Predictive value and accuracy of [18F]FDG PET/CT modified response criteria for checkpoint immunotherapy in patients with advanced melanoma.
Immune checkpoint inhibitors (ICIs) are widely used in metastatic melanoma and dramatically alter the treatment of these patients. Given the high cost and potential toxicity, a reliable method for evaluating treatment response is needed. In this study, we assessed tumor response in patients with metastatic melanoma treated with ICIs using three modified response criteria: PET Response Evaluation Criteria for Immunotherapy (PERCIMT), PET Response Criteria in Solid Tumors for up to Five Lesions (PERCIST5), and immunotherapy-modified PET Response Criteria in Solid Tumors for up to Five Lesions (imPERCIST5). Ninety-one patients with non-resectable stage IV metastatic melanoma who received ICIs were retrospectively enrolled in this study. Each patient had two [18F]FDG PET/CT scans performed before and after ICI therapy. Responses at the follow-up scan were evaluated according to PERCIMT, PERCIST5, and imPERCIST5 criteria. Patients were classified into four groups: complete metabolic response (CMR), partial metabolic response (PMR), progressive metabolic disease (PMD), and stable metabolic disease (SMD). To assess the "disease control rate," two groups have been defined based on each criterion: patients with CMR, PMR, and SMD as "disease-controlled group (i.e., responders)" and PMD as the "uncontrolled-disease group (i.e., non-responders)". The correspondence between metabolic tumor response defined by these criteria and clinical outcome was assessed and compared. The response and the disease control rates were 40.7% and 71.4%, 41.8% and 50.5%, and 54.9% and 74.7% based on the PERCIMT, PERCIST5, and imPERCIST5 criteria, respectively. PERCIMT and imPERCIST5 showed significantly different disease control rates from that of PERCIST5 (P < 0.001), whereas it was not significant between PERCIMT and imPERCIST5. Overall survival was significantly longer in the metabolic responder groups than in the non-responder groups based on PERCIMT and PERCIST5 criteria (PERCIMT: 2.48 versus 1.47 years, P = 0.003; PERCIST5: 2.57 versus 1.81 years. P = 0.017). However, according to imPERCIST5 criterion, this difference was not observed (P = 0.12). Although the appearance of new lesions can be secondary to an inflammatory response to ICIs and indicative of pseudoprogression, given the higher rate of true progression, the appearance of new lesions should be interpreted deliberately. Of the three assessed modified criteria, PERCIMT appear to provide more reliable metabolic response assessment that correlates strongly with overall patient survival.


Selective modulation of promoter recruitment and transcriptional activity of PPARgamma.
Peroxisome proliferator-activated receptor gamma (PPARgamma) is a nuclear receptor regulated by the insulin-sensitizing thiazolidinediones (TZDs). We studied selective modulation of endogenous genes by PPARgamma ligands using microarray, RNA expression kinetics, and chromatin immunoprecipitation (ChIP) in 3T3-L1 adipocytes. We found over 300 genes that were significantly regulated the TZDs pioglitazone, rosiglitazone, and troglitazone. TZD-mediated expression profiles were unique but overlapping. Ninety-one genes were commonly regulated by all three ligands. TZD time course and dose-response studies revealed gene- and TZD-specific expression kinetics. PEPCK expression was induced rapidly but PDK4 expression was induced gradually. Troglitazone EC50 values for PEPCK, PDK4, and RGS2 regulation were greater than those for pioglitazone and rosiglitazone. TZDs differentially induced histone acetylation of and PPARgamma recruitment to target gene promoters. Selective modulation of PPARgamma by TZDs resulted in distinct expression profiles and transcription kinetics which may be due to differential promoter activation and chromatin remodeling of target genes.


Expectant management versus IUI in unexplained subfertility and a poor pregnancy prognosis (EXIUI study): a randomized controlled trial.
For couples with unexplained subfertility and a poor prognosis for natural conception, is 6 months expectant management (EM) inferior to IUI with ovarian stimulation (IUI-OS), in terms of live births? In couples with unexplained subfertility and a poor prognosis for natural conception, 6 months of EM is inferior compared to IUI-OS in terms of live births. Couples with unexplained subfertility and a poor prognosis are often treated with IUI-OS. In couples with unexplained subfertility and a relatively good prognosis for natural conception (>30% in 12 months), IUI-OS does not increase the live birth rate as compared to 6 months of EM. However, in couples with a poor prognosis for natural conception (<30% in 12 months), the effectiveness of IUI-OS is uncertain. We performed a non-inferiority multicentre randomized controlled trial within the infrastructure of the Dutch Consortium for Healthcare Evaluation and Research in Obstetrics and Gynaecology. We intended to include 1091 couples within 3 years. The couples were allocated in a 1:1 ratio to 6 months EM or 6 months IUI-OS with either clomiphene citrate or gonadotrophins. We studied heterosexual couples with unexplained subfertility and a poor prognosis for natural conception (<30% in 12 months). The primary outcome was ongoing pregnancy leading to a live birth. Non-inferiority would be shown if the lower limit of the one-sided 90% risk difference (RD) CI was less than minus 7% compared to an expected live birth rate of 30% following IUI-OS. We calculated RD, relative risks (RRs) with 90% CI and a corresponding hazard rate for live birth over time based on intention-to-treat and per-protocol (PP) analysis. Between October 2016 and September 2020, we allocated 92 couples to EM and 86 to IUI-OS. The trial was halted pre-maturely owing to slow inclusion. Mean female age was 34 years, median duration of subfertility was 21 months. Couples allocated to EM had a lower live birth rate than couples allocated to IUI-OS (12/92 (13%) in the EM group versus 28/86 (33%) in the IUI-OS group; RR 0.40 90% CI 0.24 to 0.67). This corresponds to an absolute RD of minus 20%; 90% CI: -30% to -9%. The hazard ratio for live birth over time was 0.36 (95% CI 0.18 to 0.70). In the PP analysis, live births rates were 8 of 70 women (11%) in the EM group versus 26 of 73 women (36%) in the IUI-OS group (RR 0.32, 90% CI 0.18 to 0.59; RD -24%, 90% CI -36% to -13%) in line with inferiority of EM. Our trial did not reach the planned sample size, therefore the results are limited by the number of participants. This study confirms the results of a previous trial that in couples with unexplained subfertility and a poor prognosis for natural conception, EM is inferior to IUI-OS. The trial was supported by a grant of the SEENEZ healthcare initiative. The subsidizing parties were The Dutch Organisation for Health Research and Development (ZonMW 837004023, www.zonmw.nl) and the umbrella organization of 10 health insurers in The Netherlands. E.R.G. receives personal fees from Titus Health care outside the submitted work. M.G. declares unrestricted research and educational grants from Guerbet, Merck and Ferring not related to the presented work, paid to their institution VU medical centre. A.B.H. reports receiving travel and speakers fees from Nordic Pharma and Merck and he is member of the Nordic Pharma ANGEL group and of the Safety Monitoring Board of Womed. C.B.L. reports speakers fee from Inmed and Yingming, and his department receives research grants from Ferring, Merck and Guerbet paid to VU medical centre. B.W.J.M. is supported by a NHMRC Investigator grant (GNT1176437) and reports consultancy for ObsEva and Merck. M.v.W. received a grant from the Netherlands Organisation for Health Research and Development ZonMW (80-8520098-91072). F.M. received two grants from the Netherlands Organisation for Health Research and Development ZonMW (NTR 5599 and NTR 6590). The other authors report no competing interest. Dutch Trial register NL5455 (NTR5599). 18 December 2015. 26 January 2017.


Do statins improve outcomes for patients with non-small cell lung cancer? A systematic review and meta-analysis protocol.
Lung cancer is the most common neoplasm and the leading cause of cancer-related death worldwide. Non-small cell lung cancer (NSCLC), accounting for 85% of all lung cancer cases, is frequently diagnosed at an advanced and metastatic stage. In addition, survival of patients with NSCLC has not improved significantly over the recent decades. Statins are used as a cholesterol-lowering agent, but recently preclinical and clinical studies have revealed their anticancer effects. Thus, this systematic review and meta-analysis aims to clarify whether statins improve the prognosis of patients with NSCLC. We will search MEDLINE (PubMed), EMBASE, Web of Science, the Cochrane Central Register of Controlled Trials and ClinicalTrials.gov with no restriction on language. Both randomised controlled trials (RCTs) and observational cohort studies evaluating the prognostic role of statins in patients with NSCLC will be included. The primary outcome will be overall survival, and the secondary outcomes will include cancer-specific survival, disease-free survival and cancer recurrence. Two assessors will assess the RCTs using the Cochrane Collaboration's risk of bias tool and the observational cohort studies according to ROBINS-I. Publication bias will be assessed by funnel plot using the STATA software v.13.1. No ethical issues are predicted. This systematic review and meta-analysis aims to describe the prognostic effects of statins in patients with NSCLC, which would help clinicians to optimise treatment for patients with NSCLC. These findings will be published in a peer-reviewed journal and presented at national and international conferences. CRD42016047524.


Evaluation of separate vs pooled swab cultures, different media, broth enrichment and anatomical sites of screening for the detection of methicillin-resistant Staphylococcus aureus from clinical specimens.
Early identification of methicillin-resistant Staphylococcus aureus (MRSA) carriers is a major component of an MRSA control programme. The cost and laboratory workload could be markedly reduced by processing multiple swabs from one person in one culture broth (specimen pooling). We evaluated the sensitivity for MRSA detection and the growth rate of pooled swabs compared with individual processing. In total, 1254 swabs from 423 subjects (two to five swabs per subject) were submitted for detection of MRSA. Swabs were suspended in 2-mL volumes of sterile Todd-Hewitt Broth and divided into two 1-mL aliquots. One aliquot of the suspension was processed as a single specimen, and the other aliquot was mixed (pooled) with other suspensions in which swabs from the same patient were suspended. Forty-four (10%) pooled samples were positive for MRSA. Specimens from seven additional patients that were negative when pooled were positive when processed separately. There was no case where the pooled specimen was positive but the separate specimens were negative. The diagnostic sensitivity of pooled surveillance cultures compared with single cultures, when only subjects colonized by MRSA were considered, was 86% and the false-negative rate was 14%. Eighty percent of the pooled positive cultures were detected by the third day and all were detected by the fourth day. Fifty-four percent of the specimens processed separately were detected by the second day and all were detected by the fourth day. Pooling of specimens decreases the sensitivity of MRSA detection compared with processing each swab separately, particularly in swabs with a low number of colony-forming units. In all subjects whose pooled samples were negative but whose swabs examined separately were positive, the swabs examined separately were negative on primary plates and positive only after culturing in enrichment broth.


[A method for determining the phagocytic activity of polymorphonuclear leukocytes].
The proposed method is said to provide more precise and reliable information about phagocytic properties of PNL at the light-optical and electron-microscopic levels than do the other currently available and widely used techniques.


