokes that develop HT have differences in RNA expression in blood within 3 hours of stroke onset prior to treatment with thrombolytic therapy. Stroke patient blood samples were obtained prior to treatment with thrombolysis, and leukocyte RNA was assessed by microarray analysis. Strokes that developed HT (n = 11) were compared to strokes without HT (n = 33) and controls (n = 14). Genes were identified (corrected p < 0.05, fold change ≥|1.2|), and functional analysis was performed. RNA prediction of HT in stroke was evaluated using cross-validation, and in a second stroke cohort (n = 52). Ischemic strokes that developed HT had differential expression of 29 genes in circulating leukocytes prior to treatment with thrombolytic therapy. A panel of 6 genes could predict strokes that later developed HT with 80% sensitivity and 70.2% specificity. Key pathways involved in HT of human stroke are described, including amphiregulin, a growth factor that regulates matrix metalloproteinase-9; a shift in transforming growth factor-β signaling involving SMAD4, INPP5D, and IRAK3; and a disruption of coagulation factors V and VIII. Identified genes correspond to differences in inflammation and coagulation that may predispose to HT in ischemic stroke. Given the adverse impact of HT on stroke outcomes, further evaluation of the identified genes and pathways is warranted to determine their potential as therapeutic targets to reduce HT and as markers of HT risk.


Results with Zielke instrumentation for idiopathic thoracolumbar and lumbar scoliosis.
Thirty-one patients (23 adolescent, eight adult) underwent spinal fusions with Zielke instrumentation for idiopathic thoracolumbar and lumbar scoliosis. Their curves averaged 55.2 degrees. In most patients the length of fusion did not extend beyond the major curve. Correction of scoliosis was 82% in adolescents and 59% in the adults. The kyphotic component of the curves was corrected from an average of 21 degrees to 8 degrees. Correction of the spinal vertebra rotation was 46% in the adolescent patients. The tilt angle of the vertebra at the lower end of the curve was reduced significantly. The fusion rate was 93.5%. Two of the adult patients developed pseudarthrosis. Major complications were not encountered. The Zielke instrumentation system in the management of idiopathic thoracolumbar and lumbar scoliosis offers the advantages of a relatively good correction and minimal segment fusion.


Diagnosis, disclosure, and having autism: an interpretative phenomenological analysis of the perceptions of young people with autism.
Although there is extensive research examining parental experiences of assessment and diagnosis of autism, there is a paucity of research from the perspective of individuals with autism. Semi-structured interviews were conducted with nine young people with high functioning autism who were capable of providing a verbal account of their perceptions of autism and diagnosis experiences. Data were analysed using interpretative phenomenological analysis. Diagnosis, and the disclosure of the diagnosis of autism, were embedded in respondents' perceptions of "having" autism. This superordinate theme included five themes: (i) disclosure delay, (ii) providing explanations, (iii) potential effects of labelling, (iv) disruptions and opportunities, and (v) acceptance and avoidance. In relation to the existing literature, it is suggested that the effects of diagnosis, or disclosure of diagnosis, from the perspective of the person with autism be given greater consideration.


Preconception healthcare and congenital disorders: systematic review of the effectiveness of preconception care programs in the prevention of congenital disorders.
Congenital disorders are a leading cause of global burden of disease; the birth prevalence remains constant at 6%. Initiating preconception care before pregnancy may be an effective strategy to reduce congenital disorders and improve the health of reproductive-age women. Our objectives are: (1) To identify components of preconception interventions, (2) to assess the effectiveness of preconception interventions in reducing the burden of congenital disorders, and (3) to prioritize these interventions. Medline and Science Direct search terms included: preconception, pre-pregnancy, childbearing, reproduction, care, intervention, primary care, healthcare, model, program, prevention, trial, efficacy, effectiveness, congenital disorders OR abnormalities. Inclusion criteria were: (1) English, (2) human subjects, (3) women of childbearing age, (4) 1980-current data, (5) all countries, (6) experimental studies, (7) systematic reviews or meta-analysis, (8) program reports/evaluations. Data was collected and abstracted by two independent reviewers. To prioritize preconception interventions likely to have the largest impact at a population level, a ranked scoring system was created incorporating the following: (1) quality of evidence supporting the intervention, (2) effect size of the intervention, and (3) global burden of the specific congenital disease. Preconception interventions include risk screening, education, motivational counseling, disease optimization and specialist referral. The most effective interventions, based on the strength of evidence, size of impact of intervention, and disease burden are: folic acid fortification/supplementation, diabetic control, smoking and alcohol interventions, HIV management, thrombophillia screening, obesity prevention and epilepsy management. Although multiple conditions require preconception attention, only nine interventions have evidence to support their effect on congenital disorders through a randomised control trial, systematic review or meta-analysis. There is a need for more high-level research in evaluating certain preconception interventions. These findings have significant implications on planning and implementation of preconception care.


