The "Big Five" Phytochemicals Targeting Cancer Stem Cells: Curcumin, EGCG, Sulforaphane, Resveratrol and Genistein.
Cancer stem cells (CSCs) constitute a subpopulation of tumor cells that possess self-renewal and tumor initiation capacity, and the ability to give rise to the heterogeneous lineages of cancer cells that comprise the tumor. CSCs exhibit intrinsic mechanisms of resistance to virtually all conventional cancer therapeutics, allowing them to survive current cancer therapies and to initiate tumor recurrence and metastasis. Different pathways and mechanisms that confer resistance and survival of CSCs, including activation of the Wnt/β- catenin, Sonic Hedgehog, Notch, PI3K/Akt/mTOR and STAT3 signaling pathways, expression of aldehyde dehydrogenase 1 (ALDH1) and oncogenic microRNAs, and acquisition of epithelial-mesenchymal transition (EMT), have been identified recently. Certain phytochemicals, in particular curcumin, epigallocatechin-3-gallate (EGCG), sulforaphane, resveratrol and genistein have been shown to interfere with these intrinsic CSC pathways in vitro and in human xenograft mice, leading to elimination of CSCs. Moreover, recent clinical trials have demonstrated the therapeutic efficacy of five phytochemicals, alone or in combination with modern cancer therapeutics, and in various types of cancer. Since current cancer therapies fail to eradicate CSCs, leading to cancer recurrence and progression, targeting of CSCs with phytochemicals such as curcumin, EGCG, sulforaphane, resveratrol and genistein, combined with each other and/or in combination with conventional cytotoxic drugs and novel cancer therapeutics, may offer a novel therapeutic strategy against cancer.


Drug prescribing pattern for under-fives in a paediatric clinic in South-Western Nigeria.
The audit of drug prescribing pattern in under-five children is essential in identifying the various types of non-rational prescribing such as polypharmacy and irrational use of antimicrobials. The primary objective of this study was to determine the drug prescribing pattern for children aged below five years attending the paediatric outpatient clinic of a tertiary hospital in Ado-Ekiti, South-West Nigeria. A cross-sectional study was carried out using the medical records of patients who attended the paediatric clinic of the hospital between April 1 and October 30, 2013. The medical records of patients aged below five years were selected monthly for the period of the study using a regular interval ratio. Drug use indicators were assessed using the WHO guidelines on investigation of drug use in health care facilities. There were 293(55.7%) prescriptions collected from male patients and 233(44.3% from female patients with a total of 1369 prescribed drugs (2.6 ± 1.1 drugs per prescription). A total of three hundred and seventy-four (71.1%) patients had at least one antibiotic prescribed with antibiotics accounting for 28.2% of all drugs prescribed, while 13.5% of all encounters had an injection prescribed. Prescribing by generic name was done in 68.9 ± 26.5% of all prescribed medications, and 60.4% of all prescribed medications were from the latest version of the Nigerian Essential Drug List. Three hundred and twelve children (59.3%) had at least one anti-malarial drug prescribed while analgesics drugs accounted for 6.1% of all prescriptions. This study showed that over-prescription of antibiotics, significant use of injections and prescribing by generic name are real issues among Nigerian paediatric prescribers.


On the nature of sickle cell disease in the south-western province of Saudi Arabia.
Sickle cell disease (SCD) occurs at a high prevalence in different parts of Saudi Arabia. Several reports indicate that the disease follows a mild clinical course in the Saudi population of the eastern province of Saudi Arabia, while little is known about the disease in other parts of the country. This study was conducted on 53 children from the Saudi Arabian south-western province with sickle cell disease and 53 age- and sex-matched normal controls (haemoglobin AA phenotype). A statistically significant difference was encountered in the haematological parameters investigated in the two groups. The SCD patients were divided into subgroups with high and low Hb F levels, alpha- and beta-thalassaemia and glucose-6-phosphate dehydrogenase (G-6-PD) deficiency. The haematological parameters were then compared in the different sub-groups. No significant difference could be demonstrated in the haematological parameters in patients with a high or low Hb F level. In patients without thalassaemia, the red cell count, total haemoglobin and haematocrit were significantly lower, while MCV, MCH and MCHC were higher. G-6 PD deficiency existed in association with thalassaemias, and apart from a reduction in MCV and MCH, no other statistically significant difference could be demonstrated. Clinical examination revealed a severe disease with several cases suffering from the hand and foot syndrome.


2D-3D shape reconstruction of the distal femur from stereo X-ray imaging using statistical shape models.
Three-dimensional patient specific bone models are required in a range of medical applications, such as pre-operative surgery planning and improved guidance during surgery, modeling and simulation, and in vivo bone motion tracking. Shape reconstruction from a small number of X-ray images is desired as it lowers both the acquisition costs and the radiation dose compared to CT. We propose a method for pose estimation and shape reconstruction of 3D bone surfaces from two (or more) calibrated X-ray images using a statistical shape model (SSM). User interaction is limited to manual initialization of the mean shape. The proposed method combines a 3D distance based objective function with automatic edge selection on a Canny edge map. Landmark-edge correspondences are weighted based on the orientation difference of the projected silhouette and the corresponding image edge. The method was evaluated by rigid pose estimation of ground truth shapes as well as 3D shape estimation using a SSM of the whole femur, from stereo cadaver X-rays, in vivo biplane fluoroscopy image-pairs, and an in vivo biplane fluoroscopic sequence. Ground truth shapes for all experiments were available in the form of CT segmentations. Rigid registration of the ground truth shape to the biplane fluoroscopy achieved sub-millimeter accuracy (0.68mm) measured as root mean squared (RMS) point-to-surface (P2S) distance. The non-rigid reconstruction from the biplane fluoroscopy using the SSM also showed promising results (1.68mm RMS P2S). A feasibility study on one fluoroscopic time series illustrates the potential of the method for motion and shape estimation from fluoroscopic sequences with minimal user interaction.


Development of a non-target strategy for evaluation of potential biological effects of inhalable aerosols generated during purposeful room conditioning using an in vitro inhalation model.
An integrated in vitro inhalation approach was outlined to estimate potential adverse acute inhalation effects of aerosols from commercial nebulizer applications used for purposeful room conditioning such as disinfection, scenting or others. Aerosol characterization, exposure estimation and evaluation of acute biological effects by in vitro inhalation were included to generate dose-response data, allowing for determination of in vitro lowest observable adverse effect levels (LOAELs). Correlation of these to estimates of human lung deposition was included for quantitative in vitro to in vivo extrapolation approach (QIVIVE) for acute effects during human exposure. To test the proposed approach, a case study was undertaken using two realistic test materials. An acute in vitro inhalation setup with air-liquid interface A549-cells in an optimized exposure situation (P.R.I.T.® ExpoCube®) was used to expose cells and analysis of relevant biological effects (viability, mitochondrial membrane potential, stress, IL-8 release) was carried out. The observed dose-responsive effects in a sub-toxic dose-range could be attributed to the main component of one test material and its presence in the aerosol phase of the nebulized material. QIVIVE resulted in a factor of at least 256 between the in vitro LOAEL and the estimated acute human lung exposure for this test material. The case-study shows the value of the non-target in vitro inhalation testing approach especially in case of a lack of knowledge on complex product composition. It is expected that approaches like this will be of high value for product safety and environmental health in the future.


Evaluation of Healthcare Students' Knowledge on Antibiotic Use, Antimicrobial Resistance and Antimicrobial Stewardship Programs and Associated Factors in a Tertiary University in Ghana: Findings and Implications.
Antimicrobial resistance (AMR) is a major public health problem globally, and Ghana is no exception. Good knowledge regarding antibiotic use, AMR, and the concept of antimicrobial stewardship (AMS) is critical among healthcare students to curb rising AMR rates in the future. Consequently, a need to ascertain this. A cross-sectional survey was undertaken among fifth-year pharmacy, medical students and fourth (final)-year nursing and physician assistantship students at the University of Health and Allied Sciences in Ghana to assess their knowledge on antibiotic use, AMR and AMS using a web-based self-administered structured questionnaire. Descriptive statistics, Fishers' exact test, and multiple logistic regression analyses were performed. A total of 160 healthcare students were interviewed, of which 56.3% (n = 90) were male and 58.8% (n = 94) were in their fourth year of study. Good knowledge of antibiotic use, AMR, and AMS was associated with the study course (p = 0.001) and the number of years of study (p < 0.001). Overall, there were differences in the level of knowledge of antibiotics among the different healthcare students and their years of study. Efforts must now be made to enhance the curricula to ensure an improved and uniform transfer of knowledge of antibiotics, AMR, and AMS among the different healthcare students to sustain the fight against AMR in Ghana given growing concerns.