Integrated bioprocess for structured lipids, emulsifiers and biodiesel production using crude acidic olive pomace oils.
Olive pomace oil (OPO), a by-product of olive oil industry, is directly consumed after refining. The novelty of this study consists of the direct use of crude high acidic OPO (3.4-20% acidity) to produce added-value compounds, using sn-1,3-regioselective lipases: (i) low-calorie dietetic structured lipids (SL) containing caprylic (C8:0) or capric (C10:0) acids by acidolysis or interesterification with their ethyl esters, (ii) fatty acid methyl esters (FAME) for biodiesel, and (iii) sn-2 monoacylglycerols (emulsifiers), as by-product of FAME production by methanolysis. Immobilized Rhizomucor miehei lipase showed similar activity in acidolysis and interesterification for SL production (yields: 47.8-53.4%, 7 h, 50℃) and was not affected by OPO acidity. Batch operational stability decreased with OPO acidity, but it was at least three-fold in interesterification that in acidolysis. Complete conversion of OPO into FAME and sn-2 monoacylglycerols was observed after 3 h-transesterification (glycerol stepwise addition) and lipase deactivation was negligeable after 11 cycles.


Monocyte procoagulant activity in breast cancer.
Abnormalities of blood coagulation associated with neoplasia may be important in the pathogenesis of tumour spread. Most patients with advanced malignancy have evidence of activated coagulation, but the mechanisms underlying this are unclear. We have examined in vitro monocyte procoagulant activity and compared this to plasma levels of fibrinopeptide A, in 52 patients with clinically localised breast cancer. Patients with localised breast cancer and activated coagulation displayed a strong positive correlation between monocyte procoagulant activity and level of fibrinopeptide A(r = +0.86, p less than 0.001). No such relationship was demonstrated in a smaller number of patients with metastatic breast cancer. It is concluded that monocyte procoagulant activity plays an important role in coagulation activation in patients with localised breast cancer. The implications of this for adjuvant anticoagulant therapy in breast cancer are discussed.


Interaction between human evoked electrospinograms elicited by segmental and descending volleys.
Interaction between the slow negative-positive waves of human evoked electrospinograms produced by descending and segmental volleys was tested under general anaesthesia. A partial occlusion was demonstrated in these slow waves.


Ethics and fetal medicine.
Ethical issues in the clinical practice of fetal medicine are discussed, largely from the point of view of early prenatal medicine. The discussion concentrates on several aspects including the time when human life begins, the pros and cons of fetal medicine, and ethical guidelines for fetal medicine. The emphasis is placed on the importance of informed consent and an increase in genetical knowledge amongst the general public.


The treatment of schizophrenia: from premorbid manifestations to the first episode of psychosis.
To achieve the best therapeutic results in schizophrenia--like most other disorders--primary prevention is preferable to early and prompt treatment, which, in turn, is preferable to treatment of chronically established illness. Unfortunately, there currently exist no accurate markers that can provide information regarding the future course of illness and guide treatment in asymptomatic or mildly symptomatic individuals. Therefore, most treatment efforts are currently focused on patients who have already experienced their first psychotic episode. This paper reviews the efforts to identify accurate markers heralding psychotic illness, as well as treatment considerations in the early phase of the disease.


Using an existing denture to design a radiographic template for a two-implant mandibular overdenture.
This article describes a technique for modifying an existing mandibular complete denture for use as a radiographic template with a radiopaque light-activated calcium hydroxide (Ca(OH)(2)) preparation. This allows prosthetically driven treatment planning and the surgical placement of 2 implants to support the existing mandibular denture.


Acute renal failure in African children with Burkitt's lymphoma: a comparison of two treatment regimens.
The outcome for patients presenting with acute renal failure and Burkitt lymphoma (BLARF) without dialysis is poor. This was a retrospective non-randomized comparative study designed to determine the outcome of two different treatment protocols. One group of patients (TPA) received oral allopurinol, intravenous (IV) cyclophosphamide, vincristine, methotrexate, furosemide, 8.4% sodium bicarbonate, and intrathecal (IT) methotrexate; the other (TPB) alternate day IV infusion of low dose cyclosphosphamide (125 mg/m(2) x 4 doses), IT methotrexate (Days 1 and 5) and aggressive pre-emptive anti-tumor lysis syndrome therapy including oral allopurinol and calcium lactate, IV calcium gluconate, salbutamol, insulin and infusions of furosemide, sodium bicarbonate and glucose. Nine of 16 received TPA, 7 received TPB. Post chemotherapy anemia was more severe with TPA (P < 0.05). TPB patients received significantly more chemotherapy than those in TPA (P = 0.04). All 16 had tumor lysis syndrome (TLS). Six of nine patients with TPA died from this (three from other causes), two deaths in TPB were due to causes other than tumor lysis. Other evaluated outcome indices were similar in both groups. Slow IV infusion of low dose cyclophosphamide given on alternate days in addition to pre-emptive anti-TLS measures (TPB) were associated with better outcome in BLARF patients compared to a high dose multiple chemotherapy regimen (TPA).


Approach to Cataract Surgery in an Ebola Virus Disease Survivor with Prior Ocular Viral Persistence.
A 46-year-old patient with previously documented Ebola virus persistence in his ocular fluid, associated with severe panuveitis, developed a visually significant cataract. A multidisciplinary approach was taken to prevent and control infection. Ebola virus persistence was assessed before and during the operation to provide safe, vision-restorative phacoemulsification surgery.


Use of internal callus distraction in the treatment of congenital brachymetatarsia.
Congenital bilateral brachymetatarsia of the fourth metatarsal bone was treated by osteotomy and bone lengthening using the internal distraction technique. The patient was cosmetically improved. This technique has some advantages over the method of external distraction.


Effect of Holistic Nursing Intervention Combined with Humanized Nursing Intervention on Activities of Daily Living and Limb Movement Ability of Elderly Patients with Cerebral Hemorrhage after Surgery.
To improve the nursing status, activities of daily living (ADL), and limb movement ability of elderly patients with cerebral hemorrhage and to actively explore the comprehensive nursing mode that can meet the rehabilitation needs of patients. 128 elderly patients with cerebral hemorrhage admitted to our department (2019.01-2021.01) were included in the study and randomly divided into the reference group and the study group according to the numeration table, with 64 cases in each group. The reference group received routine nursing, while the study group received holistic nursing combined with humanized nursing. Statistical methods were used to analyze the nursing effect of the two intervention models. With better scores of the upper limb muscle strength, lower limb muscle strength, and total Fugl-Meyer Assessment (FMA) in both groups after nursing, the scores in the study group after nursing were higher than those in the reference group (P < 0.05). After nursing, the ADL scores of both groups were significantly improved and the score in the study group was better than that in the reference group (P < 0.05). After nursing, the degree of neurological deficit in both groups was significantly better than that before nursing and the degree in the reference group was more serious than that in the study group (P < 0.05). Compared with the reference group, the total infection rate in the study group was lower, while the nursing satisfaction was higher, with statistical significance (P < 0.05). The holistic nursing combined with humanized nursing has a remarkable effect on the postoperative intervention of elderly patients with cerebral hemorrhage. It can improve the limb movement function, enhance the ADL, reduce the degree of neurological deficit, improve the quality of life, and enhance the nursing satisfaction of patients after intervention, which is worthy of clinical popularization.


Chaos in the cardiovascular system: an update.
Rhythmic changes of blood pressure, heart rate, and other cardiovascular measures have drawn the attention of several investigators, since these oscillations can shed light onto the activity of the underlying control network. The overwhelming proportion of circulatory variations, however, are not linear, i.e., they do not consist of perfectly rhythmic components. Thus, these fluctuations are more adequately analysed by non-linear techniques, most of which are adopted from chaos theory. A spotlight issue of 'Cardiovascular Research' (Vol. 31, 1996), focused on chaos in the cardiovascular system. This current review outlines today's understanding of this field by presenting the major discoveries and developments which have taken place since then.


Molecular size distribution of somatostatin-like immunoreactivity in the cerebroventricular fluid of neurosurgical patients.
The molecular size distribution of somatostatin-like immunoreactivity (SLI) in the cerebroventricular fluid of patients with Parkinson's disease, dystonic syndromes, multiple sclerosis, basal and midline tumors, epilepsy and pain syndromes was investigated by separation with a Sephadex G-50f column and subsequent radioimmunoassay of the eluate. Marked heterogeneity of SLI was observed in most of the pools investigated. The most conspicuous feature of the elution profiles was the preponderance of the peak coeluting with synthetic somatostatin-14, whereas the peaks comigrating with synthetic somatostatin-28 and attributable to precursor-like SLI represented only minor or trace amounts of total immunoreactivity. These findings are consistent with the greater biological activity of somatostatin-14 in the human central nervous system, whereas somatostatin-28 appears to represent the more active form in the pituitary and in the intestinal mucosa. Solely in the case of brain tumor patients, some differences could be seen, resulting in an approximately equal distribution of somatostatin-14 and somatostatin-28 in two pools of ventricular fluid and by the detection of a degradation product of somatostatin-14 in another one. These observations could be explained by a lowered barrier function as a consequence of increased intracranial pressure in case of brain tumors, which is well in accordance with a markedly elevated total protein content being a sign of a lowered barrier function.


Quantification and persistence of COVID-19 virus in recently deceased individuals before and after embalming.
The COVID-19 pandemic severely affected the medical education worldwide. The infection risk for medical students and healthcare personnel who work with COVID-19 positive cadavers or tissues remains unclear. Moreover, COVID-19 positive cadavers have been rejected by medical schools, adversely impacting the continuum of medical education. Herein, the viral genome abundance in tissues from four COVID-19 positive donors before and after embalming were compared. Tissue samples were collected from the lungs, liver, spleen, and brain both pre- and postembalming. The possible presence of infectious COVID-19 was determined by inoculating human tissue homogenates onto a monolayer of human A549-hACE2 cells and observing for cytopathic effects up to 72 h postinoculation. A real- time quantitative reverse transcription polymerase chain reaction was performed to quantify COVID-19 present in culture supernatants. Fully intact viral genome sequence was possible to obtain in samples with higher levels of virus, even several days postmortem. The embalming procedure described above substantially reduces the abundance of viable COVID-19 genomes in all tissues, sometimes even to undetectable levels. However, in some cases, COVID-19 RNA can still be detected, and a cytopathic effect can be seen both pre- and postembalmed tissues. This study suggests that embalmed COVID-19 positive cadavers might be used safely with appropriate precautions followed in gross anatomy laboratories and in clinical and scientific research. Deep lung tissue is the best specimen to test for the virus. If the tests on the lung tissues are negative, there is a very low likelihood that other tissues will show positive results.