Method matters: impact of in-scenario instruction on simulation-based teamwork training.
The rationale for introducing full-scale patient simulators in training to improve patient safety is to recreate clinical situations in a realistic setting. Although high-fidelity simulators mimic a wide range of human features, simulators differ from the body of a sick patient. The gap between the simulator and the human body implies a need for facilitators to provide information to help participants understand scenarios. The authors aimed at describing different methods that facilitators in our dataset used to provide such extra scenario information and how the different methods to convey information affected how scenarios played out. A descriptive qualitative study was conducted to examine the variation of methods to deliver extra scenario information to participants. A multistage approach was employed. The authors selected film clips from a shared database of 31 scenarios from three participating simulation centers. A multidisciplinary research team performed a collaborative analysis of representative film clips focusing on the interplay between participants, facilitators, and the physical environment. After that, the entire material was revisited to further examine and elaborate the initial findings. The material displayed four distinct methods for facilitators to convey information to participants in simulation-based teamwork training. The choice of method had impact on the participating teams regarding flow of work, pace, and team communication. Facilitators' close access to the teams' activities when present in the simulation suite, either embodied or disembodied in the simulation, facilitated the timing for providing information, which was critical for maintaining the flow of activities in the scenario. The mediation of information by a loudspeaker or an earpiece from the adjacent operator room could be disturbing for team communication. In-scenario instruction is an essential component of simulation-based teamwork training that has been largely overlooked in previous research. The ways in which facilitators convey information about the simulated patient have the potential to shape the simulation activities and thereby serve different learning goals. Although immediate timing to maintain an adequate pace is necessary for professionals to engage in training of medical emergencies, novices may gain from a slower tempo to train complex clinical team tasks systematically.


Primary cellular immunodeficiencies.
Genetic defects in T-cell function lead to susceptibility to infections or to other clinical problems that are more grave than those seen in disorders resulting in antibody deficiency alone. Those affected usually present during infancy with either common or opportunistic infections and rarely survive beyond infancy or childhood. The spectrum of T-cell defects ranges from the syndrome of severe combined immunodeficiency, in which T-cell function is absent, to combined immunodeficiency disorders in which there is some, but not adequate, T-cell function for a normal life span. Recent discoveries of the molecular causes of many of these defects have led to a new understanding of the flawed biology underlying the ever-growing number of defects. Most of these conditions could be diagnosed by means of screening for lymphopenia or for T-cell deficiency in cord blood at birth. Early recognition of those so afflicted is essential to the application of the most appropriate treatments for these conditions at a very early age. The latter treatments include both transplantation and gene therapy in addition to immunoglobulin replacement. Fully defining the molecular defects of such patients is also essential for genetic counseling of family members and prenatal diagnosis.