Psychological distress and convergence of own and proxy health-related quality of life in carers of adults with an intellectual disability.
In adults with an intellectual disability, health-related quality of life (HRQoL) is often measured by proxy report. This cross-sectional study investigated whether the mental health of proxy raters impacts the way they rate HRQoL. In this study, 110 carers of adults with an intellectual disability completed measures of psychological distress (Kessler-6) and HRQoL (EQ-5D-3L) about their own HRQoL and that of the care recipient. Differences between HRQoL scores as rated by the carer about themselves and the care recipient were calculated (convergence scores) and multiple regression models were fitted to estimate the association between proxy psychological distress and convergence scores for subjective/objective HRQoL controlling for support needs of the care recipient, carer age and gender of care recipient. There was a significant association between psychological distress and subjective HRQoL convergence scores (r = .92; P = 0.03; 95%; CI: -1.76 to -0.09). There was no association between psychological distress and objective HRQoL convergence scores (r = .01; CI -0.02 to 0.001; P = 0.08). The association between psychological distress and HRQoL scores was no longer present when models did not include convergence scores. Carers experiencing more psychological distress tended to rate their own and the care recipients' subjective HRQoL more similarly. Objective HRQoL measures did not show this convergence in scores with increasing carer psychological distress. Findings differed when the analysis approach was changed, suggesting the results above require replication in future studies.


Postural Stability in Children with High Sacral Level Spina Bifida: Deviations from a Control Group.
Objective: To assess static and dynamic postural stability changes in children with high sacral level spina bifida.Methods: Thirty-five children with high sacral level spina bifida and 35 age-matched healthy controls were enrolled. Their lower extremity muscle strengths and static and dynamic postural stability parameters were measured with the use of a dynamometer and the NeuroCom Balance Master® device, respectively. Functional gait and balance were evaluated using the five times sit-to-stand test (5STS) and the 6-minute walk test (6MWT). Spinal, hip, and ankle deformities of the patient group were measured by radiologic evaluation.Results: In comparison with controls, patients were found to have lower ankle dorsiflexion and plantar-flexion strength, increased 5STS duration, and decreased 6MWT distance while both static and dynamic postural stability parameters were significantly different. Bilateral ankle muscle strengths were found to be negatively correlated with postural stability parameters. The presence of hydrocephalus or meningomyelocele in the patient group was found to have negative effects on static postural stability.Conclusion: Static and dynamic postural stability is affected even in children with high sacral level spina bifida who are expected to have best condition in this patient population. The ankle muscle strength is the main factor influencing these changes.


[Driving ability after reversal of phenylephrine 10% induced mydriasis by dapiprazole 0.5%; a prospective study on 65 eyes].
Particularly outpatients want to reach the ability for driving a car as soon as possible after pupillary dilatation. Dapiprazole is an alpha-1 adrenergic inhibitor that antagonizes the mydriatic effect of phenylephrine in a direct competitive way. The aim of this study was to determine restoration of traffic related functions after dapiprazole application in accordance with the guidelines of the German Society of Ophthalmology (DOG). 65 eyes of 33 subjects were tested (17 females, 16 males). All had driving licenses without restrictions. Before mydriasis and after reversal with dapiprazole traffic-related functions were evaluated (photopic visual acuity, mesopic vision, sensitivity to glare, colour vision, accommodation, visual field). No relevant changes of the parameter responsible for the ability to drive a motor vehicle could be shown after reaching premydriatic pupil diameters. There is evidence that reaching normal pupil diameters after reversal of mydriasis by dapiprazole is a valid sign of restoration of traffic related functions.


Odor identification in old age: demographic, sensory and cognitive correlates.
The purpose of this study was to determine correlates of odor identification in old age. One hundred and thirty-two men and women (60-91 years) were assessed in a number of tasks tapping sensory acuity (i.e., odor sensitivity, intensity discrimination, quality discrimination) and different cognitive abilities (i.e., perceptual speed, executive functioning, verbal fluency). Hierarchical regression analyses revealed that age, female sex, olfactory sensitivity, quality discrimination, cognitive speed, and verbal fluency were the most potent correlates of odor identification in general. In addition, the age-related variance in odor identification was eliminated when age-related deficits in odor sensitivity, quality discrimination, and perceptual speed were taken into account. This pattern of outcome suggests that age-related differences in these abilities underlie the well-established age impairment in odor identification.


[Betolepsy in patients with multiple lesions of major cerebral blood vessels].
The reported cases of betolepsy, namely convulsive fits and syncopal attacks induced by cough were observed in patients suffering from different pulmonary diseases, vegetovascular dystonia, and venous encephalopathy. The extracerebral mechanism occurring by the Valsalva type related to the lowering of cardiac output and blood supply to the brain was assumed to be implicated in the origin of betolepsy. In the described group of the patients with multiple occlusive lesions of the cerebral arteries, the pathogenesis of betolepsy is obviously connected with a decrease of the arteriovenous gradient of cerebral circulation to the level close to the critical one. In this case, enhancement of the cerebral blood flow attained by different approaches including reconstructive operations on cerebral vessels rather than administration of anticonvulsants is an effective means of counteracting betolepsy.


Rapid activation of p38 mitogen-activated protein kinase by corticosterone in PC12 cells.
The present study using immunoblot showed that corticosterone (B) could induce a rapid activation of p38 in PC12 cells. The dose and time response curves w ere bell shaped with a maximal activation at 10-9 mol/L and 15 min respectively. The activation was not affected by steroid nuclear receptor antagonist RU38486. Bovine serum albumin coupled B (B-BSA) could induce phosphorylation of p38. Tyrosine kinase inhibitor genistein failed to block the phosphorylation, a fact suggesting that the tyrosine kinase activity is not involved in the pathway. On the other hand, phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, could mimic the actions of B, while G 6976, a PKC inhibitor, could completely abolish the phosphorylation induced by B. These results clearly demonstrate that B activates p38 MAPK readily via a putative membrane receptor through a PKC-dependent pathway.


Pervanadate activation of intracellular kinases leads to tyrosine phosphorylation and shedding of syndecan-1.
Syndecan-1 is a transmembrane haparan sulphate proteoglycan that binds extracellular matrices and growth factors, making it a candidate to act between these regulatory molecules and intracellular signalling pathways. It has a highly conserved transmembrane/cytoplasmic domain that contains four conserved tyrosines. One of these is in a consensus sequence for tyrosine kinase phosphorylation. As an initial step to investigating whether or not phosphorylation of these tyrosines is part of a signal-transduction pathway, we have monitored the tyrosine phosphorylation of syndecan-1 by cytoplasmic tyrosine kinases in intact cells. Tyrosine phosphorylation of syndecan-1 is observed when NMuMG cells are treated with sodium orthovanadate or pervanadate, which have been shown to activate intracellular tyrosine kinases. Initial studies with sodium orthovanadate demonstrate a slow accumulation of phosphotyrosine on syndecan-1 over the course of several hours. Pervanadate, a more effective inhibitor of phosphatases, allows detection of phosphotyrosine on syndecan-1 within 5 min, with peak phosphorylation seen by 15 min. Concurrently, in a second process activated by pervanadate, syndecan-1 ectodomain is cleaved and released into the culture medium. Two phosphorylated fragments of syndecan-1 of apparent sizes 6 and 8 kDa remain with the cell after shedding of the ectodomain. The 8 kDa size class appears to be a highly phosphorylated form of the 6 kDa product, as it disappears if samples are dephosphorylated. These fragments contain the C-terminus of syndecan-1 and also retain at least a portion of the transmembrane domain, suggesting that they are produced by a cell surface cleavage event. Thus pervanadate treatment of cells results in two effects of syndecan-1: (i) phosphorylation of one or more of its tyrosines via the action of a cytoplasmic kinase(s) and (ii) cleavage and release of the ectodomain into the medium, producing a C-terminal fragment containing the transmembrane/cytoplasmic domain.