Intracellular calcium rise produced by platelet-activating factor is deactivated by fMet-Leu-Phe and this requires uninterrupted activation sequence: role of protein kinase C.
Stimulation of the neutrophils with fMet-Leu-Phe inhibits the rise in intracellular concentration of free calcium produced by the subsequent addition of platelet-activating factor. This deactivation is not observed in pertussis toxin treated cells. In addition, preincubation of the cells with the protein kinase C activator phorbol 12-myristate 13-acetate for three minutes abolishes completely the rise in calcium produced by platelet-activating factor. This inhibition is prevented by the addition of the protein kinase C inhibitor 1-(5-isoquinoline-sulfonyl)-2-methyl piperazine prior to the addition of the phorbol ester. Phorbol 12-myristate 13-acetate, at a concentration that does not produce significant inhibition, accelerates the rate of calcium removal from the cytoplasm, and this is abolished by the protein kinase C inhibitor. In contrast, the deactivation by fMet-Leu-Phe is not prevented by the protein kinase C inhibitor. The results presented here suggest that the protein kinase C system may regulate the opening by platelet-activating factor of possible plasma membrane associated pertussis toxin independent calcium channels and/or the binding of platelet-activating factor to the receptors. In addition, protein kinase C activation increases the rates of the calcium efflux pump and/or calcium sequestering by intracellular organelles. The most simple and straightforward explanation of the observed deactivation by fMet-Leu-Phe is that the addition of fMet-Leu-Phe to neutrophils stimulates the production of platelet-activating factor which then binds to and deactivates the receptors.


A Feasibility Study of Smartphone-Based Telesonography for Evaluating Cardiac Dynamic Function and Diagnosing Acute Appendicitis with Control of the Image Quality of the Transmitted Videos.
Our aim was to prove the feasibility of the remote interpretation of real-time transmitted ultrasound videos of dynamic and static organs using a smartphone with control of the image quality given a limited internet connection speed. For this study, 100 cases of echocardiography videos (dynamic organ)-50 with an ejection fraction (EF) of ≥50 s and 50 with EF <50 %-and 100 cases of suspected pediatric appendicitis (static organ)-50 with signs of acute appendicitis and 50 with no findings of appendicitis-were consecutively selected. Twelve reviewers reviewed the original videos using the liquid crystal display (LCD) monitor of an ultrasound machine and using a smartphone, to which the images were transmitted from the ultrasound machine. The resolution of the transmitted echocardiography videos was reduced by approximately 20 % to increase the frame rate of transmission given the limited internet speed. The differences in diagnostic performance between the two devices when evaluating left ventricular (LV) systolic function by measuring the EF and when evaluating the presence of acute appendicitis were investigated using a five-point Likert scale. The average areas under the receiver operating characteristic curves for each reviewer's interpretations using the LCD monitor and smartphone were respectively 0.968 (0.949-0.986) and 0.963 (0.945-0.982) (P = 0.548) for echocardiography and 0.972 (0.954-0.989) and 0.966 (0.947-0.984) (P = 0.175) for abdominal ultrasonography. We confirmed the feasibility of remotely interpreting ultrasound images using smartphones, specifically for evaluating LV function and diagnosing pediatric acute appendicitis; the images were transferred from the ultrasound machine using image quality-controlled telesonography.


High-fidelity correction of genomic uracil by human mismatch repair activities.
Deamination of cytosine to produce uracil is a common and potentially mutagenic lesion in genomic DNA. U*G mismatches occur spontaneously throughout the genome, where they are repaired by factors associated with the base excision repair pathway. U*G mismatches are also the initiating lesion in immunoglobulin gene diversification, where they undergo mutagenic processing by redundant pathways, one dependent upon uracil excision and the other upon mismatch recognition by MutS alpha. While UNG is well known to initiate repair of uracil in DNA, the ability of MutS alpha to direct correction of this base has not been directly demonstrated. Using a biochemical assay for mismatch repair, we show that MutS alpha can promote efficient and faithful repair of U*G mismatches, but does not repair U*A pairs in DNA. This contrasts with UNG, which readily excises U opposite either A or G. Repair of U*G by MutS alpha depends upon DNA polymerase delta (pol delta), ATP, and proliferating cell nuclear antigen (PCNA), all properties of canonical mismatch repair. These results show that faithful repair of U*G can be carried out by either the mismatch repair or base excision repair pathways. Thus, the redundant functions of these pathways in immunoglobulin gene diversification reflect their redundant functions in faithful repair. Faithful repair by either pathway is comparably efficient, suggesting that mismatch repair and base excision repair share the task of faithful repair of genomic uracil.


Breakage Costs in Flexible Ureteroscopy: Digital vs. Fiberoptic Modalities.
To compare the maintenance costs of digital flexible ureteroscopes (DFU) versus fiberoptic flexible ureteroscopes (FFU) to understand the long-term financial impact associated with breakage in a flexible ureteroscopy (f-URS) program. Data for breakage of FFU and DFU at an academic institution from 2019 to 2021 were obtained from our vendor (Karl Storz) and analyzed by month. Correlation test was used to evaluate significant differences in number of procedures, number of breakage events, breakage rates, and repair cost per month. Cumulative analyses were utilized to examine the number of procedures before failure (time to failure - TTF) and repair costs per procedure (RCpP). We performed a total of 2,154 f-URS, including 1,355 with FFU and 799 with DFU (P<.001). Although we found a higher number of breakage events in FFU (n=124) than DFU (n=73) (P<.001), the overall breakage rate was similar, 9.9% vs. 8.8%, respectively (P=0.86). On cumulative analysis, both modalities reached the same TTF plateau (11 cases) after 18 months. After 400 cases, the RCpP for DFU was 1.25 times higher than for FFU (P=0.04). Overall, we found no difference in overall scope breakage rates between DFU and FFU. Although there was no difference in TTF over time, at the beginning DFU displayed considerable higher durability, leading to lower RCpP. Furthermore, DFU's endurance leveled off to FFU over time, resulting in higher RCpP after 400 cases. This finding may be explained by the presence of renewed scopes after repair.


Morphological, biological, and genomic characterization of Klebsiella pneumoniae phage vB_Kpn_ZC2.
Bacteriophages (phages) are one of the most promising alternatives to traditional antibiotic therapies, especially against multidrug-resistant bacteria. Klebsiella pneumoniae is considered to be an opportunistic pathogen that can cause life-threatening infections. Thus, this study aims at the characterization of a novel isolated phage vB_Kpn_ZC2 (ZCKP2, for short). The phage ZCKP2 was isolated from sewage water by using the clinical isolate KP/08 as a host strain. The isolated bacteriophage was purified and amplified, followed by testing of its molecular weight using Pulse-Field Gel Electrophoresis (PFGE), transmission electron microscopy, antibacterial activity against a panel of other Klebsiella pneumoniae hosts, stability studies, and whole genome sequencing. Phage ZCKP2 belongs morphologically to siphoviruses as indicated from the Transmission Electron Microscopy microgram. The Pulsed Field Gel Electrophoresis and the phage sequencing estimated the phage genome size of 48.2 kbp. Moreover, the absence of lysogeny-related genes, antibiotic resistance genes, and virulence genes in the annotated genome suggests that phage ZCKP2 is safe for therapeutic use. Genome-based taxonomic analysis indicates that phage ZCKP2 represents a new family that has not been formally rated yet. In addition, phage ZCKP2 preserved high stability at different temperatures and pH values (-20 - 70 °C and pH 4 - 9). For the antibacterial activity, phage ZCKP2 maintained consistent clear zones on KP/08 bacteria along with other hosts, in addition to effective bacterial killing over time at different MOIs (0.1, 1, and 10). Also, the genome annotation predicted antibacterial lytic enzymes. Furthermore, the topology of class II holins was predicted in some putative proteins with dual transmembrane domains that contribute significantly to antibacterial activity. Phage ZCKP2 characterization demonstrates safety and efficiency against multidrug-resistant K. pneumoniae, hence ZCKP2 is a good candidate for further in vivo and phage therapy clinical applications.


Machine learning methods for estimating bent photonic crystal fiber based SPR sensor properties.
In the recent years, the use of machine learning approaches in optical devices and fibers is increasing. However, most methods concentrate on the use of Artificial Neural Network (ANN) methods due to the ability of automatically fitting to the problem. In this work, a classical non-linear regression method, namely k-Nearest Neighbor Regression (KNNR) is proposed for determining the loss characteristics of a photonic crystal fiber (PCF) based surface plasmon resonance (SPR) sensor in the presence of a bend in either x or y direction. Although KNNR is a simple method, it is very well known that in certain systems it can out-perform ANN. It is believed that PCF based structures can be a good candidate for this comparison. In order to judge the performance of different regression techniques, we have built a database that contains 1180 samples. The dataset contains PCF structure data for non-bent(straight fiber), bent in x and y-directions. Experiments show that KNNR outperforms both ANN and Linear Least Square Regression methods even when a feature space expansion method is employed. In addition, KNNR does not require any lengthy training process, allowing it to be used instantly once the training data is available. This can be exploited to complement existing simulation techniques.


Regulation of matrix vesicle phospholipid metabolism is cell maturation-dependent.
We have developed a chondrocyte culture model for assessing the regulation of matrix vesicles at two different stages of chondrogenic maturation. These chondrocytes, resting zone (RC) and growth zone (GC), retain their phenotypic markers in culture, including production of matrix vesicles with distinctive lipid compositions and enzyme activities. Isolated matrix vesicles incubated in vitro with 1,25-(OH)2D3 (1,25) or 24,25-(OH)2D3 (24,25) respond differentially. 1,25 stimulates phospholipase A2 (PA2) in GC vesicles, but not on those from RC. 24,25 inhibits PA2 in RC vesicles, but has no effect on GC. PA2 activity is required for fatty acid turnover and is the rate-limiting step in prostaglandin production. Plasma membrane phospholipids are more susceptible to the release of arachidonic acid by PA2 than are matrix vesicle phospholipids. Matrix vesicles are distinct from the plasma membrane in terms of lipid composition and arachidonic acid incorporation. 1,25 and 24,25 stimulate arachidonic acid turnover in their target cells, but by different mechanisms. 1,25 has no effect on arachidonic acid turnover in RC; however, 24,25 inhibits turnover in RC and GC. 1,25 and 24,25 also affect isolated matrix vesicle membrane fluidity. These results suggest that vitamin D metabolites modulate PA2 activity, change the composition of membrane phospholipids by altering fatty acid composition, and affect calcium transport. The effects are mediated by altering membrane fluidity and is dependent on the stage of cell differentiation.


Influence of maternal adrenalectomy on the ultrastructure of the adrenal gland in neonatal rats.
Maternal adrenalectomy at 7 or 14 days of gestation produced increased cell necrosis within zona reticularis cells on the day of birth and at 24 or 48 h after birth. Small remnants or large portions of adrenocortical cells were present within macrophages. In otherwise normal adrenocortical cells, lipid droplets were incorporated within some mitochondria. Autophagocytosis of single mitochondria was observed within adrenocortical cells. Undoubtedly ultrastructural changes represent stimulation of adrenocortical cells in neonatal rats in response to maternal adrenalectomy.


