Reappraising Preclinical Models of Separation Anxiety Disorder, Panic Disorder, and CO2 Sensitivity: Implications for Methodology and Translation into New Treatments.
Separation anxiety applies to multiple forms of distress responses seen in mammals during postnatal development, including separation from a caregiver. Childhood separation anxiety disorder is an important risk factor for developing panic disorder in early adulthood, and both conditions display an increased sensitivity to elevated CO2 concentrations inhaled from the air. By interfacing epidemiological, genetic, and physiological knowledge with preclinical animal research models, it is possible to decipher the mechanisms that are central to separation anxiety and panic disorders while also suggesting possible therapies. Preclinical research models allow for environmentally controlled studies of early interferences with parental care. These models have shown that different forms of early maternal separation in mice and rats induce elevated CO2 respiratory sensitivity, an important biomarker of separation anxiety and panic disorders. In mice, this is likely due to gene-environment interactions that affect multiple behavioural and physical phenotypes after exposure to this early adversity. Although several questions regarding the causal mechanism of separation anxiety and panic disorder remain unanswered, the identification and improved understanding of biomarkers that link these mental health conditions under the guise of preclinical research models in conjunction with human longitudinal cohort studies can help resolve these issues.


Huge cystic lymphangioma mimicking ovarian malignancy: a case report.
Lymphangiomas are rare benign congenital malformations of the lymphatic system. Clinical presentation varies from asymptomatic masses to acute abdominal pain. A 25-year-old female who presented with acute onset of abdominal distension and severe pelvic pain is presented. As a palpable mass was found on the pelvic examination and ultrasonography demonstrated a hypoechoic cystic mass, an ovarian malignancy was suspected. Exploratory laparotomy was performed and revealed cystic lymphangioma of the sigmoid colon. The clinical, radiological and pathological findings of the patient are discussed with a brief review of the literature.


Experimental cancer chemotherapy using a liver metastatic model of human colon cancer transplanted into the spleen of severe combined immunodeficient mice.
We have developed a liver metastatic model of human colon cancer using severe combined immunodeficient (SCID) mice. Liver metastases were observed in all the SCID mice on day 28 after intrasplenic injection with 5 x 10(6) dissociated tumor cells of COL-2-JCK, a human colon cancer strain serially transplanted in nude mice. When this model was applied for chemotherapeutic experiments, 5-fluorouracil (5-FU) demonstrated significant antitumor effects in preventing liver metastases, whereas the efficacy of 5-FU was limited in the currently used sc-ip chemosensitivity assay in nude mice. When the human LDH-5 isozyme was evaluated in the homogenized metastatic liver tissue of SCID mice, a good correlation was obtained between the liver tumor weights and LDH-5 isozyme, suggesting that it could be a promising quantitative indicator for metastases. This model would be useful for further studies on the treatment of liver metastases of colon cancer.


Using the collaborative intervention planning framework to adapt a health-care manager intervention to a new population and provider group to improve the health of people with serious mental illness.
Health-care manager interventions improve the physical health of people with serious mental illness (SMI) and could be widely implemented in public mental health clinics. Local adaptations and customization may be needed to increase the reach of these interventions in the public mental health system and across different racial and ethnic communities. In this study, we describe how we used the collaborative intervention planning framework to customize an existing health-care manager intervention to a new patient population (Hispanics with SMI) and provider group (social workers) to increase its fit with our local community. The study was conducted in partnership with a public mental health clinic that serves predominantly Hispanic clients. A community advisory board (CAB) composed of researchers and potential implementers (e.g., social workers, primary care physicians) used the collaborative intervention planning framework, an approach that combines community-based participatory research principles and intervention mapping (IM) procedures, to inform intervention adaptations. The adaptation process included four steps: fostering collaborations between CAB members; understanding the needs of the local population through a mixed-methods needs assessment, literature reviews, and group discussions; reviewing intervention objectives to identify targets for adaptation; and developing the adapted intervention. The application of this approach enabled the CAB to identify a series of cultural and provider level-adaptations without compromising the core elements of the original health-care manager intervention. Reducing health disparities in people with SMI requires community engagement, particularly when preparing existing interventions to be used with new communities, provider groups, and practice settings. Our study illustrates one approach that can be used to involve community stakeholders in the intervention adaptation process from the very beginning to enhance the transportability of a health-care manager intervention in order to improve the health of people with SMI.


Increased response of adipose tissue of the ob/ob mouse to the action of adrenaline after treatment with thyroxin.
The activity of the lipolytic system of the obese hyperglycemic mouse was assessed after treatment with physiological doses of thyroxin (T4). The treatment significantly increased fatty acid mobilization in response to adrenaline over the levels observed in the control mice under all conditions studied. The activities of the high- and low-Km phosphodiesterases and of adenylate cyclase were also studied. Treatment of the ob/ob mice with T4 had little effect on the activities of the cyclic AMP phosphodiesterases (high and low Km) but it partially restored the activity of adenylate cyclase, which is deficient in these animals. A correlation was found in the T4-treated obese animals between the ability of the epididymal adipose tissue to mobilize fatty acids, its ability to increase the intracellular levels of cyclic AMP, and the activity of adenylate cyclase in response to adrenaline stimulation.


The role of sclerotherapy as an adjunct to Unna's boot for treating venous ulcers: a prospective study.
Ankle ulcers associated with venous disease have been traditionally treated by Unna's compressive bandages. However, successful healing of an ulcer with this therapeutic modality is slow and tedious. The purpose of this study is to investigate whether adjunctive sclerotherapy of large venous channels near the ulcers enhances healing. Twenty-eight patients with perimalleolar venous ulceration were entered into this study. Ulcer size ranged from 6.0 to 25 cm2. All were examined for the presence of large venous channels adjacent to the ulcer bed. A portable continuous-wave Doppler supplemented digital examination of the area. Twenty-five out of 28 patients had easily detectable venous channels, and these were randomized into two groups: group I, treated by weekly changes of Unna's compressive boots; group II, treated by weekly change of Unna's compressive boots and sclerotherapy with 3% sodium tetradecyl sulfate (Sotradecol). All patients had the size of the ulcer measured during their weekly visits. Patients in group II took a mean of 2.1 weeks to exhibit 50% healing and 4.3 weeks to fully heal. Conversely patients in group I took a mean of 3.2 weeks to achieve 50% healing and 6.1 weeks to fully heal. Thus, healing was significantly faster (p less than 0.05) in group II. In conclusion, the therapeutic efficiency of Unna's compressive bandaging is significantly enhanced by the adjunctive sclerotherapy of adjacent venous channels.


Identification by nuclear magnetic resonance and mass spectrometry of a glucuronic acid conjugate of o-hydroxybenzoic acid in normal urine and uremic plasma.
An endogenous compound (included in the fraction of uremic toxins often called the "uremic middle molecules") was separated from plasma of uremic patients and urine from normal persons. As elucidated by mass spectrometry, enzymatic hydrolysis, and 1H, 13C nuclear magnetic resonance, it appears to be a conjugate of o-hydroxybenzoic acid with glucuronic acid. Its presence in urine of healthy subjects indicates its physiological character.


Neuroarchitecture of the central complex in the brain of the locust Schistocerca gregaria and S. americana as revealed by serotonin immunocytochemistry.
The central complex is a prominent structure in the insect brain, yet its anatomical organization and functional role is still poorly understood. To facilitate investigations on the physiology of the central complex, this study describes its anatomical organization in the brain of locusts (Schistocerca gregaria and Schistocerca americana) based on an investigation of serotonin immunocytochemistry. Most subdivisions of the central complex including the protocerebral bridge, several layers in the upper division of the central body, and the noduli of the central body are innervated by serotonin-immunoreactive neurons, while the lower division of the central body does not exhibit serotonin-like immunoreactivity. Several types of serotonin-immunoreactive neurons can be distinguished. A system of about 60 columnar neurons innervates the protocerebral bridge, layer III of the upper division of the central body, and the noduli. A group of 15-20 bilateral pairs of serotonin-immunoreactive neurons connects the posterior optic tubercles with the protocerebral bridge. About ten pairs of neurons with somata in the inferior protocerebrum innervate layer Ia of the upper division of the central body. In addition, large-field neurons arborize in layers Ia and Ib of the upper division of the central body and in the lateral accessory lobes. The detailed mapping of serotonin immunoreactivity provides further insight into the anatomical organization of the central complex and suggests that serotonin is a major neuroactive substance within this brain structure.


Long-term outcomes of endoscopic neck dissection in the treatment of early-stage oral cancer: a pilot study.
This study aimed to investigate the feasibility and safety of endoscopic neck dissection in the treatment of early-stage oral cancer and to evaluate the long-term clinical outcomes. Six patients with early-stage oral cancer were enrolled in this pilot study from December 2006 to May 2007. All the patients underwent endoscopic selective neck dissection (levels I-IV) of the ipsilateral neck and partial glossectomy or hemiglossectomy as the primary treatment. All endoscopic procedures were successfully performed, with important neck structures identified and preserved. All the patients survived with no persistent or recurrent disease during the 76- to 83-month follow-up. Our preliminary results indicated that endoscopic neck dissection is a technically feasible and safe technique for treating early-stage oral cancer. The oncologic indications and validation should be further confirmed in patients with clinically positive neck lymph nodes in a future study.


Midbrain activity shapes high-level visual properties in the primate temporal cortex.
Recent fMRI experiments identified an attention-related region in the macaque temporal cortex, here called the floor of the superior temporal sulcus (fSTS), as the primary cortical target of superior colliculus (SC) activity. However, it remains unclear which aspects of attention are processed by fSTS neurons and how or why these might depend on SC activity. Here, we show that SC inactivation decreases attentional modulations in fSTS neurons by increasing their activity for ignored stimuli in addition to decreasing their activity for attended stimuli. Neurons in the fSTS also exhibit event-related activity during attention tasks linked to detection performance, and this link is eliminated during SC inactivation. Finally, fSTS neurons respond selectively to particular visual objects, and this selectivity is reduced markedly during SC inactivation. These diverse, high-level properties of fSTS neurons all involve visual signals that carry behavioral relevance. Their dependence on SC activity could reflect a circuit that prioritizes cortical processing of events detected subcortically.


'Twisted plywood' structure and mineralization in the scales of a primitive living fish Amia calva.
The basal plate of the scales of Amia calva is composed of regular double twisted plywood, as in Latimeria and Dipnoan scales. However, the progressive rotation of the fibrils direction is left-handed in Amia and right-handed in the 'Sarcopterygians'. So, the similarity between these peculiar plywoods is probably the result of convergence. The basal plate of Amia scales is incompletely mineralized. There are numerous calcified ovoid corpuscles which look very like the Mandl's corpuscles of Teleost scales. The mineralization probably progresses essentially by the fusion of these corpuscles, as in Teleost scales, and would be inotropic rather than spheritic.