Recombinant versus urinary follicle stimulating hormone for ovarian stimulation in assisted reproduction.
The recent availability of recombinant follicle stimulating hormone (rFSH), with its high level of purity and batch-to-batch consistency has made it an attractive alternative to urinary FSH (uFSH) for ovarian stimulation. Several trials have compared the two preparations, but none had sufficient power to detect a clinically meaningful difference in pregnancy rates. The purpose of this study was to determine the clinical pregnancy rates per started cycle by pooling data from randomized trials which compared the use of rFSH and uFSH in treatment cycles using in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). A thorough search of the literature identified 12 trials which met the inclusion criteria. In four trials, both IVF and ICSI were performed, in seven trials only IVF was performed and in one trial only ICSI was performed. Data were extracted and pooled using the principles of meta-analysis. There was no significant heterogeneity of treatment effect across the trials. The common odds ratio and the risk difference (and their 95% confidence intervals), obtained by pooling the data using a fixed effects model, were 1.20 (1.02-1.42) and 3.7% (0.5-6.9%) respectively, in favour of rFSH. The pregnancy rate with the alpha preparation of rFSH was statistically significantly higher than with uFSH in IVF cycles. The overall conclusion from this meta-analysis is that the use of rFSH in assisted reproduction is preferred over uFSH.


Pupillary response induced by acupuncture stimulation--an experimental study.
To investigate whether acupuncture stimulation affects autonomic nerve function by measuring pupil diameters with electronic pupillography. Two studies were conducted (Experiment 1 and 2) in the Tsukuba University of Technology, Tsukuba, Japan. Experiment 1 examined the responses before, during and after acupuncture. Experiment 2 compared acupuncture and a no acupuncture control in a two period, repeated measurement crossover design. Twelve healthy male university students were recruited for Experiment 1 and nine healthy male university students for Experiment 2. The intervention was superficial acupuncture at the acupuncture point TE5. Following the insertion, gentle repetitive tapping stimulation was applied during the subject's exhalation phase in a sitting position for 90 seconds. The main outcome measures in Experiment 1 were pupil diameter, heart rate, pulse wave and blood pressure. In both experiments, pupil diameter was measured for three minutes before acupuncture stimulation, during stimulation and for three minutes after stimulation. In Experiment 1, a decrease in pupil diameter was observed after acupuncture stimulation (P=0.018) and a decrease in heart rate was observed during the stimulation (P=0.049). Moreover, a significant decrease of pulse wave amplitude was observed during and after acupuncture stimulation. In Experiment 2, a decrease in pupil diameter occurred after acupuncture stimulation (P=0.007), whereas no change occurred under the no acupuncture control. It is thought that the pupillary constriction observed after gentle, superficial acupuncture stimulation may be attributed to an increase of parasympathetic nerve function. Further study is necessary in order to further clarify the duration of the observed response and to elucidate the mechanisms behind it.


Artificial Intelligence Algorithm-Based CTA Imaging for Diagnosing Ischemic Type Biliary Lesions after Orthotopic Liver Transplantation.
The study focused on the clinical application value of artificial intelligence-based computed tomography angiography (CTA) in the diagnosis of orthotopic liver transplantation (OLT) after ischemic type biliary lesions (ITBL). A total of 66 patients receiving OLT in hospital were selected. Convolutional neural network (CNN) algorithm was used to denoise and detect the edges of CTA images of patients. At the same time, the quality of the processed image was subjectively evaluated and quantified by Hmax, Ur, Cr, and other indicators. Then, the digital subtraction angiography (DSA) diagnosis and CTA diagnosis based on CNN were compared for the sensitivity, specificity, positive predictive value, negative predictive value, and patient classification results. It was found that CTA can clearly reflect the information of hepatic aorta lesions and thrombosis in patients with ischemic single-duct injury after liver transplantation. After neural network algorithm processing, the image quality is obviously improved, the lesions are more prominent, and the details of lesion parts are also well displayed. ITBL occurred in 40 (71%) of 56 patients with abnormal CTA at early stage. ITBL occurred in only 8 (12.3%) of 65 patients with normal CTA at early stage. Early CTA manifestations had high sensitivity (72.22%), specificity (87.44%), positive predictive value (60.94%), and negative predictive value (92.06%) for the diagnosis of ITBL. It was concluded that artificial intelligence-based CTA had high clinical application value in the diagnosis of ITBL after OLT.