Spinal Serotonergic and Opioid Receptors Are Involved in Electroacupuncture-Induced Antinociception at Different Frequencies on ZuSanLi (ST 36) Acupoint.
The present study was conducted to evaluate the effect of electroacupuncture-(EAc-) induced antinociception (EAA) at different currents and frequencies in rat spinal cord. We found that naloxone (0.05 μ g i.t.) blocked EAA at different frequencies. Naltrindole (0.05 μ g i.t.) blocked EAA on the 7th day after EAc of 100 Hz. 5,7-Dihydroxytryptamine (100 μ g i.t.) significantly inhibited EAA at different frequencies on the 7th day after EAc. Pindobind (0.5 μ g i.t.), a 5-HT1A antagonist, notably attenuated EAA at different frequencies. Ketanserin (0.5 μ g i.t.), inhibited EEA at a lower frequency (<10 Hz) than at a higher frequency (100 Hz). LY-278584 (0.5 μ g i.t.) significantly inhibited EAA at a higher frequency (100 Hz) on the 7th day after EAc. The direction of effect of 8-OH-DPAT, on EAA was dependent on dosage. It had an inhibitory effect at a low dose (0.5 μ g i.t.) and a high frequency (100 Hz) but enhanced EAA at a higher dose at lower frequencies (<10 Hz). DOI (10 μ g, i.t.), did not affect EAA. These data indicate that the mechanism of EAA involves opioid receptors, and the serotonergic system, particularly, μ -, δ -opioid and 5-HT1A, 5-HT3 receptors and it is also dependent on the EAc frequency.


Parental alcohol intoxication and adverse health outcomes among offspring. A 4-year follow up HUNT study among 2399 Norwegian adolescents.
This study aimed to investigate whether experience of parental alcohol intoxication was associated with adverse health outcomes among adolescents four years later. A population-based Norwegian cohort study of 2399 adolescents who participated in the Young-HUNT1 Survey 1995-97 (T1, 13-15 years old) was followed up four years later (T2) in 2000 (Young-HUNT2, 17-19 years old). Measures were based on adolescent self-report of exposure to parental alcohol intoxication, self-rated general health, mental distress (SCL-5) and lifetime hospital admission. Multivariable logistic regression analyses, adjusted for gender, age and parental education were applied. Results show that half of the adolescents (51%) had seen parents intoxicated at age 13-15 years. Four years later, those who had reported parental alcohol intoxication at T1 had increased odds of admission to hospital with odds ratios ranging from OR1.3; CI 1.0-1.7 to OR 2.2; CI 1.3-3.9, poorer self-rated health (odds ratio ranging from 1.8;1.2-2.6 to 2.0;1.1-3.7) and more mental distress (odds ratio ranging from 1.7;1.1-2.5 to 1.9;1.0-3.6). Furthermore, the increased frequency of experience of parental alcohol intoxication are associated with higher prevalence of admission to hospital and mental distress and lower levels of self-rated health. Findings from this large, representative population of Norwegian adolescents indicate that adverse health outcomes among adolescents in a general population are related to relatively common heavy drinking behaviours among parents.


Targeting miR-126 disrupts maintenance of myelodysplastic syndrome stem and progenitor cells.
Myelodysplastic syndrome (MDS) arises from a rare population of aberrant hematopoietic stem and progenitor cells (HSPCs). These cells are relatively quiescent and therefore treatment resistant. Understanding mechanisms underlying their maintenance is critical for effective MDS treatment. We evaluated microRNA-126 (miR-126) levels in MDS patients' sample and in a NUP98-HOXD13 (NHD13) murine MDS model along with their normal controls and defined its role in MDS HSPCs' maintenance by inhibiting miR-126 expression in vitro and in vivo. Identification of miR-126 effectors was conducted using biotinylated miR-126 pulldown coupled with transcriptome analysis. We also tested the therapeutic activity of our anti-miR-126 oligodeoxynucleotide (miRisten) in human MDS xenografts and murine MDS models. miR-126 levels were higher in bone marrow mononuclear cells from MDS patients and NHD13 mice relative to their respective normal controls (P < 0.001). Genetic deletion of miR-126 in NHD13 mice decreased quiescence and self-renewal capacity of MDS HSPCs, and alleviated MDS symptoms of NHD13 mice. Ex vivo exposure to miRisten increased cell cycling, reduced colony-forming capacity, and enhanced apoptosis in human MDS HSPCs, but spared normal human HSPCs. In vivo miRisten administration partially reversed pancytopenia in NHD13 mice and blocked the leukemic transformation (combination group vs DAC group, P < 0.0001). Mechanistically, we identified the non-coding RNA PTTG3P as a novel miR-126 target. Lower PTTG3P levels were associated with a shorter overall survival in MDS patients. MiR-126 plays crucial roles in MDS HSPC maintenance. Therapeutic targeting of miR-126 is a potentially novel approach in MDS.


Prothrombotic effects of endothelin-2 (ET-2).
In this report we demonstrate that ET-2, in addition to its powerful vasoconstrictor properties and its role in hypertension, is capable of creating a hypercoagulable (prothrombotic) state. This study was facilitated by the utilization of a sensitive modified recalcification time (MRT) test that can measure coagulation at all points in the coagulation spectrum. The MRT was determined on aliquots (1 ml) of citrated blood which were added to saline (MRTS), to 10 micrograms of endotoxin (MRTE), and to 10 pg of ET-2 (MRT ET-2). The mean values of MRTS, MRTE, and MRT ET-2 were 4.3 +/- 0.8, 3.7 +/- 1.0, and 3.9 +/- 0.8 min, respectively. There was a statistically significant difference between MRTE vs. MRTS and MRT ET-2 vs. MRTS using the two-tailed t-test (p < 0.02 and p < 0.005, respectively).


The effects of hematoma on implant capsules.
Hematoma surrounding an implant is one of the many factors that have been suggested as possible causes for scar capsule contracture. In this study, experiments were designed to determine the influence of hematoma on the incidence and severity of capsule contracture in rabbits. Two implants were placed in each animal, 1 with a surrounding hematoma and 1 control. Capsules were evaluated subjectively and compared objectively with measurements of deformability, surface area, and capsule thickness. No differences were found with any of the objective criteria, which suggests that hematoma may not be a noteworthy cause of implant capsule contracture.


Complications after artificial urinary sphincter implantation in patients with or without prior radiotherapy.
To compare complications after implantation of an artificial urinary sphincter (AUS) in patients with or without prior radiotherapy (RT). Between January 2000 and December 2011, 160 patients underwent AMS 800 AUS implantation in our institution. We excluded neurological and traumatic causes, implantation on ileal conduit diversion, penoscrotal urethral cuff position and those lost to follow-up. In all, 122 patients were included in the study, 61 with prior RT and 61 without prior RT. All patients underwent the same surgical technique by two different surgeons. All AUS were implanted with a bulbar urethral cuff position. The mean (range) follow-up was 37.25 (1-126) months. In the patients without prior RT and those with prior RT, revision rates were 32.8% vs 29.5%, respectively (P = 0.59). The median time to first revision was 11.7 months. Early complications were similar in the two groups (4.9% vs 6.5%, P = 1). Erosion rates were not significantly different (4.9% vs 13.1%, P = 0.13). However, infection and explantation were more prevalent in patients with prior RT [two (3.2%) vs 10 (16.3%), P = 0.018 and three (4.9%) vs 12 (19.6%), P = 0.016, respectively]. Finally, continence rates were not significantly different [75.4% (without prior RT) vs 63.9% (with prior RT), P = 0.23]. AUS is the 'gold standard' treatment of male urinary incontinence after re-education failure in patients with or without prior RT. Our experience showed similar functional outcomes in both groups but a higher rate of major complications in the group with prior RT.


Current developments in Coot for macromolecular model building of Electron Cryo-microscopy and Crystallographic Data.
Coot is a tool widely used for model building, refinement, and validation of macromolecular structures. It has been extensively used for crystallography and, more recently, improvements have been introduced to aid in cryo-EM model building and refinement, as cryo-EM structures with resolution ranging 2.5-4 A are now routinely available. Model building into these maps can be time-consuming and requires experience in both biochemistry and building into low-resolution maps. To simplify and expedite the model building task, and minimize the needed expertise, new tools are being added in Coot. Some examples include morphing, Geman-McClure restraints, full-chain refinement, and Fourier-model based residue-type-specific Ramachandran restraints. Here, we present the current state-of-the-art in Coot usage.


Establishment of human embryonic stem cell line with a complex abnormal karyotype and the stable XIST expression.
The embryonic stem cell line chHES-3XISTa was derived from heterogeneous chHES-3 cells. chHES-3XISTa showed a new abnormal karyotype of 46,XX with 8 derivation chromosomes and expressed X inactive specific transcript (XIST) in continual culture. Tri-methylation of H3 Lys-27 (H3K27me3) punctate enrichment located in RNA Polymerase II hole was also found in all chHES-3XISTa cells. Pluripotent markers and differentiate capability in vitro were confirmed by immunochemistry staining.


Monosodium glutamate induced testicular toxicity and the possible ameliorative role of vitamin E or selenium in male rats.
Monosodium glutamate (MSG) has been recognized as flavor enhancer that adversely affects male reproductive systems. The present study was carried out to evaluate the potential protective role of vitamin E (vit E) or selenium against MSG induced oxidative stress and histopathological changes in testis tissues of rats. Mature male Wistar rats weighing 150-200 g BW were allocated to evenly twelve groups each group of ten animals, the first group was maintained as control group, the 2nd, 3rd and 4th groups were administered MSG in three different dose levels (low, medium and high) (6, 17.5 and 60 mg/kg BW), the 5th and 6th groups were given vit E in two doses (low and high) (150 and 200 mg/kg), the 7th and 8th groups were administered selenium in two doses (low and high) (0.25 and 1 mg/kg) daily via gavage for a period of 30 days. Meanwhile the 9th and 10th groups were given combinations of MSG (high dose) and vit E while, the 11th and 12th groups were given MSG (high dose) plus selenium in two recommended doses for each one. Monosodium glutamate caused an elevation in lipid peroxidation level parallel with significant decline in SOD, CAT as well as GPx activities in testis tissues. Administration of vit E or selenium to MSG-treated groups declined lipid peroxidation, increased SOD, CAT, GPx activities. Selenium or vit E significantly reduced MSG induced histopathological changes by the entire restoration of the histological structures and the testicular antioxidant status to great extent in treated rats. In conclusion, supplementation of selenium or vit E could ameliorate the MSG induced testicular toxicity to great extent and reduce the oxidative stress on testis tissues.