Social learning of an associative foraging task in zebrafish.
The zebrafish (Danio rerio) is increasingly becoming an important model species for studies on the genetic and neural mechanisms controlling behaviour and cognition. Here, we utilized a conditioned place preference (CPP) paradigm to study social learning in zebrafish. We tested whether social interactions with conditioned demonstrators enhance the ability of focal naïve individuals to learn an associative foraging task. We found that the presence of conditioned demonstrators improved focal fish foraging behaviour through the process of social transmission, whereas the presence of inexperienced demonstrators interfered with the learning of the control focal fish. Our results indicate that zebrafish use social learning for finding food and that this CPP paradigm is an efficient assay to study social learning and memory in zebrafish.


Transcriptional Architecture of Synaptic Communication Delineates GABAergic Neuron Identity.
Understanding the organizational logic of neural circuits requires deciphering the biological basis of neuronal diversity and identity, but there is no consensus on how neuron types should be defined. We analyzed single-cell transcriptomes of a set of anatomically and physiologically characterized cortical GABAergic neurons and conducted a computational genomic screen for transcriptional profiles that distinguish them from one another. We discovered that cardinal GABAergic neuron types are delineated by a transcriptional architecture that encodes their synaptic communication patterns. This architecture comprises 6 categories of ∼40 gene families, including cell-adhesion molecules, transmitter-modulator receptors, ion channels, signaling proteins, neuropeptides and vesicular release components, and transcription factors. Combinatorial expression of select members across families shapes a multi-layered molecular scaffold along the cell membrane that may customize synaptic connectivity patterns and input-output signaling properties. This molecular genetic framework of neuronal identity integrates cell phenotypes along multiple axes and provides a foundation for discovering and classifying neuron types.


Functional and Structural Changes of the Retinal Nerve Fiber Layer and Ganglion Cell Complex in Heavy Smokers.
To assess the functional and structural changes in the retinal nerve fiber layer (RNFL) and the ganglion cell complex (GCC) in heavy smokers using pattern electroretinogram (PERG), photopic negative response(PhNR) and spectral domain optical coherence tomography (SD-OCT). Sixty eyes of 30 heavy smokers (at least 15 cigarettes/day for 10 years) (study group) and 60 eyes of 30 age and gender-matched healthy non-smoker subjects (control group) were included. After full ophthalmologic examination (PERG), (PhNR) using RETI-port/scan 21 (Roland Consult, Brandenburg, Germany) and (SD-OCT using Topcon 3D OCT model 2000 FA version.8.30) were tested for all participants. Statistical analysis was performed to compare GCC, RNFL thicknesses, PERG and phNR values between groups. The mean age was 36.67±4.13 years in the study group and 36.0±4.76 years in the control group. There were no statistical significant differences between the two groups regarding intraocular pressure (p=0.43), axial length (p=0.37), and central corneal thickness (p=0.86). There were significant differences of GCC thickness values between the study group (88.4±6.6 µm) and control group (94.83±5.25 µm) (p<0.001). The mean RNFL values of study and control groups were 97.87±5.88 and 106.43±6.59 µm, respectively (p<0.001). In the study group the superior and inferior quadrants of RNFL were significantly thinner, but there were no significant differences between the two groups in nasal and temporal quadrants. There were no significant differences of PERG-P50 amplitude (p=0.49) and latency (p=0.71); however, PERG-N95 amplitude and latency showed significant differences between the two groups (p<0.001).There were significant differences of phNR amplitude and latency between the two groups (p<0.001). Multiple regression analyses demonstrated that the PhNR, PERG amplitude and latency are the most important determinants for both RNFL and GCC thicknesses. In heavy smokers RNFL thickness was decreased, the PhNR, PERG-N95 amplitudes were diminished and the implicit times were prolonged compared to non-smokers. PhNR and PERG reflect both dysfunction and loss of ganglion cells and their axons.


[Development and evaluation of ultrasound contrast agents with AS1411-conjugated nanoparticles with liquid core].
To prepare AS1411 targeted nano-ultrasonic contrast agent with liquid core, and to evaluate its ability for ultrasonic contrast enhancement and targeting MCF-7 cell in vitro. Methods: The modified solvent evaporation, self-synthesized membrane material and perfluorobrominane (PFOB) was used to form nano-ultrasonic contrast agent with PFOB core (nanoparticles, NP); then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS) catalysis was used to connect AS1411 to the surface of NP to prepare NP-AS1411. The transmission electron microscopy was chosen to check the morphology of NP-AS1411. The size, surface charge, encapsulation efficiency, biocompatibility, the contrast grey value and the stability of NP-AS1411 and NP were compared. Whether AS411 was attached to the surface of NP was checked by gel electrophoresis. Fluorescence microscopy and flow cytometry were performed to examine the targeting ability of AS1411. Results: NP-AS1411 was a shell-nuclear structure under the electron microscope. Its size was at (245.4±16.5) nm, which was larger than that of NP (P=0.05). There was no significant difference in surface charge and encapsulation efficiency between NP-AS1411 and NP (P>0.05). In the MTT experiment, the cell viability decreased significantly at high concentration of NP-AS411 (25 mg/mL) after incubation for 24 h compared with the control group (0 mg/mL ) (P<0.05). The contrast gray value of NP-AS1411 was at 86.1+ 6.7, which was significantly higher than that of deionized water (P<0.05), and equivalent to that of NP (P>0.05). The contrast grey value of AS1411-NP was 80.1±9.2 after keeping at room temperature for 24 h, which showed no obviously change comparing with that before the treatment (P>0.05). The size of NP-AS1411didn't change too (P>0.05). The results of gel electrophoresis demonstrated that the AS1411 connecting to the surface of NP was the most when the molar ratio of NP:AS1411 was at 40:1. Flow cytometry analysis confirmed that NP and NP-AS1411 were combined with MCF-7 cells separately but the fluorescence produced by the combination of NP-AS1411 and MCF-7 was more intense. Conclusion: The modified solvent evaporation and EDC/NHS catalysis could successfully prepare ultrasound contrast agents with aptamer-conjugated nanoparticles with liquid core. The targeted ultrasonic contrast agents with liquid core possess good ultrasonic contrast enhancement ability in vitro, stability and specificity as well.


Outcomes of pancreatic debridement in acute pancreatitis: analysis of the nationwide inpatient sample from 1998 to 2010.
The objective of this study was to perform a national review of patients with acute pancreatitis (AP) who undergo pancreatic debridement (PD) to evaluate for risk factors of in-hospital mortality. The Nationwide Inpatient Sample was used to identify patients with AP who underwent PD between 1998 and 2010. Risk factors for in-hospital mortality were assessed with multivariate logistic regression. From 1998 to 2010, there were 585,978 nonelective admissions with AP, of which 1,783 (.3%) underwent PD. From 1998 to 2010, the incidence of PD decreased from .44% to .25% (P < .01) and PD in-hospital mortality decreased from 29.0% to 15% (P < .05). Of patients undergoing PD, independent factors associated with increased odds of mortality were increased age (odds ratio [OR] 1.04, confidence interval [CI] 1.03 to 1.05; P < .01), sepsis with organ failure (OR 1.76, CI 1.24 to 2.51; P < .01), peptic ulcer disease (OR 1.83, CI 1.02 to 3.30; P < .05), liver disease (OR 2.27, CI 1.36 to 3.78; P < .01), and renal insufficiency (OR 1.78, CI 1.14 to 2.78; P < .05). The incidence and operative mortality of PD have decreased significantly over the last decade in the United States with higher odds of dying in patients who are older, with chronic liver, renal, or ulcer disease, and higher rates of sepsis with organ failure.


Postoperative pain experience: results from a national survey suggest postoperative pain continues to be undermanaged.
Postoperative pain can have a significant effect on patient recovery. An understanding of patient attitudes and concerns about postoperative pain is important for identifying ways health care professionals can improve postoperative care. To assess patients' postoperative pain experience and the status of acute pain management, we conducted a national study by using telephone questionnaires. A random sample of 250 adults who had undergone surgical procedures recently in the United States was obtained from National Family Opinion. Patients were asked about the severity of postsurgical pain, treatment, satisfaction with pain medication, patient education, and perceptions about postoperative pain and pain medications. Approximately 80% of patients experienced acute pain after surgery. Of these patients, 86% had moderate, severe, or extreme pain, with more patients experiencing pain after discharge than before discharge. Experiencing postoperative pain was the most common concern (59%) of patients. Almost 25% of patients who received pain medications experienced adverse effects; however, almost 90% of them were satisfied with their pain medications. Approximately two thirds of patients reported that a health care professional talked with them about their pain. Despite an increased focus on pain management programs and the development of new standards for pain management, many patients continue to experience intense pain after surgery. Additional efforts are required to improve patients' postoperative pain experience. A survey of 250 US adults who had undergone a recent surgical procedure asked about their postoperative pain experience. Approximately 80% of patients experienced pain after surgery. Of these patients, 86% had moderate, severe, or extreme pain. Additional efforts are required to improve patients' postoperative pain experience.