Thromboembolic Risk in Nonanticoagulated Patients With Atrial Fibrillation and Valvular Heart Disease.
This study sought to describe the risk of thromboembolism in nonanticoagulated atrial fibrillation patients with Evaluated Heartvalves, Rheumatic or Artificial (EHRA) Type 2 valvular heart disease (VHD) <65 or 65 to 74 years of age and with 0 or 1 non-sex comorbidity of the CHA2DS2-VASc score. A minor, but important, proportion of patients with atrial fibrillation and VHD beyond moderate-to-severe mitral stenosis and/or a mechanical prosthetic valve, so-called EHRA Type 2 VHD, have 0 or 1 coexisting non-sex comorbidities of the CHA2DS2-VASc score, and are therefore not strongly recommended oral anticoagulant therapy according to guidelines. Whether these patients are truly low risk of thromboembolism has not been investigated. This was a cohort study of 55,613 patients identified in nationwide Danish registries from 2000 to 2018, of which 1,907 patients had EHRA Type 2 VHD. Risk of thromboembolism after 1 and 5 years of follow-up were calculated. At 1 year after atrial fibrillation diagnosis, patients with EHRA Type 2 VHD had a risk of thromboembolism between 1.2% and 1.5%, according to age group (<65 or 65 to 74 years of age), and number of non-sex comorbidities of the CHA2DS2-VASc score (0 or 1). Interestingly, in patients with EHRA Type 2 VHD <65 years of age with 0 or 1 comorbidity, the risk was 1.5% (95% confidence interval: 0.7% to 2.8%) and 1.5% (95% confidence interval: 0.6% to 3.4%) at 1 year after the atrial fibrillation diagnosis. These observations suggest that in atrial fibrillation patients with EHRA Type 2 VHD, who are not currently recommended oral anticoagulant therapy according to guidelines, the risk of thromboembolism may exceed the level above which oral anticoagulation is considered beneficial.


The chemokine system and CCR5 antagonists: potential in HIV treatment and other novel therapies.
Since the recognition of human acquired immune deficiency syndrome, numerous classes of pharmacologic therapeutics have been developed to manage the disease. Current therapy includes co-administration of combinations of drugs classified by their mechanism of action as 'transcriptase inhibitors', 'protease inhibitors', 'integrase inhibitors' and the more recent 'fusion inhibitors'. This review focuses on the chemokine system and the recognition of chemokine receptors as targets for anti-human immunodeficiency virus (HIV) therapy. The FDA-approved chemokine (C-C motif) receptor 5 (CCR5) antagonist maraviroc (Selzentry) is discussed in detail, along with another compound vicriviroc, currently in clinical trials. The mechanism of action, pharmacokinetics, toxicity and current status of research on CCR5 antagonists is described. Further, potential therapeutic uses of these agents other than anti-HIV therapy are discussed.


Clinical, Epidemiologic, and Laboratory Aspects of Methicillin-Resistant Staphylococcus aureus Infections.
Oxacillin-resistant Staphylococcus aureus (abbreviated MRSA for historical reasons) is a major pathogen responsible for both hospital- and community-onset disease. Resistance to oxacillin in most clinical isolates of S. aureus is mediated by PBP2a, a penicillin-binding protein with low affinity to beta-lactams, encoded primarily by the mecA gene. Rapid and accurate methods of susceptibility testing of S. aureus isolates to identify MRSA infections are important tools to limit the spread of this organism. This review focuses on the clinical significance of MRSA infections and new approaches for the laboratory diagnosis and epidemiologic typing of MRSA strains.


Implementation of virtual microscope slides in the annual pathobiology of cancer workshop laboratory.
Virtual slides are digital facsimiles of glass microscope slides that, when viewed with a pan and zoom viewer, can emulate viewing a glass slide with a traditional microscope. Based on successful implementation of virtual slides in medical student histology and pathology courses at the University of Iowa, we developed a plan to evaluate the use of virtual slides in the American Association for Cancer Research's annual Pathobiology of Cancer Workshop. In this Workshop, nonphysician predoctoral students and postdoctoral fellows working in cancer research explore the morphological, clinical, and molecular aspects of human cancer. Over the course of a week, students examine approximately 100 glass slides in microscope laboratories, facilitated by senior cancer investigators. The goal of the present study was to evaluate virtual slides as a teaching modality in these laboratories, not as a replacement for traditional microscopy, but rather in terms of their utility in facilitating student learning as they examine glass slides with a traditional microscope. Evaluation by questionnaire indicated that virtual slides enhanced students' ability to grasp morphological features better than the traditional photomicrographs. The results of this implementation suggest that virtual slide technology may be successfully extended to other educational venues where traditional microscopy and photomicrographs are currently used.


Pressor, norepinephrine, and pituitary responses to two TRH doses in Alzheimer's disease and normal older men.
Changes in blood pressure (BP), plasma norepinephrine (NE), serum prolactin (PRL), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) associated with infusions of two thyrotropin-releasing hormone (TRH) doses (0.1 mg, 0.5 mg) were examined in 10 men with early-onset Alzheimer's disease (AD) and nine normal matched controls. During the first 5 min after TRH infusion, significant increases from baseline in systolic BP (p less than 0.001), diastolic BP (p less than 0.001), and plasma NE (p less than 0.006) occurred in the study subjects. The magnitude of the BP and NE responses did not differ significantly as a function of TRH dose (p greater than 0.3). Diastolic pressor responses to TRH were substantially blunted in AD subjects relative to controls, after both the 0.1-mg (p less than 0.003) and 0.5-mg (p less than 0.02) doses. There were trends toward attenuated responses in the AD subjects for systolic BP (p less than 0.09) and plasma NE (p less than 0.07). Significant increments in serum PRL, LH, and FSH (all p less than 0.001) also occurred after TRH, but the magnitude of the hormone responses did not differ significantly between the AD and the normal subjects (p greater than 0.18). These results suggest the possibility that TRH-evoked activation of the sympathetic nervous system (SNS), as reflected by pressor and plasma NE responses, may be attenuated in men with early-onset AD.


In vivo evidence of atherosclerotic plaque erosion and healing in patients with acute coronary syndrome using serial optical coherence tomography imaging.
The EROSION study (Effective Anti-Thrombotic Therapy Without Stenting: Intravascular Optical Coherence Tomography-Based Management in Plaque Erosion) allowed us to observe the healing process of coronary plaque erosion in vivo. The present study aimed to investigate the incidence of newly formed healed plaque and different baseline characteristics of acute coronary syndrome (ACS) patients caused by plaque erosion with or without newly formed healed plaque using optical coherence tomography (OCT). A total of 137 ACS patients with culprit plaque erosion who underwent pre-intervention OCT imaging and received no stent implantation were enrolled. Patients were stratified according to the presence or absence of newly formed healed phenotype at 1-month (137 patients) or 1-year OCT follow-up (52 patients). Patient's baseline clinical, angiographic, OCT characteristics and outcomes were compared. There were 55.5% (76/137) of patients developed healed plaque at 1 month, and 69.2% (36/52) of patients developed healed plaque at 1 year. Patients with newly formed healed plaque had larger thrombus burden, and lower degree of area stenosis (AS%) at baseline than those without, and thrombus burden and AS% were predictors of plaque healing. The healing process was accompanied by the significant increase of AS% and incidence of microchannels, and greater inflammatory response. The outcomes appeared to be similar between the two groups. Newly formed healed plaque was found in more than half of ACS patients with plaque erosion without stenting. Patients with newly formed healed plaque had lower luminal stenosis and larger thrombus burden. During healing process, luminal stenosis increased gradually.


Characterization of preclinical models of prostate cancer using PET-based molecular imaging.
Transgenic adenocarcinoma of the mouse prostate (TRAMP) mice spontaneously develop hormone-dependent and hormone-independent prostate cancer (PC) that potentially resembles the human pathological condition. The aim of the study was to validate PET imaging as a reliable tool for in vivo assessment of disease biology and progression in TRAMP mice using radioligands routinely applied in clinical practice: [(18)F]FDG and [(11)C]choline. Six TRAMP mice were longitudinally evaluated starting at week 11 of age to visualize PC development and progression. The time frame and imaging pattern of PC lesions were subsequently confirmed on an additional group of five mice. PET and [(18)F]FDG allowed detection of PC lesions starting from 23 weeks of age. [(11)C]Choline was clearly taken up only by TRAMP mice carrying neuroendocrine lesions, as revealed by post-mortem histological evaluation. PET-based molecular imaging represents a state-of-the-art tool for the in vivo monitoring and metabolic characterization of PC development, progression and differentiation in the TRAMP model.


Prenatal Maternal Factors, Intergenerational Transmission of Disease, and Child Oral Health Outcomes.
This article reviews maternal prenatal risk factors for caries in children and intergenerational transmission of caries, emphasizing early interventions for pregnant women and mother-infant pairs. A growing body of evidence focuses on maternal interventions. Studies suggest that early prenatal clinical and educational interventions are effective at reducing mother-child mutans streptococci (MS) transmission and delaying colonization and caries in young children. Dental screenings and anticipatory guidance about maternal and infant oral health should be included in prenatal care and pediatric well visits. Dental care during pregnancy is safe and recommended and can reduce maternal MS levels. Infants should visit a dentist by age 1.


Type 1 diabetic manifestations in a young man triggered by COVID-19.
COVID-19 could potentially lead to the progression of the course of undiagnosed type 1 diabetes in an infected individuals.


Manipulating Ferromagnetism in Few-Layered Cr2 Ge2 Te6.
The discovery of magnetism in 2D materials offers new opportunities for exploring novel quantum states and developing spintronic devices. In this work, using field-effect transistors with solid ion conductors as the gate dielectric (SIC-FETs), we have observed a significant enhancement of ferromagnetism associated with magnetic easy-axis switching in few-layered Cr2 Ge2 Te6 . The easy axis of the magnetization, inferred from the anisotropic magnetoresistance, can be uniformly tuned from the out-of-plane direction to an in-plane direction by electric field in the few-layered Cr2 Ge2 Te6 . Additionally, the Curie temperature, obtained from both the Hall resistance and magnetoresistance measurements, increases from 65 to 180 K in the few-layered sample by electric gating. Moreover, the surface of the sample is fully exposed in the SIC-FET device configuration, making further heterostructure-engineering possible. This work offers an excellent platform for realizing electrically controlled quantum phenomena in a single device.


Electrospun aggregation-induced emission active POSS-based porous copolymer films for detection of explosives.
Electrospun aggregation-induced emission (AIE)-active polyhedral oligomeric silsesquioxane (POSS)-based copolymer films exhibit an approximately 9-fold increase in response to explosive vapors compared to dense films although porous copolymer films have a thickness as high as 560 ± 60 nm.


