The application of human bone marrow stromal cells and poly(dl-lactic acid) as a biological bone graft extender in impaction bone grafting.
Concerns over disease transmission, high costs and limited supply have led to interest in synthetic grafts in the field of impaction bone grafting (IBG). Poly(DL-lactic acid) (PLA) grafts are attractive alternatives due to their biocompatibility, established safety and versatile manufacturing process. This study examined the potential of PLA scaffolds augmented with human bone marrow stromal cells (HBMSCs) in IBG. In vitro and in vivo studies were performed on impacted morsellised PLA seeded with HBMSC and compared to PLA alone. In vitro samples were incubated under osteogenic conditions and in vivo samples were implanted subcutaneously into severely compromised immunodeficient mice, for 4 weeks. Biochemical, histological, mechanical and 3D micro-computed tomography analyses were performed. HBMSC viability, biochemical activity and histological evidence of osteogenic cellular differentiation, post-impaction were observed in vitro and in vivo in PLA/HBMSC samples compared to impacted PLA alone. In vitro PLA/HBMSC samples demonstrated evidence of mechanical enhancement over PLA alone. In vivo studies showed a significant increase in new bone and blood vessel formation in the PLA/HBMSC constructs compared to PLA alone. With alternatives to allograft being sought, these studies have demonstrated PLA/HBMSC living composites, to be a potential prospect as a biological bone graft extender for future use in the field of IBG.


TRAUMATIC PULMONARY HERNIATION: A RARE CHEST TRAUMA MANIFESTATION.
Traumatic pulmonary hernia is an uncommon occurrence resulting from chest trauma, typically covered by the skin. Chest trauma may arise from penetrating or blunt mechanisms, with blunt trauma being more frequently observed. When lung herniation transpires, various symptoms such as chest pain, dyspnea, subcutaneous emphysema, bone crepitation, and hemoptysis (in cases of lung parenchymal damage) may manifest. We present the case of a 66-year-old woman suffering from chest pain and dyspnea after blunt chest trauma due to a fall induced by delirium following alcohol abuse.


Lymphatic drainage of the CNS: effects of lymphatic diversion/ligation on CSF protein transport to plasma.
The plasma recovery of an intraventricularly administered protein was compared before and after lymph diversion/ligation in the same conscious sheep to determine the relative roles of arachnoid villi and lymphatics in the clearance of a cerebral spinal fluid tracer. 125I-human serum albumin was injected into both lateral ventricles, and venous blood was sampled. One day later, multiple cervical vessels and the thoracic duct were cannulated for lymph collection. Uncannulated vessels were ligated. The experiment was repeated with 131I-human serum albumin as the tracer. Before lymph diversion/ligation, the time-averaged tracer transport into the plasma was 6.4 +/- 1.0%/h, with an average 6-h plasma recovery of 38.2 +/- 5.7% (percentage of injected dose). After lymph diversion/ligation, the values dropped to 2.9 +/- 0.5%/h and 17.7 +/- 2.7%, respectively. The collected lymph contained 8.7 +/- 2.6% of the tracer. No significant differences were observed in sham-operated animals. In conclusion, extracranial lymphatic vessels in sheep transport approximately one-half of the protein tracer from the cerebral spinal fluid compartment into plasma.


A multiscale coarse-grained model of the SARS-CoV-2 virion.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the COVID-19 pandemic. Computer simulations of complete viral particles can provide theoretical insights into large-scale viral processes including assembly, budding, egress, entry, and fusion. Detailed atomistic simulations are constrained to shorter timescales and require billion-atom simulations for these processes. Here, we report the current status and ongoing development of a largely "bottom-up" coarse-grained (CG) model of the SARS-CoV-2 virion. Data from a combination of cryo-electron microscopy (cryo-EM), x-ray crystallography, and computational predictions were used to build molecular models of structural SARS-CoV-2 proteins, which were then assembled into a complete virion model. We describe how CG molecular interactions can be derived from all-atom simulations, how viral behavior difficult to capture in atomistic simulations can be incorporated into the CG models, and how the CG models can be iteratively improved as new data become publicly available. Our initial CG model and the detailed methods presented are intended to serve as a resource for researchers working on COVID-19 who are interested in performing multiscale simulations of the SARS-CoV-2 virion.


Molybdenum, rhodium, and tungsten anode spectral models using interpolating polynomials with application to mammography.
Computer simulation is a convenient and frequently used tool in the study of x-ray mammography, for the design of novel detector systems, the evaluation of dose deposition, x-ray technique optimization, and other applications. An important component in the simulation process is the accurate computer-generation of x-ray spectra. A computer model for the generation of x-ray spectra in the mammographic energy range from 18 kV to 40 kV has been developed. The proposed model requires no assumptions concerning the physics of x-ray production in an x-ray tube, but rather makes use of x-ray spectra recently measured experimentally in the laboratories of the Center for Devices and Radiological Health. Using x-ray spectra measured for molybdenum, rhodium, and tungsten anode x-ray tubes at 13 different kV's (18, 20, 22, ..., 42 kV), a spectral model using interpolating polynomials was developed. At each energy in the spectrum, the x-ray photon fluence was fit using 2, 3, or 4 term (depending on the energy) polynomials as a function of the applied tube voltage (kV). Using the polynomial fit coefficients determined at each 0.5 keV interval in the x-ray spectrum, accurate x-ray spectra can be generated for any arbitrary kV between 18 and 40 kV. Each anode material (Mo, Rh, W) uses a different set of polynomial coefficients. The molybdenum anode spectral model using interpolating polynomials is given the acronym MASMIP, and the rhodium and tungsten spectral models are called RASMIP and TASMIP, respectively. It is shown that the mean differences in photon fluence calculated over the energy channels and over the kV range from 20 to 40 kV were -0.073% (sigma = 1.58%) for MASMIP, -0.145% (sigma = 1.263%) for RASMIP, and 0.611% (sigma = 2.07%) for TASMIP. The polynomial coefficients for all three models are given in an Appendix. A short C subroutine which uses the polynomial coefficients and generates x-ray spectra based on the proposed model is available on the World Wide Web at http:/(/)www.aip.org/epaps/epaps.html.


Local structure and nanoscale homogeneity of CeO2-ZrO2: differences and similarities to parent oxides revealed by luminescence with temporal and spectral resolution.
Although homogeneity at the atomic level of CeO2-ZrO2 with a Ce/Zr atomic ratio close to unity is considered to be one of the main causes for the increased total oxygen storage capacity (OSC), the characterization approaches of homogeneity remain a major challenge. We propose a simple, yet effective method, to assess both structural and compositional homogeneity of CeO2-ZrO2 by using Eu(3+) luminescence measured with time and dual spectral resolution (emission and excitation). For Eu(3+)-CeO2-ZrO2 calcined at 750 °C, the X-ray diffraction, Raman and High-Resolution Transmission Electron Microscopy data converge to a single pseudo-cubic phase. However, the evolution of Eu(3+)-delayed luminescence from cubic ceria-like to tetragonal zirconia-like emission reveals the formation of CeO2- and ZrO2-rich nanodomains and provides evidence for early phase separation. For Eu(3+)-CeO2-ZrO2 calcined at 1000 °C, the emission of Eu(3+) reveals both structural and compositional inhomogeneity. Our study identifies the differences between the local structure properties of CeO2 and ZrO2 parent oxides and CeO2-ZrO2 mixed oxide, also confirming the special chemical environment of the oxygen atoms in the mixed oxide as reported earlier by Extended X-ray Absorption Fine Structure investigations.


The cost of obesity in Hong Kong.
Obesity is now being considered a rapid growing economic burden on the healthcare system of many countries. These costs should include expenditure on prevention, as well as the investigation and treatment of obesity and its related problems. Accurate information on the cost of obesity in Hong Kong is lacking so we analysed the annual hospital discharges in a representative district hospital in Hong Kong. The estimated hospitalization costs in Hong Kong' public hospitals were HK$ 2.29 billion (US$ 0.29 billion) in 1998 and HK$ 3.36 billion (US$ 0.43 billion) in 2002 accounting for 8.2-9.8% of total public expenditure on health in Hong Kong. Based on the likelihood ratio of diabetes, hypertension and dylsipidaemia related to obesity in Hong Kong Chinese (defined as body mass index > or = 25 kg m(-2)) the estimated proportion of these conditions attributable to obesity are 10-20%. In addition, significant amounts of money are being spent by the public on obesity self management schemes of no proven benefit. In 2005, 5.5% of Hong Kong's total population was consuming proprietary drugs for obesity or its related conditions. To properly assess the overall costs of obesity, other indirect costs should also be considered such as economic losses on sick leave because of obesity and its related conditions.


MyoD cannot compensate for the absence of myogenin during skeletal muscle differentiation in murine embryonic stem cells.
myogenin (-/-) mice display severe skeletal muscle defects despite expressing normal levels of MyoD. The failure of MyoD to compensate for myogenin could be explained by distinctions in protein function or by differences in patterns of gene expression. To distinguish between these two possibilities, we compared the abilities of constitutively expressed myogenin and MyoD to support muscle differentiation in embryoid bodies made from myogenin (-/-) ES cells. Differentiated embryoid bodies from wild-type embryonic stem (ES) cells made extensive skeletal muscle, but embryoid bodies from myogenin (-/-) ES cells had greatly attenuated muscle-forming capacity. The inability of myogenin (-/-) ES cells to generate muscle was independent of endogenous MyoD expression. Skeletal muscle was restored in myogenin (-/-) ES cells by constitutive expression of myogenin. In contrast, constitutive expression of MyoD resulted in only marginal enhancement of skeletal muscle, although myocyte numbers greatly increased. The results indicated that constitutive expression of MyoD led to enhanced myogenic commitment of myogenin (-/-) cells but also indicated that committed cells were impaired in their ability to form muscle sheets without myogenin. Thus, despite their relatedness, myogenin's role in muscle formation is distinct from that of MyoD, and the distinction cannot be explained merely by differences in their expression properties.


Excitatory synaptic potentials in kainic acid-denervated rat CA1 pyramidal neurons.
Intracellular recordings were performed in the CA1 region of the rat hippocampus following an ipsilateral intraventricular injection of kainic acid. Seven days postlesion, graded bursts of up to four action potentials could be evoked by stimulation of the stratum radiatum. The evoked EPSPs underlying these bursts showed a prolonged 10-90% rise time and half-width compared to control EPSPs, an absence of a significant inhibitory phase, and an increase in magnitude and duration at depolarized resting levels. The evoked EPSPs also exhibited a significant decrease in amplitude and time course in response to D-APV (D-2-amino-5-phosphonovaleric acid; 1-20 microM), though this effect was variable from cell to cell. The prolonged time course, voltage sensitivity, and response to a selective NMDA antagonist confirmed that the major component of the EPSP in neurons from lesioned slices was mediated by NMDA receptors. The partial denervation of the CA1 area induced by the kainic acid led to both an enhanced NMDA-mediated excitatory phase and a decrease in postsynaptic inhibition, resulting in the pronounced hyperexcitability noted in the lesioned slices.