Novel paramagnetic macromolecular complexes derived from the linkage of a macrocyclic Gd(III) complex to polyamino acids through a squaric acid moiety.
Macromolecular Gd(III) complexes may find useful application as contrast agents for magnetic resonance angiography (MRA). Herein two novel systems are reported, namely Gd(DO3ASQ)3-lys16 and Gd(DO3ASQ)30-orn114. Their syntheses are based on the ability of the squaric acid moiety to act as a linker between the DO3A (1,4,7, 10-tetraazacyclododecane-1,4,7-triacetic acid) chelate moiety and the polyamino acidic chain. Moreover, the squaric acid participates in the coordination cage of the Gd(III) ion. The investigation of 1H and 17O NMR relaxation processes of solvent water nuclei allowed a detailed characterization of the systems under study. Gd(DO3ASQ)30-orn114 displays a remarkable ability to enhance the water proton relaxation rate of its solutions, and it may be considered as potential contrast agent for MRA applications.


TRPA1 receptor modulation attenuates bladder overactivity induced by spinal cord injury.
The ankyrin-repeat transient receptor potential 1 (TRPA1) has been implicated in pathological conditions of the bladder, but its role in overactive bladder (OAB) following spinal cord injury (SCI) remains unknown. In this study, using a rat SCI model, we assessed the relevance of TRPA1 in OAB induced by SCI. SCI resulted in tissue damage, inflammation, and changes in bladder contractility and in voiding behavior. Moreover, SCI caused upregulation of TRPA1 protein and mRNA levels, in bladder and in dorsal root ganglion (DRG; L6-S1), but not in corresponding segment of spinal cord. Alteration in bladder contractility following SCI was evidenced by enhancement in cinnamaldehyde-, capsaicin-, or carbachol-induced bladder contraction as well as in its spontaneous phasic activity. Of relevance to voiding behavior, SCI induced increase in the number of nonvoiding contractions (NVCs), an important parameter associated with the OAB etiology, besides alterations in other urodynamic parameters. HC-030031 (TRPA1 antagonist) treatment decreased the number and the amplitude of NVCs while the TRPA1 antisense oligodeoxynucleotide (AS-ODN) treatment normalized the spontaneous phasic activity, decreased the cinnamaldehyde-induced bladder contraction and the number of NVCs in SCI rats. In addition, the cinnamaldehyde-induced bladder contraction was reduced by exposure of the bladder preparations to HC-030031. The efficacy of TRPA1 AS-ODN treatment was confirmed by means of the reduction of TRPA1 expression in the DRG, in the corresponding segment of the spinal cord and in the bladder, specifically in detrusor muscle. The present data show that the TRPA1 activation and upregulation seem to exert an important role in OAB following SCI.


Association of Maternal Insecticide Levels With Autism in Offspring From a National Birth Cohort.
Autism is a complex neurodevelopmental disorder with a largely unknown etiology. To date, few studies have investigated prenatal exposure to toxins and risk of autism by using maternal biomarkers of exposure. Persistent organic pollutants are lipophilic halogenated organic compounds and include the insecticide dichlorodiphenyltrichloroethane (DDT), as well as its metabolite p,p'-dichlorodiphenyl dichloroethylene (p,p'-DDE), and polychlorinated biphenyls (PCBs). The objective of this study was to test whether elevated maternal levels of persistent organic pollutants are associated with autism among offspring. The investigation was derived from the Finnish Prenatal Study of Autism, a national birth cohort study based on a nested case-control design. Cases of autism among children born between 1987 and 2005 were ascertained by national registry linkages. In cases of childhood autism and matched control subjects (778 matched case-control pairs), maternal serum specimens from early pregnancy were assayed for levels of p,p'-DDE and total levels of PCBs. The odds of autism among offspring were significantly increased with maternal p,p'-DDE levels that were in the highest 75th percentile, with adjustment for maternal age, parity, and history of psychiatric disorders (odds ratio=1.32, 95% CI=1.02, 1.71). The odds of autism with intellectual disability were increased by greater than twofold with maternal p,p'-DDE levels above this threshold (odds ratio=2.21, 95% CI=1.32, 3.69). There was no association between total levels of maternal PCBs and autism. These findings provide the first biomarker-based evidence that maternal exposure to insecticides is associated with autism among offspring. Although further research is necessary to replicate this finding, this study has implications for the prevention of autism and may provide a better understanding of its pathogenesis.


Bronchial thermoplasty and the role of airway smooth muscle: are we on the right direction?
Asthma is characterized by inflammation of the airways that includes eosinophils, basal membrane thickening, epithelial sloughing, vascular changes, smooth muscle hypertrophy and hyperplasia, and mucous gland hyperplasia. Recently, there have been studies on the role of hypersensitivity and inflammation in asthma, but the role of bronchial smooth muscle remains unclear. Bronchial thermoplasty is an endoscopic procedure that is approved by the US Food and Drug Administration (FDA) for the treatment of severe refractory asthma, based on the local delivery of radio frequency at 65°C to the airways, with the aim of controlling bronchospasm through a reduction of airway smooth muscle (ASM). Several recent studies have shown significant improvement in clinical outcomes of bronchial thermoplasty for asthma, including symptom control, reduction in exacerbation and hospitalization rates, improved quality of life, and reduction in number of working days or school days lost due to asthma. Data from these recent studies have shown reduction in ASM following bronchial thermoplasty and changes in inflammation patterns. It has also been argued that bronchial thermoplasty may have modulating effects on neuroendocrine epithelial cells, bronchial nerve endings, TRPV1 nerve receptors, and type-C unmyelinated fibers in the bronchial mucosa. This may involve interrupting the central and local reflexes responsible for the activation of bronchospasm in the presence of bronchial hyperreactivity. Several questions remain regarding the use of bronchial thermoplasty, mechanism of action, selection of appropriate patients, and long-term effects. In this review, the role of ASM in the pathogenesis of asthma and the key aspects of bronchial thermoplasty are discussed, with a focus on the potential clinical effects of this promising procedure, beyond the reduction in ASM.


Effect of beta-aminopropionitrile and hyaluronic acid on repair of collagenase-induced injury of the rabbit Achilles tendon.
Collagenase was injected into the Achilles tendon of both hind legs of 10 clinically normal adult male New Zealand white rabbits. One month after induction of the injury, beta-aminoproprionitrile (BAPN) or hyaluronic acid (HA) was injected into the tendon core of the right hind leg of each rabbit, the left hind leg being left untreated. The treatment effects were evaluated by electron microscopy and analysis of the glycosaminoglycan (GAG) content of samples at 2 and 6 months post-treatment. At 2 months, collagen fibrils in tendons from both hind legs were relatively small in diameter, irregularly arranged, and interspersed with abundant active tenocytes as compared with those in normal tendon uninjured by collagenase. In the matrix, the amount of HA increased, but chondroitin-6-sulphate was eliminated. At 6 months, BAPN-treated tendons had small-diameter, regularly arranged collagen fibrils. HA-treated tendons, on the other hand, had large diameters, as well as regularly arranged collagen fibrils by comparison with non-treated tendon. The results suggest that HA, unlike BAPN, promoted healing.


Cerebrolysin for acute ischaemic stroke.
Cerebrolysin is a mixture of low-molecular-weight peptides and amino acids derived from pigs' brain tissue which has proposed neuroprotective and neurotrophic properties. It is widely used in the treatment of acute ischaemic stroke in Russia and China. To assess the benefits and risks of cerebrolysin for treating acute ischaemic stroke. We searched the Cochrane Stroke Group Trials Register (February 2009), the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library Issue 1, 2009), MEDLINE (1966 to February 2009), EMBASE (1974 to February 2009), LILACS (1982 to February 2009), Science Citation Index (1940 to February 2009), SIGLE Archive (1980 to March 2005), and a number of relevant Russian Databases (1988 to February 2009). We also searched reference lists, ongoing trials registers and conference proceedings. Randomised controlled trials comparing cerebrolysin with placebo or no treatment in patients with acute ischaemic stroke. Three review authors independently applied the inclusion criteria, assessed trial quality and extracted the data. We included one trial involving 146 participants. There was no difference in death (6/78 in the cerebrolysin group versus 6/68 in the placebo group; risk ratio (RR) 0.87, 95% confidence interval (CI) 0.29 to 2.58) or in the total number of adverse events (16.4% versus 10.3%; RR 1.62, 95% CI 0.69 to 3.82) between the treatment and control groups. There is not enough evidence to evaluate the effect of cerebrolysin on survival and dependency in people with acute ischaemic stroke. High-quality and large-scale randomised controlled trials may help to gain a better understanding of the potential value of cerebrolysin in acute ischaemic stroke.


