Perfluorooctanoic acid uptake in the mustard species Brassica juncea.
Perfluorooctanoic acid (PFOA), a surfactant, is a member of the perfluoroalkyl acids (PFAAs) family and is a contaminant of emerging concern for human and environmental health. Perfluorooctanoic acid is a persistent organic pollutant, but currently little is known about (a) the potential ecological and toxicological effects of PFOA and (b) how PFOA moves in the environment. This study uses a radiotracer (14 C-PFOA) to study the uptake and translocation of PFOA in hydroponically grown brown mustard [Brassica juncea (L.) Czern.], a representative crop species. Plants were exposed in quadruplicate over the course of 7 d (with plants sampled on Days 4 and 7) to PFOA concentrations of 0, 1, 5, 10, and 15 mg L-1 . Uptake was quantified via liquid scintillation counting of samples from the nutrient solution, roots, stems, and leaves. Transfer factors (roots to shoots) ranged from 0.15 to 4.73 kg kg-1 . Bioconcentration factors (solution to plant) ranged from 0.36 to 62.29 L kg-1 . Factors were influenced by plant compartment, day sampled, and treatment level.


Epstein-Barr virus-associated haemophagocytic lymphohistiocytosis in Wiskott-Aldrich syndrome.
A patient with Wiskott-Aldrich syndrome who developed Epstein-Barr virus-associated haemophagocytic lymphohistiocytosis (EBV-HLH) is described in this study. At 4 mo of age the patient developed fever associated with bicytopenia and splenomegaly. Analysis of a bone marrow specimen revealed extensive haemophagocytosis, and in situ hybridization for EBV of the bone marrow specimen using an EBV-encoded RNA probe was positive. Diagnosis of EBV-HLH was established and immunotherapy with HLH-94 protocol was started. HLH has been described in patients with other well-defined primary immunodeficiencies such as X-linked lymphoproliferative syndrome, Chediak-Higashi syndrome and Griscelli disease. Also, HLH was reported recently in severe combined immunodeficiency and DiGeorge syndrome. The possibility of an underlying primary immunodeficiency should be considered in paediatric patients who present with HLH during infancy.


On using sample selection methods in estimating the price elasticity of firms' demand for insurance.
We evaluate a technique based on sample selection models that has been used by health economists to estimate the price elasticity of firms' demand for insurance. We demonstrate that, this technique produces inflated estimates of the price elasticity. We show that alternative methods lead to valid estimates.


Terminal ventriculostomy in syringomyelia.
The authors report their experiences with two patients suffering from syringomyelia on whom terminal ventriculostomies were carried out. The good results which can be obtained if the patient is in a relatively good neurological condition are pointed out. The principle of terminal ventriculostomy is based on the assumption that syringomyelia is hydromyelia that has become symptomatic; with this in mind, it seems reasonable to relieve the patient's symptoms by draining the distended central canal at the lowest possible level. The father of the operation is Dr. Gardner, who has thoroughly studied syringomyelia, and proved that syringomyelia is not only an active hydromyelia but, that a "non-communicating" syringomyelia is a non-existent entity (1976, 1977).


Distal clearance margin of 1 cm or less: a safe distance in lower rectum cancer surgery.
The purpose of this study was to investigate the prognostic role of distal clearance margin (DCM) in lower rectum cancer surgery. Two-hundred-three cancer patients underwent total rectal resection, possibly followed by adjuvant chemoradiotherapy. DCM was classified as positive or negative (<1, > or =1 cm) and investigated with multivariable proportional hazard models. A total of 52 deaths, 19 local relapses, 40 distant metastases, and three second primaries were observed as first events. Five-year survival with positive, negative <1, or negative > or =1 cm DCM was 51%, 81%, and 69%, respectively (p = 0.018). The difference was significant between positive and negative DCM (p = 0.031), not between negative <1 and > or =1 cm (p = 0.106). Local and distant 5-year incidences according to DCM were 30%, 8%, and 8% (p = 0.006) and 38%, 26%, and 19% (p = 0.857), respectively. DCM, but not tumor size, is a prognostic factor after sphincter-saving surgery, which is safe whenever a negative margin is achieved.


Anti-tumor promoting activities of natural products. II. Inhibitory effects of digitoxin on two-stage carcinogenesis of mouse skin tumors and mouse pulmonary tumors.
Two-stage carcinogenesis of mouse skin papillomas induced by 7,12-dimethylbenz[a]anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA), and mouse pulmonary tumors induced by 4-nitroquinoline-N-oxide (4NQO) and glycerol, were inhibited by digitoxin (1).


An association study on the polymorphisms of dopaminergic genes with working memory in a healthy Chinese Han population.
Working memory (WM) is a highly heritable cognitive trait that is involved in many higher-level cognitive functions. In the past few years, much evidence has indicated that the reduction of dopamine activity in human brain can impair the WM system of the neuropsychiatric disorders. In this study, we hypothesized that some genes in the dopamine system were involved in the individual difference of the cognitive ability in healthy population. To confirm this hypothesis, a population-based study was performed to examine the effects of COMT, DAT (1), DRD (1), DRD (2), DRD (3), and DRD (4) on WM spans. Our results indicated there were significant associations of TaqIA and TaqIB in DRD (2) with digital WM span, respectively (χ(2) = 9.460, p = 0.009; χ(2) = 6.845, p = 0.033). On the other hand, we found a significant interaction between Ser9Gly in DRD (3) and TaqIA of DRD (2) on digital WM span (F = 3.207, p = 0.013). COMT, DAT (1) , DRD (1), and DRD (4), however, had no significant effects on digital and spatial WM spans (χ(2)<3.84, p > 0.05). These preliminary results further indicated that certain functional variants in dopamine system, such as TaqIA and TaqIB of DRD (2), were possibly involved in difference of WM in a healthy population.


Morphological patterns of anaemia among pregnant women from Sudan.
Morphological patterns of anaemia in pregnancy are considered essential for classification, diagnosis and management of patients, especially in regions with high maternal mortality like Sudan. This study evaluated morphological patterns of anaemia among pregnant women in Sudan and morphological differences across characteristics of participants. This cross-sectional study was conducted from September 2016 to February 2017. A total of 200 women were selected according to specific criteria. Laboratory tests were performed for complete blood count, blood smears were performed for morphology and vitamin B12, folate and iron levels were measured. Participants were classified as: normochromic normocytic, microcytic hypochromic, macrocytic or dimorphic. Further classification based on haemoglobin levels was also performed. A total of 116 participants (58%) had a dimorphic pattern, followed by 50 participants (25%) with a microcytic hypochromic pattern, 20 participants (10%) with a macrocytic pattern and 14 participants (7%) with a normochromic normocytic pattern. Participants with the dimorphic pattern also had low levels of iron and folate. The majority of dimorphic participants presented with mild anaemia, whereas the majority of participants with the microcytic hypochromic pattern presented with moderate or severe anaemia. A high percentage of participants in late pregnancy had the dimorphic pattern, and there were significant differences in the degree of anaemia by parity, gestational age and regular intake of haematinic supplements. The most frequent morphological pattern of anaemia in this study was dimorphic, followed by microcytic hypochromic, macrocytic and normochromic patterns. Morphological patterns appeared to predict types of vitamin and mineral deficiency and the degree of anaemia.


COAGULASE-NEGATIVE STAPHYLOCOCCUS-INDUCED FROSTED BRANCH ANGIITIS AFTER INTRAVITREAL ANTI-VASCULAR ENDOTHELIAL GROWTH FACTOR INJECTION.
To describe a case of frosted branch angiitis after intravitreal ranibizumab injection. Retrospective chart review. A patient with a history of neovascular age-related macular degeneration underwent intravitreal ranibizumab injection and subsequently developed coagulase-negative Staphylococcus endophthalmitis with findings of frosted branch angiitis. Endophthalmitis presenting as frosted branch angiitis is a rare complication after intravitreal anti-vascular endothelial growth factor injection. Early recognition is critical to optimize outcomes. To our knowledge, this is the second reported case of frosted branch angiitis as a presentation of endophthalmitis with coagulase-negative Staphylococcus.


Precursor-product relationship between rabbit type II cell lamellar bodies and alveolar surface-active material. Surfactant turnover time.
To estimate the turnover time of alveolar surfactant in New Zealand white rabbits, we injected [9,10-(3)H] palmitic acid and [2-(14)C] glycerol intravenously. From 1-48 h after injection, wer killed the animals, lavaged the lungs for alveolar surfactant with saline, and isolated the lamellar bodies by homogenization and sucrose density gradient centrifugation. Lamellar bodies and alveola surfactant had comparable phospholipid composition and surface activity. Lamellar bodies contained little DNA, no mitochondrial enzyme activity and less than 5% contaminating phospholipids from microsomal and Golgi-enriched fractions. We measured radioactivity of phosphatidylcholine, saturated phosphatidylcholine and phosphatidylglycerol for each isotope in lamellar bodies and surfactant at each time point. The plot of the integral with respect to time of the difference between lamellar body and surfactant specific activity against surfactant specific activity has a slope determined by the turnover time, and a shape which tests the precision of the precursor-product relationship. This analysis does not assume a pulse label and allows for possible recycling of tracer from surfactant to lamellar bodies. We obtained turnover times of 4-11 h. We detected an imprecise precursor-product relationship between lamellar bodies and alveolar surfactant, which is not due to experimental variability or to contamination of lamellar bodies by other subcellular fractions but may reflect imperfect mixing within surfactant compartments.


Voice restoration after total laryngopharyngectomy and cervical esophagectomy using the duckbill prosthesis.
We have presented a series of 10 patients who were successfully rehabilitated using the tracheoesophageal puncture technique for voice restoration. These patients underwent total laryngopharyngectomy or laryngopharyngoesophagectomy and reconstruction with visceral transposition (five patients), myocutaneous flaps (four patients), and skin graft and cervical flaps (one patient). All patients were able to produce adequate voice and carry on a conversation. Clinical evaluation of voice quality and acoustic analysis of voice samples indicate that the voice obtained in these patients was intelligible, and had adequate intensity, low pitch, and limited pitch variation. There were no complications related to the voice restoration procedure. It appears from our data and the limited experience reported in the literature that the tracheoesophageal prosthesis can be used successfully and safely for the speech rehabilitation of patients who undergo total laryngopharyngectomy, cervical esophagectomy, or both, regardless of the reconstructive method used.