Dose-related hepatotoxicity of 1,1,2-trichloro-1,2,2-trifluoroethane in short-term intermittent inhalation exposure in rats.
Male Wistar rats were exposed to 200, 1000 or 2000 ppm of 1,1,2-trichloro-1,2,2-trifluoroethane vapor 5 days a week 6 h daily for 1 or 2 weeks. Proliferation and vacuolisation of the smooth endoplasmic reticulum (SER) of the liver was seen electron microscopically after 1 and 2 weeks in the rats exposed to 1000 and 2000 ppm. Among the hepatic drug metabolizing enzymes, NADPH cytochrome c reductase activity showed a dose-related decrease whereas the tightly membrane-bound UDPglucuronosyltransferase exhibited a dose-dependent enhancement in its measurable activity. The overall drug oxidation reaction, 7-ethoxycoumarin O-deethylase was not affected by the 1,1,2-trichloro-1,2,2,-trifluoroethane inhalation at all, either in the liver or in the kidneys. 1,1,2-Trichloro-1,2,2-trifluoroethane binds to cytochrome P-450 with the production of a type I difference spectrum, suggesting that it may act as a substrate for this enzyme. The binding affinity is increased by phenobarbital-treatment of the rats.


NMMHC IIA triggered lipid metabolize reprogramming resulting in vascular endothelial cellular tight junction injury.
Nonmuscle myosin heavy chain IIA, played an essential role in the promotion of tight junction injury in vascular endothelial cells under oxygen glucose deprivation condition. Rat microvascular endothelial cells had been confirmed to have the susceptibility to ox-LDL stimulation under OGD condition. We proposed the hypothesis that lipid metabolic reprogramming might be the root cause for damage to RBMCs tight junction. Untargeted shotgun and targeted lipid metabolomics mass spectrometry approaches combined with principal component analysis was applied to better define the lipids contributing to the variance observed between control and different OGD time. The protein expression of tight junction of RBMCs: occludin, claudin-5, and ZO-1 were detected with immunofluorescence staining and western blot. The proof of the interaction between NMMHC IIA and SREBP1 was investigated via GST-pull down, while their specific binding fragments were also confirmed. The regulation mechanism of NMMHC IIA on SREBP1 was investigated to explore downstream regulatory signaling pathways. Untargeted and targeted shotgun lipidomics data revealed that OGD might be the conditional factor in reshaping lipid components. Mechanistic studies showed that with the increase of OGD time, PCA analysis of lipidomics obtained from RBMCs indicated their specificity in reshaping lipid components, while ≥80% major lipid components phospholipids and sphingolipids transferred from phospholipids, sphingolipids, and neutral lipids, of which neutral lipids taken the largest proportion with OGD time course. Perturbing reprogramming of lipid composition was less susceptible to OGD condition via knockdown of NMMHC IIA of vascular endothelial cells. Knockdown of NMMHC IIA could promote tight junction defense to OGD condition. NMMHC IIA could directly bind with SREBP1, then could affect sterol regulatory element binding protein-1 to adjust lipid metabolize reprogramming of RBMCs. Mechanistic studies showed that perturbing reprogramming of lipid composition could enhance tight junction damage, which was mediated by the opposing effects of NMMHC IIA.


Characterization of allergenic components from house dust mite Dermatophagoides siboney. Purification of Der s 1 and Der s 2 allergens.
Allergic reactions to house dust mites of the genus Dermatophagoides play an important role in the pathogenesis of asthma and other atopic diseases. Dermatophagoides siboney has been described as a species from Cuba. Together with D. pteronyssinus and Blomia tropicalis, it is frequently found in house dust from homes of asthmatics. The aim of this study was to investigate the allergenic composition from the house dust mite D. siboney. The characterization of D. siboney extract was performed by SDS-gPAGE and immunoblotting. Purification of individual components was performed by affinity chromatography. At least 16 components between 13 and 98 kDa stained by Coomassie Blue were found. Using a panel of 35 sera from atopic mite sensitive patients 13 components reacted to different extent with patient IgE. Two components, 25 and 14 kDa, bound to specific IgE strongly and frequently, i.e. 80 and 91% of the patients, respectively. Affinity chromatography using crossreacting monoclonal antibodies to group 1 and 2 allergens resulted in purified preparations of 25 and 14 kDa proteins, which showed IgE-binding with the majority of the human sera when tested by immuno-dot. Based on the IgE binding profile of D. siboney and on the capacity to react with crossreacting monoclonal antibodies for groups 1 and 2, it is proposed to name these two allergens, 25 and 14 kDa, Der s 1 and Der s 2, respectively.


A comparison of adenocarcinoma and squamous cell carcinoma of the cervix.
A comparison and survival analysis of 203 patients (21%) with adenocarcinoma and 756 (79%) with squamous cell cancer of the cervix for the time period 1970-1985 is reported. The mean number of new cases of adenocarcinoma (N = 12) remained the same, while squamous cell cases decreased from 57 to 32. The clinical features were compared; oral contraceptive usage, node status, diabetes, and symptoms of bleeding were not related to cell type. Nulliparity was more frequent in patients with adenocarcinoma, whereas obesity and smoking were more frequent in patients with squamous cell cancer. Survival in stage I was significantly influenced by the cell type. Patients with stage I squamous cell disease had a 90% 5-year survival, compared with 60% for adenocarcinoma (P less than .0001). Other features that influenced survival included node status (P = .001), poor differentiation of tumor histology (P = .001), diabetes (P = .001), and Papanicolaou smear interval (P = .001). Patients undergoing radical hysterectomy were analyzed separately, and adenocarcinoma cell type significantly influenced survival (P = .0008). Patients with stage II squamous cell disease had a 62% survival, compared with 47% for adenocarcinoma (P = .01); patients with stage III squamous cell disease had a 36% survival, compared with 8% for adenocarcinoma (P = .002). The percentage of adenocarcinoma has increased because of the decreased number of patients with squamous cell disease. Stage-for-stage survival is significantly decreased for patients with adenocarcinoma as opposed to squamous cell disease.


Heterodimer of A2A and Oxytocin Receptors Regulating Glutamate Release in Adult Striatal Astrocytes.
Roles of astrocytes in the modulatory effects of oxytocin (OT) in central nervous system are increasingly considered. Nevertheless, OT effects on gliotransmitter release have been neglected. In purified astrocyte processes from adult rat striatum, we assessed OT receptor (OTR) and adenosine A2A receptor expression by confocal analysis. The effects of receptors activation on glutamate release from the processes were evaluated; A2A-OTR heteromerization was assessed by co-immunoprecipitation and PLA. Structure of the possible heterodimer of A2A and OT receptors was estimated by a bioinformatic approach. Both A2A and OT receptors were expressed on the same astrocyte processes. Evidence for A2A-OTR receptor-receptor interaction was obtained by measuring the release of glutamate: OT inhibited the evoked glutamate release, while activation of A2A receptors, per se ineffective, abolished the OT effect. Biochemical and biophysical evidence for A2A-OTR heterodimers on striatal astrocytes was also obtained. The residues in the transmembrane domains 4 and 5 of both receptors are predicted to be mainly involved in the heteromerization. When considering effects of OT in striatum, modulation of glutamate release from the astrocyte processes and of glutamatergic synapse functioning, and the interaction with A2A receptors on the astrocyte processes should be taken into consideration.


Outcome of meningitis among children less than 2-y-old in Haryana.
Incidence of neurological complications and disability following meningitis among children less than 2 y in a community setting in Haryana was assessed. Cases were enrolled from hospitals of in Yamunanagar district. They were first assessed for disability or neurological complications at their home by health workers 1 y after hospital discharge using standard WHO ten questionnaire screening tool, and then by medical officers; finally they were examined by a pediatrician. Eighty one children could be contacted from a total of 91 meningitis cases. Among these 16 children had died (case fatality ratio 19.7 %). Among 65 survivors, disability was observed in 11 (17 %), and neurological complications were found in 24 (37 %) children. Microcephaly (8.6 %), seizures (6.7 %), cerebral palsy, hemiparesis and development deficit (some with hearing impairment, and language delay) were the major disabilities/neurological complications.


Plant development: timing when to flower.
The time to flowering in Arabidopsis is controlled by many genes; recently, progress has been made in the cloning of a number of such genes. One of them, the FCA gene, turns out to encode an RNA binding protein.


Application of aortic allograft in trachea transplantation.
The use of tracheal implants for tracheal reconstruction remains a challenge in thoracic medicine due to the complex structure of the trachea in mammalian organisms, including smooth muscles, cartilage, mucosa, blood vessels, cilia, and other tissues, and the difficulty in achieving tracheal regeneration using implants from either allografts or synthetic biomaterials. This project used the Lee-Sung strain pig, a swine breed local to Taiwan, as the experimental subject. The aorta of the pig was harvested, decellularized to form the scaffold, and transplanted into the trachea of allogeneic pigs together with growth factors. Postoperative physiological function and tissue changes were observed. The postoperative physiological parameters of the LSP were monitored, and they were sacrificed after a certain period to observe the pathological changes in the tracheal epithelial cells and cartilages. Overall, six LSP tracheal transplantations were performed between March 4, 2020, and March 10, 2021. These included aortic patch anastomosis for pig 1 and aortic segmental anastomosis for pigs 2-6. The shortest and longest survival periods were 1 day and 147 days, respectively. Excluding the pig that survived for only 1 day due to a ruptured graft anastomosis, all other subjects survived for over 1 month on average. In this study, we grafted a decellularized porcine aorta into a recipient pig with a tracheal defect. We found cryopreservation of the allogeneic aorta transplantation was a feasible and safe method for the management of airway disease, and immunosuppressants were unnecessary during the treatment course.