Platelet-Inspired Nanocells for Targeted Heart Repair After Ischemia/Reperfusion Injury.
Cardiovascular disease is the leading cause of mortality worldwide. While reperfusion therapy is vital for patient survival post-heart attack, it also causes further tissue injury, known as myocardial ischemia/reperfusion (I/R) injury in clinical practice. Exploring ways to attenuate I/R injury is of clinical interest for improving post-ischemic recovery. A platelet-inspired nanocell (PINC) that incorporates both prostaglandin E2 (PGE2)-modified platelet membrane and cardiac stromal cell-secreted factors to target the heart after I/R injury is introduced. By taking advantage of the natural infarct-homing ability of platelet membrane and the overexpression of PGE2 receptors (EPs) in the pathological cardiac microenvironment after I/R injury, the PINCs can achieve targeted delivery of therapeutic payload to the injured heart. Furthermore, a synergistic treatment efficacy can be achieved by PINC, which combines the paracrine mechanism of cell therapy with the PGE2/EP receptor signaling that is involved in the repair and regeneration of multiple tissues. In a mouse model of myocardial I/R injury, intravenous injection of PINCs results in augmented cardiac function and mitigated heart remodeling, which is accompanied by the increase in cycling cardiomyocytes, activation of endogenous stem/progenitor cells, and promotion of angiogenesis. This approach represents a promising therapeutic delivery platform for treating I/R injury.


A Model for Incident Review Committees in Behavioral Health Settings.
Despite the fact that incident review committees have been a key component of quality improvement in behavioral health settings for decades, specific models of how these committees are structured and operate are not well described. We present a model for an incident review committee that has been implemented in 2 large, academic acute care psychiatric hospitals. We believe the model not only permitted us to efficiently and effectively review untoward incidents, but that it also provided an approach to calibrating standards of care for the institution, engaging physicians in an interdisciplinary effort, promulgating a culture of quality review and improvement throughout the organization, promoting continuity and sustainability of the incident review process, and, most importantly, driving beneficial change in clinical practice. Demonstration of the effectiveness of this model requires formal investigation.


Ge-rich SiGe-on-insulator for waveguide optical modulator application fabricated by Ge condensation and SiGe regrowth.
We have numerically analyzed plasma dispersion effect in a Ge-rich SiGe layer for optical modulator applications. Since strain induces reduction in effective masses of electron and hole, we expect enhanced plasma dispersion effect in a strained Ge-rich SiGe layer. The plasma dispersion effects of Si(0.15)Ge(0.85) on Si(0.2)Ge(0.8) for hole and electron are expected to be approximately 3.0 and 1.5 times larger than those of Si. To realize Ge-rich SiGe-based waveguide optical modulators, we have also investigated the fabrication procedure of SiGe-on-insulator (SGOI) wafers. We have successfully fabricated Ge-rich SGOI wafers without any thick SiGe buffer layers by using Ge condensation in conjunction with the SiGe regrowth technique. We have evaluated the SGOI by Raman spectroscopy, atomic force microscopy (AFM), reflected high energy electron diffraction (RHEED) and transmission electron microscopy (TEM). Ge-rich SiGe waveguides have been fabricated on the SGOI wafer. The propagation loss was found to be approximately 13 dB/mm, which can be reduced to be below 2 dB/mm by optimizing the Ge condensation process. We expect that strained SiGe grown on the fabricated SGOI exhibits more than 2.3 times higher plasma dispersion than Si in case of a carrier injection type, suitable for high-performance waveguide optical modulators.


Cerebrovascular arterial dissection in children and young adults.
Dissection of the cervical and intracranial portions of the carotid and vertebrobasilar arteries may occur spontaneously or in association with trauma or other underlying risk factors. Astute recognition allows appropriate diagnostic and therapeutic protocols to be instituted. Dissection of extracranial carotid and vertebral arteries differ from their intracranial counterparts, both in terms of presentation and prognosis. Limited data are available regarding the most effective medical treatment of dissection in children and young adults. Interventional neuroradiology may offer a new approach in difficult cases. A multicenter controlled clinical trial is needed to assess the most effective treatment.


A review of telemedicine cost-effectiveness studies.
As interest in telemedicine grows, many of its proponents and vendors increasingly suggest that it is now time to move to full-scale implementation of telemedicine services in a variety of contexts throughout the world, and question the need for further evidence of its utility and cost-effectiveness. We have reviewed the published literature relating to cost-effectiveness studies in telemedicine and have identified some important weaknesses. Ten recommendations regarding the design of economic evaluations of telemedicine are suggested.


Attenuation of pro-inflammatory cytokines and oxidative stress by misoprostol in renal ischemia/reperfusion in rats.
The ischemia/reperfusion (I/R) process alters metabolic pathways, releasing reactive oxygen species and pro-inflammatory cytokines that cause tissue necrosis and activate cellular apoptotic pathways. Misoprostol (MSP) is a prostaglandin E1 analog that has demonstrated a cytoprotective role in the I/R process. The study objective was to evaluate the effects of MSP on the regulation of pro-inflammatory and oxidative stress mediators in an I/R-induced acute kidney injury rat model. Wistar rats were divided into 3 groups. Sham and I/R were given 1 mL/day of physiological solution; MSP+I/R was given intragastric MSP (300 μg/kg) for 3 days. For I/R and MSP+IR, the renal hilum was clamped for 45 min, followed by 15 h of reperfusion. Renal function tests, pro-inflammatory cytokines, mediators of oxidative stress, and histological analysis were evaluated. Pro-inflammatory cytokine activity was significantly attenuated in the MSP+I/R group. However, there was no statistically significant difference between Sham and MSP. Regarding antioxidant activity, MSP+I/R showed a significant decrease in these mediators compared with Sham and I/R. Histologically, scarce medullary necrosis was observed with a preserved renal cortex in the MSP group.


Membrane cytochromes of Escherichia coli chl mutants.
The cytochromes present in the membranes of Escherichia coli cells having defects in the formate dehydrogenase-nitrate reductase system have been analyzed by spectroscopic, redox titration, and enzyme fractionation techniques. Four phenotypic classes differing in cytochrome composition were recognized. Class I is represented by strains with defects in the synthesis or insertion of molybdenum cofactor. Cytochromes of the formate dehydrogenase-nitrate reductase pathway are present. Class II strains map in the chlC-chlI region. The cytochrome associated with nitrate reductase (cytochrome bnr) is absent in these strains, whereas that associated with formate dehydrogenase (cytochrome bfdh) is the major cytochrome in the membranes. Class III strains lack both cytochromes bfdh and bnr but overproduce cytochrome d of the aerobic pathway even under anaerobic conditions in the presence of nitrate. Class III strains have defects in the regulation of cytochrome synthesis. An fdhA mutant produced cytochrome bnr but lacked cytochrome bfdh. These results support the view that chlI (narI) is the structural gene for cytochrome bnr and that chlC (narG) and chlI(narI) are in the same operon, and they provide evidence of the complexity of the regulation of cytochrome synthesis.


The role of estrogen deficiency in skin ageing and wound healing.
The links between hormonal signalling and lifespan have been well documented in a range of model organisms. For example, in C. elegans or D. melanogaster, lifespan can be modulated by ablating germline cells, or manipulating reproductive history or pregnenolone signalling. In mammalian systems, however, hormonal contribution to longevity is less well understood. With increasing age human steroid hormone profiles change substantially, particularly following menopause in women. This article reviews recent links between steroid sex hormones and ageing, with special emphasis on the skin and wound repair. Estrogen, which substantially decreases with advancing age in both males and females, protects against multiple aspects of cellular ageing in rodent models, including oxidative damage, telomere shortening and cellular senescence. Estrogen's effects are particularly pronounced in the skin where cutaneous changes post-menopause are well documented, and can be partially reversed by classical Hormone Replacement Therapy (HRT). Our research shows that while chronological ageing has clear effects on skin wound healing, falling estrogen levels are the principle mediator of these effects. Thus, both HRT and topical estrogen replacement substantially accelerate healing in elderly humans, but are associated with unwanted deleterious effects, particularly cancer promotion. In fact, much current research effort is being invested in exploring the therapeutic potential of estrogen signalling manipulation to reverse age-associated pathology in peripheral tissues. In the case of the skin the differential targeting of estrogen receptors to promote healing in aged subjects is a real therapeutic possibility.


Emergence of rifampin-resistant Rhodococcus equi in an infected foal.
To investigate the emergence of rifampin resistance in Rhodococcus equi strains isolated from foals and their environment in Japan, we compared the in vitro antimicrobial susceptibilities to rifampin of 640 isolates from 64 infected foals and 98 soil isolates from their horse-breeding farms. As a control, 39 human isolates from patients with and without AIDS were also tested for susceptibility to rifampin. All of the isolates showed rifampin sensitivity, except isolates from one infected foal and two patients with AIDS that showed rifampin resistance. To investigate the emergence of rifampin-resistant R. equi in the infected foal, which had received rifampin monotherapy for a month before euthanasia, 99 isolates of R. equi from the lesions and 20 isolates from the intestinal contents of the one foal with rifampin-resistant organisms were analyzed for rifampin susceptibilities, pathogenicities, and ribotypes. Of the 99 isolates from the lesions, all of which were virulent R. equi strains containing a virulence plasmid with a size of 85 or 90 kb, 90 (91%) isolates were rifampin resistant (MIC, > or = 12.5 microg/ml). On the other hand, of the 20 isolates from the intestinal contents, 11 (55%) isolates showed rifampin resistance (MIC, > or = 25 microg/ml), and 5 of them were avirulent R. equi strains. Among these 101 rifampin-resistant R. equi isolates with and without virulence plasmids characterized by ribotyping, 58 were type I, 20 were type II, 11 were type III, and 12 were type IV. These results demonstrated that at least eight different rifampin-resistant R. equi strains emerged concurrently and respectively from the different lesions and intestinal contents of the infected foal.