The regional function of cGAS/STING signal in multiple organs: One of culprit behind systemic lupus erythematosus?
The systemic dysregulation of adaptive and innate immunity have been identified as major hallmark of systemic lupus erythematosus (SLE) pathogenesis that predominantly affects women. Patients with SLE develop heterogeneous clinical manifestations which involve of multiple organ damage including renal, spleen, nervous system, joints and hematopoietic organs. A high rate of cell death, e.g., NETosis, and clearance deficiencies by myeloid cells led to increased cell debris and accumulation of endogenous nucleic acids, and the presence of anti-nuclear antibodies (ANAs) derived from immune response can break of self-tolerance and exacerbate SLE pathology. Currently, the nucleic acid receptors, such as Toll-like receptors, RIG-I-like receptors, AIM2-like receptors and IFI 200-family have been uncovered to be potential predisposing causes for SLE via triggering interferon (IFN) response and maturation of IL-1β. Notably, as the newly found DNA sensor, cyclic GMP-AMP synthase (cGAS) can activate the stimulator of interferon genes (STING), which plays a pivotal role in DNA/RNA sensing pathway, for type I IFN and other inflammatory cytokines induction including IL-6 and attributes to STING-associated inflammatory disorders. Interestingly, the elevated levels of IFN-α/β and IFN-stimulated genes were found in SLE patients than healthy individuals. Given this, we propose a hypothesis that the cGAS-STING pathway in multiple organs function versatile and can facilitate overall disease progression of SLE though impertinent cytosolic self-DNA sensing.


End-of-life care research with bereaved informal caregivers--analysis of recruitment strategy and participation rate from a multi-centre validation study.
One essential element of research is the successful recruitment of participants. However, concerns are obvious regarding the ethical implications of involving terminally ill and even dying patients and their informal caregivers as research participants. This study aims to illustrate central issues encountered when recruiting bereaved informal caregivers for a questionnaire validation study on the quality of dying and death. Between July 2012 and November 2013, informal caregivers of deceased inpatients who were treated at two palliative care units in Germany were invited to participate in a questionnaire validation study. Informal caregivers were called by a trained researcher at the end of the fourth week after death at the earliest and by the sixteenth week after death at the latest and asked to participate in a face-to-face interview in their private home. The overall participation rate of all eligible informal caregivers was 76.1% (226/297). The mean burden score was 2.5 (NRS from 0=no burden to 10=maximum burden; n=221). Higher burden scores (≥4) were associated with emotional and burdensome memories (n=34) being invoked throughout the interview. Severe or maximum burden scores (≥7) were stated by 13.2% of participants. The average time between the associated patient's death and the informal caregiver's interview was 57.3 days (range 26-176 days, median 49.5 days). 5.3% of all 226 interviews were not completed due to different reasons. Participants' comments on the way in which the study was conducted gave insight into their motivation to take part in the study and their evaluation of the interview situation. The recruitment strategy can be recommended to other researchers developing research with bereaved carers. The burden caused by study participation was acceptable to the researchers carrying out this research, although a small group of participants experienced high levels of burden which should be anticipated and appropriate support services offered. Family caregivers are willing to support end-of-life care research, have different motivations for participation and even reported benefits from participation. Nevertheless, study designs have to take into account and ease the potential burden of interviews for caregivers experiencing grief.


The true cost of child abuse at a level 1 pediatric trauma center.
Child physical abuse (CPA) is a significant cause of morbidity and mortality. Children who sustain CPA consume significant healthcare resources. We hypothesized that the costs to care for children who sustain for children with CPA-type injuries are greater than the costs to care for children who sustain accidental injuries. All confirmed CPA patients between the ages of 0 and 19 years old, who were admitted to a level 1 pediatric trauma center between January 2010 and September 2018, were retrospectively reviewed. We compared outcomes, including mortality, length of stay (LOS), diagnostic work-up, and overall cost using propensity matching between CPA and accidentally injured trauma patients. Patients were matched based on injury severity score (ISS). The CPA cohort (n = 595) was younger (1.31 +/- 1.96 years, p < 0.0001) than the accidental trauma patients (8.6 +/-5.54 years). The majority of the CPA patients had Medicaid coverage (75.1%), when compared to accidental trauma patients (37.5%; p < 0.0001). CPA patients had longer ICU LOS (2.43 days; p < 0.0001), increased ventilation days (2.57 days; p < 0.0001), and longer hospital LOS (6.56 days; p = 0.0004). The overall mortality rate for CPA patients was higher than accidental trauma patients (9.9% vs. 1.2%; p < 0.0001). The median hospital cost was significantly higher for those with CPA ($18,000) than accidental trauma ($10,100; p < 0.0001). The costs to care for children who sustain CPA-type injuries are significantly greater than the costs to care for children who sustain accidental trauma. Better screening tools, more provider education and broader community outreach efforts are needed to reduce the societal and economic costs associated with child physical abuse. Treatment. Level III.


[Oncological outcome of radical cystectomy for BCG failure compared to primary invasive disease].
BCG therapy deeply modified prognosis of high-risk non muscle invasive (NMI) urothelial carcinomas. However, these tumors remain potentially lethal. The objective of this study was to compare oncological outcome of radical cystectomy (RC) for BCG failure to primary invasive (PI) tumors. RC performed between 2001 and 2011 were retrospectively reviewed. Clinicopathological and follow-up data were compared between RC performed for: NMI high-grade recurrence under BCG therapy (ReNMI); MI recurrence (≥ T2) under BCG therapy (ReMI); primary invasive tumors (PI). The three groups were defined according to tumor status on last TUR before RC. All NMI high-grade bladder tumors at diagnosis had maintenance BCG immunotherapy. Two hundred patients were included, 155 PI, 21 ReNMI et 24 ReMI. Median follow up was 42 months (1.74-135.9). Mean BCG instillations number was 8 ± 4.2 versus 9.5 ± 4.3 for ReNMI and ReMI respectively (P=0.24). Upstaging (≥ pT2) occurred in 33% of ReNMI. The rate of pN+ was 24%, 42% and 30% for the ReNMI, ReMI et PI respectively (P=0.39). No differences were observed between the groups for lymphovascular invasion, extracapsular extension if pN+, soft tissue surgical margins and adjuvant chemotherapy. 5-year cancer specific survival (CSS) was 48% for the ReNMI, 18% for the ReMI and 47% for the PI (P=0.02). Progression to muscle invasion under BCG therapy was an independent pejorative prognostic factor for CSS (P=0.05). BCG failure led to poor prognosis, particularly when tumors progressed to muscle invasion. Recurrent NMI high-grade tumors seemed to have comparable prognosis than PI tumors because of the high amount of upstaging and nodal invasion. BCG failure is a therapeutic emergency.


Immunological changes following electroconvulsive therapy in multiple sclerosis.
Electroconvulsive therapy (ECT) is a well-established treatment option in case of treatment-resistant depression (TRD). Only a few cases of ECT in depressed patients with multiple sclerosis (MS) were reported so far suggesting efficacy for the treatment of severe depression in MS, while data on possible neurological deterioration remained unclear. In this case study we report on a case of a middle-aged man with MS. He was on dimethyl fumarate for relapse prevention since 2019 and without signs of active disease in a recent cerebral MRI. He suffered from treatment-resistant severe bipolar depression and thus received a total of 14 ECT sessions. We changed from right-unilateral to bilateral stimulation technique after the 7th session. We rated depression severity and measured biomarkers of neurodegeneration and inflammation before and after the ECT series to determine the impact of ECT on tolerance, response and neurobiology. The ECT series was tolerated well without neurological deterioration and any new neurological symptoms. The seizure quality was sufficient on average. We saw partial response corresponding to an improvement of about 35% in BDI-II and MADRS. The concentration of inflammation and neurodegeneration biomarkers was low both pre-treatment and post-treatment with increases from pre- to post ECT mainly in the CCL-2 pathway. In our patient with TRD and MS ECT was safe and feasible. We did not see any neurobiological signs of disease activation of MS or neurodegeneration during the course of ECT, which may even be beneficial as it led to increase in the neuroprotective CCL-2 pathway in the presented patient.


Generalized Bose-Einstein phase transition in large-m component spin glasses.
It is proposed to understand finite dimensional spin glasses using a 1/m expansion, where m is the number of spin components. It is shown that this approach predicts a replica symmetric state in finite dimensions. The point about which the expansion is made, the infinite-m limit, has been studied in the mean-field limit in detail and has a very unusual phase transition, rather similar to a Bose-Einstein phase transition but with N(2/5) macroscopically occupied low-lying states.


Developmental programming: prenatal testosterone excess disrupts anti-Müllerian hormone expression in preantral and antral follicles.
To investigate the impact of prenatal T excess on the expression of key ovarian regulators implicated in follicular recruitment and persistence using a large animal model of polycystic ovarian syndrome (PCOS). Interventional, animal model study. Academic research unit. A total of 25 female fetuses, 14 prepubertal female, and 24 adult female Suffolk sheep. Prenatal T treatment. Immunohistochemical determination of expression of anti-Müllerian hormone (AMH), kit ligand, and growth differentiation factor 9 (GDF9) in fetal, prepubertal, and adult ovarian tissues. Prenatal T treatment reduced the AMH protein expression in granulosa cells (GC) of preantral follicles and increased its expression in antral follicles compared with age-matched adult controls. These differences were not evident in prepubertal animals. Protein expression of GDF9 and kit ligand was not altered at any of the developmental time points studied. Prenatal T exposure is associated with changes in AMH expression in preantral and antral follicles in adult ovaries, similar to findings in women with PCOS. These findings indicate that abnormal folliculogenesis in PCOS may be at least in part mediated by changes in AMH expression.


The effect of exercise on bone mass of osteoporotic patients on fluoride treatment.
The study was conducted in order to determine the effect of exercise on osteoporotic patients with a mean age of 65 years. The majority of these patients had been on sodium fluoride for at least 2 years. Thirty-eight patients were assigned to either the hospital or home group. Nineteen were in the former group, which performed the 1 hour exercise twice weekly in the Hospital. The patients in the latter group were taught the exercises and were expected to continue them at home. The exercise protocol consisted of muscle-strengthening and aerobic activities. Bone mineral mass was determined by neutron activation analysis, which measured total calcium in the trunk and pelvis. It is expressed as a calcium bone index (CaBI), with a normal range of 0.75 to 1.2. The level of physical performance was determined by calculating the maximum oxygen uptake, VO2max, attained by a graded exercise test on the treadmill. Calcium bone index and VO2max were obtained before and after the 12.5 month study. Prior to the study, the hospital and home groups had low calcium bone index values (0.68 and 0.66 respectively). Both groups showed a significant increase in calcium bone index (p less than 0.001 for the hospital group, p less than 0.01 for the home group) after the study. When compared to the home group the hospital group was found to have a significantly higher post-study calcium bone index value (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)


Vesicovaginal fistula repair with rectus abdominus myofascial interposition flap.
Complex, recurrent vesicovaginal fistulas (VVFs) can be very challenging to repair and often require interposition of nonirradiated, well-vascularized tissue between the urinary system and vagina. We report our experience using a rectus abdominus myofascial (RAM) interposition flap for VVF repair. A retrospective analysis was performed to identify patients who had undergone VVF repair with RAM interposition. Data were collected focusing on preoperative patient characteristics, etiology of VVF, intraoperative parameters, including surgical techniques, and postoperative patient outcomes. We used a RAM interposition flap for VVF repair in 5 patients. All VVFs had developed postoperatively; no patient had received radiotherapy. VVF developed after total abdominal hysterectomy (TAH) or radical cystectomy in 3 and 2 cases, respectively. Both cases of VVF after radical cystectomy occurred in conjunction with orthotopic diversion (neobladder-vaginal fistula). In 3 patients with post-TAH VVF, a total of five previous failed repairs were attempted before RAM interposition. In 1 patient with a neobladder-vaginal fistula, who had received adjuvant chemotherapy, RAM interposition failed, and the patient ultimately required cutaneous urinary diversion after two subsequent failed attempts at repair (68 months of follow-up). The remaining 4 patients (80%) had no evidence of recurrent VVF or voiding abnormalities at a mean follow-up of 19 months (range 8 to 32). Rectus abdominus muscle can be a successful interposition flap during repair of complex, recurrent VVF. In our experience, this has been successful in most cases, particularly in younger patients with nonmalignant processes.