Morbidity in Pregnant Women with a Prosthetic Heart Valve.
Women with a prosthetic heart valve are perceived to be at higher risk for adverse outcomes, but their absolute and relative risk of experiencing maternal morbidity and cardiac complications is largely unknown. The objective of the study was to determine the risk of maternal morbidity and cardiac complications in women with a prior heart valve replacement, compared with matched counterparts without known cardiac disease. A retrospective population-based matched cohort study was completed in the province of Ontario, Canada, where there is universal health care. Included were all women of child-bearing age who had bioprosthetic or mechanical replacement of the mitral or aortic valve, April 1994 to March 2016 (valve replacement group). Those in the valve replacement group, and who had at least 1 birth, were 1:4 matched to a community comparison group without heart disease and who also had at least 1 birth. Matching was by maternal age at cohort entry, year of cohort entry, geographic area, income level, and age at first birth. Maternal outcomes included severe maternal morbidity, all-cause mortality, and cardiac morbidity as well as a prolonged hospital length of stay >7 days. Relative risks and 95% confidence intervals were further adjusted for age at birth and immigration status. There were 90 live births among the 64 women in the valve replacement group and 404 live births among the 253 women in the matched community comparison group. There were no stillbirths. Severe maternal morbidity occurred in 13 pregnancies (14.4%) in the valve replacement group and 6 (1.5%) in the community comparison group (adjusted relative risk, 9.73, 95% confidence interval, 3.70-25.59); there were no maternal deaths. The corresponding rates of prolonged hospital length of stay were 37.8% and 18.8% (adjusted relative risk, 2.33, 95% confidence interval, 1.48-3.67). Pregnant women who had aortic or mitral valve replacement were more likely to experience severe maternal morbidity, as well as prolonged hospital length of stay, than matched counterparts without heart disease. This information can enhance shared decision making about the timing of valve replacement and pregnancy planning in young and middle-aged women. To determine the absolute and relative risk of maternal morbidity and cardiac complications in women with prior heart valve replacement, a retrospective population-based matched cohort study was completed in the province of Ontario, Canada, where there is universal health care. Included were all women of child-bearing age who had bioprosthetic or mechanical replacement of the mitral or aortic valve, April 1994 to March 2016 (valve replacement group). Those in the valve replacement group, and who had at least one birth, were 1:4 matched to a community comparison group without heart disease and who also had at least 1 birth. There were 90 live births among the 64 women in the valve replacement group and 404 live births among the 253 women in the matched community comparison group. Severe maternal morbidity occurred in 13 pregnancies (14.4%) in the valve replacement group and 6 (1.5%) in the community comparison group (adjusted relative risk, 9.73); there were no maternal deaths. The corresponding rates of prolonged hospital length of stay were 37.8% and 18.8% (adjusted relative risk, 2.33). In summary, pregnant women who had an aortic or mitral valve replacement were more likely to experience severe maternal morbidity, as well as prolonged hospital length of stay, than matched counterparts without heart disease.


Coming Out to Parents in Lesbian and Bisexual Women: The Role of Internalized Sexual Stigma and Positive LB Identity.
The experience of "coming out" (CO) to parents is often a crucial event in the lives of lesbian and bisexual (LB) women, associated with lower internalized sexual stigma (ISS) and higher positive LB identity. Few studies have compared the experiences of LB women in the CO process. Rather, most prior research has either: (1) not addressed bisexuality or eliminated bisexual individuals from the analysis; (2) combined bisexual women and bisexual men in the same sexual orientation group; or (3) examined bisexual participants alongside lesbian women and gay men, using a single monolithic measure. Thus, the present research aimed at investigating the role of ISS and positive LB identity in inhibiting or encouraging CO to parents in a sample of 241 lesbian women (M age = 27.61, SD = 7.19) and 186 bisexual women (M age = 25.23, SD = 5.81), aged 18-40 years. Most participants reported that they had already revealed their sexual orientation to their mother (69%) and their father (52%). More lesbian women had CO to both their mother and their father than had bisexual women. These lesbian women reported lower levels of ISS and higher levels of LB positive identity relative to bisexual women. On average, CO to mothers occurred at age 20 (SD = 5.54), while CO to fathers occurred at age 22 (SD = 5.63). LB women did not differ in the average age of CO to mothers or fathers, or in parental reactions to CO. Finally, ISS was found to affect the process of CO to both parents via positive identity (bootstrapping estimate = -0.26, SE = 0.08, 95% CI = -0.43, 0.11), whereas sexual orientation was not found to moderate the path from ISS to CO to both parents. The present study contributes to our understanding of the differences between LB women when developing their sexual orientation, highlighting the relevance of a positive LB identity for CO to parents. Research and clinical implications and directions for future research are discussed.


Thermal enhancement of gangue-cemented paste backfill with graphite and silica sand: an experimental investigation.
Thermal conductivity of mine backfill is vital for understanding the geothermal energy extraction potential in deep underground mines. Geothermal energy extraction efficiency could be improved with higher thermal conductivity of backfill. Thermal conductivities of gangue-cemented paste backfill enhanced with graphite and silica sand were investigated through laboratory tests in this article. The thermal conductivity measurement was conducted using the Hot Disk Thermal Constants Analyzer. Effects of graphite and silica sand on the thermal conductivity, slump, and compressive strength of gangue-cemented paste backfill were analyzed. The results highlighted the favorable influence of graphite on the thermal performance of backfilling materials, with adverse effect on the mechanical behavior and negative impact on the flowability. Thermal conductivity of gangue-cemented paste backfill markedly increased with graphite ratio, while more water is consumed for the workable slurry with the increased graphite ratio, which also leads to the degradation of compressive strength. The reasonable graphite content should be determined with the expected thermal enhancement and acceptable compressive strength. The addition of silica sand can effectively enhance the thermal conductivity of GCPB without degrading the compressive performance and flow performance. This study would provide a new insight into the design of backfill considering the thermal conductivity and lay a foundation for the efficient geothermal energy in deep mines.


Direct evidence of microbiological water quality changes on bacterial quantity and community caused by plumbing system.
Drinking water quality deteriorates from treatment plant to customer taps, especially in the plumbing system. There is no direct evidence about what the differences are contributed by plumbing system. This study compared the water quality in the water main and at customer tap by preparing a sampling tap on the water main. The biomass was quantified by adenosine triphosphate (ATP) and the microbial community was profiled by 454 pyrosequencing. The results showed that in distribution pipes, biofilm contributed >94% of the total biomass, while loose deposits showed little contribution (< 2%) because of the low amount of loose deposits. The distribution of biological stable water had minor effects on the microbiocidal water quality regarding both quantity (ATP 1 ng/L vs. 1.7 ng/L) and community of the bacteria. Whereas the plumbing system has significant contribution to the increase of active biomass (1.7 ng/L vs. 2.9 ng/L) and the changes of bacterial community. The relative abundance of Sphingomonas spp. at tap (22%) was higher than that at water main (2%), while the relative abundance of Pseudomonas spp. in tap water (15%) was lower than that in the water from street water main (29%). Though only one location was prepared and studied, the present study showed that the protocol of making sampling tap on water main offered directly evidences about the impacts of plumbing system on tap water quality, which makes it possible to distinguish and study the processes in distribution system and plumbing system separately.


Association analysis between FASN genotype and milk traits in Mediterranean buffalo and its expression among different buffalo tissues.
Fatty acid synthase (FASN) is a multifunctional protein that catalyzes the synthesis of long-chain saturated fatty acid. In this study, we identified the single nucleotide polymorphisms (SNPs), and their association with milk traits in Mediterranean buffalo, and the expression of FASN gene in different tissues was measured. Nine SNPs (g.-1640G > A, g.-1099C > T, g.1095C > A, g.3221G > A, g.4762G > A, g.5299G > A, g.7164G > A, g.7272 T > C, and g.8927 T > C) were identified by DNA pooled sequencing and then genotyped. Seven identified SNPs except g.3221G > A and g.8927 T > C were found significantly associated with both fat and protein percentage, and also the g.7164G > A and g.8927 T > C had significant association with peak milk yield and protein percentage, respectively. One haplotype block was successfully constructed by linkage disequilibrium (LD) analysis and it showed a significant association with both fat percentage and protein percentage. Expression of FASN gene was found in almost all the buffalo tissues including mammary gland, heart, liver, spleen, lung, kidney, uterus, and ovary, and to be highest in lung and mammary gland. Our findings suggest that polymorphisms in the buffalo FASN gene are associated with milk production traits and can be used as a candidate gene for milk traits and marker-assisted selection in buffalo breeding program.


The Association of the Oxford Classification Score with Longitudinal Estimated Glomerular Filtration Rate Decline in Patients with Immunoglobulin A Nephropathy: A Mixed-Method Study.
The Oxford Classification score, which predicts renal outcomes for immunoglobulin A nephropathy (IgAN), is widely used in clinical practice. Nevertheless, the relationship between these markers and longitudinal changes in renal function are poorly understood. This was a population-based retrospective cohort study of 280 adults with biopsy-proven primary IgAN from 2011 to 2018. We used generalized additive mixed models to control for traditional kidney disease risk factors to analyze the associations between Oxford Classification MEST-C scores (mesangial hypercellularity, M; endocapillary hypercellularity, E; segmental glomerulosclerosis, S; tubular atrophy/interstitial fibrosis, T; crescents, C) and longitudinal changes in the estimated glomerular filtration rate (eGFR) after renal biopsy. The median eGFR was 78.2 mL/min/1.73 m2 at baseline, and then it decreased on average by 1.3 mL/min/1.73 m2 per year in the entire cohort. In adjusted models, compared with patients without relative lesions, the presence of T > 50% (T2) (-5.7; 95% confidence interval [CI], -9.5 to -2.0 mL/min/1.73m2 per year) was associated with the fastest eGFR decline. S present (S1) (-2.9; 95% CI, -4.6 to -1.1 mL/min/1.73m2 per year) and C > 25% glomeruli (C2) (-3.4; 95% CI, -6.4 to -0.5 mL/min/1.73m2 per year) also demonstrated steeper eGFR declines. However, we found no association between M > 0.5 (M1), E present (E1), T 26%-50% (T1), and C present ≥ 1 glomerulus (C1), and progressive eGFR decline (p > 0.05). The Oxford Classification scores, S1, T2, and C2, were independently associated with the longitudinal decreases in renal function in patients with IgAN. These findings suggested therapies targeted at improving early damage to these lesions might be essential to delay renal progression.


A dimeric fluorescent protein yields a bright, red-shifted GEVI capable of population signals in brain slice.
A bright, red-shifted Genetically Encoded Voltage Indicator (GEVI) was developed using a modified version of the fluorescent protein, tdTomato. Dimerization of the fluorescent domain for ArcLight-type GEVIs has been shown to affect the signal size of the voltage-dependent optical signal. For red-shifted GEVI development, tdTomato was split fusing a single dTomato chromophore to the voltage sensing domain. Optimization of the amino acid length and charge composition of the linker region between the voltage sensing domain and the fluorescent protein resulted in a probe that is an order of magnitude brighter than FlicR1 at a resting potential of -70 mV and exhibits a ten-fold larger change in fluorescence (ΔF) upon 100 mV depolarization of the plasma membrane in HEK 293 cells. Unlike ArcLight, the introduction of charged residues to the exterior of dTomato did not substantially improve the dynamic range of the optical signal. As a result, this new GEVI, Ilmol, yields a 3-fold improvement in the signal-to-noise ratio compared to FlicR1 despite a smaller fractional change in fluorescence of 4% per 100 mV depolarization of the plasma membrane. Ilmol expresses well in neurons resolving action potentials in neuronal cultures and reporting population signals in mouse hippocampal acute brain slice recordings. Ilmol is the brightest red-shifted GEVI to date enabling imaging with 160-fold less light than Archon1 for primary neuron recordings (50 mW/cm2 versus 8 W/cm2) and 600-fold less light than QuasAr2 for mouse brain slice recordings (500 mW/cm2 versus 300 W/cm2). This new GEVI uses a distinct mechanism from other approaches, opening an alternate engineering path to improve sensitivity and speed.