Myocardial adenosine triphosphate content as a measure of metabolic and functional myocardial protection in children undergoing cardiac operation.
In an effort to quantitate the metabolic and functional response to global myocardial ischemia as a prelude to specific interventions to improve myocardial protection in children, the following data were collected. Twenty children (age, 1.16 +/- 0.3 years) underwent repair of congenital intracardiac malformations using aortic cross-clamping and cold potassium cardioplegia (ischemic time, 56.1 +/- 4.5 minutes). Metabolic protection was assessed by measuring the myocardial adenosine triphosphate (ATP) content by microbioluminescence. Before and after ischemia 10-mg myocardial samples were obtained from the left ventricular apex using a Tru-cut biopsy needle. In 15 patients, postoperative ventricular function was measured by radionuclide ventriculography at 72 to 96 hours following operation. Five of 6 patients with a postischemic ATP level less than 40% of control (26.3 +/- 2.8) had a left ventricular ejection fraction (EF) lower than 55% (50.3 +/- 2.3). Seven of 9 patients with an ATP level greater than 40% of the preischemic level (98.0 +/- 14.4) had a normal EF (61.8 +/- 2.9; p less than 0.04). Two other patients with postischemic ATP levels lower than 40% of control died of low cardiac output and had no postoperative ventricular function studies. Thus, of 7 patients with postischemic ATP levels lower than 40% of preischemic levels, 2 died and 5 had depressed left ventricular function. These data support the concept that low postischemic ATP levels correlate with death or poor postoperative ventricular function, and indicate that this variable will be useful to assess future improvements in myocardial protection during pediatric cardiac operations.


PEAR1 suppresses the proliferation of pulmonary microvascular endothelial cells via PI3K/AKT pathway in ALI model.
Activation of the proliferation of pulmonary microvascular endothelial cells (PMVECs) is a key step in the recovery of the integrity of endothelial monolayer, which helps to alleviate acute lung injury (ALI). Platelet endothelial aggregation receptor-1 (PEAR1), expressed on endothelial cells, was reported to inhibit the proliferation of vascular endothelial cells and angiogenesis. However, little is known about its role and mechanism in vascular endothelial disorders in ALI. The aim of this study was to investigate the impact of PEAR1 on the proliferation of pulmonary microvascular endothelial cells in ALI. We tested the expression level of PEAR1 in the lungs of WT mice in ALI model induced by intestinal IR. Primary human pulmonary microvascular endothelial cells (HPMECs) were stimulated by 1 mg/L LPS in vitro. We synthesized siPEAR1 and Flag-PEAR1 plasmid to verify the role of PEAR1 on regulating the proliferation of HPMECs under LPS condition and to explore related signaling pathways. The expression level of PEAR1 significantly increased in ALI induced by intestinal IR. PEAR1 knockdown enhanced the proliferation level of HPMECs, which, however, was inhibited by PEAR1 overexpression. PEAR1 knockdown activated PI3K/AKT pathway both in steady state and under LPS condition. PI3K inhibitor, LY294002, reversed the increasing proliferation level and cell progression of HPMECs induced by PEAR1 knockdown after LPS challenge. PEAR1 acts as a negative regulator in the proliferation of HPMECs in ALI model via the PI3K/AKT pathway.


Comparison of EQ-5D, HUI, and SF-36-derived societal health state values among spine patient outcomes research trial (SPORT) participants.
To compare societal values across health-state classification systems and to describe the performance of these systems at baseline in a large population of persons with confirmed diagnosis of intervertebral disc herniation (IDH), spinal stenosis (SpS), or degenerative spondylolisthesis (DS). We compared values for EQ-5D (York weights), HUI (Mark 2 and 3), SF-6D, and the SF-36-derived estimate of the Quality of Well Being (eQWB) score using signed rank tests. We tested each instrument's ability to discriminate between health categories and level of symptom satisfaction. Correlations were assessed with Spearman rank correlations. We evaluated ceiling and floor effects by comparing the proportion at the highest and the lowest possible score for each tool. In addition, we compared proportions at the highest and lowest levels by dimension. The number of unique health states assigned was compared across instruments. We calculated the difference between those who were very dissatisfied and all others. Mean values ranged from 0.39 to 0.63 among 2097 participants ages 18-93 (mean age 53, 47% female) with significant differences in pair-wise comparisons noted for all systems. Correlations ranged from 0.30 to 0.78. Although all systems showed statistically significant differences in health state values when baseline comparisons were made between those who were very dissatisfied with their symptoms and those who were not, the magnitude of this difference ranged widely across systems. Mean differences (95% CI) between those very dissatisfied and all others were 0.30 (0.269, 0.329) for EQ-5D, 0.22 (0.190, 0.241) for HUI(3), 0.18 (0.161, 0.201) for HUI(2), 0.11 (0.095, 0.117) for SF-6D, 0.04 (0.039, 0.049) for eQWB, and 0.07 (0.056, 0.077) for VAS (with transformation applied to group means). Differences in preference-weighted health state classification systems are evident at baseline in a population with confirmed IDH, SpS, and DS. Caution should be used when comparing health state values derived from various systems.


Does the Presence of Type 2 Diabetes or Metabolic Syndrome Impact Reduction in Waist Circumference During Weight Loss?
Our aim in this study was to compare the change in waist circumference given the same degree of weight loss in patients who meet the criteria for metabolic syndrome or type 2 diabetes and those who do not meet these criteria. Because visceral adiposity is a key feature of both conditions and intra-abdominal adipocytes show higher lipolytic activity, we sought to determine whether changes in waist circumference differed in individuals with and without these conditions. The Ottawa Hospital Weight Management Clinic offers a course in lifestyle modification and uses 12 weeks of total meal replacement. We compared the decrease in waist circumference between patients with metabolic syndrome or diabetes and those without these conditions who had lost a similar amount of weight using measurements from the first 6 weeks of meal replacement. We evaluated 3,559 patients who attended the program between September 1992 and April 2015. The patient population was largely Caucasian and of European descent and all meetings were face to face. The mean weight loss for men was 15.1±20.2 kg, and the mean weight loss for women was 9.7±2.4 kg. There were no significant differences in decrease in waist circumference between those with and without metabolic syndrome in both men (11.7±3.9 cm vs 11.4±3.8 cm, p=0.48) and women (9.0±3.6 cm vs 9.1±3.7 cm, p=0.26). Our results show that, given the same degree of weight loss, patients with and without diabetes or metabolic syndrome experience a similar change in waist circumference.


An investigation of pyruvate metabolism in patients with cerebellar and spinocerebellar degeneration.
This study extends previous observations of pyruvate metabolism in the spino-cerebellar degenerations by screening for abnormalities of pyruvate oxidation using the rise in blood pyruvate after an oral glucose load and examining the activity of the lipoamide dehydrogenase (LAD) moeity of the pyruvate dehydrogenase complex in the serum of 31 patients with Friedreich's ataxia, hereditary spastic ataxia and primary cerebellar degeneration. Serum LAD activity was significantly reduced in 10 Friedreich's ataxia patients when compared to controls and to 10 patients with spastic ataxia, thus confirming previous studies. Two patients with Friedreich's ataxia and 2 with primary cerebellar degeneration had abnormal blood pyruvate curves after oral glucose loading. The findings suggest that abnormal pyruvate oxidation occurs in some cases of Friedreich's ataxia and primary cerebellar degeneration and that the abnormality of pyruvate metabolism is not necessarily reflected in the serum LAD activity of these patients. The relevance of these findings to the heterogeneity of the hereditary ataxias is discussed.


Novel pressurized solvent extraction vessels for the analysis of polychlorinated biphenyl congeners in avian whole blood.
Persistent organic pollutants remain a serious threat to many food-chain systems. New pollutants continue to emerge. The present study has created novel extraction vessels which are compatible with readily available commercial instrumentation to validate the analysis of one class of persistent organic pollutants, polychlorinated biphenyls (PCBs), in avian blood. The volumes used can be reasonably sampled without sacrificing individuals, or comprising breeding or migratorial success. The procedure consists of the pressurized solvent extraction (PSE) of analytes in a novel PSE extraction vessel. The new extraction cell contains a 38-cm long, coiled, re-packable, in situ clean-up column. Lipid elimination, using Florisil, occurs within the coiled region of the extraction vessel, eliminating the requirement for post extraction clean-up. For development, 0.2 g samples of chicken whole blood have been used. Extract volumes are reduced from (30 to 10) cm(3), compared to unmodified systems. The new PSE vessel with its integrated clean-up method showed satisfactory performance for the analysis of ten environmentally relevant PCB congeners in chicken whole blood samples with recoveries in the range of (70-130)%. Detection limits using gas chromatography coupled with large volume injection ion-trap mass spectrometry (GC-LVI-ITMS-MS) were in the range of (0.05-0.5) ng g(-1). The relative standard deviations for all congeners investigated were better than 5%. This is the first PSE validation to have been conducted on unaltered whole blood samples.


NrCAM secreted by endometrial stromal cells enhances the progestin sensitivity of endometrial cancer cells through epigenetic modulation of PRB.
Progestin is one of the main hormone treatment regimens for early-stage estrogen receptor- and progesterone receptor (PR)-positive endometrial cancer (EC). However, the response rate of EC to progestins is unsatisfactory. Investigating the mechanisms related to progestin treatment could help improve treatment efficacy. Studies have demonstrated that normal endometrial stromal cells (ESCs) increase the inhibitory effect of progestin on EC cell proliferation via paracrine signaling, but the mechanisms involved remain unclear. In this study, we found that ESCs had different morphological features between progestin-sensitive and -insensitive EC tissues. ESCs presented typical decidualization changes in progestin-sensitive cases, while they remained slim in progestin-insensitive EC lesions, indicating no response. Furthermore, ESCs enhanced the inhibitory effect of medroxyprogesterone acetate (MPA) on EC cell proliferation by secreting neuron cell adhesion molecule (NrCAM). MPA treatment enhanced NrCAM secretion by ESCs. EC xenografts in BALB/C nude mice demonstrated that MPA combined with NrCAM had an increased tumor inhibitory effect compared with MPA or NrCAM alone. Mechanistically, MPA upregulated NrCAM expression in ESCs through PR. Specifically, NrCAM increased PR expression in EC cells through TET1-induced hydroxymethylation of the PRB gene promoter region. These findings indicate that NrCAM or NrCAM combined with progestins could be a new EC treatment.