Successful Treatment of Combined Aspergillus and Cytomegalovirus Abscess in Brain and Lung After Liver Transplant for Toxic Fulminant Hepatitis.
Invasive aspergillosis is one of the most important and fatal complications after liver transplant, especially in patients with involvement of the central nervous system. We present a case of a patient who developed cerebral and pulmonary aspergillosis, coinfected with cytomegalovirus, after liver transplant for toxic fulminant hepatitis. The patient was treated successfully with neurosurgical intervention and voriconazole. Voriconazole is considered more effective in cerebral aspergillosis than other anti-fungal agents due to the greater penetration into central nervous system and higher cerebrospinal fluid and brain tissue levels.


Occult interpositional rotator cuff - an extremely rare case of traumatic rotator cuff tear.
Traumatic interposition of a rotator cuff tendon in the glenohumeral joint without recognizable glenohumeral dislocation is an unusual complication after shoulder trauma. Here we report the clinical and imaging presentations of a 17-year-old man with trapped rotator cuff tendons in the glenohumeral joint after a bicycle accident. The possible trauma mechanism is also discussed.


Influences of dielectric constant and scan rate on hysteresis effect in perovskite solar cell with simulation and experimental analyses.
In this work, perovskite solar cells (PSCs) with different transport layers were fabricated to understand the hysteresis phenomenon under a series of scan rates. The experimental results show that the hysteresis phenomenon would be affected by the dielectric constant of transport layers and scan rate significantly. To explain this, a modified Poisson and drift-diffusion solver coupled with a fully time-dependent ion migration model is developed to analyze how the ion migration affects the performance and hysteresis of PSCs. The modeling results show that the most crucial factor in the hysteresis behavior is the built-in electric field of the perovskite. The non-linear hysteresis curves are demonstrated under different scan rates, and the mechanism of the hysteresis behavior is explained. Additionally, other factors contributing to the degree of hysteresis are determined to be the degree of degradation in the perovskite material, the quality of the perovskite crystal, and the materials of the transport layer, which corresponds to the total ion density, carrier lifetime of perovskite, and the dielectric constant of the transport layer, respectively. Finally, it was found that the dielectric constant of the transport layer is a key factor affecting hysteresis in perovskite solar cells.


Synthesis and antibacterial activity of dual-action agents of a beta-lactam antibiotic with cytotoxic agent mitozolomide or temozolomide.
Dual-action agents 5a-f and 12a-f, a beta-lactam antibiotic combined with a cytotoxic agent, mitozolomide (Meto) or temozolomide (Temo), were synthesised. The antibacterial activity (MICs) of the dual-action agents has been determined against a panel of bacteria including several beta-lactamase producing strains. The tests showed 5a-f active against the non-beta-lactamase producing methicillin sensitive Staphylococcus aureus (MSSA) strains, however, little synergistic effect between the beta-lactam and the cytotoxic agent was observed. 12a-f demonstrated some synergistic effect against bacteria. 12a, in particular, is active against ampicillin resistant (beta-lactamase-producing) strains of Serratia marcescens.


Para-Hisian Pacing: New Insights of an Old Pacing Maneuver.
More than 2 decades ago, para-Hisian pacing was introduced to assess the pattern of retrograde conduction during electrophysiological studies. Although there is no ideal maneuver for every patient and condition, para-Hisian pacing is a valuable and handy strategy to differentiate between retrograde conduction over the atrioventricular node and the accessory pathways. The dynamic behavior of para-Hisian pacing, in a region with unique anatomical features, can produce various activation patterns and intriguing electrophysiological phenomena. Although the demonstration of a retrograde nodal activation pattern during para-Hisian pacing does not rule out the presence of an accessory pathway, evidence of retrograde conduction over an accessory pathway does not prove its active role in the culprit tachycardia. Multipolar His bundle recordings, detailed atrial mapping, and recognition of the truly captured structures and the impact of temporal changes of autonomic tone or pacing rates, are essential keys for accurate interpretation of this maneuver that may ultimately guide judicious catheter ablation of the arrhythmic substrate. This review aims to summarize the practical usefulness and potential pitfalls of the para-Hisian pacing maneuver, focusing on the interpretation of electrocardiograms and intracardiac recordings.


The dual lives of bidirectional promoters.
The sequencing of the human genome led to many insights into gene organization and structure. One interesting observation was the high frequency of bidirectional promoters characterized by two protein encoding genes whose promoters are arranged in a divergent or "head-to-head" configuration with less than 2000 base pairs of intervening sequence. Computational estimates published by various groups indicate that nearly 10% of the coding gene promoters are arranged in such a manner and the extent of this bias is a unique feature of mammalian genomes. Moreover, as a class, head-to-head promoters appear to be enriched in specific categories of gene function. Here we review the structure, composition, genomic properties and functional classifications of genes controlled by bidirectional promoters and explore the biological implication of these features. This article is part of a Special Issue entitled: Chromatin in time and space.


Tailoring the Microstructure of a Solid Oxide Fuel Cell Anode Support by Calcination and Milling of YSZ.
In this study, the effects of calcination and milling of 8YSZ (8 mol% yttria stabilized zirconia) used in the nickel-YSZ anode on the performance of anode supported tubular fuel cells were investigated. For this purpose, two different types of cells were prepared based on a Ni-YSZ/YSZ/Nd2NiO4+δ-YSZ configuration. For the anode preparation, a suspension was prepared by mixing NiO and YSZ in a ratio of 65:35 wt% (Ni:YSZ 50:50 vol.%) with 30 vol.% graphite as the pore former. As received Tosoh YSZ or its calcined form (heated at 1500 °C for 3 hours) was used in the anode support as the YSZ source. Electrochemical results showed that optimization of the fuel electrode microstructure is essential for the optimal distribution of gas within the support of the cell, especially under electrolysis operation where the performance for an optimized cell (calcined YSZ) was enhanced by a factor of two. In comparison with a standard cell (containing as received YSZ), at 1.5 V and 800 °C the measured current density was -1380 mA cm(-2) and -690 mA cm(-2) for the cells containing calcined and as received YSZ, respectively. The present study suggests that the anode porosity for improved cell performance under SOEC is more critical than SOFC mode due to more complex gas diffusion under electrolysis mode where large amount of steam needs to be transfered into the cell.


Trypanosoma brucei: Inhibition of cathepsin L is sufficient to kill bloodstream forms.
The lysosomal cysteine protease activity of Trypanosoma brucei comprises a cathepsin B enzyme (TbCATB) and a cathepsin L enzyme (TbCATL). Inhibition of the cysteine protease activity is lethal to bloodstream-form trypanosomes but it was not entirely clear which of the two enzymes are essential for survival of the parasites. Here we show that the vinyl sulfone compound LU-102 selectively inhibits TbCATL without affecting TbCATB and the proteasomal trypsin-like activity within trypanosomes. Therefore, the trypanocidal activity displayed by LU-102 can be attributed solely to the inhibition of TbCATL demonstrating that this enzyme is essential to the survival of T. brucei.


Permeability of the mouse gallbladder to blood-borne horseradish peroxidase.
Horseradish peroxidase (HRP) was administered intravenously to mice by bolus injection. The subsequent uptake and fate of the HRP by the lateral and basal cell surfaces of resting and stimulated gallbladder epithelial cells was followed by light and electron microscopy. At 10 min after injection, HRP was visible in the lamina propria of the gallbladder and within 20 min of injection, HRP had permeated the basement membrane and had entered the lateral intercellular space, extending as far as the apical tight junction. Over the following 30 min, there was evidence of vesicular epithelial HRP uptake and 1 h after injection, HRP was visible in epithelial secretory granules within the lumen of the gallbladder and apical transport vesicles. These data provide evidence of a blood-to-bile transport pathway which could represent an important route of entry to bile by various blood-borne macromolecules.


Amino bearing core-shell structured magnetic covalent organic framework nanospheres: Preparation, postsynthetic modification with phenylboronic acid and enrichment of monoamine neurotransmitters in human urine.
Covalent organic frameworks (COFs) have been increasingly employed in separation science, including sample preparation. Herein we fabricated the amino bearing core-shell structured COFs nanospheres [Fe3O4@TpBD(NH2)2], and a novel magnetic boronate affinity adsorbent was synthesized by postsynthetic modification of the Fe3O4@TpBD(NH2)2 with 2-formylphenylboronic acid. The magnetic boronate affinity adsorbent possesses fast magnetic response and high binding capacity up to 1037 μmol g-1 for dopamine. Besides, it was used as an adsorbent for extraction of urinary monoamine neurotransmitters at neutral pH. A method for detection of the monoamine neurotransmitters was developed by coupling the magnetic solid phase extraction with high-performance liquid chromatography-fluorescence detection. Under the optimized conditions, a good analytical method was obtained in the linear dynamic range of 2-200 ng mL-1 with R2 between 0.9917 and 0.9966, with low limit of detection (0.31-0.54 ng mL-1) and limit of quantification (1.04-1.80 ng mL-1). The recoveries of the monoamine neurotransmitters were in the range of 86.3-115%, with relative standard deviations of 2.34-10.5% (intra-day) and 2.84-14.4% (inter-day). The method was successfully applied to the determination of the monoamine neurotransmitters in human urine samples. This work is of great importance for preparing functionalized core-shell structured magnetic covalent organic framework nanospheres, it also demonstrates the feasibility of the functionalized magnetic COFs as adsorbents in sample pretreatment.