Loss of iron-sulfur clusters from biotin synthase as a result of catalysis promotes unfolding and degradation.
Biotin synthase (BioB) is an S-adenosylmethionine radical enzyme that catalyzes addition of sulfur to dethiobiotin to form the biotin thiophane ring. In vitro, Escherichia coli BioB is active for only one turnover, during which the [2Fe-2S]2+ cluster is destroyed, one sulfide from the cluster is incorporated as the biotin thiophane sulfur, while Fe2+ ions and the remaining S2- ion are released from the protein. The present work examines the fate of the protein following the loss of the FeS clusters. We examine the quaternary structure and thermal stability of active and inactive states of BioB, and find that loss of either the [4Fe-4S]2+ or [2Fe-2S]2+ clusters results in destabilization but not global unfolding of BioB. Using susceptibility to limited proteolysis as a guide, we find that specific regions of the protein appear to be transiently unfolded following loss of these clusters. We also examine the in vivo degradation of biotin synthase during growth in low-iron minimal media and find that BioB is degraded by an apparent ATP-dependent proteolysis mechanism that sequentially cleaves small fragments starting at the C-terminus. BioB appears to be resistant to degradation and capable of multiple turnovers only under high-iron conditions that favor repair of the FeS clusters, a process most likely mediated by the Isc or Suf iron-sulfur cluster assembly systems.


Vesicle transporter GOLT1B mediates the cell membrane localization of DVL2 and PD-L2 and promotes colorectal cancer metastasis.
Colorectal cancer (CRC) is the third most diagnosed and second leading cause of cancer death worldwide. Hallmark proteins processing is usually dysregulated in cancers. Finding key regulatory molecules is of great importance for CRC metastasis intervention. GOLT1B is a vesicle transport protein which is involved in cytosolic proteins trafficking. However, its role in cancer has never been addressed. CRC cell lines and subcutaneous xenograft animal model were utilized to investigate the biological function of GOLT1B. Patients samples were used to validate the correlation between GOLT1B and clinical outcome. In vivo targeted delivery of GOLT1B-siRNA was investigated in PDX (Patient derived tumor xenograft) model. We found that GOLT1B was highly expressed in CRC, and was an independent prognostic marker of overall survival (OS) and progression free survival (PFS). GOLT1B could promote CRC metastasis in vitro and in vivo. GOLT1B overexpression could increase DVL2 level and enhance its plasma membrane translocation, which subsequently activated downstream Wnt/β-catenin pathway and increase the nuclear β-catenin level, hence induce epithelial-mesenchymal transition (EMT). In addition, GOLT1B could also interact with PD-L2 and increase its membrane level. Co-culture of GOLT1B-overexpresed CRC cells with Jurkat cells significantly induced T cells apoptosis, which might further promote cancer cell the migration and invasion. Further, targeted delivery of GOLT1B siRNA could significantly inhibit tumor progression in GOLT1B highly expressed PDX model. Taken together, our findings suggest that the vesicle transporter GOLT1B could promote CRC metastasis not only by assisting DVL2 translocation and activating Wnt/β-catenin pathway, but also facilitating PD-L2 membrane localization to induce immune suppression. Targeted inhibition of GOLT1B could be a potential therapeutic strategy for CRC treatment.


Ectopic ACTH syndrome associated with ovarian steroid-cell tumor.
Several cases of ovarian neoplasms resulting in Cushing's syndrome due to ectopic secretion of ACTH or ectopic secretion of F have been reported. Tumors producing ACTH include adenocarcinoma, androblastoma, Sertoli cell carcinoma, carcinoid tumor and teratoma. Cortisol secretion has been reported in ovarian steroid cell tumor (unclassified steroid cell tumor). We present a case of a 19-year-old woman with Cushing's syndrome in course of an ovarian steroid cell tumor with ectopic ACTH production. To our knowledge, it is the first reported case of ACTH secreting ovarian steroid cell tumor causing Cushing's syndrome.


Circadian rhythm of preferred temperature in fish: Behavioural thermoregulation linked to daily photocycles in zebrafish and Nile tilapia.
Ectothermic vertebrates, e.g. fish, maintain their body temperature within a specific physiological range mainly through behavioural thermoregulation. Here, we characterise the presence of daily rhythms of thermal preference in two phylogenetically distant and well-studied fish species: the zebrafish (Danio rerio), an experimental model, and the Nile tilapia (Oreochromis niloticus), an aquaculture species. We created a non-continuous temperature gradient using multichambered tanks according to the natural environmental range for each species. Each species was allowed to freely choose their preferred temperature during the 24h cycle over a long-term period. Both species displayed strikingly consistent temporal daily rhythms of thermal preference with higher temperatures being selected during the second half of the light phase and lower temperatures at the end of the dark phase, with mean acrophases at Zeitgeber Time (ZT) 5.37 h (zebrafish) and ZT 12.5 h (tilapia). Interestingly, when moved to the experimental tank, only tilapia displayed consistent preference for higher temperatures and took longer time to establish the thermal rhythms. Our findings highlight the importance of integrating both light-driven daily rhythm and thermal choice to refine our understanding of fish biology and improve the management and welfare of the diversity of fish species used in research and food production.


Clustering of early cortical responses to median nerve stimulation from average and single trial MEG and EEG signals.
Median nerve electrical stimulation (MNES) produces early and strong averaged magnetoencephalography (MEG) or electroencephalography (EEG) signals, despite considerable single trial (ST) variability, demonstrated in separate MEG and EEG studies. Here, simultaneous MEG/EEG recordings are used to assess whether same or different aspects of ST variability are influencing EEG and MEG. Clustering techniques provided groupings for the ST timeseries for cortical responses to MNES derived from one modality. These groupings were applied to the corresponding ST timeseries derived from the other modality to quantify the similarity in variability captured by MEG and EEG signals. Estimates of early cortical activity elicited by MNES derived from MEG and EEG signals were very similar, provided ongoing mu rhythm was removed. Similarity between EEG and MEG estimates included both results based on average signals and measures of ST variability. Either MEG or EEG can provide a robust measure of the early cortical activity elicited by MNES as well as of its variability. Reliable indices of early cortical responses to MNES can be derived from either MEG or EEG data. These indices can be based on average signals, as is routinely done with clinical EEG, but it could also rely on hitherto little utilized measures of ST variability.


Sonography in male infertility: a useful yet underutilized diagnostic tool.
To assess the utility of comprehensive sonographic examination including scrotal sonography, Testicular Doppler and Transrectal Ultrasound (TRUS) to evaluate the male reproductive system and differentiate between obstructive (OG) and non-obstructive (NOG) causes of azoospermia. 30 infertile men with azoospermia and 30 control subjects with normospermia underwent sonographic evaluation. FNAC/biopsy findings were used for assigning a final diagnosis of obstructive or non-obstructive azoospermia. Qualitative and quantitative imaging parameters were retrospectively compared between the groups using Chi-square/Fisher's exact test and unpaired t-test, respectively. P < 0.05 was considered significant. Ectasia of rete testis/epididymal tubules, altered epididymal echogenicity, dilated terminal vas deferens were significantly more common in OG while inhomogeneous testicular echo-texture and reduced testicular vascularity were more common in NOG (P < 0.05). Testicular volume and epididymal head size were significantly higher in OG than in NOG and controls (18.2 ml/10 mm Vs 8.2 ml/7.2 mm and 13.4 ml/8.8 mm respectively; P < 0.05); while Resistive Index (RI) of intra-testicular vessels was higher in NOG as compared to OG and controls (0.65 vs 0.54 and 0.52 respectively; P < 0.05). On ROC curve analysis, cut-off values of testicular volume (AUC: 0.939; P < 0.001), epididymal head size (AUC: 0.772; P = 0.001) and testicular RI (AUC: 0.761; P = 0.001) to differentiate between the groups were 12.1 ml (sensitivity-94.4%; specificity-83.3%), 9 mm (sensitivity-66.7%; specificity-71%) and 0.62 (sensitivity-62%; specificity-100%) respectively. Comprehensive sonographic evaluation can be used to differentiate obstructive from non-obstructive infertility and should be routinely incorporated in the diagnostic workup of infertile men with azoospermia.


Lumbar ligamentum flavum burden: Evaluating the role of ATTRwt amyloid deposition in ligamentum flavum thickness at all lumbar levels.
Wild-type transthyretin (ATTRwt) amyloid deposition has been found in the ligamentum flavum (LF) of patients undergoing spinal stenosis surgery. Our group previously reported that ATTRwt amyloid is associated with an increased lumbar ligamentum flavum thickness at symptomatic levels that required surgery. A comprehensive evaluation of LF thickness at asymptomatic levels in addition to symptomatic, treated levels has never been performed in ATTRwt patients. In this study, we compare the total LF thickness of all lumbar levels (lumbar LF burden) in ATTRwt and non-ATTRwt patients. We retrospectively identified 177 patients who underwent lumbar spine surgery. Ligamentum flavum thickness of 885 lumbar levels was measured on T2-weighted axial MRI. Amyloid presence was confirmed through Congo red staining of specimens, and subtype of ATTRwt was confirmed using mass-spectrometry and gene sequencing. Of the 177 patients, 30 (16.9%) were found to have ATTRwt in the ligamentum flavum. One hundred and fifty ATTRwt levels and 735 non-ATTRwt levels were measured by four different reviewers, with an intraclass coefficient (ICC) of 0.79. Mean ligamentum flavum thickness was 4.64 (±1.31) mm in the ATTRwt group and 3.99 (±1.45) mm in the non-ATTRwt group (p < 0.001). The lumbar LF burden (sum of ligamentum flavum thickness at all lumbar levels) for ATTRwt patients was 23.22 (±4.48) mm, and for non-ATTRwt patients was 19.96 (±5.49) mm (p = 0.003) CONCLUSION: The lumbar LF burden is greater in patients with ATTRwt amyloid compared to non-ATTRwt patients. This supports prior evidence that ATTRwt amyloid deposition might be associated with increased LF thickness and lumbar stenosis. This potential association requires more research and could be an important finding, as medications have recently become available that can treat patients with ATTRwt amyloid deposition.