[Therapeutic problems in disorders of sex development].
The compatibility between genetic, gonadal, genital, somatic and psychic sex should be present for the proper sexual development. If there is no such compatibility, disorders of sex development (DSD) appear. Medical procedure in such cases leads to many problems which mainly come from the lack of sufficient knowledge about the pathophysiology of the disorders. The main difficulties met by diagnostic and therapeutic team are: determination of the official sex, prediction of gender identity, hormonal activity of gonads and fertility, as well as the decision to undertake surgical procedures involving the genitals and gonads. Disorders of sex development lead also to psychological problems of patients and their families, because they disturb the proper social functioning.


Targeted delivery of curcumin for treating type 2 diabetes.
Type 2 diabetes is a chronic condition in which cells have reduced insulin signalling, leading to hyperglycemia and long-term complications, including heart, kidney and liver disease. Macrophages activated by dying or stressed cells, induce the transcription factor nuclear factor kappa-B leading to the production of pro-inflammatory cytokines including TNF and IL-6. These inflammatory macrophages in liver and adipose tissue promote insulin resistance, and medications which reduce inflammation and enhance insulin signalling improve glucose control. Curcumin is an anti-oxidant and nuclear factor kappa-B inhibitor derived from turmeric. A number of studies have shown that dietary curcumin reduces inflammation and delays or prevents obesity-induced insulin resistance and associated complications, including atherosclerosis and immune mediate liver disease. Unfortunately dietary curcumin is poorly absorbed by the digestive system and undergoes glucuronidation and excretion rather than being released into the serum and systemically distributed. This confounds understanding of how dietary curcumin exerts its beneficial effects in type 2 diabetes and associated diseases. New improved methods of delivering curcumin are being developed including nanoparticles and lipid/liposome formulations that increase absorption and bioavailability of curcumin. Development and refinement of these technologies will enable cell-directed targeting of curcumin and improved therapeutic outcome.


[Hyperphenylalaninaemia with normal phenylalanine-hydroxylase activity and a deficiency of tetrahydrobiopterin and dihydropteridine reductase].
Two cases of hyperphenylalaninemia with a normal activity of phenylalanine hydroxylase are described. No activity of DHP reductase was found in the first case, having very high biopterin levels in basal conditions and after intravenous perfusion of phenylalanine. In the other case, the DHP reductase activity is normal but plasma and urinary levels of the reduced forms of biopterin are largely lowered and do not increase during the phenylalanine load. Early substitutive treatment with L-dopa and 5-HTP in one of the cases avoided the development of the "progressive neurological illness unresponsive to dietary treatment" characterizing two variants. This raises the question of a liver biopsy in order to assay the hydroxylation enzyme activities when screening hyperphenylalaninemia whatever the type.


Hemoglobin levels and coronary artery disease.
Anemia is a risk factor for adverse cardiovascular disease outcomes. However, there is limited information concerning the association of hemoglobin concentration and new onset of clinically recognized coronary artery disease (CAD). An historical cohort study was conducted with patients from Veterans Affairs medical centers. Baseline hemoglobin determinations were evaluated with respect to CAD using data from records of 25,622 subjects with no known heart disease. Coronary artery disease was identified from a new diagnosis based on the International Classification of Diseases, Ninth Edition, coding or a new prescription for nitroglycerin. Models were adjusted for age, sex, body mass index, smoking, systolic blood pressure, diastolic blood pressure, fasting glucose, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, creatinine clearance, and use of statin or beta-blocker. Among the cohort, 5297 (20.7%) subjects developed CAD over 73,895 person-years of follow-up. Compared with control hemoglobin levels of 15.0 to 17.0 g/dL, the multivariable-adjusted risk of CAD increased with lower hemoglobin levels: an adjusted hazard ratio (HR) of 1.47 and 95% confidence interval (CI) of 1.18 to 1.84 for hemoglobin levels of 9.0 to 11.0 g/dL; an HR of 1.34 and 95% CI of 1.20 to 1.49 for 11.0 to 13.0 g/dL; and an HR of 1.07 and 95% CI of 1.01 to 1.13 for 13.0 to 15.0 g/dL. Hemoglobin levels > or = 17.0 g/dL were also associated with increased risk for CAD (adjusted HR 1.22, 95% CI 1.08-1.37). Hemoglobin levels > or = 17 or < 15 g/dL are independently associated with increase risk for new cardiac events.


Deoxycoformycin (pentostatin): clinical pharmacology, role in the chemotherapy of cancer, and use in other diseases.
Pentostatin (2'-deoxycoformycin, dCF) is a purine nucleoside analog and a product of the fermentation of Streptomyces antibioticus. It is a tight-binding inhibitor of adenosine deaminase (ADA), an enzyme essential in the cellular metabolism of purines. Children with congenital absence of ADA suffer from atrophy of lymphoid tissues and severe combined immune deficiency (SCID) syndrome. It was hypothesized that pentostatin would be lymphocytotoxic and this proved to be true; this finding prompted its investigation in lymphoid neoplasms. It was anticipated that pentostatin would be most active in neoplasms with high intracellular concentrations of ADA, e.g. acute lymphocytic leukemia (ALL), particularly of the T-cell variety. Although pentostatin proved to be active in ALL, large doses were required and major toxic effects outweighed therapeutic benefits. By contrast, pentostatin proved to be exceptionally active in hairy cell leukemia (HCL), a B-cell neoplasm with low intracellular concentrations of ADA. Pentostatin has since been shown to possess activity in chronic lymphocytic leukemia, prolymphocytic leukemia, cutaneous T-cell lymphomas, adult T-cell lymphoma-leukemia, and low grade non-Hodgkin's lymphomas. It potentiates the activity of vidarabine against viruses and against the cells of acute myeloid leukemia. Pentostatin is inactive in melanoma and renal carcinoma, but has not been adequately evaluated in other solid tumors. The toxic effects of pentostatin include renal failure, central nervous system (CNS) depression, immunosuppresion, keratoconjunctivitis, and opportunistic infections. In the absence of pre-existing bone marrow compromise, pentostatin produces only mild myelosuppression. Aside from its use as an antineoplastic agent, pentostatin has potential applications as an immunosuppressive drug, as an antiviral agent, as an antimalarial compound, and in the protection of cells of the CNS from damage induced by ischemia and anoxia. Clinical studies with pentostatin are ongoing, and its roles in the management of neoplastic and non-neoplastic diseases have yet to be fully defined.


Psychosocial impact of hidradenitis suppurativa: a qualitative study.
Hidradenitis suppurativa influences patients' lives in many ways. It is therefore necessary to focus on the effects of the disease on daily life in order better to define patient-related outcomes in hidradenitis suppurativa studies. Interviews were conducted with 12 patients with hidradenitis suppurativa. Initial single interviews were followed by semi-structured and structured qualitative focus group interviews in order to improve the richness of the data and obtain in-depth understanding of the impact of the topics. Important topics were found to relate to aspects of interpersonal contact, especially in relation to smell and appearance, various emotional reactions, and feelings of lack of control. It was found that hidradenitis suppurativa has a great emotional impact on patients and promotes isolation due to fear of stigmatization. Shame and irritation are frequent and relate to smell, scars, itching and pain. Quality of life is adversely affected and professional support is needed.


Literature Review: Herbal Medicine Treatment after Large-Scale Disasters.
Large-scale natural disasters, such as earthquakes, tsunamis, volcanic eruptions, and typhoons, occur worldwide. After the Great East Japan earthquake and tsunami, our medical support operation's experiences suggested that traditional medicine might be useful for treating the various symptoms of the survivors. However, little information is available regarding herbal medicine treatment in such situations. Considering that further disasters will occur, we performed a literature review and summarized the traditional medicine approaches for treatment after large-scale disasters. We searched PubMed and Cochrane Library for articles written in English, and Ichushi for those written in Japanese. Articles published before 31 March 2016 were included. Keywords "disaster" and "herbal medicine" were used in our search. Among studies involving herbal medicine after a disaster, we found two randomized controlled trials investigating post-traumatic stress disorder (PTSD), three retrospective investigations of trauma or common diseases, and seven case series or case reports of dizziness, pain, and psychosomatic symptoms. In conclusion, herbal medicine has been used to treat trauma, PTSD, and other symptoms after disasters. However, few articles have been published, likely due to the difficulty in designing high quality studies in such situations. Further study will be needed to clarify the usefulness of herbal medicine after disasters.


An integrated membrane system for the biocatalytic production of 3'-sialyllactose from dairy by-products.
An integrated membrane system was investigated for the production of 3'-sialyllactose by an engineered sialidase using casein glycomacropeptide (CGMP) and lactose as substrates. CGMP was purified by ultrafiltration (UF) to remove any small molecules present and then an enzymatic membrane reactor (EMR) was used to separate the product and reuse the enzyme. A PLCC regenerated cellulose membrane was found to be the most suitable for both the UF purification and EMR. Subsequently, nanofiltration (NF) was conducted to increase the purity of the 3'-sialyllactose by removing the excess lactose present. The NTR7450 membrane outperformed others in NF due to its high retention of 3'-sialyllactose (98%) and relatively low rejection of lactose (40%). The lactose in the permeate could be concentrated by the NF45 membrane and recycled into the EMR. The described integrated membrane system enables a more economic and efficient enzymatic production of 3'-sialyllactose.


Aliiroseovarius marinus sp. nov., isolated from seawater.
A novel Gram-stain-negative, rod-shaped, non-spore-forming, non-flagellated, strictly aerobic strain, designated A6024T, was isolated from coastal seawater near Rizhao, PR China (119.61° E 35.47° N). The organism grew optimally at 28 °C, in pH 6.0-7.0 and in the presence of 3.0 % (w/v) NaCl. The strain required seawater or artificial seawater for growth and NaCl alone did not support growth. Strain A6024T contained ubiquinone 10 as the sole respiratory quinone and C18 : 1 ω7c (75.2 %) as the most abundant fatty acid. The predominant polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, one unidentified aminolipid and two unidentified lipids. The DNA G+C content of strain A6024T was 59.9 mol%. The results of phylogenetic analysis based on 16S rRNA gene sequences showed that the novel strain was related most closely to Aliiroseovarius halocynthiae MA1-10T, Aliiroseovarius pelagivivens GYSW-22T and Aliiroseovarius crassostreae CV919-312T with 98.3, 97.6 and 97.4 % sequence similarities, respectively. The calculated average nucleotide identity values and DNA-DNA hybridization values between strain A6024T and the phylogenetically related Aliiroseovarius species were in the range 76.0-85.6 % and 19.6-29.4 %, respectively. On the basis of the results of polyphasic analyses, strain A6024T represents a novel species of the genus Aliiroseovarius, for which the name Aliiroseovarius marinus sp. nov. is proposed. The type strain is A6024T (=KCTC 72114T=MCCC 1K03595T).