A haplotype constituted of four MMP-2 promoter polymorphisms (-1575G/A, -1306C/T, -790T/G and -735C/T) is associated with coronary triple-vessel disease.
Vascular lesion development is associated with an accumulation of extracellular matrix proteins within the vessel wall. The proteins are degraded by matrix metalloproteinases (MMPs). There is also evidence indicating a participation of the MMPs in the weakening of atherosclerotic plaque that predisposes to lesion disruption. The aim of the study was to test an association among haplotypes of four single nucleotide MMP-2 promoter polymorphisms and the angiographically confirmed coronary triple-vessel disease (TVD). Incidence of haplotypes of four MMP-2 promoter polymorphisms (-1575G/A, -1306C/T, -790T/G and -735C/T) determined by PCR reactions with restriction analyses in 187 patients with coronary TVD (153 men, 34 women, age median 65 years) was compared to 196 control subjects without clinical signs of coronary heart disease (131 men and 65 women, age median 60 years). The incidence of two similar haplotypes was found to be different between patients and healthy subjects. The haplotype GCTC was more frequent in the TVD patients (P=0.01) though the haplotype GCGC was identified only in healthy subjects (P=0.001). Interestingly, the GCTC is the most frequent polymorphic haplotype composed of four promoter SNPs localized in the MMP-2 gene (53% in healthy subjects vs. 66% in patients with TVD) and the haplotype GCGC is the least frequent polymorphic one (4.4% in healthy subjects vs. 0% in patients with TVD). Two different MMP-2 promoter haplotypes differing only in -790T/G allele are significantly more or less frequent in coronary TVD compared to non-ischemic persons. Thus, the -790T/G MMP-2 genotype might be used as a genetic marker representing MMP-2 promoter variability for the TVD with odds ratio for TT and TG genotypes 2.59, 95% confidential interval 1.21-5.55, P=0.009. The analysis of promoter MMP-2 gene variability could help us to understand individual susceptibility to MMP inhibitor treatment of the coronary artery disease.


Identification and characterization of cancer initiating cells from BRCA1 related mammary tumors using markers for normal mammary stem cells.
It is hypothesized that cancer stem cells arise either from normal stem cells or from progenitor cells that have gained the ability to self-renew. Here we determine whether mammary cancer stem cells can be isolated by using antibodies that have been used for the isolation of normal mammary stem cells. We show that BRCA1 mutant cancer cell lines contained a subpopulation of CD24+CD29+ or CD24+CD49f+ cells that exhibited increased proliferation and colony forming ability in vitro, and enhanced tumor-forming ability in vivo. The purified CD24+CD29+ cells could differentiate and reconstitute the heterogeneity found in parental cells when plated as a monolayer. Under low-attachment conditions, we detected "tumorspheres" only in the presence of double positive cells, which maintained their ability to self-renew. Furthermore, CD24+CD29+ cells could form tubular structures reminiscent of the mammary ductal tree when grown in three-dimensional cultures, implying that these cancer cells maintain some of the characteristics of the normal stem cells. Nevertheless, they could still drive tumor formation since as low as 500 double positive cells immediately after sorting from BRCA1 mutant primary tumors were able to form tumors with the same heterogeneity found in the original tumors. These data provide evidence that breast cancer stem cells originate from normal stem cells and advance our understanding of BRCA1-associated tumorigenesis with possible implications for future cancer treatment.


Acute upper gastrointestinal bleeding: a population-based, five-year follow-up study.
Data on long-term rebleeding risk and mortality in acute upper gastrointestinal bleeding (AUGIB) patients are scarce and comparison to controls are lacking. Aimsof the study were to assess long-term prognosis of AUGIB patients and compare to controls. A population-based retrospective case-control study conducted at the National University Hospital of Iceland and included all patients who underwent endoscopy in 2010-2011. AUGIB was defined as haematemesis or coffee ground vomiting leading to hospitalization or occurring in a hospitalized patient. Controls underwent endoscopy in 2010-2011, matched for sex/age. Rebleeding was defined as AUGIB >14 days up to five years after index bleeding. Overall, 303 patients had AUGIB, mean age 67 (±18), controls66 years (±19), females, 51 and 46%, respectively. The five-year rebleeding rate for AUGIB patients was 13% (95%CI 9-17%), higher than the rate of bleeding events in controls, 3% (95%CI 1-5%; log-rank <0.001), hazard ratio (HR) 6.0 (95%CI 2.4-15) when correcting for comorbidities, NSAID's, PPI's and antithrombotics. The mortality of AUGIB patients at end of follow-up was higher when compared to controls, 39% (95%CI 49-33%) vs. 26% (95%CI 30-21%), log-rank <0.001, comorbidity-adjusted HR 1.4 (1.1-1.9). A subanalysis of non-variceal AUGIB yielded similar results in regard to rebleeding and mortality rates. AUGIB patients were at 6-fold risk of rebleeding compared to bleeding events in controls at five years of follow-up. Five-year mortality was higher in AUGIB patients when compared to controls even when correcting for age and comorbidities, suggesting that an episode of AUGIB indicates serious frailty.


Febrile spontaneous abortion and the IUD.
A case-control study was done to determine the risk and pathogenesis of febrile spontaneous abortion for intrauterine device (IUD) wearers compared to non-wearers. Four groups of women coming to a large city hospital between 1970 and 1975 were compared: 1) women presenting with febrile spontaneous abortion, 2) women presenting with afebrile spontaneous abortion, 3) women presenting with afebrile spontaneous abortion which subsequently became febrile, and 4) women delivering a live infant. Pregnant women with an IUD in situ had a 5-fold higher risk of both febrile and afebrile spontaneous abortion compared to pregnant women without an IUD in situ. Women who had a spontaneous abortion that shifted from afebrile to febrile were somewhat more likely to be IUD wearers than the other 2 spontaneous abortion groups. In our study population, the increased risk of febrile spontaneous abortion for IUD wearers appeared primarily due to the increased risk of the spontaneous abortion event itself, rather than a primary IUD-related infection causing febrile spontaneous abortion.


Timing and Type of Venous Thromboembolic Prophylaxis in Isolated Severe Liver Injury Managed Non-Operatively.
The optimal timing and type of pharmacological venous thromboembolic prophylaxis (VTEp) after severe liver injury selected for nonoperative management (NOM) are controversial. The aim of this study was to assess the effect of timing and type of VTEp in severe liver injuries selected for NOM. ACS-TQIP database study (2013-17) including patients with blunt isolated severe liver injuries (AIS ≥ 3), selected for NOM, who received VTEp with either unfractionated heparin (UH) or low-molecular-weight heparin (LMWH). Patients who underwent laparotomy or angiointervention within 24 h or prior to the initiation of VTEp were excluded. The study population was stratified according to the timing of VTEp ≤ 48 h (EP) and > 48 h (LP) groups. Univariate and multivariate analyses were used to identify differences between the groups. A total of 4074 patients was included in the study. 2004 (49.2%) received EP and 2070 (50.8%) LP. Patients with more severe injuries were more likely to receive LP than an EP [ISS 24 (19-29) vs 22 (17-27), p < 0.001]. On multivariate analysis (correcting for age, gender, comorbidities, blood pressure, GCS, ISS, type of VTEp), LP was identified as an independent risk factor for thromboembolic events (OR 1.52, p = 0.032) and mortality (OR 2.49, p = 0.031). LMWH was independently associated with lower mortality (OR 0.36, p = 0.007), compared to UH. EP did not increase the risk of laparotomy or angiointervention after starting VTEp, compared to LP (p = 0.992). Early VTEp (≤ 48 h) is safe and independently associated with fewer thromboembolic events and a lower mortality after isolated severe liver injuries managed nonoperatively. LMWH was independently associated with improved outcomes when compared with UH.


Sex-specific differences in colorectal cancer: A multicenter retrospective cohort study.
Due to sex-specific differences in the incidence and clinical and histopathological characteristics of colorectal cancer (CRC), understanding the impact of sex on CRC may suggest sex-targeted strategies for screening, treatment, and prevention, leading to improved prognosis of CRC. However, there have been few studies investigating the sex-specific differences in CRC in the Republic of Korea. We aimed to assess sex differences in CRC in the Republic of Korea. This was a retrospective, multicenter, cohort study of patients diagnosed with CRC between January 2012 and December 2013 at nine hospitals. Patients who had an uncertain CRC stage, were diagnosed with other cancers within 5 years, had carcinoma in situ, non-epithelial cancer, or primary cancer other than CRC, were excluded. Factors associated with overall survival or progression-free survival were investigated using Cox regression analysis. Cumulative probability of metachronous lesions was compared using the Kaplan-Meier estimator survival analysis and we compared the survival curves of each group using a log-rank test. Outcomes were compared using the chi-square, Fisher's exact, or Student's t-test, as appropriate. Three thousand one hundred and forteen patients (1999 men, 1315 women) were included. There was no significant difference in the age at onset between men and women. The proportion of patients diagnosed through regular health check-ups, and asymptomatic at time of diagnosis, was higher in men (48.9% men vs. 42.0% women, p < .001). Rectal cancers were more common in men (38.8% men vs. 31.8% women, p < .001). Right colon cancers were more common in women (31.4% women vs. 22.7% men, p < .001). KRAS mutations were found in 109/317 (34.4%) women and 112/480 (23.3%) men. Overall CRC survival and progression-free survival were similar in both sexes. Sex differences in CRC may be due to the biological and social-behavioral differences between the sexes. They should be considered during screening, diagnosis, and treatment of CRC for better outcomes.


Differential localization of acetylcholinesterase in neuronal and non-neuronal cells.
Acetylcholinesterase (AChE) expression is regulated in cell types at the transcriptional and translational levels. In this study, we characterized and compared AChE catalytic activity, mRNA, protein expression, and protein localization in a variety of neuronal (SH-SY5Y neuroblastoma and primary cerebellar granule neurons (CGN)) and non-neuronal (LLC-MK2, HeLa, THP-1, and primary astrocytes) cell types. All cell lines expressed AChE catalytic activity; however the levels of AChE-specific activity were higher in neuronal cells than in the non-neuronal cell types. CGN expressed significantly more AChE activity than SH-SY5Y cells. All cell lines analyzed expressed AChE protein at equivalent levels, as well as mRNA splice variants. Localization of AChE was characterized by immunofluorescence and confocal microscopy. SH-SY5Y, CGN, and nerve-growth factor-differentiated PC-12 cells exhibited a pattern of AChE localization characterized as diffuse in the cytoplasm and punctate staining along neurites and on the plasma membrane. The localization in HeLa, LLC-MK2, fibroblasts, and undifferentiated PC-12 cells was significantly different than in neuronal cells-AChE was intensely localized in the perinuclear region, without staining near or on the plasma membrane. Based on the evidence presented here, we hypothesize that the presence of AChE protein doesn't correlate with catalytic activity, and the diffuse cytoplasmic and plasma membrane localization of AChE is a property of neuronal cell types.