Endogenous proteolytic enzymes--a study of their impact on cod (Gadus morhua) muscle proteins and textural properties in a fermented product.
The aim of this study was to investigate endogenous proteolytic activities in a cod product and their impact on muscle proteins and textural properties during fermentation and storage. The result of specific proteolytic activities showed that cathepsins, especially cathepsin B, had the highest activities during fermentation and storage. SDS-PAGE indicated more degradation of myofibrillar proteins by cathepsin L than other proteases and that the hydrolysis by cathepsins was pronounced in the last stage of fermentation. Texture analysis showed that cathepsins had a negative impact on gel strength and this impact increased in the last stage of fermentation. However the product still had a firm texture. During storage (4 °C) for one week, no significant changes were seen in the gel strength. In conclusion, cathepsins had more impact on muscle proteins and textural properties than other proteases during fermentation but had little impact on gel strength during storage at 4 °C.


Endogenous expression and localization of HIS-72::mTurquoise2 in C. elegans.
To generate a non-red/green fluorescent fusion histone protein in C. elegans, we have generated a C-terminal mTurquoise2-tagged HIS-72 at the endogenous locus using CRISPR. We found that HIS-72::mTurquoise2 localizes in a similar pattern to the previously published HIS-72::GFP strain.


Quantitatively characterizing the textural features of sonographic images for breast cancer with histopathologic correlation.
In this study, quantitative characterization of sonographic image texture and its correlation with histopathologic findings was developed for facilitating clinical diagnosis. A statistical feature matrix was applied to quantify the texture difference (ie, the dissimilarity) of the sonographic images for malignant and benign breast tumors. Thirty-three patients were recruited for this study. Imaging was performed on a commercially available sonographic imaging system in clinical use. The parameters used for image acquisition were kept the same during clinical examination. On the basis of dissimilarity values, 3 phenomena were noted in the relatively large malignancies studied. First, stellate carcinoma showed the least dissimilarity on sonographic images; second, circumscribed carcinoma showed the most dissimilarity; and third, malignant tissue mixed with fibrous and cellular parts (dense lymphocyte infiltration and prominent intraductal tumors) had dissimilarity values in between. Image textures with smaller dissimilarity values (especially for those values <4.4 in our study) are likely to be stellate carcinoma. From the experimental results, it is shown that the cellular and fibrous content with spatial distribution of breast masses determine the dissimilarity values on sonographic images. The dissimilarity may be used to quantitatively represent the image texture and is well correlated with the histopathologic description.


Between Cyberplace and Cyberspace: the researcher's role in virtual setting research.
Disciplines such as Internet Research, the Psychology of Cyberspace and the Social Psychology of Cyberplaces call for an epistemological reflection not merely on the universe of objects they deal with, but also, and perhaps especially, on the research settings used to investigate them. With this work, we intend to make a contribution to the debate on three issues: psychosocial interpretation of the new environments, the "mediated" nature of the researcher-setting-study object relationship, and cyberplaces as settings for mediated interaction research.


Antihypertensive effect and pharmacokinetics of nitrendipine in children.
Nitrendipine, a new calcium-channel antagonist, was used to treat 25 children (aged 6 months to 17 years) with severe hypertension. Systolic and diastolic blood pressures (mean +/- SEM) fell from 148 +/- 2/99 +/- 2 mm Hg to 128 +/- 4/77 +/- 3 mm Hg after 24 hours and to 121 +/- 2/75 +/- 2 mm Hg after 2 weeks. No further reductions in systolic or diastolic blood pressure were observed after continued therapy. Transient reflex tachycardia occurred during the first week of therapy. Other adverse effects were uncommon and included headaches, flushing, palpitations, and edema. Pharmacokinetic parameters were estimated at steady state after an oral dose of 0.56 +/- 0.04 mg/kg in 13 children. Although absolute oral bioavailability could not be determined, estimates of the area under the plasma concentration versus time curve, the apparent peak serum concentration, and the apparent time at which the peak serum concentration occurred indicated that both the rate of absorption and oral bioavailability are variable. Coadministration of nitrendipine with food decreased the rate of absorption and may have reduced oral bioavailability. A relationship between age and the apparent plasma elimination half-life of nitrendipine was not observed. Nitrendipine, 0.25 to 0.5 mg/kg per dose administered orally every 6 to 12 hours, appeared to be an effective and safe treatment for resistant hypertension in infants and children.


Factors associated with mortality among moderate and severe patients with COVID-19 in India: a secondary analysis of a randomised controlled trial.
Large data on the clinical characteristics and outcome of COVID-19 in the Indian population are scarce. We analysed the factors associated with mortality in a cohort of moderately and severely ill patients with COVID-19 enrolled in a randomised trial on convalescent plasma. Secondary analysis of data from a Phase II, Open Label, Randomized Controlled Trial to Assess the Safety and Efficacy of Convalescent Plasma to Limit COVID-19 Associated Complications in Moderate Disease. 39 public and private hospitals across India during the study period from 22 April to 14 July 2020. Of the 464 patients recruited, two were lost to follow-up, nine withdrew consent and two patients did not receive the intervention after randomisation. The cohort of 451 participants with known outcome at 28 days was analysed. Factors associated with all-cause mortality at 28 days after enrolment. The mean (SD) age was 51±12.4 years; 76.7% were males. Admission Sequential Organ Failure Assessment score was 2.4±1.1. Non-invasive ventilation, invasive ventilation and vasopressor therapy were required in 98.9%, 8.4% and 4.0%, respectively. The 28-day mortality was 14.4%. Median time from symptom onset to hospital admission was similar in survivors (4 days; IQR 3-7) and non-survivors (4 days; IQR 3-6). Patients with two or more comorbidities had 2.25 (95% CI 1.18 to 4.29, p=0.014) times risk of death. When compared with survivors, admission interleukin-6 levels were higher (p<0.001) in non-survivors and increased further on day 3. On multivariable Fine and Gray model, severity of illness (subdistribution HR 1.22, 95% CI 1.11 to 1.35, p<0.001), PaO2/FiO2 ratio <100 (3.47, 1.64-7.37, p=0.001), neutrophil lymphocyte ratio >10 (9.97, 3.65-27.13, p<0.001), D-dimer >1.0 mg/L (2.50, 1.14-5.48, p=0.022), ferritin ≥500 ng/mL (2.67, 1.44-4.96, p=0.002) and lactate dehydrogenase ≥450 IU/L (2.96, 1.60-5.45, p=0.001) were significantly associated with death. In this cohort of moderately and severely ill patients with COVID-19, severity of illness, underlying comorbidities and elevated levels of inflammatory markers were significantly associated with death. CTRI/2020/04/024775.


Impact of an educational training program on pharmacists' documentation practice at a teaching hospital.
This study aims to determine the impact of an educational training program on the quantity and quality of the pharmacists' documentation practice at the Winchester District Memorial Hospital. This study is a part of an evaluation for continuous quality improvement, performed applying a pre- and post-test model. The primary endpoint was the number of Pharmacists' Patient-Care Records in patient charts with the listed action codes. Charts of 80 patients were surveyed at three points (two months before and one and three months after the sessions) for 240 charts. Additionally, a pre- and post-educational questionnaire was administered using case scenarios and the Pharmacists' Patient-Care Record code system. The number of charts containing Pharmacists' Patient-Care Records with Action Codes listed is 70%, 73%, and 64% in the first, second, and third chart review rounds, respectively. According to the pre- and post-educational session questionnaire, the average score per pharmacist is 47% and 73%, respectively. The initial estimate of the documentation practice is 50% or less, which is an underestimation. Furthermore, the quantity of pharmacists' documentation is not significantly affected by the educational sessions; however, the quality is improved.


Molecular characteristic of mcr-1 producing Escherichia coli in a Chinese university hospital.
Colistin has been considered as a last-line treatment option in severe infections caused by multidrug-resistant (MDR) gram-negative pathogens. However, the emergence of the mobile colistin resistance gene (mcr-1) has challenged this viewpoint. The aim of this study is to explore the prevalence of mcr-1 in Escherichia coli (E. coli) in a Chinese teaching hospital, and investigate their molecular characteristics. A total of 700 E. coli isolates were used to screen mcr-1 by PCR and sequencing in a Chinese university hospital from August 2014 to August 2015. Susceptibility test of mcr-1-producing isolates was determined by Vitek -2 Compact system. 26 virulence factors (VFs), phylogenetic groups, Multi-locus sequence typing (MLST), and DNA Fingerprinting (ERIC-PCR) of strains were investigated by PCR. Four (0.6%) mcr-1 producing E. coli isolates were found in this study. The results of antibiotic susceptibility test showed that all four isolates were resistant to colistin, ciprofloxacin, levofloxacin, cefazolin, and trimethoprim/sulfamethoxazole, and were susceptible to amikacin, ertapenem and imipenem. In addition, all 4 isolates exhibited high-level resistance to aztreonam, cefotaxime and gentamicin. The numbers of VFs contained in mcr-1 positive isolates were no more than 4 in our study. MLST result demonstrated that these isolates were assigned to two sequence types: ST156 and ST167. The result of phylogenetic analysis showed that four mcr-1-positive isolates belong to two phylogenetic groups: A and B1 group. ERIC-PCR showed that four mcr-1 positive strains were categorized into three different genotypes. Our study demonstrated a low prevalence of mcr-1 in E. coli clinical isolates in a Chinese teaching hospital, and we have gained insights into the molecular characteristics of these mcr-1-positive strains. Increasing the surveillance of these infections, as well as taking effective infection control measures are urgently needed to take to control the transmission of mcr-1 gene.


Voltage dependent anion channel and its interaction with N-acetyl-L-Cysteine (NAC) under oxidative stress on planar lipid bilayer.
Mitochondria are the major source of Hydrogen Peroxide (H2O2), a reactive oxygen species, in the cells. The reactive oxygen species generated by the mitochondria oxidize major proteins including Voltage Dependent Anion Channel (VDAC). We were interested to know how the effect of H2O2 is countered by antioxidants present around the mitochondria. N-Acetyl-l-Cysteine (NAC) is a naturally existing antioxidant in the cells. Keeping this in view, the modulatory effect of antioxidant NAC on H2O2 oxidized VDAC has been investigated through in vitro electrophysiological studies. First, the effect of H2O2 and NAC was studied on independently incorporated single-channel VDAC. It was observed that NAC suppresses VDAC conductance with a half-maximal inhibitory concentration (IC50) of ∼1.04 μM. In contrast, H2O2 enhances VDAC conductance. Later, oxidative stress was induced by H2O2 on VDAC increased conductance with half-maximal effective concentration (EC50) of ∼302 nM. An application of 1 μM NAC on H2O2 treated (300 nM) VDAC reversed the effect of oxidation. In the next step, NAC and H2O2 were added in reverse order. When oxidative stress was induced using H2O2, reduction in conductance by NAC was 4.5 ± 0.404 nS. The change in conductance is nearly 6.3%. However, if antioxidant NAC was incubated first followed by H2O2 treatment, the conductance of VDAC was 3.09 ± 0.27 nS. The change in conductance is near 33%. Both H2O2 and NAC also affected various conducting states of VDAC. In-silico studies indicated the binding of NAC at Lysine and Glutamic acid of VDAC. Hence, NAC was found to be effective in protection of VDAC against H2O2-induced oxidative stress due to its strong binding.