Cognitive and behavioral effects of brief seizures in mice.
Comorbidities associated with epilepsy greatly reduce patients' quality of life. Since antiepilepsy drugs show limited success in ameliorating cognitive and behavioral symptoms, there is a need to better understand the mechanisms underlying epilepsy-related cognitive and behavioral impairments. Most prior research addressing this problem has focused on chronic epilepsy, wherein many factors can simultaneously impact cognition and behavior. The purpose of the present study was to develop a testing paradigm using mice that can provide new insight into how short-term biological changes underlying acute seizures impact cognition and behavior. In Experiment 1, naïve C57BL/6J mice were subjected to either three brief, generalized electroconvulsive seizure (ECS) or three sham treatments equally spaced over the course of 30 min. Over the next 2 h, mice were tested in a novel object recognition paradigm. Follow-up studies examined locomotor activity immediately before and after (Experiment 2), immediately after (Experiment 3), and 45 min after (Experiment 4) a set of three ECS or sham treatments. Whereas results demonstrated that there was no statistically significant difference in recognition memory acquisition between ECS and sham-treated mice, measures of anxiety-like behavior were increased and novel object interest was decreased in ECS-treated mice compared with that in sham. Interestingly, ECS also produced a delayed inhibitory effect on locomotion, decreasing open-field activity 45-min posttreatment compared to sham. We conclude that a small cluster of brief seizures can have acute, behaviorally relevant effects in mice, and that greater emphasis should be placed on events that take place before chronic epilepsy is established in order to better understand epilepsy-related cognitive and behavioral impairments. Future research would benefit from using the paradigms defined above to study the effects of individual seizures on mouse cognition and behavior.


Effect of pattern of preceding inspiration on FEV1 in asthmatic children.
In adults, both peak expiratory flow (PEF) and forced expiratory volume in one second (FEV1) are significantly influenced by the time course of the inspiration preceding the forced expiration. The aim of this study was to evaluate the effects of three different inspiratory manoeuvres on PEF, FEV1, and forced vital capacity (FVC) in asthmatic children. Twenty five symptomless asthmatic children performed forced expiration preceded by three different inspiratory manoeuvres, which consisted of: a rapid inspiration with a 2 s end-inspiratory breathhold (Manoeuvre No. 1); a rapid inspiration without an end-inspiratory breathhold (Manoeuvre No. 2); and a slow inspiration lasting about 5 s with an end-inspiratory breathhold of at least 4 s (Manoeuvre No. 3). All manoeuvres were performed in a randomly assigned sequence each morning for three consecutive days. In each session, the manoeuvres were repeated three times and the highest value was chosen. Both FVC and FEV1 obtained with Manoeuvre No. 3 were significantly lower than the corresponding values obtained with Manoeuvre Nos. 1 and 2. The mean (SD) FVC values were 2.76 (0.66) L with Manoeuvre No. 1, 2.67 (0.58) L with Manoeuvre No. 2 and 2.52 (0.52) L with Manoeuvre No. 3. The corresponding values of FEV1 were 2.25 (0.53), 2.22 (0.53) and 2.07 (0.44) L, respectively. By contrast, the values of PEF, obtained with a portable peak flow meter, were similar with the three different inspiratory manoeuvres. The results of this study show that in symptomless asthmatic children the preceding inspiratory manoeuvre may influence forced vital capacity and forced expiratory volume in one second. Hence, in order to reduce variability due to interference by physiological factors and so improve reproducibility of pulmonary function tests, the inspiratory manoeuvres must be accurately standardized.


Health Care Provider Perceptions of Using Social Media to Improve Sexual Health for Adolescent and Young Adult Cancer Survivors.
This study explored (1) health care provider (HCP) perceived barriers and facilitators to social media communication about sexual health with adolescent and young adult survivors, and (2) strategies that can help HCPs navigate social media use for this purpose. Thematic analysis of 11 semistructured HCP interviews resulted in four themes and suggests that social media platforms offer a promising avenue to foster sexual health communication but that resources and training are needed to improve HCP capacity to use this approach ethically and effectively. Future studies are needed to determine specific communication strategies and whether these strategies would lead to improved outcomes.


Cytomegalovirus pneumonia in transplant patients: CT findings.
Our goal was to assess the CT findings of cytomegalovirus (CMV) pneumonia in transplant patients. The study included 10 transplant patients who had chest CT scan and pathologically proven isolated pulmonary CMV infection. Five patients had bone marrow transplant and five had solid organ transplant. The CT scans were retrospectively reviewed for pattern and distribution of disease and the CT findings compared with the findings on open lung biopsy (n = 9) and autopsy (n = 1). Nine of 10 patients had parenchymal abnormalities apparent at CT and 1 had normal CT scans. The findings in the nine patients included small nodules (n = 6), consolidation (n = 4), ground-glass attenuation (n = 4), and irregular lines (n = 1). The nodules had a bilateral and symmetric distribution and involved all lung zones. The consolidation was most marked in the lower lung zones. The CT findings of CMV pneumonia in transplant patients are heterogeneous. The most common patterns include small nodules and areas of consolidation.


Different regulation of eosinophil activity in Crohn's disease compared with ulcerative colitis.
The aim of this investigation was to study the involvement of eosinophil and neutrophil granulocytes in different stages of Crohn's disease (CD) and ulcerative colitis (UC). Biopsy samples were taken from the right flexure of the colon and from the rectum in patients with active (n=12) and inactive colonic CD (n=7), patients with active (n=33) and inactive UC (n=24), and from control subjects (n=11). Cell suspensions from biopsies and blood were analyzed by flow cytometry with regards to activation markers and viability. Immunohistochemistry was used to evaluate cell number and degranulation. Blood eosinophils were cultured with Th1 and Th2 cytokines, and the expression of activity markers was assessed by flow cytometry. Eosinophil number, viability, and activity were increased during active CD and UC compared with controls. The activity, assessed as CD44 expression, tended to diminish during inactive CD but was increased further in quiescent UC. Neutrophil number and activity were increased only during inflammation in both diseases. Culture of blood eosinophils with IL-5 and IL-13 caused increased CD44 expression, whereas IL-5 and IFN-gamma induced elevated CD69 expression. We observed different patterns of eosinophil activation in CD and UC, with the highest CD44 expression during quiescent UC. Our in vitro experiments with recombinant cytokines suggest that the diverse mechanisms of eosinophil activation in CD and UC are a result of different cytokine milieus (Th1 vs. Th2). In contrast, neutrophil activation reflects the disease activity in CD and UC, irrespective of Th cell skewing.


[Relationship in cardiovascular reactivity to mental stress and early involvement of target organs in non-treated mild arterial hypertension. Hospitalet Study].
It has been previously reported that an exaggerated response of blood pressure to mental stress tasks is associated to an increased cardiovascular risk. The objective of this cross-sectional study, with sequential inclusion of subjects who met the inclusion criteria, was to asses the relationships between the response of blood pressure and heart rate to two different mental stress tasks and early target organ-damage, defined as abnormalities in the echocardiogram and/or an increase of urinary albumin excretion (UAE) or microalbuminuria, in untreated mild hypertensive subjects. Two hundred and eleven subjects aged 18 to 65 years (56% males) with mild hypertension (SBP between 140-180 mmHg and/or DBP between 90-105 mmHg) were included in Hospitalet Study. One hundred and thirty seven of them accepted to participate in the study of cardiovascular reactivity. Two different tasks were applied in the same day: a stressful interview (SI) and a mental arithmetic stress tasks (MAST). An echocardiogram of good quality was obtained in 123 cases (89.8%) and 24 h UAE was measured in 108 cases (78.8%). The prevalence of left ventricular hypertrophy was 21.1% (95% CI = 14.3-29.4) and the prevalence of microalbuminuria was 15.7% (95% CI = 9.4-24.4%). After adjusting for the baseline blood pressure, a significant correlation was observed between increase of systolic BP during SI and UAE (r = 0.21; p = 0.03) and between increase of diastolic BP during SI and relative wall thickness (r = 0.32; p < 0.005). When we analyzed the changes of BP during MAST, a significant correlation was observed between increase of diastolic BP (adjusted for baseline diastolic BP) and left atrial size (r = 0.21; p = 0.02). We did not find any significant correlation between the increases of BP (systolic or diastolic) during MAST or increases of heart rate during both tasks and left ventricular mass index or UAE. A weak correlation was observed between cardiovascular reactivity of blood pressure during mental stress tasks and early target organ damage in mild hypertension. We did not find any relationship between the response to heart rate during the tasks and early target organ damage.