The Causes and Consequences of Nonenzymatic Protein Acylation.
Thousands of protein acyl modification sites have now been identified in vivo. However, at most sites the acylation stoichiometry is low, making functional enzyme-driven regulation in the majority of cases unlikely. As unmediated acylation can occur on the surface of proteins when acyl-CoA thioesters react with nucleophilic cysteine and lysine residues, slower nonenzymatic processes likely underlie most protein acylation. Here, we review how nonenzymatic acylation of nucleophilic lysine and cysteine residues occurs; the factors that enhance acylation at particular sites; and the strategies that have evolved to limit protein acylation. We conclude that protein acylation is an unavoidable consequence of the central role of reactive thioesters in metabolism. Finally, we propose a hypothesis for why low-stoichiometry protein acylation is selected against by evolution and how it might contribute to degenerative processes such as aging.


Anti-proliferative, apoptosis inducing, and antioxidant potential of Callistemon lanceolatus bark extracts: an in vitro and in silico study.
The present study reports anticancer and antioxidant activities of Callistemon lanceolatus bark extracts. Anticancer activity was studied against MDA-MB-231 cells. Antioxidant assessment of the chloroform and methanol extracts showed considerable free radical scavenging, metal ion chelating, and reducing power potential. Chloroform extract exhibited potent inhibition of cancer cell proliferation in MTT assay (IC50 9.6 μg/ml) and promoted programmed cell death. Reactive oxygen species (ROS) generation, mitochondria membrane potential (MMP) disruption ability, and nuclear morphology changes were studied using H2-DCFDA, JC-1, and Hoechst dyes, respectively, using confocal microscopy. Apoptotic cells exhibited fragmented nuclei, increased ROS generation, and altered MMP in dose- and time-dependent manner. Chloroform extract upregulated the BAX-1 and CASP3 mRNA expression coupled with downregulation of BCL-2 gene. Further, in silico docking of phytochemicals present in C. lanceolatus with anti-apoptotic Bcl-2 protein endorsed apoptosis by its inhibition and thus corroborated the experimental findings. Obatoclax, a known inhibitor of Bcl-2 was used as a reference compounds.


Increased expression of plasminogen activator inhibitor-1 (PAI-1) is associated with depression and depressive phenotype in C57Bl/6J mice.
Plasminogen activator inhibitor 1 (PAI-1), which is elevated in numerous disease states, has been implicated as a stress-related protein involved in the pathogenesis of depression. We measured PAI-1 in the plasma of healthy and depressed individuals and assessed plasminogen activator (PA) expression and regulation by PAI-1 in cultured normal human astrocytes (NHA). Elevated plasma PAI-1 levels were found in depressed patients. Brain tissues from depressed individuals also showed stronger expression of hippocampal PAI-1 by confocal imaging in comparison to healthy individuals. Using a lipopolysaccharide-induced inflammatory model of depression in mice, we measured PAI-1 in murine plasma and brain, by ELISA and immunohistochemistry, respectively. Similar elevations were seen in plasma but not in brain homogenates of mice exposed to LPS. We further correlated the findings with depressive behavior. Ex vivo experiments with NHA treated with proinflammatory cytokines implicated in the pathogenesis of depression showed increased PAI-1 expression. Furthermore, these studies suggest that urokinase-type plasminogen activator may serve as an astrocyte PA reservoir, able to promote cleavage of brain-derived neurotrophic factor (BDNF) during stress or inflammation. In summary, our findings confirm that derangements of PAI-1 variably occur in the brain in association with the depressive phenotype. These derangements may impede the availability of active, mature (m)BDNF and thereby promote a depressive phenotype.


The synthesis of glucose and ammonia by kidney tubules isolated from suckling and early-weaned lambs.
Glucose and ammonia production were examined in kidney tubules isolated from suckling and early-weaned lambs, on days 10-30 after birth, with abrupt weaning occurring at day 14. There were no differences in the rates of glucose or ammonia production for a given substrate by tubules isolated from any of the lambs, regardless of age or stage of weaning. The preferred substrates for gluconeogenesis were glycerol = lactate greater than propionate = pyruvate = fructose = proline greater than alanine greater than glutamate greater than glutamine greater than aspartate greater than glycine greater than serine, and for ammoniagenesis were glutamine much greater than alanine greater than aspartate much greater than serine greater than glycine = glutamate = proline.


Susceptibility to erythromycin of anaerobes of the genera Bacteroides, Fusobacterium, Sphaerophorus, Veillonella, Clostridium, Corynebacterium, Peptococcus, Peptostreptococcus.
The minimal inhibitory concentrations (MIC) of erythromycin were determined by broth dilution tests for 313 anaerobic strains, most of which were clinical isolates. All the gram-positive anaerobes tested (84 Peptococcaceae, including 21 Peptostreptococcus anaerobius and 15 Peptococcus variabilis; 65 Corynebacterium acnes and 29 Clostridium strains, including 13 C. perfringens) were sensitive (MIC values 0.012 through 3.12 microgram erythromycin/ml); so were 111 cultures of gram-negative anaerobes (52 Bacteroides fragilis, 12 B. thetaiotaomicron, 7 B. vulgatus, 13 B. oralis, 4 B. melaninogenicus, 10 Sphaerophorus necrophorus, 2 Veillonella sp., 11 members of other species). Erythromycin at concentrations of 6.25 through 200.0 microgram/ml was active against 24 strains (1 B. fragilis, 4 Fusobacterium fusiforme, 9 Sph. freundi, 10 Sph. varius). The present results are compared to the limited number of reports existing with regard to the susceptibility of anaerobes to erythromycin.


KLF9 suppresses cell growth and induces apoptosis via the AR pathway in androgen-dependent prostate cancer cells.
Kruppel-like factors (KLFs) play an important role in many biological processes including cell proliferation, differentiation and development. Our study showed that the level of KLF9 is lower in PCa cell lines compared to a benign prostate cell line; the androgen-independent cell line PC3 expresses significantly lower KLF9 than the androgen-dependent cell line, LNCaP. Forced overexpression of KLF9 suppressed cell growth, colony formation, and induced cell apoptosis in LNCaP cells. We also found that KLF9 expression was induced in response to apoptosis caused by flutamide, and further addition of dihydrotestosterone antagonized the action of flutamide and significantly decreased KLF9 expression. Furthermore, activation of the androgen receptor (AR) was inhibited by the overexpression of KLF9. Our research shows that KLF9 is lower in androgen-independent cell lines than in androgen-dependent cell lines; Overexpression of KLF9 dramatically suppresses the proliferation, anchorage-independent growth, and induces apoptosis in androgen-dependent cells; KLF9 inhibition on prostate cancer cell growth may be acting through the AR pathway. Our results therefore suggest that KLF9 may play a significant role in the transition from androgen-dependent to androgen-independent prostate cancer and is a potential target of prevention and therapy.


[Substantiation of the system of thrombus formation prevention in treatment of patients with hip joint pathology against the background of arterial insufficiency of the lower extremities].
To evaluate effectiveness of electroneurostimulation of the shin muscles as a constituent part of the prevention of TEC in the hip joint pathology at the hospital stage of treatment. With this purpose, there was conducted the extended comparative analysis of the parameters of the blood coagulation system in 27 patients with pathology of the hip joints and the stated concomitant vascular insufficiency. The comparison group comprised 50 patients with uncomplicated pathology of the hip joint who underwent hip joint replacement surgery. In the patients of both groups the product dabigatran etexilate in the standard dosage (220 mg/day) was used for specific prevention. The authors managed to confirm qualitatively and quantitatively the effectiveness of the new variant of non-specific prevention in patients with a high risk of TEC development. Similar changes of the parameters of blood coagulation system were obtained at the use of electroneurostimulation of the shin muscles against the background of the use of the direct anticoagulant dabigatran etexilate. The numbers of thromboses in the group of patients with degenerative and dystrophic diseases of the hip joint with and without arterial insufficiency of the lower extremities are quite comparable (3.7 and 6%). Recanalization of the veins of the lower extremities in every individual case was achieved at the use of therapeutic doses of dabigatran etexilate (300 mg/day). The use of dabigatran etexilate permitted to minimize the risk of deep venous thrombosis (DVT) at hip interventions and contributed to recanalization of veins in recent thromboses in the postoperative period.


Metagenomic evidence for taxonomic dysbiosis and functional imbalance in the gastrointestinal tracts of children with cystic fibrosis.
Cystic fibrosis (CF) results in inflammation, malabsorption of fats and other nutrients, and obstruction in the gastrointestinal (GI) tract, yet the mechanisms linking these disease manifestations to microbiome composition remain largely unexplored. Here we used metagenomic analysis to systematically characterize fecal microbiomes of children with and without CF, demonstrating marked CF-associated taxonomic dysbiosis and functional imbalance. We further showed that these taxonomic and functional shifts were especially pronounced in young children with CF and diminished with age. Importantly, the resulting dysbiotic microbiomes had significantly altered capacities for lipid metabolism, including decreased capacity for overall fatty acid biosynthesis and increased capacity for degrading anti-inflammatory short-chain fatty acids. Notably, these functional differences correlated with fecal measures of fat malabsorption and inflammation. Combined, these results suggest that enteric fat abundance selects for pro-inflammatory GI microbiota in young children with CF, offering novel strategies for improving the health of children with CF-associated fat malabsorption.


Human T-cell leukemia virus-1-positive cell line established from a patient with small cell lung cancer.
A stable cell line, KHM-3S, was established from a patient with small cell lung cancer (SCLC), who had a high serum level of soluble interleukin 2 receptors (sIL2-R) and was seropositive for human T cell leukemia virus (HTLV)-1. KHM-3S cells were positive for IL2-R (Tac) and NKH-1, but negative for other lymphocytic markers such as OKT 11, OKT 4, OKT 8, T cell receptor (WT 31), B 1, and B 4. Moreover, the KHM-3S cells were negative for leukocyte common antigen and strongly positive for neuron-specific enolase (NSE). Secretion of sIL2-R and NSE by the KHM-3S line was detected by an enzyme-linked immunosorbent assay. Rearrangement of the T cell receptor gene and monoclonal HTLV-1 integration were found by Southern blot analysis of KHM-3S DNA. However, Northern blot analysis showed no T cell receptor mRNA. KHM-3S may be useful for studies on the role of HTLV-1 in carcinogenesis and IL2-R expression in SCLC.