Activation of the cisplatin and transplatin complexes in solution with constant pH and concentration of chloride anions; quantum chemical study.
The thermodynamics of cisplatin and transplatin hydration is studied within the model of constant pH solution. Several implicit solvation models were chosen for the determination of pK(a) and pK constants of the hydration reactions. The polarizable dielectric model (DPCM), integral equation formalism polarizable model (IEFPCM), and polarizable conductor model (CPCM) were combined with the 'united atom model for Hartree-Fock' (UAHF) method for cavity construction and the B3LYP/6-31++G(2dp,2pd) level of calculations for the determination of electronic energies. The results were compared with the COSMO-RS and SM8 model developed by Truhlar (with M06 and MPWX functionals and the charge model CM4). The RMS difference between experimental and calculated pK(a) values of cis/transplatin, water, HCl, and NH (4) (+) was used to evaluate accuracy of calculations. The DPCM model was confirmed to perform the best. The predicted pK(a) constants were used in Legendre transformation for the estimation of the ΔG' energies in the constant-pH model. The dependence of the pK constant on pH is plotted and compared with experimental value at pH=7.4. The influence of various chloride concentrations on the molar fractions of dissolved forms of cisplatin is examined for the DPCM model. The increased ratio of cisplatin active aqua-forms is clearly visible for 4 mM chloride solution in comparison with 104 mM Cl(-) concentration.


New insights for rapid evaluation of bactericidal activity: a semi-automated bioluminescent ATP assay.
A new assay, much more rapid and efficient than the existing standardized tests, is introduced for the evaluation of bactericidal activity of chemical disinfectants and antiseptics under simulated practical conditions of use. The bactericidal activity of biocides was quantified using a novel semi-automated assay based on the European Norm (EN) standard suspension tests but determining bacterial cell viability by intracellular adenosine tri-phosphate (ATP) content quantification instead of traditional culture-based microbiological techniques. The new test was validated by comparison to the standard suspension tests EN 1276 and EN 13727. During the validation, the linearity of the ATP detection system, limit of detection, specificity, sensitivity, relative accuracy and precision (repeatability and reproducibility) were determined. The validation study showed that the new assay evaluates the activity of biocides as well as the EN standard suspension tests, but it allows a large number of test conditions to be efficiently analysed. The new test can therefore be applied to accurately establish the lowest active concentration (MBCs) of disinfectants or antiseptics under simulated practical conditions of use and to compare the susceptibility of a large number of strains and conditions via inactivation curves. This is not possible in any reasonably practicable way with the EN standards considering the time and cost required for each determination.


Maximizing the value of genomics in the drug discovery and development process.
Genomics and all the associated technologies it has spawned have fundamentally changed the way research and development organizations carry out the work they do in the early stages of discovery. However, successful organizations must move from product concept through to registration as efficiently as possible. In order to achieve this, the early "basic" science must be combined with the clinical perspective from the start. Furthermore, since the genomics industry was first established in the early 1990s it has evolved from tool and data suppliers to drug development companies. It is therefore likely that we will see technology advances in genomics become more limited in the years ahead.


Advances in rectal cancer surgery in Japan.
A review of advancement of rectal cancer surgery in Japan is presented. The standard operation for rectal cancer was altered in the 1960s from abdominoperineal resection to the pull-through technique and the handsewn anterior resection in the 1970s, and it became the stapled anterior resection in the 1980s. Today, more than 75 percent of rectal cancers are treated with sphincter-preserving anterior resections, and the remaining 20 percent by abdominoperineal resections. Colonic J-pouch is used with anastomoses involving very low anterior rectal resection for cancers. In the late 1970s, a method of dissecting extended pelvic nodes was adopted to decrease local recurrence. However, extended dissection has been applied to only T3 and T4 cancers of the lower rectum because of postoperative dysfunction of pelvic organs. This was caused by injury to the pelvic nerve plexus, thus lowering the quality of life of the patients. Since the middle of the 1980s, the autonomic nerve-preserving operation attracted surgeons' attention because it prevented these dysfunctions from occurring as a result of the treatment of cancer in the upper rectum and for T1 or T2 cancers in the lower rectum. In this article, recent advances in rectal cancer surgery in Japan are reviewed.


To teach and to learn in day surgery. The role of residents.
Our Division of Surgery is a University Department. Residents perform in Day Surgery theatre room with tutors of proven experience. This study looks to explore ways in which senior tutors in Day Surgery can contribute to teach. Infact the School of postgraduate Surgery obliges postgraduate surgeons to carry out a number of minor and medium size operations. This study was carried out in 2006. In this year were performed n.116 Inguinal Hernia repairs with method by Trabucco. Residents performed N.51 Inguinal Hernia repairs as first surgeons, assisted by a tutor with proven experience. All patients were discharged home on the day of their operations. The post-operative morbidity rate was the same of that encountered during surgery performed by experienced surgeons. Two recurrences were reported: one patient operated by a resident and one patient operated by an experienced colleague. In Day Surgery it is not possible to do a teaching programme. The opportunity to see patients' pre-operatively and follow them through surgery discharge on the same day is unique. The residents can contribute to quality of Day Surgery and learn. Day Surgery Units could play an increased role in medical education.


Comparison of Three-year outcomes after primary percutaneous coronary intervention in patients with left ventricular ejection fraction <40% versus ≥ 40% (from the HORIZONS-AMI trial).
Left ventricular (LV) dysfunction and multivessel disease (MVD) have been associated with greater mortality after ST-segment elevation myocardial infarction. The aim of this study was to evaluate the impact of LV dysfunction and MVD in patients with ST-segment elevation myocardial infarctions treated with primary percutaneous coronary intervention (PCI). Patients from the Harmonizing Outcomes With Revascularization and Stents in Acute Myocardial Infarction (HORIZONS-AMI) trial treated with primary PCI in whom baseline LV function was assessed using left ventriculography were included in this study. Early and late (3-year) outcomes were examined in groups of patients with reduced (<40%) and preserved (≥ 40%) LV ejection fractions (LVEFs), further stratified by the presence of MVD. A total of 2,430 patients were included. Patients with reduced LVEFs were older; were more likely to be women; were more likely to have histories of myocardial infarction, PCI, and heart failure; and were more likely to present in heart failure. Patients with reduced LVEFs had greater 30-day (8.9% vs 0.9%, hazard ratio 9.81, 95% confidence interval 5.23 to 18.42, p <0.0001) and 3-year (17.1% vs 3.7%, hazard ratio 5.03, 95% confidence interval 3.37 to 7.50, p <0.0001) mortality. Among patients with LVEFs <30% (n = 45), 30% to 40% (n = 157), 40% to 50% (n = 373), 50% to 60% (n = 659), and ≥ 60% (n = 1,196), 3-year mortality was 29.4%, 13.5%, 6.4%, 3.8%, and 2.9%, respectively (p for trend <0.0001). MVD was associated with greater mortality in patients with preserved but not reduced LVEFs. By multivariate analysis, LV dysfunction was the strongest predictor of 30-day and 3-year mortality. In conclusion, the presence of LV dysfunction as assessed on baseline left ventriculography in patients who undergo primary PCI in the contemporary era is a powerful predictor of early and late mortality, regardless of the extent of coronary artery disease.


Burst bicycle exercise facilitates diagnosis of latent long QT syndrome.
The aim of this study was to develop a diagnostic technique to detect latent long QT syndrome. Asymptomatic patients with genetically diagnosed long QT syndrome (LQTS) may have a normal resting QT interval, yet remain at risk for cardiac events. We hypothesized that the QT response during a novel burst exercise protocol simulating clinical events might distinguish patients with "latent LQTS" from healthy subjects. A burst bicycle protocol was performed on 31 healthy subjects and 31 patients with LQTS (13 LQT2, 3 LQT1, 15 unknown genotype). The bicycle exercise protocol involved sudden maximal exertion against a fixed workload (200 W) for 1 minute. Digitized 12-lead eletrocardiograms were acquired every 10 seconds at baseline for 1 minute, during 1 minute of burst exercise, and for 5 minutes during recovery. Patients with LQTS were segregated according to whether the baseline QTc was normal (< or = 440 milliseconds, n = 13) or abnormal (> 440 milliseconds, n = 18). During exercise, the QTc increased to a greater extent in the group with latent LQTS (DeltaQTc 98 +/- 36 milliseconds) in comparison with controls (DeltaQTc 65 +/- 19 milliseconds, P < .01) and those with baseline QTc prolongation (DeltaQTc 17 +/- 70 milliseconds, P < .01). In patients with a normal baseline QTc, a DeltaQTc > 85 milliseconds had a sensitivity of 85% and a specificity of 86% for LQTS (P = .0004). Marked QTc prolongation during burst bicycle ergometry provides potentially diagnostic information for patients with normal baseline QTc and suspected LQTS.


Synchrotron phase-contrast microtomography of coprolites generates novel palaeobiological data.
Coprolites (fossil faeces) reveal clues to ancient trophic relations, and contain inclusions representing organisms that are rarely preserved elsewhere. However, much information is lost by classical techniques of investigation, which cannot find and image the inclusions in an adequate manner. We demonstrate that propagation phase-contrast synchrotron microtomography (PPC-SRμCT) permits high-quality virtual 3D-reconstruction of coprolite inclusions, exemplified by two coprolites from the Upper Triassic locality Krasiejów, Poland; one of the coprolites contains delicate beetle remains, and the other one a partly articulated fish and fragments of bivalves.


Comparison of the efficacy of the pulsed bipolar system and conventional bipolar electrosurgery in laparoscopically assisted vaginal hysterectomy.
We compared the safety and effectiveness of the pulsed bipolar system (PlasmaKinetic; Gyrus Medical, Maple Grove, MN) and conventional electrosurgery (Kleppinger bipolar forceps; Richard Wolf Instruments, Vernon Hills, IL) in laparoscopically assisted vaginal hysterectomy (LAVH). In this prospective, nonrandomized study, 62 women with benign gynecologic diseases scheduled for LAVH were divided into two groups: one group underwent LAVH with the pulsed bipolar system and the second group underwent LAVH with conventional electrosurgery. Outcome measures for both groups were compared in terms of length of operative time, amount of blood loss, requirement of blood transfusion, and length of hospital stay. The mean operative time (87.6+/-28.1 minutes vs. 93.4+/-20.2 minutes, P=0.368), blood loss (196.8+/-143.7 mL vs. 253.2+/-125.8 mL, P=0.105), and blood transfusion rate (3.2% vs. 6.5%, P=1.0) were slightly greater in the conventional electrosurgery group than those in the pulsed bipolar system group, although these differences were not statistically significant. The mean length of hospital stay was similar in both groups (3 days). No patients developed serious complications related either to conventional electrosurgery or to the pulsed bipolar system. Our findings indicate that the pulsed bipolar system is as safe and effective as conventional electrosurgery, and may offer an alternative option for patients undergoing LAVH.