Graded levels of Eimeria challenge altered the microstructural architecture and reduced the cortical bone growth of femur of Hy-Line W-36 pullets at early stage of growth (0-6 wk of age).
An experiment was carried out to evaluate the impact of mixed Eimeria challenge on skeletal health of Hy-Line W-36 pullets. A total of 540, 16-day-old pullets were randomly allocated into 5 treatment groups, including a nonchallenged control. A mixed Eimeria species solution containing 50,000 E. maxima, 50,000 E. tenella, and 250,000 E. acervulina oocysts per mL was prepared and challenged to 1 group as a high-dose treatment. The 2-fold serial dilution was done to prepare the medium-high (25,000 E. maxima; 25,000 E. tenella; 125,000 E. acervulina), the medium-low (12,500 E. maxima; 12,500 E. tenella; 62,500 E. acervulina), and the low (6,250 E. maxima; 6,250 E. tenella; 31,250 E. acervulina) dose treatments which were challenged to 3 corresponding groups, respectively. The mineral apposition rate (MAR) was measured from 0 to 14 d post inoculation (DPI) and 14 to 28 DPI using calcein injection. The microstructural architecture of the femur was analyzed using the Skyscan X-ray microtomography (microCT) on 6, 14, and 28 DPI. The results showed that the MAR decreased linearly with an increase in the challenged dose (P < 0.05) during 0 to 14 DPI. The results of microCT revealed that cortical and total BMD, BMC, bone volume (BV), and bone volume as a fraction of tissue volume (BV/TV) of femur decreased both linearly (P < 0.05). Conversely, the total number of pores increased linearly with an increase in challenge dosages on 6 and 14 DPI. Trabecular BMD, BV, BV/TV, trabecular number, and trabecular thickness decreased linearly with an increase in the challenge dosages (P < 0.05) on 6 DPI. Furthermore, Eimeria infection significantly increased the number of osteoclasts and osteoclastic activity (P = 0.001). The result of this study suggests that the mixed Eimeria challenge negatively impacts the quality of skeletal health in a linear or quadratic manner with an increase in the concentration of Eimeria oocysts. The negative impact on long bone development might be due to malabsorption, nutrient deficiency during the infection, along with oxidative stress/inflammation disrupting the balance of osteoblastic and osteoclastic cells and their functions.


Adenosine A2A receptor mediates hypnotic effects of ethanol in mice.
Ethanol has extensive effects on sleep and daytime alertness, causing premature disability and death. Adenosine, as a potent sleep-promoting substance, is involved in many cellular and behavioral responses to ethanol. However, the mechanisms of hypnotic effects of ethanol remain unclear. In this study, we investigated the role of adenosine in ethanol-induced sleep using C57BL/6Slac mice, adenosine A2A receptor (A2AR) knockout mice, and their wild-type littermates. The results showed that intraperitoneal injection of ethanol (3.0 g/kg) at 21:00 decreased the latency to non-rapid eye movement (NREM) sleep and increased the duration of NREM sleep for 5 h. Ethanol dose-dependently increased NREM sleep, which was consistent with decreases in wakefulness in C57BL/6Slac mice compared with their own control. Caffeine (5, 10, or 15 mg/kg), a nonspecific adenosine receptor antagonist, dose-dependently and at high doses completely blocked ethanol-induced NREM sleep when administered 30 min prior to (but not after) ethanol injection. Moreover, ethanol-induced NREM sleep was completely abolished in A2AR knockout mice compared with wild-type mice. These findings strongly indicate that A2AR is a key receptor for the hypnotic effects of ethanol, and pretreatment of caffeine might be a strategy to counter the hypnotic effects of ethanol.


[Acute toxoplasmic myelitis in the newborns].
The present paper describes 3 patients with congenital toxoplasmosis. The spinal cord was extremely affected. Differential diagnoses were established with poliomyelitis and neonatal hypoxia. Histopathological studies revealed toxoplasmosis.


The SMYD3-MTHFD1L-formate metabolic regulatory axis mediates mitophagy to inhibit M1 polarization in macrophages.
SMYD3 (protein 3 containing SET and MYND structural domains) belongs to the SMYD methylesterase family and is a histone lysine methyltransferase that promotes gene transcription mainly by catalysing the trimethylation of lysine at position 4 of histone subunit 3 (H3K4me3). Studies have shown that SMYD3 plays a key role in tumour cell proliferation and differentiation; however, its role in macrophage polarization is unclear. We screened the M1- and M2-polarized macrophage differential histone modifying enzyme using bioinformatics analysis. The SMYD3 overexpression plasmid was transfected into M1 macrophages, and the SMYD3-regulated target gene was analysed by RNA-seq and ChIP-Seq. The effect of knocking down MTHFD1L on M1 polarization and the change of the intracellular metabolite formic acid content were investigated. M1 macrophages were stimulated with different concentrations of formic acid (2, 10 and 40 mM) to detect the expression of M1-related genes, ROS production, and changes in the expression of the mitophagy-related proteins LC3, PINK1 and p-Parkin. Here, we used bioinformatics to analyse SMYD3, a histone methyltransferase associated with M1 polarization; overexpression of SMYD3 significantly suppressed the LPS/IFN-γ-induced M1 phenotype in macrophages. RNA-seq analysis demonstrated that SMYD3 significantly activated the one-carbon folate metabolic pathway in M1 macrophages. In addition, we used ChIP-seq analysis to identify methylenetetrahydrofolate dehydrogenase 1-like (MTHFD1L) as the target gene of the transcriptional activation by SMYD3 through H3K4me3 histone modification. Activation of MTHFD1L causes the accumulation of the intracellular metabolite formate. Exogenous stimulation with different concentrations of formate increased the expression of key genes involved in the mitochondrial respiratory chain complex, ROS production, and the expression of autophagy-related proteins LC3, PINK1, and p-Parkin and suppressed the expression of M1-related genes. Our study demonstrates that SMYD3 regulates the activity of the mitochondrial metabolic enzyme MTHFD1L through H3K4me3 histone methylation modification, promotes formate synthesis and induces mitophagy, which inhibits M1 polarization in macrophages.


Adsorption of Pb²⁺, Cd²⁺, Cu²⁺ and Cr³⁺ onto titanate nanotubes: competition and effect of inorganic ions.
Adsorption of Pb(2+), Cd(2+), Cu(2+) and Cr(3+) from aqueous solutions onto titanate nanotubes (TNTs) in multiple systems was systematically studied. Particular attention was paid to competitive adsorption and the effect of inorganic ions. TNTs showed large adsorption capacity for the four heavy metals, with the mechanism of ion-exchange between metal ions and H(+)/Na(+) located in the interlayers of TNTs. Binary or quaternary competitive adsorption indicated that the adsorption capacity of the four heavy metals onto TNTs followed the sequence of Pb(2+) (2.64 mmol g(-1)) ≫ Cd(2+) (2.13 mmol g(-1)) > Cu(2+) (1.92 mmol g(-1)) ≫ Cr(3+) (1.37 mmol g(-1)), which followed the reverse order of their hydration energies. Moreover, inorganic ions including Na(+), K(+), Mg(2+) and Ca(2+) inhibited the adsorption of heavy metals on TNTs, because they competed for adsorption sites, decreased the activity of heavy metal ions, and promoted the aggregation of TNTs. However, Al(3+) and Fe(3+) generally enhanced adsorption because the resulting hydroxyl-Al/Fe intercalated or coated TNTs could also capture metal ions. Furthermore, minor effect of inorganic ions on adsorption of Pb(2+) resulted from its strong affinity to TNTs. Difficult desorption and small inhibiting effect by Na(+), K(+), Mg(2+) and Ca(2+) on adsorption of Cr(3+) was due to the formed stable complex of HOCr(OTi)₂ ≡ with TNTs. Present study indicated potential applications of TNTs in wastewater treatment for heavy metals.


Bdnf mRNA splice variants differentially impact CA1 and CA3 dendrite complexity and spine morphology in the hippocampus.
Brain-derived neurotrophic factor (BDNF) is an activity-dependent neurotrophin critical for neuronal plasticity in the hippocampus. BDNF is encoded by multiple transcripts with alternative 5' untranslated regions (5'UTRS) that display activity-induced targeting to distinct subcellular compartments. While individual Bdnf 5'UTR transcripts influence dendrite morphology in cultured hippocampal neurons, it is unknown whether Bdnf splice variants impact dendrite arborization in functional classes of neurons in the intact hippocampus. Moreover, the contribution of Bdnf 5'UTR splice variants to dendritic spine density and shape has not been explored. We analyzed the structure of CA1 and CA3 dendrite arbors in transgenic mice lacking BDNF production from exon (Ex) 1, 2, 4, or 6 splice variants (Bdnf-e1, -e2, -e4, and -e6-/- mice) and found that loss of BDNF from individual Bdnf mRNA variants differentially impacts the complexity of apical and basal arbors in vivo. Consistent with the subcellular localization studies, Bdnf Ex2 and Ex6 transcripts significantly contributed to dendrite morphology in both CA1 and CA3 neurons. While Bdnf-e2-/- mice showed increased branching proximal to the soma in CA1 and CA3 apical arbors, Bdnf-e6-/- mice showed decreased apical and basal dendrite complexity. Analysis of spine morphology on Bdnf-e6-/- CA1 dendrites revealed changes in the percentage of differently sized spines on apical, but not basal, branches. These results provide further evidence that Bdnf splice variants generate a spatial code that mediates the local actions of BDNF in distinct dendritic compartments on structural and functional plasticity.


Pancreatic pleuropericardial effusions presenting as tumor of the lung.
A 45-year-old man presented with what was thought to be a mass in the left upper lung and a pericardial friction rub. He was subsequently discovered to have a loculated pleural effusion and pericardial effusion associated with chronic pancreatitis. This is the first instance we were able to find of pancreatitis mimicking bronchogenic carcinoma with pericardial metastasis.


Plant and mouse EB1 proteins have opposite intrinsic properties on the dynamic instability of microtubules.
Most eukaryotic cells contain microtubule filaments, which play central roles in intra-cellular organization. However, microtubule networks have a wide variety of architectures from one cell type and organism to another. Nonetheless, the sequences of tubulins, of Microtubule Associated proteins (MAPs) and the structure of microtubules are usually well conserved throughout the evolution. MAPs being known to be responsible for regulating microtubule organization and dynamics, this raises the question of the conservation of their intrinsic properties. Indeed, knowing how the intrinsic properties of individual MAPs differ between organisms might enlighten our understanding of how distinct microtubule networks are built. End-Binding protein 1 (EB1), first described as a MAP in yeast, is conserved in plants and mammals. The intrinsic properties of the mammalian and the yeast EB1 proteins have been well described in the literature but, to our knowledge, the intrinsic properties of EB1 from plant and mammals have not been compared thus far. Here, using an in vitro assay, we discovered that plant and mammalian EB1 purified proteins have different intrinsic properties on microtubule dynamics. Indeed, the mammalian EB1 protein increases microtubules dynamic while the plant EB1 protein stabilizes them.