Interleukin-1β plays key roles in LPA-induced amplification of LPA production in neuropathic pain model.
Lysophosphatidic acid (LPA) is a bioactive lipid mediator that exerts a wide range of biological actions. In recent decades, LPA has been demonstrated as an important initiator of neuropathic pain based on the mechanisms of LPA-induced feed-forward LPA amplification. In this study, we examined the possible involvement of interleukin (IL)-1β in such LPA production. Intrathecal (i.t.) LPA injection rapidly increased the expression of IL-1β mRNA in the spinal dorsal horn as early as 0.5 h after injection, and the level reached peak at 2 h. Through a developed quantitative mass spectrometry for detecting LPA species, the elevated levels of 18:1, 16:0, and 18:0 LPA in the spinal dorsal horn were observed at 3 h after 18:1 LPA injection and this elevation was completely blocked by the pretreatment of IL-1β-neutralizing antibody. Moreover, enzyme assay experiments showed that LPA (i.t.) significantly activated calcium-independent phospholipase A2 (iPLA2) and cytosolic phospholipase A2 (cPLA2) in the spinal dorsal horn at 1 and 2 h, respectively, and these biochemical changes were also significantly inhibited by IL-1β-neutralizing antibody. Similarly, IL-1β-neutralizing antibody reversed LPA-induced neuropathic pain-like behavior. These findings suggest that the early release of IL-1β is involved in LPA-induced amplification of LPA production, which underlies the initial mechanisms of LPA-induced neuropathic pain.


Accessibility of cell surface thiols in human lymphocytes is altered by ionophores or OKT-3 antibody.
The accessibility of cell surface sulfhydryl groups in human peripheral lymphocytes was investigated with 5,5'-dithiobis-(2-nitrobenzoic acid) in the presence and absence of ionophore antibiotics and the monoclonal antibody, OKT-3. Only a few accessible protein thiols have been found on the cells as demonstrated by labeling with a fluorescent non-penetrating thiol-marker, monobromotrimethyl-ammoniobimane and the subsequent gel electrophoretic analysis of the protein pattern. Difference spectrophotometric measurement of thiol-DTNB reaction revealed that ionophores altering the transmembrane potential induced an enhanced cell surface thiol-exposure on the minute time scale. The effect showed a dependence on the external concentration of the cations. The binding of monoclonal antibody, OKT-3, directed against T3 complexes, resulted in a similar, concentration-dependent increase of thiol-accessibility. These data are interpreted as early membrane-effects of ionophores and the specific antibody including changes in the conformational equilibrium or vertical displacements of certain membrane proteins. These events are likely to be coupled to the changes in the transmembrane potential of the lymphocytes.


Improved adsorption and degradation performance by S-doping of (001)-TiO2.
In this work, sulfur-doped (S-doped) TiO2 with the (001) face exposed was synthesized by thermal chemical vapor deposition at 180 or 250 °C using S/Ti molar ratios R S/Ti of 0, 0.5, 1, 2, 3, 4 and 5. The S-doped samples synthesized at 250 °C exhibit a significantly improved photocatalytic performance. More precisely, S-doping has the following effects on the material: (1) S can adopt different chemical states in the samples. Specifically, it exists in the form of S2- replacing O2- at a ratio of R S/Ti = 1 and also in the form of S6+ replacing Ti4+ at R S/Ti ≥ 2. As a result, S-doping causes a lattice distortion, because the ionic radii of S2- and S6+ differ from that of the O2- and Ti4+ ions. (2) S-doping increases the adsorption coefficient A e for methylene blue (MB) from 0.9% to 68.5% due to the synergistic effects of the oxygen vacancies, increased number of surface chemical adsorption centers as a result of SO4 2- adsorption on the TiO2 surface and the larger pore size. (3) S-doping increases the MB degradation rate from 6.9 × 10-2 min-1 to 18.2 × 10-2 min-1 due to an increase in the amount of •OH and •O2- radicals.


Anesthesia Machine and New Modes of Ventilation.
Mechanical ventilation is ubiquitous in the operating room. This article explores the anesthesia machine as a ventilator, examining its unique features and differences from ventilators designed for long-term use. It will describe standard and nonstandard modes of ventilation. The reader will develop a more nuanced understanding of how to tailor ventilation and oxygenation strategies based on patient and anesthetic scenarios as well as with the assistance of new technologies.


Orthotopic liver transplantation in the presence of partial or total portal vein thrombosis: problems in diagnosis and management.
From January 1989 to May 1992, 70 orthotopic liver transplantations (OLT) were performed in 69 patients, 53 of whom had cirrhosis (77%). Eleven patients (16%) had preoperative partial or total portal vein thrombosis (PVT). Ten of these patients had cirrhosis of various causes. PVT was total in three cases and partial in eight. Total PVT was detected preoperatively in all three cases. By contrast, partial PVT was diagnosed preoperatively in only three of the eight cases. In the five other cases of partial PVT, the obstruction was discovered intraoperatively during dissection of the portal vein. Surgical management of PVT consisted of phlebothrombectomy in ten cases followed by usual end-to-end portal anastomosis in nine cases and anastomosis of the graft's portal vein to the splenomesenteric confluence in one case. Atypical anastomosis of the graft's portal vein to a dilated choledocal vein was performed in one case of total PVT. There were no deaths or complications related to the presence of preoperative PVT or to its management. One patient died postoperatively of primary graft nonfunction at day 5. One patient had arterial thrombosis 3 months after OLT and was successfully retransplanted. Two patients died of recurrent carcinoma 3 and 7 months after OLT. Eight patients are alive 4 to 39 months after OLT. We conclude from this series that (1) the prevalence of preoperative PVT among patients transplanted for advanced cirrhosis may be high (19% of the cirrhotics in this series); (2) PVT is often partial and so difficult to diagnose preoperatively; (3) PVT, even when total, can be managed successfully during surgery and does not seem to affect survival.


Trends in childhood illness and treatment in Australian general practice, 1971-2001.
To determine changes in morbidity and management of disease in children in Australian general practice. A comparative study of general practice consultations in children under 15 years, using data from cross-sectional general practice surveys (1990-91 and 2000-01), and a descriptive comparison with a similar study from 1971. Relative rates of management (rate/100 general practice encounters) of the most common children's problems and treatments. Problems with significantly higher management rates in 2000-01 compared with 1990-91 included vaccination (11.1 v 7.6 per 100 encounters in 1990-91) and contact/allergic dermatitis (3.1 v 2.5). Those managed significantly less often in 2000-01 v 1990-91 included acute otitis media (7.7 v 9.4), asthma (5.4 v 8.8), tonsillitis (4.4 v 6.0), acute bronchitis (3.8 v 5.3) and gastroenteritis (1.7 v 2.7). Asthma management rates rose from 2.4% of all problems managed in 1971 to 7.2% in 1990-91, then fell in 2000-01 to 4.6%. More frequent rates of counselling and advice in 2000-01 (28.4% of encounters v 22.9% in 1990-91) were associated with a decrease in rates of prescribing and supply of medication (56.6% of encounters v 64.3% in 1990-91). Antibiotic prescribing declined significantly (from 33.8 per 100 encounters in 1990-91 to 25.2 in 2000-01), as did prescribing of respiratory medications (from 15.5 to 9.9 per 100 encounters), while prescribing of vaccines and systemic corticosteroids doubled (from 9.6 to 18.8 per 100 encounters, and from 0.6 to 1.2, respectively). (All comparisons between 1990-91 and 2000-01 are significant at P < 0.01.) These findings point to the emergence of a generation of Australian children who are generally well vaccinated and are less likely to present to GPs with "traditional" childhood illnesses.


The melibiose permease system of Escherichia coli K12.
The melibiose permease system of E. coli K12 has been explored using a strain deficient in lactose permease: 300 P. The accumulation of 1-S-methyl-beta-D-thiogalactopyranoside (TMG) was observed. The uptake system was inducible by melibiose and a number of analogs at 30 degrees C. At higher temperatures the differential rate of synthesis decreases until becoming negligible at 42 degrees C. The uptake tends toward a steady state which corresponds to an accumulation several hundredfold over the sugar concentration in the medium. At a given temperature the steady state level was proportional to the initial rate of uptake whatever the degree of induction and the substrate concentration. Lowering the temperature decreased the initial rate of uptake but increased the steady state level. This uptake system was pH dependent with better efficiency at pH 8. It was also dependent on the presence of sodium or lithium ions active at 5 mM whereas potassium at 170 mM enable only about half maximal uptake. The uptake in a medium with choline chloride was less than one fifth of optimal activity. Addition of Li+ brought about half maximal activation at approximately 0.5 mM. The activation consists mainly in a decrease of apparent Km. The emphasis of this study was put on the similarities and differences with lactose permease which is able to transport the same sugar to approximately the same extent. Inducer specificities and substrate specificities were compared and a method of measuring the two activities in the same cells was devised.


Cost-effectiveness of sevelamer versus calcium carbonate plus atorvastatin to reduce LDL in patients with chronic renal insufficiency with dyslipidemia and hyperphosphatemia.
We conducted a cost-effectiveness analysis to compare costs and clinical outcomes of sevelamer versus calcium carbonate plus atorvastatin for treatment of dyslipidemia in patients with chronic renal insufficiency. The model was from the third-party payer perspective. Efficacy and adverse event rates for each regimen were obtained from published clinical trials. Drug costs were based on average wholesale prices; monitoring costs were based on Medicare reimbursement rates. Our model suggests that the combination of calcium carbonate plus atorvastatin is substantially more cost-effective than sevelamer in reducing low-density lipoprotein (LDL) in these patients. One-way sensitivity analyses were performed to assess if 25% and 50% price reductions in sevelamer affected overall cost-effectiveness results. A 50% sevelamer price reduction was less expensive than combination therapy but remained less cost-effective. A two-way sensitivity analysis on the probability that a patient achieves the goal of a 35% LDL reduction resulted in calcium carbonate plus atorvastatin remaining more cost-effective. Further cost-effectiveness studies are necessary to corroborate our data.


Gamibojungikki-tang decreases immobility time on the forced swimming test and increases interferon-gamma production from MOLT-4 cells.
Gamibojungikki-tang (GBIT) has been used for the purpose of development of physical strength in Korea. We investigated the anti-immobility effect of GBIT on the forced swimming test (FST) and then measured the blood biochemical parameters related to fatigue, glucose (Glc); blood urea nitrogen (BUN); lactic dehydrogenase (LDH); creatine kinase (CK) and total protein (TP). GBIT (0.01, 0.1, 1 g/kg) was orally administered to mice for 7 days. After 7 days, the immobility time was significantly decreased in the GBIT-administration group (105.0+/-12.1 s for 1 g/kg) in comparison with the control group (152.3+/-16.2 s). The contents of Glc and TP in the blood serum were significantly increased in GBIT-administration group (1g/kg) compared with control group, while LDH was significantly decreased. Surface phenotyping of spleen cells by FACS analysis revealed an increasing tendency of CD4+ and CD8+ number, without statistical significance. In addition, GBIT (0.01-1 mg/ml) increased the interferon-gamma and interlukin-2 levels in MOLT-4 T-cells. These results suggest that GBIT may be useful in the immune function improvement.