In vitro quantitative determination of phospholipid adducts of chloroform intermediates in hepatic and renal microsomes from different rodent strains.
We have comparatively studied in vitro the oxidative and reductive pathways of chloroform metabolism in hepatic and renal microsomes of rodent strains used for carcinogenicity testing (B6C3F1 mice, Osborne Mendel and Sprague Dawley rats). To this aim we exploited the regioselective binding of phosgene to phospholipid (PL) polar heads and of dichloromethyl radical to PL fatty acyl chains, using a method based on the chemical transmethylation of PL adducts, followed by phase partitioning of the resulting products (De Biasi et al., 1992). The analysis of results let us to conclude at first that a (14)C label partitioning by 89.2 (±6.5)% or 13.7 (±5.0)% in the aqueous phase is typical of the PL adduct with phosgene (PL-PHOS) or with dichloromethyl radical (PL-RAD), respectively. Metabolism of 0.1 mM CHCl(3) was mainly oxidative in all the samples, being hepatic microsomes more active than renal ones by about one order of magnitude and levels of CHCl(3)-derived PL adducts in B6C3F1 mouse liver microsomes higher than in rat samples. At 5 mM CHCl(3), total levels of PL adducts in renal microsomes reached levels almost similar to those found in liver microsomes. However, while B6C3F1 mouse kidney microsomes produced both reactive metabolites, similarly as the hepatic samples, Osborne Mendel rat kidney microsomes bioactivated CHCl(3) only reductiveiy, producing the radical. The relevance of this finding depends on the fact that phosgene is known to be the major cause of CHCl(3) toxicity, based on data with the rat liver and mouse liver and kidney, while nephrotoxicity in rats occurs with minimal production of COCl(2). Chloroform reductive bioactivation may therefore provide a reasonable explanation for the toxicity of chloroform to the rat kidney. The same finding may be of interest in elucidating the metabolic reasons of the chloroform-induced kidney tumors in Osborne Mendel rats.


[Simultaneous detection of T-, B- and D-rosette forming lymphocytes and null cells in humans].
For the identification of T-, B-, and D-rosette-forming lymphocytes and the "null" cells in the human peripheral blood a simultaneous reaction of rosette-forming cells with the use of zymosan-complement, sheep red blood cells, fixation and staining of smears in extraction of the lymphocytes tested with the aid of verographin may be recommended.


Mechanistic correlation between mitochondrial permeability transition pores and mitochondrial ATP dependent potassium channels in ischemia reperfusion.
Mitochondrial dysfunction is one of the fundamental causes of ischemia reperfusion (I/R) damage. I/R refers to the paradoxical progression of cellular dysfunction and death that occurs when blood flow is restored to previously ischemic tissues. I/R causes a significant rise in mitochondrial permeability resulting in the opening of mitochondrial permeability transition pores (MPTP). The MPTP are broad, nonspecific channels present in the inner mitochondrial membrane (IMM), and are known to mediate the deadly permeability alterations that trigger mitochondrial driven cell death. Protection from reperfusion injury occurs when long-term ischemia is accompanied by short-term ischemic episodes or inhibition of MPTP from opening via mitochondrial ATP dependent potassium (mitoKATP) channels. These channels located in the IMM, play an essential role in ischemia preconditioning (PC) and protect against cell death by blocking MPTP opening. This review primarily focuses on the interaction between the MPTP and mitoKATP along with their role in the I/R injury. This article also describes the molecular composition of the MPTP and mitoKATP in order to promote future knowledge and treatment of diverse I/R injuries in various organs.


Research on Beers Criteria and STOPP/START Criteria based on the FDA FAERS database.
Inappropriate medication criteria for the elderly have played an important role in ensuring the safety of medications for the elderly. Too few drugs included in the criteria cannot guarantee the safety of medication for the elderly. Too many drugs included in the criteria will result in less selective medication for the elderly. This paper uses real-world data to evaluate the relationship between antihypertensive drugs and falls, so as to provide references for experts and scholars to revise the criteria of potentially inappropriate medications for the elderly and clinical safe medication. We use the US Food and Drug Administration Adverse Event Reporting System (FDA FAERS) to evaluate the association between specific antihypertensive drugs in six categories (alpha-1 receptor blockers (α-1 blockers), calcium channel blockers (CCBs), angiotensin-converting enzyme inhibitors (ACEIs), angiotensin II receptor blockers (ARBs), beta-receptor blockers (β-blockers), and diuretics) and falls by data mining algorithms, including the reporting odds ratio (ROR), the proportional reporting ratio (PRR), Medicines and Healthcare Products Regulatory Agency (MHRA), and the empirical Bayes geometric mean (EBGM) and compared with the relevant drugs included in the Beers Criteria and STOPP/START Criteria. There are a total of 5,157,172 co-occurrences found in 973,447 reports aged 65 years or older from 2016 to 2019 in the FDA FAERS database, and the number of co-occurrences of falls is 5917 for the six categories of 51 antihypertensive drugs. Four kinds of mining methods overlap detection of 12 kinds of positive signal drugs, none of which are not included in the Beers Criteria and 7 drugs are included in the STOPP/START Criteria; 1-3 kinds of mining methods overlap detection of positive signal drugs, a total of 12 kinds, and one drug is included in the Beers Criteria and 5 drugs are included in the STOPP/START Criteria; 22 drugs have fall adverse events, but no positive signal is detected, and 13 drugs are included in STOPP/START Criteria; and 5 drugs have no fall adverse events and 3 drugs are included in the STOPP/START Criteria. The FAERS database was used to confirm the potential connection between some antihypertensive drugs and fall adverse events through data mining algorithms. The Beers Criteria did not clearly indicate the antihypertensive drugs that caused falls, and the antihypertensive drugs included in the STOPP/START Criteria were too extensive and did not include β-blockers and diuretics. It is recommended that experts and scholars use real-world data (such as FAERS, EudraVigilance, WHO VigiBase, and so on) to further explore the relationship between specific antihypertensive drugs and falls in the elderly, so as to revise and improve the criteria for inappropriate medications for the elderly.


The history of electron ionization in LC-MS, from the early days to modern technologies: A review.
This review article traces the history of the use of liquid chromatography coupled with mass spectrometry (LC-MS) using electron ionization (EI) from the first attempts up to the present day. At the time of the first efforts to couple LC to MS, 70 eV EI was the most common ionization technique, typically used in gas chromatography-mass spectrometry (GC-MS) and providing highly reproducible mass spectra that could be collated in libraries. Therefore, it was obvious to transport this dominant approach to the early LC-MS coupling attempts. The use of LC coupled to EI-MS is challenging mainly due to restrictions related to high-vacuum and high-temperature conditions required for the operation of EI and the need to remove the eluent carrying the analyte before entering the ion source. The authors will take readers through a journey of about 50 years, showing how through the succession of different attempts it has been possible to successfully couple LC with EI-MS, which in principle appear to be incompatible.


All-Solid-State Thin Film μ-Batteries for Microelectronics.
Continuous advances in microelectronics and micro/nanoelectromechanical systems enable the use of microsized energy storage devices, namely solid-state thin-film μ-batteries. Different from the current button batteries, the μ-battery can directly be integrated on microchips forming a very compact "system on chip" since no liquid electrolyte is used in the μ-battery. The all-solid-state battery (ASSB) that uses solid-state electrolyte has become a research trend because of its high safety and increased capacity. The solid-state thin-film μ-battery belongs to the family of ASSB but in a small format. However, a lot of scientific and technical issues and challenges are to be resolved before its real application, including the ionic conductivity of the solid-state electrolyte, the electrical conductivity of the electrode, integration technologies, electrochemical-induced strain, etc. To achieve this goal, understanding the processing of thin films and fundamentals of ion transfer in the solid-state electrolytes and hence in the μ-batteries becomes utmost important. This review therefore focuses on solid-state ionics and provides inside of ion transportation in the solid state and effects of chemistry on electrochemical behaviors and proposes key technology for processing of the μ-battery.


Solubilization and partial purification of hyaluronate synthetase from oligodendroglioma cells.
Hyaluronate synthetase was solubilized with digitonin from crude membranes of mouse oligodendroglioma cells. Detergent extraction was carried out in 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid-buffered saline with an optimal digitonin to protein ratio (w/w) of 0.7-0.8. The solubilized synthetase was partially purified approximately 230-fold by gel filtration and ion-exchange chromatography. The solubilized enzyme displayed similar properties to membrane-bound enzyme: (a) it synthesized high molecular weight hyaluronate which eluted in the void volume of a Sepharose CL-2B column; (b) the apparent Km values obtained for UDP-GlcUA and UDP-GlcNAc were 50 and 100 microM, respectively; and (c) treatment of intact cells with hyaluronidase prior to extraction with digitonin resulted in a 3-fold increase in solubilized synthetase activity. Furthermore, gel filtration chromatography of the solubilized hyaluronidase-treated synthetase complex showed that it was smaller than the solubilized untreated synthetase complex, due to shorter nascent-bound hyaluronate. The solubilized synthetase was shown to be associated with hyaluronate in the form of a complex. Both hyaluronidase-treated and -untreated synthetase-hyaluronate complexes after solubilization were adsorbed by an affinity matrix using the hyaluronate binding domain of rat chondrosarcoma proteoglycan as ligand. This solubilized active enzyme preparation should allow the identification and characterization of the components of the hyaluronate-synthetase complex.


A randomized trial demonstrating successful boosting responses following simultaneous aerosols of measles and rubella (MR) vaccines in school age children.
The reactogenicity and immunogenicity of combined measles and rubella (MR) booster vaccination, via aerosol and subcutaneous routes, was assessed in 562 healthy children. Rates of rubella seroconversion and geometric mean titers (GMT) were similar for both routes. Rates of measles PN seroconversion, GMT and measles ELISA post-vaccination seropositivity and seroconversion rate were each higher for aerosol vaccine (54%, 3928 IU/l, 99.6 and 98.8%), than for subcutaneous vaccine (7%, 866 IU/l, 92.2 and 82.4%) (P<0.01). Reactogenicity was higher for subcutaneous vaccine (P<0.05). This study demonstrates that aerosol vaccine was more immunogenic for measles antibodies, and equally immunogenic for rubella antibodies. Aerosol vaccine was less reactogenic.