Comparison of the aortic valve calcium content in the bicuspid and tricuspid stenotic aortic valve using non-enhanced 64-detector-row-computed tomography with prospective ECG-triggering.
The aim of our study was to compare the calcium content measured by non-enhanced multidetector-row-computed tomography (MDCT) between patients with significant stenosis of bicuspid (BAV) and tricuspid aortic valve (TAV). Another aim of our study was to assess the accuracy of the non-enhanced MDCT to distinguish BAV and TAV based on the calcified plaque morphology, and to compare the results with the transesophageal echocardiography. A retrospective analysis of prospectively collected data was performed. Consecutive patients with symptomatic aortic stenosis (AS) admitted to hospital for evaluation before valve surgery underwent clinical evaluation, transthoracic and transesophageal echocardiography, and non-enhanced examination with the 64-detector-row CT using prospective ECG triggering with data acquisition in diastolic phase. The data acquisition started at 55% of the R-R interval. The patients were examined in the supine position in mild inspiration. Data were evaluated using dedicated software for calcium scoring, the volume of calcifications and calcium content were obtained. Thirty-seven patients (20 males, age 48-83 years) were enrolled. BAV was present in 13 patients, TAV in 24 patients. The calcium score in patients with severe AS (mean gradient >50 mmHg) was higher than in those with moderate AS (1123+/-616 mg versus 634+/-475, P=0.011). Significant correlation between the calcium scores and transaortic gradients was found (r=0.53, P=0.002). The patients with BAV did not differ significantly from those with TAV in the AS severity (58+/-13 versus 53+/-20 mmHg), nor in the valve calcium score (1168+/-717 versus 795+/-530 mg, P=0.093). The overall sensitivity to detect BAV in patients with calcified severe AS was 0.923 (12/13) and specificity 0.958 (23/24). The overall accuracy was 0.945 (35/37). We observed higher calcium score in patients with severe AS than with moderate AS. However, no difference in aortic valve calcium score between BAV and TAV was found. Thus, in our sample, the aortic valve calcium score correlated with AS severity, not with aortic valve morphology. Based on the calcified plaque space relationship, the aortic BAV and TAV could be distinguished in most cases.


Generation of micro-droplet arrays by dip-coating of biphilic surfaces; the dependence of entrained droplet volume on withdrawal velocity.
Droplet array chips were realized using an alignment-free fabrication process in silicon. The chips were textured with a homogeneous nano-scale surface roughness but were partially covered with a self-assembled monolayer of perfluorodecyltrichlorosilane (FDTS), resulting in a super-biphilic surface. When submerged in water and withdrawn again, microliter sized droplets are formed due to pinning of water on the hydrophilic spots. The entrained droplet volumes were investigated under variation of spot size and withdrawal velocity. Two regimes of droplet formation were revealed: at low speeds, the droplet volume achieved finite values even for vanishing speeds, while at higher speeds the volume was governed by fluid inertia. A simple 2D boundary layer model describes the behavior at high speeds well. Entrained droplet volume could be altered, post-fabrication, by more than a factor of 15, which opens up for more applications of the dip-coating technique due to the significant increase in versatility of the micro-droplet array platform.


Iris melanoma outcomes based on the Cancer Genome Atlas (TCGA) classification in 78 consecutive patients.
The Cancer Genome Atlas (TCGA) classification of genetic alterations in uveal melanoma is widely used for prognostication. We present novel observations on the impact of TCGA Group specifically for iris melanoma. This was a retrospective cohort study at a tertiary referral ocular oncology center. All patients with a diagnosis of iris melanoma who underwent genetic evaluation and assessment for TCGA classification between 20 November 1995 and 5 April 2021 were included. The main outcome measures were visual acuity, secondary glaucoma, tumor recurrence, melanoma-related metastasis and death per TCGA group. There were a total of 78 patients included. The mean patient age was 49.6 years (median 53.0, range 3.0-85.0), mean tumor basal diameter was 6.7 mm (median 6.0, range 1.5-22.0), and mean tumor thickness was 2.6 mm (median 2.5, range 0.5-8.5). Cytology results confirmed iris melanoma (93%) or were inconclusive (7%). The TCGA groups included Group A (n = 36, 46%), Group B (n = 7, 9%), Group C (n = 34, 44%), and Group D (n = 1, 1%). There was no statistically significant difference in outcomes of visual acuity, tumor thickness reduction, secondary glaucoma, tumor recurrence, melanoma-related metastasis or death per individual TCGA group (A vs. B vs. C vs. D) and per bimodal comparison (A/B vs. C/D). In this analysis, iris melanoma was classified as TCGA group A or B in 55% and as C or D in 45%. The TCGA classification was not predictive of melanoma-related metastasis or death.


The Efficacy and Frequency of Self-monitoring of Blood Glucose in Non-insulin-Treated T2D Patients: a Systematic Review and Meta-analysis.
Self-monitoring of blood glucose (SMBG) is a useful tool in diabetes management, but its efficacy and optimal application in type 2 diabetes (T2D) patients treated without insulin have been controversial. We aimed to evaluate the efficacy of SMBG in controlling blood glucose levels in non-insulin-treated T2D patients and to determine the optimal frequency and the most appropriate population to benefit from SMBG. Eligible publications from January 2000 to April 2022 were retrieved from PubMed, Embase, Cochrane Library, and ClinicalTrials.gov databases. Randomized controlled trials comparing SMBG with no SMBG or structured SMBG (S-SMBG, SMBG with defined timing and frequency of glucose measurements) were included. Meta-analyses and sub-analyses were performed to assess the efficacy, optimal frequency, and most appropriate population for SMBG. Risk of bias was assessed regarding randomization, allocation sequence concealment, blinding, incomplete outcome data, selective outcome reporting, and other biases. Twenty-two studies involving 6204 participants were identified, including 17 comparing SMBG with no SMBG and 4 comparing SMBG with S-SMBG. SMBG reduced HbA1c (MD -0.30%, 95% CI -0.42 to -0.17) compared with no SMBG, and S-SMBG performed better than SMBG (MD -0.23%, 95% CI -0.38 to -0.07). Subgroup analyses showed that HbA1c control was better with SMBG at 8-11 times weekly (MD -0.35%, 95% CI -0.51 to -0.20) compared with other frequencies and with lifestyle adjustments (MD -0.37%, 95% CI -0.50 to -0.23) than with no adjustments. No significant differences in HbA1c were observed between baseline HbA1c subgroups (≤ 8% and > 8%, P = 0.63) and between diabetes duration subgroups (≤ 6 years and > 6 years, P = 0.72), respectively. SMBG was effective for controlling HbA1c in non-insulin-treated T2D patients, although lacking detailed monitoring design. Better outcomes were seen with SMBG at 8-11 times weekly and lifestyle adjustment based on SMBG results. PROSPERO (CRD42021285604).


Targeting resuscitation to normalization of coagulating status: Hyper and hypocoagulability after severe injury are both associated with increased mortality.
The prevalence and impact of hypercoagulability (hypo) in severely injured patients early after injury remains unclear. We hypothesize that the predominant phenotype of postinjury coagulopathy is hypercoagulability (hyper) and it is associated with increased mortality. Blood samples from 141 healthy volunteers assayed with thrombelastography (TEG) were used to identify thresholds of hypo and hypercoagulability (above 95th/below the 5thpercentile) in four TEG indices. These cutoffs were subsequently evaluated in severely injured trauma patients (ISS>15) from two level 1 trauma centers. 2540 patients with a median ISS of 25 were analyzed. Normal TEG was present in 36% of patients. Hyper was found in 38% of patients, with mixed (11%) and hypo (15%) being less common. Compared to normal coagulation patients and after controlling for age, sex, blood pressure, and injury hyper (0.013), mixed (p < 0.001) and hypo (p < 0.001) were all independent predictors of mortality. These data support the ongoing need for goal directed resuscitation in trauma patients, it appears the optimal resuscitation strategy should be targeted towards normalization of coagulation status as both early hyper and hypocoagulability are associated with increased mortality.


Heterologous humoral immunity to human and zoonotic coronaviruses: Aiming for the achilles heel.
Coronaviruses are evolutionarily successful RNA viruses, common to multiple avian, amphibian and mammalian hosts. Despite their ubiquity and potential impact, knowledge of host immunity to coronaviruses remains incomplete, partly owing to the lack of overt pathogenicity of endemic human coronaviruses (HCoVs), which typically cause common colds. However, the need for deeper understanding became pressing with the zoonotic introduction of three novel coronaviruses in the past two decades, causing severe acute respiratory syndromes in humans, and the unfolding pandemic of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This renewed interest not only triggered the discovery of two of the four HCoVs, but also uncovered substantial cellular and humoral cross-reactivity with shared or related coronaviral antigens. Here, we review the evidence for cross-reactive B cell memory elicited by HCoVs and its potential impact on the puzzlingly variable outcome of SARS-CoV-2 infection. The available data indicate targeting of highly conserved regions primarily in the S2 subunits of the spike glycoproteins of HCoVs and SARS-CoV-2 by cross-reactive B cells and antibodies. Rare monoclonal antibodies reactive with conserved S2 epitopes and with potent virus neutralising activity have been cloned, underscoring the potential functional relevance of cross-reactivity. We discuss B cell and antibody cross-reactivity in the broader context of heterologous humoral immunity to coronaviruses, as well as the limits of protective immune memory against homologous re-infection. Given the bidirectional nature of cross-reactivity, the unprecedented current vaccination campaign against SARS-CoV-2 is expected to impact HCoVs, as well as future zoonotic coronaviruses attempting to cross the species barrier. However, emerging SARS-CoV-2 variants with resistance to neutralisation by vaccine-induced antibodies highlight a need for targeting more constrained, less mutable parts of the spike. The delineation of such cross-reactive areas, which humoral immunity can be trained to attack, may offer the key to permanently shifting the balance of our interaction with current and future coronaviruses in our favour.


Pancreas transplantation at Mayo: I. Patient selection.
From October 1987 to December 1988, 59 patients underwent assessment for combined kidney and pancreas transplantation or pancreas transplantation after receiving a kidney allograft. We report our criteria for accepting candidates for transplantation, the results of the selection process, and the clinical and laboratory profile of those patients who underwent transplantation. Of the overall group, 22 patients (37%) were approved medically, 3 of whom were awaiting financial approval. Of the 59 patients, 15 (25%) were not approved for the transplantation program for medical reasons; in addition, 16 patients declined participation and 3 were not accepted because of lack of financial resources. Medical reasons for exclusion from pancreas transplantation were coronary artery disease in six patients, severe peripheral vascular disease in six patients, other medical problems in two patients, and noncompliance in one patient. Thus, many patients who underwent assessment for pancreas transplantation did not enter the program because of medical, financial, or personal preference reasons. In most cases, the medical reason for exclusion from pancreas transplantation was a cardiovascular disorder.


Dimensionality reduction via path integration for computing mRNA distributions.
Inherent stochasticity in gene expression leads to distributions of mRNA copy numbers in a population of identical cells. These distributions are determined primarily by the multitude of states of a gene promoter, each driving transcription at a different rate. In an era where single-cell mRNA copy number data are more and more available, there is an increasing need for fast computations of mRNA distributions. In this paper, we present a method for computing separate distributions for each species of mRNA molecules, i.e. mRNAs that have been either partially or fully processed post-transcription. The method involves the integration over all possible realizations of promoter states, which we cast into a set of linear ordinary differential equations of dimension [Formula: see text], where M is the number of available promoter states and [Formula: see text] is the mRNA copy number of species j up to which one wishes to compute the probability distribution. This approach is superior to solving the Master equation (ME) directly in two ways: (a) the number of coupled differential equations in the ME approach is [Formula: see text], where [Formula: see text] is the cutoff for the probability of the jth species of mRNA; and (b) the ME must be solved up to the cutoffs [Formula: see text], which must be selected a priori. In our approach, the equation for the probability to observe n mRNAs of any species depends only on the the probability of observing [Formula: see text] mRNAs of that species, thus yielding a correct probability distribution up to an arbitrary n. To demonstrate the validity of our derivations, we compare our results with Gillespie simulations for ten randomly selected system parameters.