Bovine papillomavirus E5 oncoprotein binds to the activated form of the platelet-derived growth factor beta receptor in naturally occurring bovine urinary bladder tumours.
Studies regarding the functions of the bovine papillomavirus (BPV) E5 oncoprotein in vivo are lacking and no E5-mediated mechanism underlying epithelial carcinogenesis is known. We have shown that BPV-2 DNA is present in the majority of naturally occurring urinary bladder tumours of cattle and that E5 is expressed in the cancer cells. Here we show that the interaction between the platelet-derived growth factor (PDGF) beta receptor and BPV E5, described in vitro in cultured cells, takes place in vivo in bovine urinary bladder cancers. In these cancers, E5 and PDGF beta receptor colocalize, as shown by confocal microscopy, and physically interact, as shown by coimmunoprecipitation. Furthermore, the PDGF beta receptor associated with E5 is highly phosphorylated, suggesting the functional activation of the receptor upon E5 interaction. Our results demonstrate, for the first time, that E5-PDGF beta receptor interaction occurs during the natural history of bovine urinary bladder tumours, suggesting an important role for E5 in carcinogenesis. Finally, the system provides a suitable animal model of papillomavirus-associated cancer to test therapeutic vaccination against E5. Successful bladder tumour regression would provide a valuable model for therapeutic vaccination against papillomavirus-associated tumours.


STI PCR: An efficient method for amplification and de novo synthesis of long DNA sequences.
Despite continuous improvements, it is difficult to efficiently amplify large sequences from complex templates using current PCR methods. Here, we developed a suppression thermo-interlaced (STI) PCR method for the efficient and specific amplification of long DNA sequences from genomes and synthetic DNA pools. This method uses site-specific primers containing a common 5' tag to generate a stem-loop structure, thereby repressing the amplification of smaller non-specific products through PCR suppression (PS). However, large target products are less affected by PS and show enhanced amplification when the competitive amplification of non-specific products is suppressed. Furthermore, this method uses nested thermo-interlaced cycling with varied temperatures to optimize strand extension of long sequences with an uneven GC distribution. The combination of these two factors in STI PCR produces a multiplier effect, markedly increasing specificity and amplification capacity. We also developed a webtool, calGC, for analyzing the GC distribution of target DNA sequences and selecting suitable thermo-cycling programs for STI PCR. Using this method, we stably amplified very long genomic fragments (up to 38 kb) from plants and human and greatly increased the length of de novo DNA synthesis, which has many applications such as cloning, expression, and targeted genomic sequencing. Our method greatly extends PCR capacity and has great potential for use in biological fields.


Procalcitonin as a predictor of early antibiotic treatment failure in patients with gram-negative bloodstream infections caused by urinary tract infections.
We retrospectively evaluated whether initial procalcitonin (PCT) levels can predict early antibiotic treatment failure (ATF) in patients with gram-negative bloodstream infections (GN-BSI) caused by urinary tract infections from January 2018 to November 2019. Early ATF was defined as the following: (1) hemodynamically unstable or febrile at Day 3; (2) the need for mechanical ventilation or continuous renal replacement therapy at Day 3; (3) patients who died within 3 days (date of blood culture: Day 0). The study included 189 patients; 42 showed early ATF. Independent risk factors for early ATF were initial admission to the intensive care unit (odds ratio: 7.735, 95% confidence interval: 2.567-23.311; P < 0.001) and PCT levels ≥30 ng/mL (odds ratio: 5.413, 95% confidence interval: 2.188-13.388; P < 0.001). Antibiotic factors were not associated with early ATF. Initial PCT levels may be helpful to predict early ATF in these patients.


Cytogenotoxic potential of a hazardous material, polystyrene microparticles on Allium cepa L.
Plastic pollution represents a global concern for the biodiversity conservation, ecosystem and public health. The polystyrene is one of the dominant pollutants in both terrestrial and aquatic ecosystem. This work measured the hazardous nature of 100 nm micropolystyrene (MPS) using 25, 50, 100, 200, and 400 mg/L concentrations in terms of oxidative stress, morphotoxicity and cytogenotoxicity in Allium cepa. The results were compared with the positive control (PC) (400 mg/L chlorpyrifos). MPS significantly (p < 0.05) reduced the root length while induced the production of hydroxyl, superoxide radicals with a concomitant increase in DPPH scavenging activity and lipid peroxidation as compared to the negative control. The significant decrease in mitotic index with respect to the negative control (MI: 23.855 ± 5.336 %; lowest MI: 3.88 ± 1.042 %) showed the cytotoxic nature of MPS. Genotoxicity was assessed by various chromosomal and nuclear aberrations. The highest 3.029 ± 0.403 % (PC: 3.09 ± 0.535 %) chromosomal abnormality index and 2.31 ± 0.338 % (PC: 1.178 ± 0.095 %) nuclear abnormality index were observed. MPS down-regulated the expression of plant CDKA encoding gene: cdc2, an important cell cycle regulator. The overall results indicated that MPS could induce cytogenotoxicity through the exacerbation of ROS production and inhibition of cdc2.


Natural infant-directed speech facilitates neural tracking of prosody.
Infants prefer to be addressed with infant-directed speech (IDS). IDS benefits language acquisition through amplified low-frequency amplitude modulations. It has been reported that this amplification increases electrophysiological tracking of IDS compared to adult-directed speech (ADS). It is still unknown which particular frequency band triggers this effect. Here, we compare tracking at the rates of syllables and prosodic stress, which are both critical to word segmentation and recognition. In mother-infant dyads (n=30), mothers described novel objects to their 9-month-olds while infants' EEG was recorded. For IDS, mothers were instructed to speak to their children as they typically do, while for ADS, mothers described the objects as if speaking with an adult. Phonetic analyses confirmed that pitch features were more prototypically infant-directed in the IDS-condition compared to the ADS-condition. Neural tracking of speech was assessed by speech-brain coherence, which measures the synchronization between speech envelope and EEG. Results revealed significant speech-brain coherence at both syllabic and prosodic stress rates, indicating that infants track speech in IDS and ADS at both rates. We found significantly higher speech-brain coherence for IDS compared to ADS in the prosodic stress rate but not the syllabic rate. This indicates that the IDS benefit arises primarily from enhanced prosodic stress. Thus, neural tracking is sensitive to parents' speech adaptations during natural interactions, possibly facilitating higher-level inferential processes such as word segmentation from continuous speech.


Development of local injectable, bone-targeting nanocarriers of triptolide for treatment of bone-only metastasis.
Triptolide (TP) is known for its diverse pharmacological activities but also its delivery and toxicity issues. This study aimed at exploiting TP's anticancer effects at lower risk of systemic toxicity by developing local-injectable "bone-targeting TP nanoparticle" (TPN) for bone-only metastasis treatment. The lipid/oil-based TPNs decorated with alendronate (ALE) achieved size of 70.4-111.2 nm with good dispersion stability. The drug encapsulation efficiency reached 97 % and drug release profiles were in biphasic, controlled manner lasting for 5 days in medium with serum proteins and calcium. TPNs were more cytotoxic than free TP against MDA-MB-231 breast cancer cells (IC50: 16.40 ± 0.80 nM vs 25.45 ± 1.83 nM, P < 0.05) but less cytotoxic against MC3T3-E1 osteoblasts (P < 0.05). When combined with paclitaxel or docetaxel, low dose TPN (containing 10 nM) significantly increased the effectiveness of the two chemotherapy drugs against MDA-MB-231 (IC50 values decreased from 7.3 nM to 2.5 nM for docetaxel; from 4.6 nM to 1.1 nM), indicating potent chemosensitization effects. Retardation of in vitro cancer cell migration by TPN was also observed in the standard scratch assay. ALE decoration significantly enhanced the TPN affinity for both calcium hydroxyapatite and porcine bone chip models, which led to enhancement in TP retention in the bones up to 8.1-fold versus free drug. Overall, TPN demonstrated good potential as a local-injectable, bone-targeted nanotherapy tailored for eradication of bone-only metastasis at reduced risk of systemic toxicity.


Whole-exome sequencing identifies a novel compound heterozygous mutation of ANKS6 gene in a Chinese nephronophthisis patient.
Nephronophthisis (NPHP) is an autosomal recessive cystic kidney disease that leads to renal failure in childhood or adolescence. NPHP and the related syndromes have been termed 'ciliopathies' because most NPHP gene products localize to the cilium or its associated structures. Here, we report a 2-year and 11-month-old Chinese girl with end-stage renal disease (ESRD), severe anemia, thrombocytopenia and myocardial hypertrophy. We performed trio-whole-exome sequencing to identify the causative variant of this patient. We identified an unreported compound heterozygous mutation (c.2420dupT, p.Thr808Aspfs*2 and c.1973-1G > A) in ANKS6 in the proband. The frameshift mutation c.2420dupT of ANKS6 was inherited from the proband's unaffected father and the splicing mutation c.1973-1G > A of ANKS6 was inherited from the proband's unaffected mother. Homozygous mutation in ANKS6 leads to NPHP16 (OMIM#615382) and this is the first case with a compound heterozygous mutation in the NPHP16 gene. We have identified a patient with ANKS6 variants in the East-Asian population for the first time. This case report expands the clinical and genetic spectra of NPHP and emphasizes the usefulness of whole-exome sequencing for genetic diagnosis of kidney disease.


L-DOPA treatment in MPTP-mouse model of Parkinson's disease potentiates homocysteine accumulation in substantia nigra.
One of the intermediates of methionine cycle, the homocysteine (Hcy), elevates in plasma of Parkinson's disease (PD) patients undergoing L-DOPA (3,4-dihydroxyphenylalanine) therapy and has been regarded as a risk factor of the disease. Several evidences pointed out that Hcy causes degeneration of dopaminergic neurons. In rodent, elevated level of Hcy in brain or infusion of the same directly into the substantia nigra (SN) potentiates dopaminergic neurodegeneration. However, the influence of L-DOPA therapy on the levels of Hcy in dopamine-rich regions of the brain (striatum and SN) of experimental models of PD is not known. The present study, for the first time, tested the hypothesis that L-DOPA treatment in experimental mouse model of PD potentiates Hcy accumulation in the dopamine-rich regions of the brain. We found a significant elevation of Hcy level in nigrostriatum in naïve as well as parkinsonian mice as a result of chronic L-DOPA treatment. Interestingly, L-DOPA treatment significantly elevates Hcy level in nigra but not in striatum of parkinsonian mice, when compared with L-DOPA naïve group. However, there is no significant decrease in the number of dopaminergic neurons in SN region in the parkinsonian mice given L-DOPA treatment. Thus, the present study demonstrates that L-DOPA treatment potentiates the level of Hcy in the SN without causing aggravated neurodegeneration in parkinsonian mice model.