Flood management: prediction of microbial contamination in large-scale floods in urban environments.
With a changing climate and increased urbanisation, the occurrence and the impact of flooding is expected to increase significantly. Floods can bring pathogens into homes and cause lingering damp and microbial growth in buildings, with the level of growth and persistence dependent on the volume and chemical and biological content of the flood water, the properties of the contaminating microbes, and the surrounding environmental conditions, including the restoration time and methods, the heat and moisture transport properties of the envelope design, and the ability of the construction material to sustain the microbial growth. The public health risk will depend on the interaction of these complex processes and the vulnerability and susceptibility of occupants in the affected areas. After the 2007 floods in the UK, the Pitt review noted that there is lack of relevant scientific evidence and consistency with regard to the management and treatment of flooded homes, which not only put the local population at risk but also caused unnecessary delays in the restoration effort. Understanding the drying behaviour of flooded buildings in the UK building stock under different scenarios, and the ability of microbial contaminants to grow, persist, and produce toxins within these buildings can help inform recovery efforts. To contribute to future flood management, this paper proposes the use of building simulations and biological models to predict the risk of microbial contamination in typical UK buildings. We review the state of the art with regard to biological contamination following flooding, relevant building simulation, simulation-linked microbial modelling, and current practical considerations in flood remediation. Using the city of London as an example, a methodology is proposed that uses GIS as a platform to integrate drying models and microbial risk models with the local building stock and flood models. The integrated tool will help local governments, health authorities, insurance companies and residents to better understand, prepare for and manage a large-scale flood in urban environments.


Early development of negative and positive affect: Implications for ADHD symptomatology across three birth cohorts.
High levels of early emotionality (of either negative or positive valence) are hypothesized to be important precursors to early psychopathology, with attention-deficit/hyperactivity disorder (ADHD) a prime early target. The positive and negative affect domains are prime examples of Research Domain Criteria (RDoC) concepts that may enrich a multilevel mechanistic map of psychopathology risk. Utilizing both variable-centered and person-centered approaches, the current study examined whether levels and trajectories of infant negative and positive emotionality, considered either in isolation or together, predicted children's ADHD symptoms at 4 to 8 years of age. In variable-centered analyses, higher levels of infant negative affect (at as early as 3 months of age) were associated with childhood ADHD symptoms. Findings for positive affect failed to reach statistical threshold. Results from person-centered trajectory analyses suggest that additional information is gained by simultaneously considering the trajectories of positive and negative emotionality. Specifically, only when exhibiting moderate, stable or low levels of positive affect did negative affect and its trajectory relate to child ADHD symptoms. These findings add to a growing literature that suggests that infant negative emotionality is a promising early life marker of future ADHD risk and suggest secondarily that moderation by positive affectivity warrants more consideration.


Atopic dermatitis management with tacrolimus ointment (Protopic®).
Tacrolimus ointment is the first of a new class of non-steroidal topical immunomodulators indicated for the treatment of atopic dermatitis. Topical tacrolimushas been subject to an extensive clinical development program involving more than 16,000 patients. A clinical trial program, including vehicle-controlled studies, short- and long-term comparative studies and long-term safety studies, has investigated tacrolimus 0.1%and 0.03%ointment for the treatment of atopic dermatitis in adults and children aged 24 months and older. Tacrolimusmonotherapy is rapidly effective, resulting in clinical improvements within three days of starting therapy, and produces a progressive increase in efficacy that is sustained during long-term treatment. Tacrolimus treats the signs and symptoms of atopic dermatitis, reduces the incidence of flares, and offers the potential for long-term disease control. No major safety concerns have been reported to date. Tacrolimusointment is generally well tolerated, the primary adverse events being mild to moderate and transient application-site reactions: skin burning, pruritus and erythema. Tacrolimus oint-ment is a significant advance in dermatology and provides physicians with an alternative to conventional topical corticosteroid therapy.


[E.S.BOTKIN, A SPECIMEN OF FIDELITY TO PROFESSIONAL DUTY AND OATH OF LOYALTY, AN EXAMPLE OF COURAGE AND HONOUR OF THE RUSSIAN DOCTOR].
Evgeny Sergeevch Botkin, son of the legendary Russian internist Sergey Petrovich Botkin, was a court physician for Tsar Nikolai II. After Nikolai abdicated the throne on 15 March (2 March in the Julian calendar) 1917, E.S. Botkin felt it was his duty to accompany the Romanovs into exile to Siberia and continued to selflessly treat the crown prince Aleksey, other members of the Romanov family and all those who applied for his advice. He was shot together with the Romanovs in the basement room of the Ipatiev house, Ekaterinburg, remaining loyal to professional duty and the word given to the Emperor.


A pharmacological and clinical review on topiramate, a new antiepileptic drug.
Topiramate is a new antiepileptic drug which has recently become available in the United States and in a number of European countries. Pharmacological studies suggest that its mode of action is multifactorial and involves blockade of voltage-dependent sodium channels, potentiation of GABAergic transmission and inhibition of excitatory pathways through an action at AMPA receptor sites. Carbonic anhydrase inhibiting properties have also been demonstrated but they are considered not to be relevant to anticonvulsant activity. Topiramate is well absorbed from the gastrointestinal tract, peak plasma levels being usually attained in 2-3 hours. The drug is negligibly (9-17%) bound to plasma proteins and is eliminated partly by renal excretion in unchanged form and partly by oxidation and hydrolysis. In healthy volunteers, the half-life is about 20-30 hours, but elimination rate is accelerated in patients taking concomitant enzyme inducing drugs such as phenytoin, carbamazepine and barbiturates. Topiramate has no major effects on plasma levels of concurrent anticonvulsants, except for a rise in plasma phenytoin in occasional patients. In double-blind add-on trials in refractory partial epilepsy, a significant reduction in seizure frequency has been demonstrated in over 40% of topiramate-treated patients (vs about 10% of those treated with placebo), a response rate which compares favourably with that observed with other new antiepileptic drugs. Dosages found to be effective in add-on controlled trials range between 200 and 1000 mg day-1, although most patients are likely to benefit from receiving 400 mg day-1 or less. Preliminary data suggest that topiramate may be effective also in generalized epilepsies, but this needs to be confirmed in prospective studies. The most common adverse effects of topiramate are CNS-related and include dizziness, fatigue, visual disturbances, ataxia, mental slowing and impaired concentration. Paresthesias, anorexia, weight loss and increased risk of nephrolithiasis have been also reported. Many of these effects are related to dose and/or to rate of dose titration. Based on these data, topiramate appears to be a valuable new drug, whose main current indication is in the add-on management of refractory partial and secondarily generalized seizures. Studies on its potential-value as monotherapy are in progress.


Adaptation of membrane-associated determinants is an outgrowth of their metabolism.
The quantity of receptors expressed by specialized cells for hormones, foreign antigenic substances, and other chemical stimuli diminishes after they associate with ligands, correlating with a specific reduction in the responsiveness of the cell to further stimulation. Ligand-induced adaptation of membrane receptors may be akin to antigenic modulation--the reversible disappearance of membrane-associated determinants stimulated by specific antibodies. Antigenic modulation has been studied extensively in the thymus-leukemia (TL) system of mouse leukemias. Antibody-induced changes in the quantities of TL antigens expressed by ASL-1 and RADA-1 cells, independently arising leukemia cell lines of strain A mice, are outgrowths of their metabolic turnover. After interacting with TL antibodies, the rate of TL antigen disappearance from the membrane increases while the rate of antigen synthesis remains unchanged. Antiserum with specificities for two membrane-associated determinants of ASL-1 cells, TL and a tumor-associated antigen, leads to modulation of TL antigens alone; the tumor antigen persists. Selectivity of modulation is taken as an indication that complex cellular controls govern the affect of antiserum on the expression of membrane antigens. To detect the presence of regulatory controls governing the expression of membrane determinants, stable somatic hybrids of ASL-1 murine leukemia cells and LM(TK)- cells, a sustained mouse cell line, were prepared and antiserum affects on membrane antigen expression were investigated. The metabolic half-lives of each of three antigen determinants investigated was distinct from the others examined. The hybrid cells have lost their capacity to mudulate TL antigens. TL antigens of hybrid cells, unlike those of parental ASL-1 cells, continue to be expressed in the presence of high titers of TL antiserum. Similarities between antigenic modulation and down regulation exist, among which are the fate of receptor-ligand complexes, changes in their metabolism after binding to ligand, and the dependence of the reactions upon continued sources of cellular energy.