Vulnerabilities in mIDH2 AML confer sensitivity to APL-like targeted combination therapy.
Although targeted therapies have proven effective and even curative in human leukaemia, resistance often ensues. IDH enzymes are mutated in ~20% of human AML, with targeted therapies under clinical evaluation. We here characterize leukaemia evolution from mutant IDH2 (mIDH2)-dependence to independence identifying key targetable vulnerabilities of mIDH2 leukaemia that are retained during evolution and progression from early to late stages. Mechanistically, we find that mIDH2 leukaemia are metastable and vulnerable at two distinct levels. On the one hand, they are characterized by oxidative and genotoxic stress, in spite of increased 1-carbon metabolism and glutathione levels. On the other hand, mIDH2 leukaemia display inhibition of LSD1 and a resulting transcriptional signature of all-trans retinoic acid (ATRA) sensitization, in spite of a state of suppressed ATRA signalling due to increased levels of PIN1. We further identify GSH/ROS and PIN1/LSD1 as critical nodes for leukaemia maintenance and the combination of ATRA and arsenic trioxide (ATO) as a key therapeutic modality to target these vulnerabilities. Strikingly, we demonstrate that the combination of ATRA and ATO proves to be a powerfully synergistic and effective therapy in a number of mouse and human mIDH1/2 leukemic models. Thus, our findings pave the way towards the treatment of a sizable fraction of human AMLs through targeted APL-like combinatorial therapies.


Hepatoprotective activity of raspberry ketone is mediated via inhibition of the NF-κB/TNF-α/caspase axis and mitochondrial apoptosis in chemically induced acute liver injury.
Raspberry Ketone (RK) is a natural phenolic compound which is marketed nowadays as a popular weight-reducing remedy, with reported antioxidant and anti-inflammatory activities. However, its biological activity is not fully elucidated. Hepatotoxicity is the leading cause of acute liver failure in Europe and North America, and its management is still challenging. Therefore, this study aimed to assess the therapeutic detoxification activity of RK against liver injury in vivo and to explore the underlying mechanisms using carbon tetrachloride (CCl4)-induced hepatotoxicity as a model. First, a dose-response study using 4 different doses, 25, 50, 100, and 200 mg kg-1 day-1, of RK was conducted. RK was administered for 5 days as a pretreatment, followed by a single dose of CCl4 (1 ml kg-1, 1 : 1 v/v CCl4 : olive oil). The RK dose of 200 mg kg-1 showed the greatest protective effect and was selected for further investigations. CCl4 hepatotoxicity was confirmed by elevation of liver enzymes, and histopathological examination. CCl4-induced oxidative stress was evident from increased lipid peroxidation measured as thiobarbituric acid reactive substances (TBARS) along with depleted superoxide dismutase (SOD), reduced glutathione (GSH), and total antioxidant capacity (TAC). Increased oxidative stress was associated with increased cytochrome c expression with subsequent activation of caspase-9 and caspase-3, in addition to DNA fragmentation reflecting apoptosis. CCl4 also induced the expression of inflammatory cytokines (NF-κB and TNF-α). Interestingly, RK hepatoprotective activity was evident from the reduction of liver enzymes, and maintenance of hepatocyte integrity and microstructures as evaluated by histopathological examination using H and E, and transmission electron microscopy. The antioxidant activity of RK was demonstrated by the increase of TAC, SOD, and GSH, with a concomitant decrease of the TBARS level. Moreover, RK pretreatment inhibited CCl4-induced upregulation of inflammatory mediators. RK antiapoptotic activity was indicated by the reduction of the expression of cytoplasmic cytochrome-C, a decrease of caspases, and inhibition of DNA fragmentation. In conclusion, this study demonstrates that RK is a promising hepatoprotective agent. The underlying mechanisms include antioxidant, anti-inflammatory, and anti-apoptotic activities. This is the first study reporting RK hepatoprotective activity in acute hepatic injury and approves its antiapoptotic effect in the liver.


Development of cardiac troponin-I biosensor based on boron nitride quantum dots including molecularly imprinted polymer.
The cardiac Troponin-I (cTnI) is one of the subunits of cardiac troponin complexes and a pivotal biochemical marker of acute myocardial infarction (AMI). Due to its myocardial specificity, cTnI is widely used for the diagnosis of AMI diseases. In this study, a novel imprinted biosensor approach based on boron nitride quantum dots (BNQDs) was presented for cTnI detection in plasma samples. Various characterization methods such as scanning electron microscope (SEM), transmission electron microscope (TEM), x-ray diffraction (XRD), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used for all characterizations of nanomaterials. After the characterization analysis, cTnI imprinted electrode was developed in the presence of 100.0 mM pyrrole containing 25.0 mM cTnI. After that, the analytical studies of cTnI in plasma samples were performed by using cTnI imprinted biosensor. The results of the study have revealed that 0.01-5.00 ng mL-1 and 0.0005 ng mL-1 were found as the linearity range and the detection limit (LOD). Moreover, the selectivity of cTnI imprinted glassy carbon electrode (GCE) was investigated for plasma sample analysis in the presence of other nonspecific and specific proteins including cardiac myoglobin (MYG), bovine serum albumin (BSA) and cardiac troponin T (cTnT), respectively. Furthermore, the prepared biosensor was examined in terms of stability, repeatability, reproducibility and reusability. Finally, the imprinted biosensor was applied to the plasma samples having high recovery.


Examination of the factor structure of the Schizotypal Personality Questionnaire among British and Trinidadian adults.
Much debate in schizotypal research has centred on the factor structure of the Schizotypal Personality Questionnaire (SPQ), with research variously showing higher-order dimensionality consisting of two to seven dimensions. In addition, cross-cultural support for the stability of those factors remains limited. Here, we examined the factor structure of the SPQ among British and Trinidadian adults. Participants from a White British subsample (n = 351) resident in the UK and from an African Caribbean subsample (n = 284) resident in Trinidad completed the SPQ. The higher-order factor structure of the SPQ was analysed through confirmatory factor analysis, followed by multiple-group analysis for the model of best fit. Between-group differences for sex and ethnicity were investigated using multivariate analysis of variance in relation to the higher-order domains. The model of best-fit was the four-factor structure, which demonstrated measurement invariance across groups. Additionally, these data had an adequate fit for two alternative models: (a) 3-factor and (b) modified 4-factor model. The British subsample had significantly higher scores across all domains than the Trinidadian group, and men scored significantly higher on the disorganised domain than women. The four-factor structure received confirmatory support and, importantly, support for use with populations varying in ethnicity and culture.


RegPredict: an integrated system for regulon inference in prokaryotes by comparative genomics approach.
RegPredict web server is designed to provide comparative genomics tools for reconstruction and analysis of microbial regulons using comparative genomics approach. The server allows the user to rapidly generate reference sets of regulons and regulatory motif profiles in a group of prokaryotic genomes. The new concept of a cluster of co-regulated orthologous operons allows the user to distribute the analysis of large regulons and to perform the comparative analysis of multiple clusters independently. Two major workflows currently implemented in RegPredict are: (i) regulon reconstruction for a known regulatory motif and (ii) ab initio inference of a novel regulon using several scenarios for the generation of starting gene sets. RegPredict provides a comprehensive collection of manually curated positional weight matrices of regulatory motifs. It is based on genomic sequences, ortholog and operon predictions from the MicrobesOnline. An interactive web interface of RegPredict integrates and presents diverse genomic and functional information about the candidate regulon members from several web resources. RegPredict is freely accessible at http://regpredict.lbl.gov.


Effect of radioactive iodine therapy on cytokine production in Graves' disease: transient increases in interleukin-4 (IL-4), IL-6, IL-10, and tumor necrosis factor-alpha, with longer term increases in interferon-gamma production.
Spontaneous and mitogen-stimulated production of interleukin-4 (IL-4), IL-6, IL-10, IL-12, interferon-gamma (IFNgamma), and tumor necrosis factor-alpha were evaluated by enzyme-linked immunospot assay of peripheral blood mononuclear cells from patients with Graves' disease immediately before and at 4, 17, and 59 days after treatment with radioactive iodine. Patients had significantly reduced IL-4 and IFNgamma production before treatment compared with healthy controls. Both cytokines were increased to normal levels by day 17 after treatment, and IFNgamma remained at normal levels on day 59, whereas IL-4 returned to subnormal levels at this time. IL-12 production was initially normal and was not significantly altered by therapy. IL-6, IL-10, and tumor necrosis factor-alpha were also normal before radiotherapy, but increased significantly on day 17, returning to pretreatment levels by day 59. Thus, radioiodine treatment induced a transient increase in both proinflammatory and antiinflammatory cytokines and a more prolonged increase in IFNgamma production, the latter representing a definite shift toward a type 1 cytokine profile.


Testing for Within × Within and Between × Within Moderation using Random Intercept Cross-Lagged Panel Models.
Random-Intercept Cross-Lagged Panel Models allow for the decomposition of measurements into between- and within-person components and have hence become popular for testing developmental hypotheses. Here, we describe how developmental researchers can implement, test and interpret interaction effects in such models using an empirical example from developmental psychopathology research. We illustrate the analysis of Within × Within and Between × Within interactions utilising data from the United Kingdom-based Millennium Cohort Study within a Bayesian Structural Equation Modelling framework. We provide annotated Mplus code, allowing users to isolate, estimate and interpret the complexities of within-person and between person dynamics as they unfold over time.


How to successfully influence laboratory test utilization.
In 1994, we discovered that our hospital was higher than the 75th percentile in the average number of laboratory tests performed per inpatient discharge compared with other teaching hospitals with a case mix index of greater than 1.35. Because of the diversity and volume of tests offered, no single project would have significantly affected the number of tests ordered per patient. Pathologists together with the administrative director and section managers identified areas of potential improvement, noting tests that were high volume, expensive, difficult to perform, or of questionable medical benefit. We relied on a combination of administrative changes and physician education initiatives to influence physician test-ordering behavior. The impact of these initiatives was measured by reviewing monthly revenue and usage reports before and after changes were implemented. Each of these initiatives helped to gradually decrease the average number of inpatient tests per discharge from 44 in the first quarter of 1994 to 29 in the third quarter of 1999. Compared with our peer group, our hospital's rank has steadily improved from the 75th to the 15th percentile.