Physicochemical properties of flavodoxin from Desulfovibrio vulgaris.
Reductive titration curves of flavodoxin from Desulfovibrio vulgaris displayed two one-electron steps. The redox potential E-2 for the couple oxidized flavodoxin/flavodoxin semiquinone was determined by direct titration with dithionite. E-2 was -149 plus or minus 3 mV (pH 7.78, 25 degrees C). The redox potential E-1 for the couple flavodoxin semiquinone/fully reduced flavodoxin was deduced from the equilibrium concentration of these species in the presence of hydrogenase and H-2. E-1 was -438 plus or minus 8 mV (pH 7.78, 25 degrees C). Light-absorption and fluorescence spectra of flavodoxin in its three redox states have been recorded. Both the rate and extent of reduction of flavodoxin semiguinone with dithionite were found to depend on pH. An equilibrium between the semiquinone and hydroquinone forms occurred at pH values close to the neutrality, even in the presence of a large excess of dithionite, suggesting an ionization in fully reduced flavodoxin with a pK-a = 6.6. The association constants K for the three FMN redox forms with the apoprotein were deduced from the value of K (K = 8 times 10-7 M-1) measured with oxidized EMN at pH 7.0. Oxidized flavodoxin was found to comproportionate with the fully reduced protein (k-comp = 4.3 times 10-3 M-1 times s-1, pH 9.0, 22 degrees C) and with reduced free FMN (K-comp = 44 M-1 times s-1, pH 8.1, 20 degrees C). Fast oxidation of reduced flavodoxin occurred in the presence of O-2. Slower oxidation of semiquinone was dependent on pH in a drastic way.


Catalytic asymmetric synthesis of pyrroloindolines via a rhodium(II)-catalyzed annulation of indoles.
Herein we report the synthesis of pyrroloindolines via a catalytic enantioselective formal [3+2] cycloaddition of C(3)-substituted indoles. This methodology utilizes 4-aryl-1-sulfonyl-1,2,3-triazoles as carbenoid precursors and the rhodium(II)-tetracarboxylate catalyst Rh2(S-PTAD)4. A variety of aryl-substituted pyrroloindolines were prepared in good yields and with high levels of enantioinduction.


Familial biochemical and clinical correlates of alcoholics with low platelet monoamine oxidase activity.
This study substantiates previous reports that low platelet MAO activity is associated with alcoholism. The results also indicate that low platelet MAO activity in alcoholic probands is associated with a higher prevalence of psychiatric hospitalization in first-degree family members as well as alcoholism and suicide attempts among alcoholics. Psychiatrically ill family members of the alcoholic probands with low platelet MAO activity also demonstrate low enzyme activity whereas the well family members have normal enzyme activity.


Mechanism of Capacitive Coupling of Proximal Electromagnetic Sources With Biological Bodies.
This study investigates the absorption of the induced E-field in homogeneous biological tissue exposed to highly localized field sources in proximity of the body, such as the charged tips of antennas, where E-field coupling dominates. These conditions are relevant for compliance testing of modern mobile phones where exposure is evaluated at small separation between radiators and the body. We derive an approximation that characterizes the decay of the induced E-field in the tissue as a function of distance. The absorption is quantified in terms of the local specific absorption rate (SAR) at the tissue surface as a function of the charge at the antenna tip. The approximation is based on the analytical evaluation of the E-fields of a charged disk under quasi-static conditions. We validate this approximation using full-wave simulations of dipoles. We demonstrate that the coupling mechanism of the E-field is dominated by the perpendicular field component and that wave propagation need not be considered for the characterization of the exposure. The surface SAR decreases approximately with the fourth power of the distance and with the square of the ratio of the permittivities of the tissue and free-space. The approximation predicts the induced maximum E-field with an accuracy of better than 1.5 dB. © 2022 Bioelectromagnetics Society.


Mapping of atmospheric heavy metal deposition in Guangzhou city, southern China using archived bryophytes.
Atmospheric heavy metal contamination is becoming a serious threat to environmental and human health in Chinese megacities. This study evaluated the concentrations of arsenic (As), cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn) and Pb isotopic compositions in herbarium and native bryophytes collected from Guangzhou from 1932 to 2018. Relatively low mean metal concentrations were measured for bryophytes collected in the 1930s. The highest mean concentrations of Cd (0.72 ± 0.32 mg/kg), Cu (28.1 ± 9.8 mg/kg), Pb (125.9 ± 62.4 mg/kg) and Zn (273 ± 130 mg/kg) were found in the bryophytes from 1979 to 2000, following the commencement of the Reform and Opening-Up Program in 1978. The mean Pb concentrations (74.7 ± 6.3 mg/kg) decreased sharply from 2001 onwards, following the cessation of leaded petrol across the Chinese mainland in 2000. However, these values are still higher than those in 1950-1978, corresponding to a significant increase in atmospheric Pb emissions from coal combustion, nonferrous metal smelting and motor vehicle petrol consumption in China in the 2000s. The lead isotopic ratios of bryophyte archives (206Pb/207Pb 1.141-1.229, 208Pb/207Pb 2.376-2.482) indicate that lithogenic input and anthropogenic input arising from leaded petrol and industrial emissions have been the main sources of atmospheric heavy metal deposition in the city of Guangzhou over the past 85 years. Herbarium bryophyte can be utilised to reconstruct temporal and spatial shifts in atmospheric heavy metal deposition to better understand and manage the current air quality in Chinese megacities.


Heme as a target for protection against doxorubicin-induced apoptosis in H9c2 cardiomyocytes.
Heme homeostasis is of vital importance to many biological processes associated with cell redox activity. However, the role of heme in the doxorubicin (DOX)-induced cardiotoxicity is still not clear. The aim of the present study was to test the hypothesis that heme is related to the DOX-induced oxidative stress and inhibition of heme expression may protect H9c2 cardiomyocytes against DOX-induced cardiotoxicity. For the evaluation of heme changing under doxorubicin treatment, H9c2 cells were treated with 0.5, 1, 2, and 4 mg/mL doxorubicin respectively. H9c2 cells were divided into 5 groups: Control group (cells were cultured without intervention), DOX group (cells were treated with 2 mg/mL doxorubicin for 6 h), Heme depletion+DOX group (cells were cultured with heme-depleted serum media, 0.5 mM succinylacetone and 2 mg/mL doxorubicin), Heme group (cells were treated with 30 μM heme), and Heme depletion+DOX+Heme group. Apoptotic cells were detected by flow cytometry with Annexin V-FITC/PI. The intracellular oxidant levels were measured by DCFH-DA fluorescence. The levels of heme were detected by ELISA. Doxorubicin significantly increased intracellular heme level from 5013 ± 187 ng/mL to the highest level of 11,720 ± 107 ng/mL, as well as the intracellular oxidants and cell apoptosis rate elevated by the increase of doxorubicin concentration. Heme depletion can significantly suppress the DOX-induced apoptosis from 39.8 ± 0.5% to 20.8 ± 0.5% (p < 0.001). Re-supplemented with exogenous heme partially but significantly restored the DOX-induced apoptosis. Heme plays an important role in doxorubicin toxicity-induced cardiomyocyte injury. By appropriate reduction in the accumulation of free heme in cardiomyocytes, doxorubicin-induced cardiotoxicity may be alleviated.


Temperature responses in rats after acute and chronic administrations of caffeine.
Acute administrations (IP) of caffeine produced dose-dependent changes in the body temperature of rats. Low doses (12.5 and 25 mg/kg) induced hyperthermia soon after drug administration, while high doses (50 and 100 mg/kg) produced maximal hypothermia approximately 2 hr later. The acute effects of caffeine were also dependent on ambient temperature. The hyperthermic and hypothermic responses were attenuated and blocked, respectively, in rats maintained at 32 degrees C. At 4 degrees C, the hypothermic response was exacerbated, and the hyperthermic response was absent. Tolerance rapidly developed to the hypothermic actions of caffeine when rats were administered the drug over 28 days. Following the emergence of tolerance, hyperthermia was observed in rats given 50 mg/kg of caffeine. The hyperthermic responses to the low doses of caffeine were not altered by the frequency of drug administration. Both the acute and chronic effects of caffeine on thermoregulation are not unlike those found for morphine. Possible mechanisms of action underlying the thermoregulatory effects of caffeine are discussed.


C1 binding by mouse IgM. The effect of abnormal glycosylation at position 402 resulting from a serine to asparagine exchange at residue 406 of the mu-chain.
We have previously shown that IgM-Asn406, a mutant IgM which has asparagine in place of the serine which is normally found at position 406, also has an abnormally glycosylated mu-chain and is defective in complement-dependent cytolysis. Here we show by analyzing cyanogen bromide fragments from normal and mutant mu-chains that the site of abnormal glycosylation is at the neighboring position, Asn402. The cytolytic defect was shown to be due to impaired C1 binding. At physiological ionic strength, the C1 binding defect was estimated to be 12-fold, which correlates well with the measured defect in cytolytic activity; also, the severity of the defect in C1 binding by the mutant protein decreases with decreasing ionic strength. Kinetic studies showed that the difference in affinities is due to a proportional difference in the association rate for C1q. By comparing IgM made in the presence and absence of deoxymannojirimycin, we show further that the defect in cytolytic activity derives mostly from the abnormal oligosaccharide.


Investigating drug safety in pregnancy based on the German Pharmacoepidemiological Research Database (GePaRD): A proof-of-concept analysis on the association between valproate and spina bifida.
Large health-care databases are increasingly used for research on drug utilization and safety in pregnancy. For the German Pharmacoepidemiological Research Database (GePaRD), covering ~20% of the German population, algorithms have been developed to identify pregnancies, to estimate their date of onset and to link mothers to their babies. Using this methodology, we aimed to conduct a proof-of-concept analysis on the known association between valproate (VPA) exposure in early pregnancy and spina bifida in the exposed child. We identified all pregnancies in GePaRD between 2006 and 2016 in women aged 12 to 50 years. To each VPA dispensation of these women, an exposure period was assigned, based on the dispensation date and the number of defined daily doses in the dispensed package. A pregnancy was classified as exposed to VPA in the critical time window if this exposure period overlapped with the first 55 days of pregnancy. Risk ratios were calculated for spina bifida in live births and induced abortions comparing VPA-exposed ones to all pregnancies. Overall, we identified 1 271 384 pregnancies fulfilling the inclusion criteria. Of these, 668 pregnancies (0.053%) were classified as exposed to VPA in the critical time window regarding spina bifida. An induced abortion accompanied by a diagnosis of spina bifida was observed in one of the VPA-exposed pregnancies (0.15%) and in 154 of all pregnancies (0.012%), yielding a risk ratio of 12.4 (95% confidence interval [CI]: 1.7-88.2). Out of 775 875 pregnancies ending in a live birth, 366 (0.047%) were classified as VPA exposed. A diagnosis of spina bifida was coded in 3 of 366 VPA-exposed live births (0.82%) and in 260 of all live births (0.03%), yielding a relative risk of 24.5 (95% CI: 7.9-76.0). Our proof-of-concept analysis based on GePaRD showed a strong association between intrauterine exposure to VPA and occurrence of spina bifida. The results are plausible and consistent with the literature, supporting the suitability of GePaRD and the developed algorithms to conduct studies on drug safety in pregnancy.