Assessing the potential for prevention or earlier detection of on-site monitoring findings from randomised controlled trials: Further analyses of findings from the prospective TEMPER triggered monitoring study.
Clinical trials should be designed and managed to minimise important errors with potential to compromise patient safety or data integrity, employ monitoring practices that detect and correct important errors quickly, and take robust action to prevent repetition. Regulators highlight the use of risk-based monitoring, making greater use of centralised monitoring and reducing reliance on centre visits. The TEMPER study was a prospective evaluation of triggered monitoring (a risk-based monitoring method), whereby centres are prioritised for visits based on central monitoring results. Conducted in three UK-based randomised cancer treatment trials of investigational medicine products with time-to-event outcomes, it found high levels of serious findings at triggered centre visits but also at visits to matched control centres that, based on central monitoring, were not of concern. Here, we report a detailed review of the serious findings from TEMPER centre visits. We sought to identify feasible, centralised processes which might detect or prevent these findings without a centre visit. The primary outcome of this study was the proportion of all 'major' and 'critical' TEMPER centre visit findings theoretically detectable or preventable through a feasible, centralised process. To devise processes, we considered a representative example of each finding type through an internal consensus exercise. This involved (a) agreeing the potential, by some described process, for each finding type to be centrally detected or prevented and (b) agreeing a proposed feasibility score for each proposed process. To further assess feasibility, we ran a consultation exercise, whereby the proposed processes were reviewed and rated for feasibility by invited external trialists. In TEMPER, 312 major or critical findings were identified at 94 visits. These findings comprised 120 distinct issues, for which we proposed 56 different centralised processes. Following independent review of the feasibility of the proposed processes by 87 consultation respondents across eight different trial stakeholder groups, we conclude that 306/312 (98%) findings could theoretically be prevented or identified centrally. Of the processes deemed feasible, those relating to informed consent could have the most impact. Of processes not currently deemed feasible, those involving use of electronic health records are among those with the largest potential benefit. This work presents a best-case scenario, where a large majority of monitoring findings were deemed theoretically preventable or detectable by central processes. Caveats include the cost of applying all necessary methods, and the resource implications of enhanced central monitoring for both centre and trials unit staff. Our results will inform future monitoring plans and emphasise the importance of continued critical review of monitoring processes and outcomes to ensure they remain appropriate.


A quantitative, immunochemical demonstration of the postnatal development of neurotensin in the medial preoptic area of the rat.
The postnatal ontogeny of neurotensin (NT) in the medial preoptic area (MPO) of the rat was quantitatively investigated using immunocytochemical and neurochemical techniques. On postpartum day 0, NT-like immunoreactive neurons could not be identified within the MPO by peroxidase-antiperoxidase histochemistry and, in fact, did not appear until 9 days postpartum. After this age, the number of NT cells in the MPO increased ontogenetically, as did concentrations of NT in MPO extracts measured by radioimmunoassay and enzyme-linked immunosorbent assay. In organotypic MPO explants derived from neonatal rats, NT-like immunoreactivity was detected only after the explants were maintained 14 days in vitro, after which, the number of NT-like immunoreactive somata increased as the time in vitro was extended. These findings indicate that a substantial amount of differentiation occurs postnatally in the MPO both in vivo and in vitro and, more specifically, that the ontogenetic expression of NT may be an intrinsic property of the MPO.


SHON expression predicts response and relapse risk of breast cancer patients after anthracycline-based combination chemotherapy or tamoxifen treatment.
SHON nuclear expression (SHON-Nuc+) was previously reported to predict clinical outcomes to tamoxifen therapy in ERα+ breast cancer (BC). Herein we determined if SHON expression detected by specific monoclonal antibodies could provide a more accurate prediction and serve as a biomarker for anthracycline-based combination chemotherapy (ACT). SHON expression was determined by immunohistochemistry in the Nottingham early-stage-BC cohort (n = 1,650) who, if eligible, received adjuvant tamoxifen; the Nottingham ERα- early-stage-BC (n = 697) patients who received adjuvant ACT; and the Nottingham locally advanced-BC cohort who received pre-operative ACT with/without taxanes (Neo-ACT, n = 120) and if eligible, 5-year adjuvant tamoxifen treatment. Prognostic significance of SHON and its relationship with the clinical outcome of treatments were analysed. As previously reported, SHON-Nuc+ in high risk/ERα+ patients was significantly associated with a 48% death risk reduction after exclusive adjuvant tamoxifen treatment compared with SHON-Nuc- [HR (95% CI) = 0.52 (0.34-0.78), p = 0.002]. Meanwhile, in ERα- patients treated with adjuvant ACT, SHON cytoplasmic expression (SHON-Cyto+) was significantly associated with a 50% death risk reduction compared with SHON-Cyto- [HR (95% CI) = 0.50 (0.34-0.73), p = 0.0003]. Moreover, in patients received Neo-ACT, SHON-Nuc- or SHON-Cyto+ was associated with an increased pathological complete response (pCR) compared with SHON-Nuc+ [21 vs 4%; OR (95% CI) = 5.88 (1.28-27.03), p = 0.012], or SHON-Cyto- [20.5 vs. 4.5%; OR (95% CI) = 5.43 (1.18-25.03), p = 0.017], respectively. After receiving Neo-ACT, patients with SHON-Nuc+ had a significantly lower distant relapse risk compared to those with SHON-Nuc- [HR (95% CI) = 0.41 (0.19-0.87), p = 0.038], whereas SHON-Cyto+ patients had a significantly higher distant relapse risk compared to SHON-Cyto- patients [HR (95% CI) = 4.63 (1.05-20.39), p = 0.043]. Furthermore, multivariate Cox regression analyses revealed that SHON-Cyto+ was independently associated with a higher risk of distant relapse after Neo-ACT and 5-year tamoxifen treatment [HR (95% CI) = 5.08 (1.13-44.52), p = 0.037]. The interaction term between ERα status and SHON-Nuc+ (p = 0.005), and between SHON-Nuc+ and tamoxifen therapy (p = 0.007), were both statistically significant. SHON-Nuce+ in tumours predicts response to tamoxifen in ERα+ BC while SHON-Cyto+ predicts response to ACT.


[Concentration of plasma glycosphingolipids in acute hepatitis (author's transl)].
There are mainly four neutral glycosphingolipids in human blood plasma: Monohexosyl, dihexosyl, trihexosyl and tetrahexosyl ceramide. In patients with viral hepatitis (n=21) during the acute phase all four fractions of plasma glycosphingolipids were elevated compared to healthy subjects (n=23). With the exception of trihexosyl ceramide all fractions demonstrated statistically striking elevations in the acute phase of viral hepatitis. Simultaneously in the acute phase of hepatitis triglycerides and cholesterol in serum were significantly increased. It is concluded the elevation of glycosphingolipid levels in plasma is a metabolic consequence of hyperlipoproteinemia. Furthermore it is supposed that a part of plasma glycosphingolipids is synthezised de novo by the liver like VLDL.


Successful pregnancy after transabdominal cervical evacuation of cervical pregnancy.
Cervical pregnancy was managed by transabdominal cervical evacuation. Three years later, the woman was delivered of a healthy male infant by cesarean section.


[Transvaginal mesh in repair of pelvic organs prolapse as a minimally invasive surgical procedure].
Prolapse of genital organs with or without urinary stress incontinention is the most often health problem in the elderly female population tending to increase with ageing. The aim of this study was to assess the perioperative complications and short-term outcomes of prolaps repair using transvaginal polypropylene mesh (Prolift system, Gynecare, Ethicon, USA). A retrospective study was conducted evaluating 96 women from September 2006 to January 2010 who underwent vaginal repair with implatation of a soft mesh manufactured by Gynecare, Ethicon, USA. All the patients had a stage 3 or stage 4 prolapse according to the POP-Q system of ICS. Total mesh was used in 12 (13%) patients isolated anterior mesh in 52 (54%) patients and isolated posterior mesh in 32 (33%) patients. We reported one intra-operative bladder injury and no other serious complications. At 3 months, all 96 patients were available for follow-up. Vaginal erosion occured in 9 (9.3%) patients, shrinkage of mesh in 6 (6.2%) patients and de novo urinary incontinence in 5 (5.2%) patients. Failure rate was 6.25% (recurrent prolapse stage 3 or 4 even asymptomatic). Our study suggests that transvaginal polypropylene mesh applied with a tension-free technique is a safe and effective method with low intraoperative complications and low morbidity rates. However, some complications are serious and require highly specialised management.


The novel approach to the biomonitor survey using one- and two-dimensional Kohonen networks.
To compare the applicability of the leaves of horse chestnut (Aesculus hippocastanum) and linden (Tilia spp.) as biomonitors of trace element concentrations, a coupled approach of one- and two-dimensional Kohonen networks was applied for the first time. The self-organizing networks (SONs) and the self-organizing maps (SOMs) were applied on the database obtained for the element accumulation (Cr, Fe, Ni, Cu, Zn, Pb, V, As, Cd) and the SOM for the Pb isotopes in the leaves for a multiyear period (2002-2006). A. hippocastanum seems to be a more appropriate biomonitor since it showed more consistent results in the analysis of trace elements and Pb isotopes. The SOM proved to be a suitable and sensitive tool for assessing differences in trace element concentrations and for the Pb isotopic composition in leaves of different species. In addition, the SON provided more clear data on seasonal and temporal accumulation of trace elements in the leaves and could be recommended complementary to the SOM analysis of trace elements in biomonitoring studies.


Experiences of nursing students of Evidence-Based Practice Education according to Rogers' Diffusion of Innovation Model: A Directed Content Analysis.
Evidence based practice (EBP) education is essential in promoting of clinical care, but an effective educational strategy for teaching EBP in nursing faculties is not available. The aim of this study was to explore the experiences of nursing students of EBP Education according to Rogers' Diffusion of Innovation Model. This qualitative study was carried out using a directed content analysis method and purposeful sampling. Data were collected until saturation by fourteen semi-structured face-to-face individual interviews and two focus group discussions with nursing students from two nursing faculties in Tehran, Iran. Rogers' Model was used in this study. Data were classified into five themes and 11 categories according to the Rogers's Model. Themes and main categories were knowledge (educational enrichment, new strategy for education), persuasion (internalization of education, improvement of motivation), decision (acceptance, use in the future), implementation (objectivity, consolidation of learning) and confirmation (learning and teaching, achieving a goal, self-confidence). EBP Education, based on the teaching strategy of Rogers's Model, leads to an improved EBP learning. All the necessary steps for a better education of it are included in this educational approach which can be used to teach any new subject like EBP.