Electrophysiological and behavioral effects of group III metabotropic glutamate receptors on pallidal neurons in normal and parkinsonian rats.
The globus pallidus plays a critical role in movement regulation. Glutamate being an important excitatory neurotransmitter modulates the activity of pallidal neurons through both ionotropic and metabotropic glutamate receptors (mGluRs). Morphological studies have shown that group III mGluRs are generally located presynaptically in the globus pallidus. Up to now, little is known about the in vivo electrophysiological effects of group III mGluRs on the pallidal neurons. This study investigated the electrophysiological and behavioral effects of group III mGluRs on pallidal neurons in both normal and 6-hydroxydopamine (6-OHDA) lesioned parkinsonian rats. Micropressure ejection of group III mGluR agonist, L-2-amino-4-phosphonobutyrate (L-AP4), increased or decreased the firing rate of pallidal neurons in both normal and parkinsonian rats. The L-AP4-induced excitatory effects on the lesioned side of parkinsonian rats (117.4 ± 17.2%) were stronger than that in normal rats (64.3 ± 10.1%). While the proportion of neurons that were unresponsive to L-AP4 on the lesioned side of parkinsonian rats (50%) was more than that of normal rats (13%). Unilateral microinjection of L-AP4 into the globus pallidus induced a contralateral dystonic posturing in the presence of systemic haloperidol administration. The selective group III mGluRs antagonist, (RS)-α-cyclopropyl-4-phosphonophenylglycine, had no effect on pallidal neurons when used alone and could block both L-AP4-induced electrophysiological and behavioral effects. Combining electrophysiological and behavioral findings, we concluded that activation of group III mGluRs modulate the activity of pallidal neurons under both normal and parkinsonian state.


Reclassification of Bisgaard taxon 37 and taxon 44 as Psittacicella melopsittaci gen. nov., sp. nov., Psittacicella hinzii sp. nov. and Psittacicella gerlachiana sp. nov. within Psittacicellaceae fam. nov. of the order Pasteurellales.
Bacteria isolated from lesions as well as apparently normal tissues of psittacine birds have previously been reported as taxon 37 and taxon 44 of Bisgaard. 16S rRNA gene sequence comparisons revealed a distant relationship to members of Pasteurellaceae at the species, genus and family levels. The polar lipid profile consisted of the major components phosphatidylethanolamine and phosphatidylglycerol. A new family Psittacicellaceae fam. nov. is proposed with the type genus Psittacicella gen. nov. The new genus Psittacicella includes the type species Psittacicella melopsittaci sp. nov. with type strain B96/4T (=CCUG 70858T=DSM 105476T), Psittacicella hinzii sp. nov. with type strain 111T (=CCUG 52861T=CCM 8842T) and Psittacicella gerlachiana sp. nov. with type strain EEAB3T1T (=CCUG 70857T=DSM 105477T). In addition to the major polar lipids, strain 111T possessed the non-identified aminophospholipids APL1 and APL2 and trace amounts of four lipids (L1-L4) whereas strain B94/4T showed the minor unidentified aminophospholipids APL3 and APL2 and trace amounts of unidentified lipid L3. These results demonstrate that strain B96/4T can be distinguished from 111T based on presence/absence of the unidentified lipids APL1 and APL3. The total polar lipid profile of strain EEAB3T1T differed from B96/4Tonly in one minor lipid. Strain B96/4T can further be distinguished from 111T by acid formation from trehalose and raffinose and the α-glucosidase test. Strains 111T and EEAB3T1T can be separated based on acid formation from trehalose and the α-glucosidase test. Strains B96/4T and EEAB3T1T can be separated by acid formation from raffinose and eight signature indels in the RpoB protein.


The COVID-19 crisis silver lining: interprofessional education to guide future innovation.
Globally, the advent and rapid spread of the COVID-19 virus has created significant disruption to health professions education and practice, and consequently interprofessional education, leading to a model of learning and practicing where much is unknown. Key questions for this ongoing evolution emerge for the global context leading to reflections on future directions for the interprofessional education field and its role in shaping future practice models. Health professions programs around the world have made a dramatic shift to virtual learning platforms in response to closures of academic institutions and restrictions imposed on learners accessing practice settings. Telemedicine, slow to become established in many countries to date, has also revolutionized practice in the current environment. Within the state of disruption and rapid change is the awareness of a silver lining that provides an opportunity for future growth. Key topics explored in this commentary include reflection on the application of existing competency frameworks, consideration of typology of team structures, reconsideration of theoretical underpinnings, revisiting of core dimensions of education, adaptation of interprofessional education activities, and the role in the future pandemic planning. As an international community of educators and researchers, the authors consider current observations relevant to interprofessional education and practice contexts and suggest a response from scholarship voices across the globe. The current pandemic offers a unique opportunity for educators, practitioners, and researchers to retain what has served interprofessional education and practice well in the past, break from what has not worked as well, and begin to imagine the new.


Differential effects of platelets and platelet inhibition by ticagrelor on TLR2- and TLR4-mediated inflammatory responses.
Platelets and platelet-monocyte interaction play an important role in inflammation. Both pro- and anti-inflammatory effects of platelet inhibition have been reported in animal models. This study aimed to investigate the effect of platelets and platelet inhibition by the new P2Y12 receptor antagonist ticagrelor on monocyte function, as assessed by cytokine responses to Toll-like Receptor (TLR) ligands. In a set of in vitro experiments, peripheral blood mononuclear cells (PBMC) incubated with the TLR2 ligand Pam3CSK4 produced less cytokines in the presence of platelets, whereas platelets increased the production of cytokines when PBMC were exposed to TLR4 ligand lipopolysaccharide (LPS). These effects of platelets were dependent on direct platelet-leukocyte aggregation and for the Pam3CSK4-induced response, on phagocytosis of platelets by monocytes. In a double blind, placebo-controlled crossover trial in healthy volunteers, a single oral dosage of 180 mg ticagrelor reduced platelet-monocyte complex (PMC) formation. This was associated with an increase in pro-inflammatory cytokines in blood exposed to Pam3CSK4, but a decrease in these cytokines in blood exposed to LPS. These findings show that platelets differentially modulate TLR2- and TLR4-mediated cytokine responses of PBMC. Through inhibition of platelet-leukocyte interaction, P2Y12 receptor antagonists may either exert a pro- or anti-inflammatory effect during infections depending on the TLR primarily involved.


Comorbidity measures for use with administrative data.
This study attempts to develop a comprehensive set of comorbidity measures for use with large administrative inpatient datasets. The study involved clinical and empirical review of comorbidity measures, development of a framework that attempts to segregate comorbidities from other aspects of the patient's condition, development of a comorbidity algorithm, and testing on heterogeneous and homogeneous patient groups. Data were drawn from all adult, nonmaternal inpatients from 438 acute care hospitals in California in 1992 (n = 1,779,167). Outcome measures were those commonly available in administrative data: length of stay, hospital charges, and in-hospital death. A comprehensive set of 30 comorbidity measures was developed. The comorbidities were associated with substantial increases in length of stay, hospital charges, and mortality both for heterogeneous and homogeneous disease groups. Several comorbidities are described that are important predictors of outcomes, yet commonly are not measured. These include mental disorders, drug and alcohol abuse, obesity, coagulopathy, weight loss, and fluid and electrolyte disorders. The comorbidities had independent effects on outcomes and probably should not be simplified as an index because they affect outcomes differently among different patient groups. The present method addresses some of the limitations of previous measures. It is based on a comprehensive approach to identifying comorbidities and separates them from the primary reason for hospitalization, resulting in an expanded set of comorbidities that easily is applied without further refinement to administrative data for a wide range of diseases.


Elimination of hepatitis C virus RNA in infected human hepatocytes by adenovirus-mediated expression of ribozymes.
Hepatitis C virus (HCV), a positive-strand RNA virus, is the major infectious agent responsible for causing chronic hepatitis. Currently, there is no vaccine for HCV infection, and the only therapy for chronic hepatitis C is largely ineffective. To investigate new genetic approaches to the management of HCV infection, six hammerhead ribozymes directed against a conserved region of the plus strand and minus strand of the HCV genome were isolated from a ribozyme library, characterized, and expressed from recombinant adenovirus vectors. The expressed ribozymes individually or in combination were efficient at reducing or eliminating the respective plus- or minus-strand HCV RNAs expressed in cultured cells and from primary human hepatocytes obtained from chronic HCV-infected patients. This study demonstrates the potential utility of ribozyme therapy as a strategy for the treatment of hepatitis C virus infection.


Cloning and characterization of goldfish activin type IIB receptor.
We have cloned a full length cDNA coding for the activin type IIB receptor (GactRIIB) from the goldfish ovary. GactRIIB shares 73 and 70% amino acid identity in the extracellular domain, and 78 and 80% identity in the intracellular domain with the type IIB receptors of the mouse and Xenopus respectively. The intracellular domain of GactRIIB contains two serine kinase consensus sequences, DFKSRN and GTRRYMAPE, in agreement with the reports in other vertebrates that serine/threonine phosphorylation is involved in activin signal transduction. The identity of GactRIIB was confirmed by transient expression in the COS cells followed by activin binding. Iodinated human activin A bound to the GactRIIB-transfected cells and the binding could be completely inhibited by unlabeled activin. Affinity labeling revealed a band of about 85 kDa, which is in agreement with the reported type II receptors in other vertebrates. Together with the fact that activin is expressed in the goldfish ovary, the cloning of activin receptors from the ovary suggests paracrine and autocrine roles for activin in the goldfish ovarian functions.


Changes in T2-weighted MRI of supinator muscle, pronator teres muscle, and extensor indicis muscle with manual muscle testing.
[Purpose] In order to detect muscle activity with manual muscle testing, T2-weighted magnetic resonance (T2w-MR) images were detected by a 0.2 T compact MRI system. [Subjects and Methods] The subjects were 3 adult males. Transverse T2-weighted multi-slice spin-echo images of the left forearm were measured by a 39 ms echo-time with a 2,000 ms repetition time, a 9.5 mm slice thickness, 1 accumulation and a total image acquisition time of 4 min 16 s. First, T2w-MR images in the resting condition were measured. Then, manipulative isometric contraction exercise (5 sec duration) to the supinator muscle, the pronator teres muscle or the extensor indicis muscle was performed using Borg's rating of perceived exertion (RPE) scale of 15-17. The T2w-MR images were measured immediately after the exercise. [Results] T2w-MR image intensities increased significantly in the supinator muscle, the pronator teres muscle and the extensor indicis muscle after the exercise. However, the image intensities in the rest of the muscle did not change. [Conclusion] Using T2w-MR images, we could detect muscle activity in a deep muscle, the supinator muscle, and a small muscle, the extensor indicis muscle. These results also support the reliability of the manual muscle testing method.