Temporal and spatial patterns of transforming growth factor-beta 1 expression in developing rat molars.
Regulatory peptides of the TGF-beta family affect various aspects of embryonic development. Recent immunolocalization and in situ hybridization studies have demonstrated a specific time- and tissue-dependent expression of TGF-beta 1 in the developing mouse embryo. The purpose of this study was to evaluate the distribution of TGF-beta 1 within rat molars at different stages of development, using a well-characterized antibody, highly specific for TGF-beta 1, and immunohistochemical methods of detection. TGF-beta 1 was immunolocalized intensely within the ectodermally derived stellate reticulum and the mesenchyme of the dental papilla at the bell stage of development. Marked immunostaining was also evident in the papillary layer and the reduced dental organ subjacent to ameloblasts in the differentiation and secretory phases of amelogenesis. During the formation of coronal tissues and in the pre-eruptive phase, immunoreactive TGF-beta 1 was localized conspicuously within the dental follicle overlying the tooth germ. This temporospatial pattern of expression of TGF-beta 1 appears to correlate with specific events in morphogenesis, histogenesis and cytodifferentiation during tooth development.


Characterization of a New Protein Family Associated With the Silica Deposition Vesicle Membrane Enables Genetic Manipulation of Diatom Silica.
Diatoms are known for their intricate, silicified cell walls (frustules). Silica polymerization occurs in a compartment called the silica deposition vesicle (SDV) and it was proposed that the cytoskeleton influences silica patterning through the SDV membrane (silicalemma) via interactions with transmembrane proteins. In this work we identify a family of proteins associated with the silicalemma, named SAPs for Silicalemma Associated Proteins. The T. pseudonana SAPs (TpSAPs) are characterized by their motif organization; each contains a transmembrane domain, serine rich region and a conserved cytoplasmic domain. Fluorescent tagging demonstrated that two of the TpSAPs were localized to the silicalemma and that the intralumenal region of TpSAP3 remained embedded in the silica while the cytoplasmic region was cleaved. Knockdown lines of TpSAP1 and 3 displayed malformed valves; which confirmed their roles in frustule morphogenesis. This study provides the first demonstration of altering silica structure through manipulation of a single gene.


Is the beagle dog an appropriate experimental animal for extrapolating data to humans on organ distribution patterns of U, Th, and Pu?
Concentrations and organ distribution patterns of alpha-emitting isotopes of U (238U and 234U), Th (232Th, 230Th, and 228Th), and Pu (239,240Pu) were determined for beagle dogs of our colony. The dogs were exposed to environmental levels of U and Th isotopes through ingestion (food and water) and inhalation to stimulate environmental exposures of the general human population. The organ distribution patterns of these radionuclides in beagles are compared to patterns in humans to determine if it is appropriate to extrapolate organ content data from beagles to humans. The results indicated that approximately 80% of the U and Th accumulated in bone in both species. The organ content percentages of these radionuclides in soft tissues such as liver, kidney, etc. of both species were comparable. The human lung contained higher percentages of U and Th than the beagle lung, perhaps because the longer life span of humans resulted in a longer exposure time. If the U and Th content of dog lung is normalized to an exposure time of 58 y and 63 y, median ages of the U and Th study populations, respectively, the lung content for both species is comparable. The organ content of 239,240Pu in humans and beagles differed slightly. In the beagle, the liver contained more than 60%, and the skeleton contained less than 40% of the Pu body content. In humans, the liver contained approximately 37%, and the skeleton contained approximately 58% of the body content. This difference may have been due to differences in the mode of intake of Pu in each species or to differences in the chemical form of Pu. In general, the results suggest that the beagle may be an appropriate experimental animal from which to extrapolate data to humans with reference to the percentage of U, Th, and Pu found in the organs.


Incidental appendectomy?--Yes.
A consecutive group of 100 patients over the age of 40 years, who were operated upon for histologically proven acute appendicitis, were reviewed. Although there were no deaths, 36 had complications, of which 30 were related to wound infection. Thirty-one patients were found at operation to have perforation. Of these 100 patients, 13 could have had their appendices removed previously at the time of an elective laparotomy. The morbidity of incidental appendectomy was reviewed in 200 randomly selected patients, also over the age of 40 years, who underwent either elective hysterectomy or cholecystectomy. Half had an incidental appendectomy and the other half did not. The indication for appendectomy was the preference of the surgeon. There was no important difference in operative or postoperative problems in the two groups of patients. The authors conclude that incidental appendectomy is a worthwhile and safe procedure unless there are specific contraindications.


[Visual nonverbal set in preschool- and schoolchildren].
Visual set for perception of unequal circles was formed in 5-6-year-old preschool- (n = 30) and schoolchildren (n = 43). Children of 5.3-6.1 years old formed a less rigid set than 9-10-year-olds. The reaction time to a probe stimulus significantly depended on the set stability and stage. The role of structural maturation of the frontal cortical areas in set formation and stability is discussed. The factor of set rigidity was shown to be significant for cognitive activity.


Direct activation of inward rectifier potassium channels by PIP2 and its stabilization by Gbetagamma.
Inward rectifier K+ channels, which modulate electrical activity in many cell types, are regulated by protein kinases, guanine-nucleotide-binding proteins (G proteins) and probably actin cytoskeleton. Generation of phosphatidylinositol 4,5-bisphosphate (PIP2) by ATP-dependent lipid kinases is known to activate inward rectifier K+ channels in cardiac membrane patches. Here we report that several cloned inward rectifier K+ channels directly bind PIP2, and that this binding correlates with channel activity. Application of ATP or PIP2 liposomes activates the cloned channels. Stabilized by lipid phosphatase inhibitors, PIP2 antibodies potently inhibit each channel with a unique rate (GIRK1/4 approximately GIRK2 >> IRK1 approximately ROMK. Consistent with the faster dissociation of PIP2 from the GIRK channels, the carboxy terminus of GIRK1 binds 3H-PIP2 liposomes more weakly than does that of IRK1 or ROMK1. Mutation of a conserved arginine to glutamine at position 188 reduces the ability of ROMK1 to bind PIP2 and increases its sensitivity to inhibition by PIP2 antibodies. Interactions between GIRK channels and PIP2 are modulated by the betagamma subunits of the G protein (Gbetagamma). When GIRK1/4 channels are allowed to run down completely, they are not activated by addition of Gbetagamma alone, but application of PIP2 activates them in minutes without Gbetagamma and in just seconds with Gbetagamma. Finally, coexpression of Gbetagamma with GIRK channels slows the inhibition of K+ currents by PIP2 antibodies by more than 10-fold. Thus Gbetagamma activates GIRK channels by stabilizing interactions between PIP2 and the K+ channel.


Clinical Significance of Biliary Dilatation and Cholelithiasis after Subtotal Gastrectomy.
The well-organized study to support that increased cholelithiasis and bile duct dilatation can occur after gastrectomy has not been reported. The aim of this study was to determine the incidence of cholelithiasis and the degree of common bile duct (CBD) dilatation in patients undergoing subtotal gastrectomy, compared to those undergoing endoscopic treatment for gastric cancer. Patients who diagnosed with gastric cancer and received treatment at six academic referral centers were investigated for the incidence and time of cholelithiasis and the degree of CBD dilatation after treatment by analysis of 5-year follow-up CTs. The operation group underwent subtotal gastrectomy without vagotomy, while in the control group endoscopic treatment was administered for gastric cancer. A total of 802 patients were enrolled in 5-year analysis (735 patients in the operation group and 67 patients in the control group). Cholelithiasis occurred in 47 patients (6.39%) in the operation group and 3 patients (4.48%) in the control group (p=0.7909). The incidences of cholelithiasis were 4.28% in Billoth-I and 7.89% in Billoth-II (p=0.0487). The diameter of proximal CBD and distal CBD increased by 1.11 mm and 1.41 mm, respectively, in the operation group, compared to 0.4 mm and 0.38 mm, respectively, in the control group (pœ0.05). Patients with increased CBD dilatation more than 5 mm showed statistically significant increases in alkaline phosphatase and gamma-glutamyltransferase. The incidence of cholelithiasis was not increased due to subtotal gastrectomy without vagotomy, but the incidence was higher after Billoth-II compared to Billoth-I. In addition, significant change in the CBD diameter was observed after subtotal gastrectomy.