Where are we today in split liver transplantation?
Results of surgical innovations using partial liver grafts from deceased donors have improved the availability of transplantable organs. However, current data on outcomes after split liver transplantation (SLT) are conflicting. This article reviews the current state of SLT, focusing on long-term outcomes and predictors for patient and graft survival after SLT. The conventional SLT has been proven to be a durable life-saving procedure. Early results for full left-right SLT for two adults are promising but this technique had not showed efficacy for wide application. Predictors of diminished patient survival after SLT included the use of split grafts in critically ill recipients (model for end-stage liver disease score >30), retransplant patients, cold ischemia time more than 10 h, and the performance of SLT in low-volume liver transplant centers. Conventional SLT performed in specialized centers resulted in long-term survival outcomes comparable with whole-organ liver transplantation. Full left-right SLT for two adults remains experimental. Splitting of the liver is an effective approach to expand the donor pool and remains an untapped resource for patients in need of liver transplantation. Split graft-to-recipient pairing is crucial for optimal organ allocation and survival outcomes after liver transplantation.


Financial Toxicity in Breast Implant-Associated Anaplastic Large Cell Lymphoma.
Financial toxicity is a growing concern due to its considerable effects on medical adherence, quality of life, and mortality. The cost associated with breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) is substantial from diagnosis to treatment, including adjuvant therapy and surgery. This study aims to assess the prevalence of financial toxicity in BIA-ALCL patients. We performed a cross-sectional, survey-based study on women with confirmed cases of BIA-ALCL from December 2019 to March 2023. The primary study outcomes were financial toxicity measured by Comprehensive Score for Financial Toxicity (COST) score and patient-reported financial burden measured by the responses to the Evaluation of the Financial Impact of BIA-ALCL survey. Lower COST scores signify higher financial toxicity. Responses were linked to patient data extracted from the medical records. Thirty-two women treated for confirmed BIA-ALCL were included. Patients were all White and were diagnosed at a median age of 51 years (range, 41-65 years). The mean COST score was 27.9 ± 2.23. Lower COST scores were associated with receipt of radiotherapy (P = 0.033), exceeding credit card limits (P = 0.036), living paycheck to paycheck (P = 0.00027), requiring financial support from friends and family (P = 0.00044), and instability in household finances (P = 0.034). Financial toxicity is prevalent in BIA-ALCL patients and has a substantial impact on patient reported burden. Insurance denial is frequent for patients with a prior history of cosmetic augmentation. Risk assessments and cost discussions should occur throughout the care continuum to minimize financial burden.


Polarized uterine epithelial cells preferentially present antigen at the basolateral surface: role of stromal cells in regulating class II-mediated epithelial cell antigen presentation.
To study Ag presentation in the female reproductive tract, DO11.10 TCR transgenic mice specific for the class II MHC-restricted OVA(323-339) peptide and non-transgenic BALB/c mice were used. We report here that freshly isolated uterine epithelial cells, uterine stromal, and vaginal APCs present OVA and OVA(323-339) peptide to naive- and memory T cells, which is reduced when cells are incubated with Abs to CD80 and 86. To determine whether polarized primary epithelial cells present Ags, uterine epithelial cells were cultured on cell inserts in either the upright or inverted position. After reaching confluence, as indicated by high transepithelial resistance (>2000 ohms/well), Ag presentation by epithelial cells incubated with memory T cells and OVA(323-339) peptide placed on the basolateral surface (inverted) was 2- to 3-fold greater than that seen with epithelial cells in contact with T cells and peptide on the apical surface (upright). In contrast, whereas freshly isolated epithelial cells process OVA, polarized epithelial cells did not. When epithelial cells grown upright on inserts were incubated with T cells and OVA(323-339) peptide, coculture with either hepatocyte growth factor or conditioned stromal medium increased epithelial cell Ag presentation (approximately 90% higher than controls). These studies indicate that uterine stromal cells produce a soluble factor(s) in addition to a hepatocyte growth factor, which regulates epithelial cell Ag presentation. Overall, these results demonstrate that polarized epithelial cells are able to present Ags and suggest that uterine stromal cells communicate with epithelial cells via a soluble factor(s) to regulate Ag presentation in the uterus.