Alterations in lectin binding in the proximal and distal colon of Sprague-Dawley rats with 1,2 dimethylhydrazine administration.
Sequential changes in rhodamine or fluorescein isothiocyanate-conjugated lectin binding of proximal and distal colonic crypts were studied during and after the administration of the 1,2 dimethylhydrazine (DMH). Five adult its unexposed to DMH or vehicle served as baseline controls. Tissue from normal appearing colon and tumor tissue was incubated with Ulex europaeus agglutinin 1 (UEA), Arachis hypogaea (PNA), Dolichos biflorus (DBA) and Griffonia simplicafolia 1 (GSA1). Distinct regional differences were noted in the baseline controls. UEA, PNA and DBA binding were absent in the distal colonic crypt cells. In the proximal colon minimal UEA and PNA binding was noted in the lower crypt whereas DBA binding was intense. GSA1 binding was diffuse in the upper and lower crypt of both regions. During carcinogenesis a progressive increase in PNA binding was noted in normal appearing colonic crypts from both regions. A progressive increase in PNA binding in proximal and distal colonic tumors was noted over time. Similar to normal tissue, DBA bound markedly to proximal colon tumors but was absent in most distal colonic tumors. UEA stained all proximal tumors intensely at all time points. In distal colonic tumors, UEA staining was diminished at 30 weeks compared to tumors analyzed at 16, 22 and 26 weeks. Mucin depletion was also a feature of tumor tissue compared to adjacent normal and hyperplastic glands. This study documents the region specific changes in lectin binding in normal and neoplastic colonic crypts induced by DMH.


Hemograms of gnotobiotic and conventional infant baboons (Papio cynocephalus).
Five baboons delivered by cesarean section were placed in sterile isolation and maintained in this environment for varying periods of time. Four of these neonates were inoculated at birth with viral material and used in special studies. The fifth animal was not inoculated and remained germfree for 11 1/2 mo. Hematologic observations were made on all these subjects. The animals reared in sterile isolators had stable erythrocyte values as compared with those of their conventional counterpart. Conventionally-reared baboons had higher leukocyte counts than the infants in a sterile environment, and the lymphocyte numbers of isolator-reared infants increased at a slower rate than the conventional infants.


[Postoperative bleeding after tooth extractions in patients controlled with warfarin--a clinico-statistical study on the factors influencing postoperative bleeding].
There are many reports about tooth extractions of patients taking warfarin, however PT-INR level is used to examine the postoperative bleeding in patients. To investigate other factors, postoperative bleeding, age, gender, PT-INR level, combined use of anti-platelet drugs, conditions of extracted tooth, a number of tooth extractions at a treatment, methods of management for warfarin therapy, degree of the alveolar bone loss and size of radiolucency of apical region were examined in this study. To apply Mann-Whitney U-test and chi2-test, ninety-three patients (38 male and 55 female) who took warfarin and visited our clinic for tooth extractions from April 1994 to November 2002 were classified into 2 groups: One group showed hemostasis by the next day (77 patients), the other showed the continuous bleeding after the next day (16 patients). These analyses indicated that PT-INR level, a number of tooth extractions at a treatment, methods of management for warfarin therapy, and size of radiolucency of apical region influenced postoperative bleeding. In addition, stepwise logistic regression analysis was applied to all of the factors, obtained from 77 patients out of 93 patients. This data showed that PT-INR level, a number of tooth extractions at a treatment and methods of management for warfarin therapy influenced postoperative bleeding. These results suggest that before the tooth extractions not only PT-INR level but methods of management for warfarin therapy and size of wound could be important to control the postoperative bleeding in warfarin taking patients.


Shrinkage of microstructures produced by two-photon polymerization of Zr-based hybrid photosensitive materials.
An investigation of the shrinking behaviour of a zirconium-based sol-gel composite micro-structured by two-photon polymerization is presented and a simple, straightforward methodology allowing the evaluation of shrinkage is suggested. It is shown that volume reduction is directly related to the average laser power (irradiation dose) used for the microfabrication and becomes a critical issue near the polymerization threshold. It is demonstrated that this shrinkage can be employed beneficially to improve the structural resolution. This is demonstrated by the presence of stopbands in the photonic crystal nanostructures fabricated with controlled volume reduction. Well above the polymerization threshold, the studied material exhibits remarkably low shrinkage. Therefore, no additional effort for the pre-compensation of distortion and for the improvement of structural stability is required.


Actin interaction and regulation of cyclin-dependent kinase 5/p35 complex activity.
Cyclin-dependent kinase 5 (Cdk5) plays a critical role during neurodevelopment, synaptic plasticity, and neurodegeneration. Cdk5 activity depends on association with neuronal proteins p35 and p25, a proteolytic product of p35. Cdk5 regulates the actin cytoskeletal dynamics that are essential for neuronal migration, neuritic growth, and synaptogenesis. However, little is known about the interaction of actin and Cdk5 and its effect on neuronal Cdk5 activity. In a previous study, we observed that Cdk5/p35 activity is negatively correlated with co-immunoprecipitated F-actin (filamentous actin) amounts in the mouse brain, and suggested that F-actin inhibits the formation of the Cdk5/p35 complex [Journal of Neuroscience (2008) vol. 28, p. 14511]. The experiments reported here were undertaken to elucidate the relationship between actin and the formation of the Cdk5/p35 complex and its activity. Instead of an F-actin-mediated inhibition, we propose that G-actin (globular actin) in the F-actin preparations is responsible for inhibiting Cdk5/p35 and Cdk5/p25 kinase activity. We found that F-actin binds to p35 but not p25 or Cdk5. We have shown that G-actin binds directly to Cdk5 without disrupting the formation of the Cdk5/p35 or Cdk5/p25 complexes. G-actin potently suppressed Cdk5/p35 and Cdk5/p25 activity when either histone H1 or purified human tau protein were used as substrates, indicating a substrate-independent inhibitory effect of G-actin on Cdk5 activity. Finally, G-actin suppressed the activity of Cdk5 immunoprecipitated from wild type and p35-deficient mouse brain, suggesting that G-actin suppresses endogenous Cdk5 activity in a p35-independent manner. Together, these results suggest a novel mechanism of actin cytoskeletal regulation of Cdk5/p35 activity.


The association between hemogram parameters and the development of contrast-induced nephropathy in patients presenting with non-ST-elevation myocardial infarction.
Hemogram parameters such as mean platelet volume (MPV), neutrophil/lymphocyte ratio (NLr), red cell distribution width and platelet distribution width are widely used inflammatory indicators to assess prognosis in various cardiovascular diseases. In this study, we aimed to investigate the role of hemogram parameters to predict the development of contrast-induced nephropathy (CIN) in patients presenting with non-ST segment elevation myocardial infarction (non-STEMI) and treated with percutaneous coronary intervention (PCI). All pateints who underwent PCI with a diagnosis of non-STEMI between 2017 and 2020 in our center were included retrospectively in this study. A total of 387 patients were included in this retrospective study. Advanced age (p=0.001, β:0.005, OR [95 % CI]: 0.002-0.007), diabetes mellitus (p=0.013, β:0.205, OR [95 % CI]: 0.150-0.260), congestive heart failure (p=0.009, β:0.095, OR [95 % CI]: 0.024-0.166), volume of contrast medium (p=0.008, β:0.241, OR [95 % CI]: 0.184-0.392), MPV (p=0.02, β:0.047, OR [95 % CI]: 0.028-0.065) and NLr (p=0.001, β:0.052, OR [95 % CI]: 0.040-0.063) were found as independent risk factors associated with CIN development according to multivariate logistic regression analysis. A cut off value of 5.5 for NLr was associated with 79.6 % sensitivity and 79.5 % specificity and 9.05 for MPV was associated with 64.1 % sensitivity and 58.7 % specificity in prediction of CIN development. Hematological parameters, assessed by routine blood count analysis may serve as a promising and useful marker for CIN especially when used in combination with traditional risk factors. MPV and NLr were demonstrated as predictors of CIN development in non-STEMI patients who were treated with PCI in our study.


Feeding ecology of King George whiting Sillaginodes punctatus (Perciformes) recruits in seagrass and unvegetated habitats. Does diet reflect habitat utilization?
This study investigated the feeding ecology of King George whiting Sillaginodes punctatus recruits to determine how diet composition varies between habitat types (seagrass and unvegetated habitats), and between sites separated by distance. Broad-scale sampling of seagrass and unvegetated habitats at nine sites in Port Phillip Bay (Australia) indicated the diet composition varied more by distance into the bay than by habitat. Near the entrance to the bay the diet was dominated by harpacticoids and gammarid amphipods, in the middle reaches of the bay the diet was completely dominated by harpacticoids, while at sites furthest into the bay, mysids and crab zoea were also important. Abundances of prey in guts was significantly higher between 1000 and 2200 hours compared with other times, indicating diurnal feeding. Laboratory determined gut evacuation rate (based on an exponential model) was estimated to be -0·54. Daily rations were highly variable among sites and habitat types. Sillaginodes punctatus recruits consumed much higher quantities of prey on unvegetated habitat than seagrass habitat at some middle reach sites; with prey consumption of harpacticoid copepods on unvegetated habitat approaching 3000 individuals per day at one site. The results of this study provide insight into why habitat associations of S. punctatus recruits within mosaics of seagrass and unvegetated habitat show high spatial variation.


Renal tubular uptake of protein: effect of molecular charge.
The effect of molecular charge of proteins on proximal tubular reabsorption was evaluated in the rat. Native and two cationized forms of albumin, native and anionized lysozyme, and native and anionized cytochrome c were iodinated with 125I. The different forms of each type of protein were alternately microinfused into the same site of proximal convoluted tubules in vivo. Tubular reabsorption was determined as the difference between the amounts of TCA-precipitable radioactivity infused and recovered in the urine. At low concentration of albumin 5 times more cationized than anionic albumin and 2.7 times more cationic than anionized lysozyme were reabsorbed by the proximal tubule. At two of four concentrations, proximal tubular uptake of cationic cytochrome c exceeded that of anionized cytochrome c. Uptake of cationic cytochrome c exceeded that of cationic lysozyme; however, the difference in uptake between native cationic and anionized species of the two proteins was much greater for lysozyme than for cytochrome c. The data reveal that a higher isoelectric point significantly enhances proximal tubular reabsorption of albumin, lysozyme, and cytochrome c and that proteins with similar molecular weight and isoelectric point are not necessarily reabsorbed to the same degree. This suggests that in addition to total molecular charge the molecular configuration and/or distribution of electrical charges on teh protein surface determine protein binding by the luminal membrane and subsequent endocytosis by the proximal tubule.


Human immunodeficiency virus type 1 central DNA flap: dynamic terminal product of plus-strand displacement dna synthesis catalyzed by reverse transcriptase assisted by nucleocapsid protein.
To terminate the reverse transcription of the human immunodeficiency virus type 1 (HIV-1) genome, a final step occurs within the center of the proviral DNA generating a 99-nucleotide DNA flap (6). This step, catalyzed by reverse transcriptase (RT), is defined as a discrete strand displacement (SD) synthesis between the first nucleotide after the central priming (cPPT) site and the final position of the central termination sequence (CTS) site. Using recombinant HIV-1 RT and a circular single-stranded DNA template harboring the cPPT-CTS sequence, we have developed an SD synthesis-directed in vitro termination assay. Elongation, strand displacement, and complete central flap behavior were analyzed using electrophoresis and electron microscopy approaches. Optimal conditions to obtain complete central flap, which ended at the CTS site, have been defined in using nucleocapsid protein (NCp), the main accessory protein of the reverse transcription complex. A full-length HIV-1 central DNA flap was then carried out in vitro. Its synthesis appears faster in the presence of the HIV-1 NCp or the T4-encoded SSB protein (gp32). Finally, a high frequency of strand transfer was shown during the SD synthesis along the cPPT-CTS site with RT alone. This reveals a local and efficient 3'-5' branch migration which emphasizes some important structural fluctuations within the flap. These fluctuations may be stabilized by the NCp chaperone activity. The biological implications of the RT-directed NCp-assisted flap synthesis are discussed within the context of reverse transcription complexes, assembly of the preintegration complexes, and nuclear import of the HIV-1 proviral DNA to the nucleus toward their chromatin targets.


Effects of choline-deprivation on paracetamol- or phenobarbital-induced rat liver metabolic response.
Choline is an essential nutrient that seems to be involved in a wide variety of metabolic reactions and functions in both humans and rodents. Various pathophysiological states have been linked to choline deprivation (CD). The aim of the present study was to determine the effect of CD upon biochemical, histological and metabolic alterations induced by drugs that affect hepatic functional integrity and various drug metabolizing systems via distinct mechanisms. For this purpose, paracetamol (ACET) or phenobarbital (PB) were administered to male Wistar rats that were fed with standard rodent chow (normally fed, NF) or underwent dietary CD. The administration of ACET increased the serum aspartate aminotransferase levels in NF rats, while CD restricted this increase. On the other hand, ACET suppressed alkaline phosphatase levels only in CD rats. Moreover, CD prevented the PB-induced increase of the mitotic activity of hepatocytes. The administration of ACET down-regulated CYP1A2 and CYP2B1 expression in CD rats, while up-regulating them in NF rats. The administration of PB suppressed CYP1A2 apoprotein levels in CD rats, whereas the drug had no effect on NF rats. The PB-induced up-regulation of CYP2B, CYP2E1 and CYP1A1 isozymes was markedly higher in CD than in NF rats. In addition, PB increased glutathione-S-transferase activity only in CD rats. Hepatic glutathione content (GSH) was suppressed by ACET in NF rats, whereas the drug increased GSH in CD rats. Our data suggest that CD has a significant impact on the hepatic metabolic functions, and in particular on those related to drug metabolism. Thus, CD may modify drug effectiveness and toxicity, as well as drug-drug interactions, particularly those related to ACET and PB.


May Measurement Month 2017: an analysis of blood pressure screening results from Australia-South-East Asia and Australasia.
Increased blood pressure (BP) is the single biggest contributing risk factor to the global disease burden. May Measurement Month (MMM) is a global initiative of the International Society of Hypertension aimed at raising awareness of high BP. In Australia, hypertension affects around six million adults and continues to remain the greatest attributable cause of cardiovascular mortality and morbidity (48.3%), stroke deaths (28%), and kidney disease (14%). An opportunistic cross-sectional survey was carried out during May 2017 predominantly in capital cities across Australia which included adult volunteers. Blood pressure measurement, the definition of hypertension and statistical analysis followed the standard MMM protocol. Additional information obtained included anthropometric data and responses to questionnaires on demographic, lifestyle, and environmental factors. Data were collected from 3817 individuals. After multiple imputation, of the 3758 individuals for whom a mean of the second and third BP reading was available, 1188 (31.2%) had hypertension. Of 3213 individuals not receiving antihypertensive treatment, 591 (18.4%) were hypertensive, and 239 (40.1%) of the 596 individuals receiving treatment had uncontrolled BP. Adjusted BP was higher in association with antihypertensive medication, cerebrovascular disease, smoking, and alcohol consumption. Blood pressure was higher when measured on the right arm and on Tuesdays. MMM17 was one of the largest BP screening campaigns undertaken in Australia using standardized BP measurements. In line with previous surveys, around one-third of screened adults had hypertension and approximately 40% of treated individuals remained uncontrolled. These results suggest that opportunistic screening can identify significant numbers with raised BP.


Langmuir-blodgett films of a functionalized molecule with cross-sectional mismatch between head and tail.
A functionalized surfactant has been investigated as floating monolayers by synchrotron x-ray diffraction and as bilayers transferred to solid supports by the Langmuir-Blodgett technique through atomic force microscopy. The transfer process is accompanied by an increase of the unit cell area (about 17 percent) and by an increase of the average domain diameter of nanometer-scale domains (about three times). The unit cell area of the floating monolayer corresponds to close packing of the head groups and a noncharacteristic packing of the tifted alkyl chains. The larger unit cell area of the bilayer film is consistent with a particular ordered packing of the alkyl chains, leaving free space for the head groups.


Elimination of malignant clonogenic cells from human bone marrow using multiple myeloid cell-specific monoclonal antibodies and complement.
Single or combined monoclonal antibodies (McAbs) Zh53, Zh820, and Zh2-1 have been used to eliminate malignant clonogenic cells from human bone marrow. The test of cytotoxicity showed that all of these McAbs could express high specific cytotoxic action against HL-60 cells and were selectively complement-dependent cytotoxic to various types of fresh leukemic cells. Clonogenic assay detected that single treatment with antibody and rabbit complement (RC) could reduce clonogenic units of HL-60 cells by more than 2 logs and two treatments reduced clonogenic units by more than 4 logs. However, combination of 2 McAbs could reduce clonogenic units by 4-5 logs. The data suggest that multiple treatments with McAbs and RC or a combination of 2 McAbs are more effective than a single treatment in eliminating clonogenic tumor cells. Treatment of normal human bone marrow with Zh53, Zh2-1 and RC did not produce a loss of normal CFU-GM, but treatment with Zh820 reduced the clonic units of normal CFU-GM by 24%.


Comparative sensitivity and specificity of imaging modalities in staging bladder cancer prior to radical cystectomy: a systematic review and meta-analysis.
The detection of lymph node metastases in bladder cancer has a significant impact on treatment decisions. Multiple imaging modalities are available to clinicians including magnetic resonance imaging, computed tomography and positron emission tomography. We aimed to investigate the utility of alternate imaging modalities on pre-cystectomy imaging in bladder cancer for the detection of lymph node metastases. We performed systematic search of Web of Science (including MEDLINE), EMBASE and Cochrane libraries in accordance with the PRISMA statement. Studies comparing lymph node imaging findings with final histopathology were included in our analysis. Sensitivity and specificity data were quantified using patient-based analysis. A true positive was defined as a node-positive patient on imaging and node positive on histopathology. Meta-analysis of studies was performed using a mixed-effects, hierarchical logistic regression model. Our systematic search identified 35 articles suitable for inclusion. MRI and PET have a higher sensitivity than CT while the specificity of all modalities was similar. The summary MRI sensitivity = 0.60 (95% CI 0.44-0.74) and specificity = 0.91 (95% CI 0.82-0.96). Summary PET/CT sensitivity = 0.56 (95% CI 0.49-0.63) and specificity = 0.92 (95% CI 0.86-0.95). Summary CT sensitivity = 0.40 (95% CI 0.33-0.49) and specificity = 0.92 (95% CI 0.86-0.95). MRI and PET/CT provides superior sensitivity compared to CT for detection of positive lymph nodes in bladder cancer prior to cystectomy. There is variability in the accuracy that current imaging modalities achieve across different studies. A number of other factors impact on detection accuracy and these must be considered.


Proteolysis Targeting Chimeras (PROTACs) Based on Celastrol Induce Multiple Protein Degradation for Triple-Negative Breast Cancer Treatment.
The pursuit of single drugs targeting multiple targets has become a prominent trend in modern cancer therapeutics. Natural products, known for their multi-targeting capabilities, accessibility, and cost-effectiveness, hold great potential for the development of multi-target drugs. However, their therapeutic efficacy is often hindered by complex structural modifications and limited anti-tumor activity. In this study, we present a novel approach using celastrol (CST)-based Proteolysis Targeting Chimeras (PROTACs) for breast cancer therapy. Through rational design, we have successfully developed compound 6a, a potent multiple protein degrader capable of selectively degrading GRP94 and CDK1/4 in tumor cells via the endogenous ubiquitin-proteasome system. Furthermore, compound 6a has demonstrated remarkable inhibitory effects on cell proliferation and migration, and induction of apoptosis in 4T1 cells through cell cycle arrest and activation of the Bcl-2/Bax/cleaved Caspase-3 apoptotic pathway. In vivo administration of compound 6a has effectively suppressed tumor growth with an acceptable safety profile. Our findings suggest that the CST-based PROTACs described herein can be readily extended to other natural products, offering a potential avenue for the development of natural product-based PROTACs for cancer treatment.


Effect of nicotinic acid and dipyridamole on tissue blood flow in peripheral vascular disease.
The effect of 2-weeks' treatment with daily doses of 150 mg nicotinic acid, 300 mg dipyridamole and placebo, in random order, on muscle blood flow and skin temperature gradient in the legs was measured in 6 patients with advanced arteriosclerotic peripheral vascular disease. Mean resting skin temperature gradient and muscle blood flow were uninfluenced by any of these treatments. Dipyridamole, but not nicotinic acid, caused a significant increase in exercise muscle blood flow compared to placebo (p less than 0.01). This improvement was maintained after 2-months' continuous treatment at the end of the trial. It is concluded that dipyridamole may be of value in the medical management of peripheral vascular disease.


Neuroprotective effects of SMTP-44D in mice stroke model in relation to neurovascular unit and trophic coupling.
Stachybotrys microspora triprenyl phenol (SMTP)-44D has both anti-oxidative and anti-inflammatory activities, but its efficacy has not been proved in relation to the pathological changes of neurovascular unit (NVU) and neurovascular trophic coupling (NVTC) in ischemic stroke. Here, the present study was designed to assess the efficacies of SMTP-44D, moreover, compared with the standard neuroprotective reagent edaravone in ischemic brains. ICR mice were subjected to transient middle cerebral artery occlusion (tMCAO) for 60 min, SMTP-44D (10 mg/kg) or edaravone (3 mg/kg) was intravenously administrated through subclavian vein just after the reperfusion, and these mice were examined at 1, 3, and 7 d after reperfusion. Compared with the vehicle group, SMTP-44D treatment revealed obvious ameliorations in clinical scores and infarct volume, meanwhile, markedly suppressed the accumulations of 4-HNE, 8-OHdG, nitrotyrosine, RAGE, TNF-α, Iba-1, and cleaved caspase-3 after tMCAO. In addition, SMTP-44D significantly prevented the dissociation of NVU and improved the intensity of NAGO/BDNF and the number of BDNF/TrkB and BDNF/NeuN double positive cells. These effects of SMTP-44D in reducing oxidative and inflammatory stresses were similar to or stronger than those of edaravone. The present study demonstrated that SMTP-44D showed strong anti-oxidative, anti-inflammatory, and anti-apoptotic effects, moreover, the drug also significantly improved the NVU damage and NVTC in the ischemic brain.


Exposure to diphtheria toxin during the juvenile period impairs both inner and outer hair cells in C57BL/6 mice.
Diphtheria toxin (DT) administration into transgenic mice that express the DT receptor (DTR) under control of specific promoters is often used for cell ablation studies in vivo. Because DTR is not expressed in mice, DT injection has been assumed to be nontoxic to cells in vivo. In this study, we demonstrated that DT application during the juvenile stage leads to hearing loss in wild-type mice. Auditory brainstem response measurement showed severe hearing loss in C57BL/6 mice administered DT during the juvenile period, and the hearing loss persisted into adulthood. However, ototoxicity did not occur when DT was applied on postnatal day 28 or later. Histological studies demonstrated that hearing loss was accompanied by significant degeneration of inner and outer hair cells (HCs), as well as spiral ganglion neurons. Scanning electron microscopy showed quick degeneration of inner HCs within 3days and gradual degeneration of outer HCs within 1week. These results demonstrated that DT has ototoxic action on C57BL/6 mice during the juvenile period, but not thereafter, and the hearing loss was due to degeneration of inner and outer HCs by unknown DT-related mechanisms.


In-solution Y-chromosome capture-enrichment on ancient DNA libraries.
As most ancient biological samples have low levels of endogenous DNA, it is advantageous to enrich for specific genomic regions prior to sequencing. One approach-in-solution capture-enrichment-retrieves sequences of interest and reduces the fraction of microbial DNA. In this work, we implement a capture-enrichment approach targeting informative regions of the Y chromosome in six human archaeological remains excavated in the Caribbean and dated between 200 and 3000 years BP. We compare the recovery rate of Y-chromosome capture (YCC) alone, whole-genome capture followed by YCC (WGC + YCC) versus non-enriched (pre-capture) libraries. The six samples show different levels of initial endogenous content, with very low (< 0.05%, 4 samples) or low (0.1-1.54%, 2 samples) percentages of sequenced reads mapping to the human genome. We recover 12-9549 times more targeted unique Y-chromosome sequences after capture, where 0.0-6.2% (WGC + YCC) and 0.0-23.5% (YCC) of the sequence reads were on-target, compared to 0.0-0.00003% pre-capture. In samples with endogenous DNA content greater than 0.1%, we found that WGC followed by YCC (WGC + YCC) yields lower enrichment due to the loss of complexity in consecutive capture experiments, whereas in samples with lower endogenous content, the libraries' initial low complexity leads to minor proportions of Y-chromosome reads. Finally, increasing recovery of informative sites enabled us to assign Y-chromosome haplogroups to some of the archeological remains and gain insights about their paternal lineages and origins. We present to our knowledge the first in-solution capture-enrichment method targeting the human Y-chromosome in aDNA sequencing libraries. YCC and WGC + YCC enrichments lead to an increase in the amount of Y-DNA sequences, as compared to libraries not enriched for the Y-chromosome. Our probe design effectively recovers regions of the Y-chromosome bearing phylogenetically informative sites, allowing us to identify paternal lineages with less sequencing than needed for pre-capture libraries. Finally, we recommend considering the endogenous content in the experimental design and avoiding consecutive rounds of capture, as clonality increases considerably with each round.


Ambulatory blood pressure monitoring can unmask hypertension in patients with psoriasis vulgaris.
Psoriasis vulgaris is one of the most prevalent chronic, inflammatory skin disorders. Patients with psoriasis have excess risk of essential hypertension. Masked hypertension (MH), defined as normal office blood pressure (BP) with elevated ambulatory BP (ABPM), has been drawing attention recently due to its association with increased risk of developing sustained hypertension, cardiovascular morbidity, and mortality. The aim of this study was to investigate the prevalence of MH in psoriatic patients. On hundred and ten middle-aged, normotensive, non-obese patients with psoriasis vulgaris and 110 age- and sex-matched normotensive controls were included in the study. ABPM was performed in all participants over a 24-h period. The clinical severity of the disease was determined according to current indexes. The prevalence of MH among subjects with psoriasis vulgaris was 31.8% and increased compared to control subjects (p<0.01). Predictors of MH in patients with psoriasis vulgaris were detected as male sex, smoking, obesity-related anthropometric measures, and disease activity. Male sex, waist circumference, and diffuse psoriatic involvement were detected as independent predictors of MH. MH is prevalent in patients with psoriasis vulgaris. Assessment with ABPM and close follow-up for development of hypertension is reasonable.


Breast reconstruction following excision of phylloides tumor.
There are few papers published on breast reconstruction after excision of phylloides tumor. Six patients who had reconstruction of the breast following excision of phylloides tumor are described. All underwent wide excision or subcutaneous mastectomy followed by immediate or delayed reconstruction with implants or autologous tissue. The mean follow-up was 5 years (range, 2.5-7 years). One patient died of metastases; the others survived without evidence of recurrence. The etiology, incidence, diagnosis, and treatment of these tumors are discussed. The aesthetic results in these patients is also described.


Ganglioside GQ1b-induced terminal differentiation in cultured mouse keratinocytes. Phosphoinositide turnover forms the onset signal.
Investigations were undertaken to see whether mouse keratinocyte differentiation was elicited by gangliosides. Among the gangliosides tested only GQ1b, a tetrasialoganglioside containing two disialosyl residues, induced keratinocyte differentiation, as indicated by the formation of cornified envelopes, enhancement of transglutaminase activity and suppression of DNA synthesis. Upon stimulation with GQ1b the mass content of Ins(1,4,5)P3 and the intracellular Ca2+ levels were markedly enhanced in a time- and dose-dependent manner, whereas no significant changes were observed with other gangliosides, thereby indicating activation of phospholipase C for the onset of keratinocyte differentiation. Furthermore, only GQ1b promoted the translocation of protein kinase C (PKC) from cytosol to membrane. Inhibition of PKC with H-7 or down-regulation of the enzyme by prolonged pre-treatment with phorbol 12,13-dibutyrate greatly suppressed transglutaminase activity and formation of cornified envelopes induced by GQ1b. These results demonstrate that the tetrasialoganglioside GQ1b generates the initial differentiation signal in mouse keratinocytes through phosphoinositide turnover, and also suggest that PKC activation may act at certain, as yet unidentified, stages of differentiation processes.


Alpha and beta phases of 4-aminoquinoline-2-carboxylic acid monohydrate.
The structures of two phases of the title compound, C10H8N2O2.H2O, are reported. While the organic molecule has very nearly the same molecular geometry in the two phases, the hydrogen-bonding patterns are quite different. In each case, however, a three-dimensional network of hydrogen bonds is formed. In the alpha phase, six N-H ... O hydrogen bonds have donor-acceptor distances ranging from 2.681 (2) to 3.206 (2) A, while three O-H...O hydrogen bonds range from 2.837 (2) to 3.173 (2) A, and in the beta phase, four N-H ... O hydrogen bonds range from 2.668 (2) to 3.038 (2) A, while two O-H...O hydrogen bonds are 2.951 (3) and 2.959 (3) A in length.


A usability assessment of AHLTA in ambulatory clinics at a military medical center.
The investigators conducted a formal usability evaluation of the military's electronic health record known as Armed Forces Health Longitudinal Technology Application (AHLTA). Seventeen providers from different specialties in the ambulatory setting were interviewed and observed at a military medical center. Data were analyzed by human factors experts. Observations and interviews yielded four major usability findings: (1) limited AHLTA use during the actual encounter, (2) difficulties in obtaining situational awareness of the patient, (3) work-arounds with nonintegrated systems, and (4) frustrations in the use of the structured documentation. This assessment is congruent with usability concerns voiced in other military health system (MHS) and nonmilitary clinical systems. Improving the usability of future MHS clinical systems could lead to improved clinical decision making, patient safety, and increased information accuracy.


Atrial natriuretic peptide predicts disease progression and digital ulcers development in systemic sclerosis patients.
Systemic sclerosis (SSc) is an autoimmune disease characterized by micro/macrovascular damage due to the underlying fibrosis. Markers able to predict the progression of cardiovascular damage, including digital ulcers, in SSc are warranted. We aimed at characterizing the relevance of N-terminal proatrial natriuretic peptide (NT-proANP) and N-terminal probrain natriuretic peptide plasma levels in relation to cardiovascular damage and digital ulcers in a cohort of Italian SSc patients. Seventy patients were enrolled (64 women and six men; mean age 56.7 ± 14 years) with a disease duration of 11.1 ± 8.3 years. Clinical, instrumental (nailfold videocapillaroscopy, ECG, transthoracic echocardiography, pulmonary function test with diffusion lung CO), NT-proANP and N-terminal probrain natriuretic peptide plasma levels measurement were performed at baseline. The clinical follow-up lasted 24 months. The statistical approach used to achieve the study objectives included multivariate analysis, receiver operating characteristic curve, Kaplan-Meier and Cox regression analyses. Both NT-proNPs levels correlated with systolic pulmonary arterial pressure, but only the NT-proANP level correlated with right heart dimension. Both NT-proNPs levels were higher in patients experiencing events at follow-up but only the NT-proANP level significantly predicted the progression of cardiovascular damage, including development of pulmonary arterial hypertension (PAH). NT-proANP levels were higher in patients with digital ulcers and strongly predicted their development. Our results show that the NT-proANP plasma level significantly correlates with disease progression such as new onset of PAH, worsening of pulmonary hypertension and development of digital ulcers in a cohort of SSc Italian patients. If future studies will confirm our findings, the plasma NT-proANP level could be used in clinical practice as a novel sensitive marker for PAH and digital ulcers development in SSc.