TOZAL Study: an open case control study of an oral antioxidant and omega-3 supplement for dry AMD.
The primary objective of this prospective study was to measure the change from baseline in visual function--Best-Corrected Visual Acuity (BCVA) via the Early Treatment Diabetic Retinopathy Study (ETDRS) chart, contrast sensitivity, central 10 degree visual fields and retinal imaging (angiograms and photographs) at 6 months in subjects with atrophic (dry) age-related macular degeneration treated with a targeted nutritional supplement. 37 mixed gender patients with a mean age of 76.3 +/- 7.8 years were enrolled at 5 independent study sites and received standard of care with a novel formulation of a nutritional supplement. Results were compared to a placebo cohort constructed from the literature that was matched for inclusion and exclusion criteria. A paired t-test was used to test a null hypothesis and a two-sided alpha level of 0.05 was used to determine statistical significance. 76.7% of subjects receiving the nutritional supplement demonstrated stabilization or improvement of BCVA at 6 months. Subjects gained an average of 0.0541 logMAR or one-half of a line of visual acuity (VA) over the 6-month period. There was a statistically significant improvement in VA from baseline with P = .045. The results provide strong evidence that the treatment being studied produces an improvement in VA. Treatment with this unique nutritional supplement increased VA above the expected baseline decrease in the majority of patients in this population with dry macular degeneration. The results of the TOZAL study agree with the LAST and CARMIS studies and are predictive for positive visual acuity outcomes in the AREDS II trial. However, patients will likely require supplementation for longer than 6 months to effect changes in additional visual parameters.


Adverse reaction of Parasika Yavani (Hyoscyamus niger Linn): Two case study reports.
Adverse drug reaction (ADR) is an unpleasant reaction related to the use of medicine at its therapeutic dose. Ayurveda is well aware of such adverse reactions. Parasika Yavani (Hyoscyamus niger Linn.) is an Ayurvedic drug effectively used in many psychological disorders, if not used judiciously it causes adverse reactions. In present study two cases of ADR on the usage of Parasika Yavani are reported. Churna in capsule form given in different dosage forms (500 mg once a day, 250 mg twice a day, 250 mg once a day) in Chittodwega (generalised anxiety disorder). 500mg capsule was given to many patients in the study, but no adverse reactions were noticed except in above given two cases. So, in these two cases, the dose was tapered down to 250 mg twice a day, and then to 250 mg once a day to avert the adverse reactions and to fix the therapeutic dose in such individuals (250 mg once a day). On analysis, these two individuals were found to be of Pitta Prakriti. Parasika Yavani is found to increase Pitta and triggers the establishment of ADRs. So, while administering therapeutic dosage, a physician should be vigilant. In the current study, it is observed that 500 mg of Parasika Yavani powder in Pitta Prakriti individuals triggered ADRs while 250 mg once a day was safe. It was also observed that Kapha and Vata Prakriti, patients did not develop any adverse reactions.


Emi1 class of proteins regulate entry into meiosis and the meiosis I to meiosis II transition in Xenopus oocytes.
Xenopus oocytes are arrested at the G2/prophase boundary of meiosis I and enter meiosis in response to progesterone. A hallmark of meiosis is the absence of DNA replication between the successive cell division phases meiosis I (MI) and meiosis II (MII). After the MI-MII transition, Xenopus eggs are locked in metaphase II by the cytostatic factor (CSF) arrest to prevent parthenogenesis. Early Mitotic Inhibitor 1 (Emi1) maintains CSF arrest by inhibiting the ability of the Anaphase Promoting Complex (APC) to direct the destruction of cyclin B. To investigate whether Emi1 has an earlier role in meiosis, we injected Xenopus oocytes with neutralizing antibodies against Emi1 at G2/prophase and during the MI-MII transition. Progesterone-treated G2/prophase oocytes injected with anti-Emi1 antibody fail to activate Maturation Promoting Factor (MPF), a complex of cdc2/cyclin B, and the MAPK pathway, and do not undergo germinal vesicle breakdown (GVBD). Injection of purified Delta90 cyclin B protein or blocking anti-Emi1 antibody with purified Emi1 protein rescues these meiotic processes in Emi1-neutralized oocytes. Acute inhibition of Emi1 in progesterone treated oocytes immediately after GVBD causes rapid loss of cdc2 activity with simultaneous loss of cyclin B levels and inactivation of the MAPK pathway. These oocytes decondense their chromosomes and enter a DNA replication phase instead of progressing to MII. Prior ablation of Cdc20, addition of methyl-ubiquitin, or addition of nondestructible Delta90 cyclin B rescues the MI-MII transition in Emi1-inhibited oocytes.


The IL23R R381Q gene variant protects against immune-mediated diseases by impairing IL-23-induced Th17 effector response in humans.
IL-23 and Th17 cells are key players in tissue immunosurveillance and are implicated in human immune-mediated diseases. Genome-wide association studies have shown that the IL23R R381Q gene variant protects against psoriasis, Crohn's disease and ankylosing spondylitis. We investigated the immunological consequences of the protective IL23R R381Q gene variant in healthy donors. The IL23R R381Q gene variant had no major effect on Th17 cell differentiation as the frequency of circulating Th17 cells was similar in carriers of the IL23R protective (A) and common (G) allele. Accordingly, Th17 cells generated from A and G donors produced similar amounts of Th17 cytokines. However, IL-23-mediated Th17 cell effector function was impaired, as Th17 cells from A allele carriers had significantly reduced IL-23-induced IL-17A production and STAT3 phosphorylation compared to G allele carriers. Our functional analysis of a human disease-associated gene variant demonstrates that IL23R R381Q exerts its protective effects through selective attenuation of IL-23-induced Th17 cell effector function without interfering with Th17 differentiation, and highlights its importance in the protection against IL-23-induced tissue pathologies.


Quorum-sensing regulation of virulence factors in bacterial biofilm.
Chronic polymicrobial wound infections are often characterized by the presence of bacterial biofilms. They show considerable structural and functional heterogeneity, which influences the choice of antimicrobial therapy and wound healing dynamics. The hallmarks of biofilm-associated bacterial infections include elevated antibiotic resistance and extreme pathogenicity. Biofilm helps bacteria to evade the host defense mechanisms and persist longer in the host. Quorum-sensing (QS)-mediated cell signaling primarily regulates biofilm formation in chronic infections and plays a major role in eliciting virulence. This review focuses on the QS mechanisms of two major bacterial pathogens, Staphylococcus aureus and Pseudomonas aeruginosa and explains how they interact in the wound microenvironment to regulate biofilm development and virulence. The review also provides an insight into the treatment modalities aimed at eradicating polymicrobial biofilms. This information will help us develop better diagnostic modalities and devise effective treatment regimens to successfully manage and overcome severe life-threatening bacterial infections.


Meningoencephalocele formation after nasal septoplasty and management of this complication.
Patients undergoing nasal septoplasty can face many complications. Some of these complications are rare but their results are life threatening. Being aware of this complication could prevent further problems such as enlargement of the bone and dural defect, herniation of the meninges and brain tissue through the defect by pulsation of the brain and ascending infection. With early diagnosis, a less aggressive method could be used to treat this complication. A 50-year-old woman was admitted our hospital with the complaint of loss of consciousness. Her Glasgow coma score was 7 on admission. She had no lateralizing signs, but had nuchal rigidity. Blood pressure was 200/110mm Hg, the respiratory pattern was apneic, complete blood count revealed 12000 leucocytes/mm3 and arterial blood gases showed respiratory acidosis and other biochemical parameters were within normal limits. Computerized cranial tomography (CCT) showed diffuse brain edema without evidence of other signs. Lumbar puncture was performed revealing purulent and highly viscous cerebrospinal fluid (CSF). Late diagnosis and late repair of arachnoidodural tearing could lead to life-threatening complications, and cases with meningitis and larger defects may require more extensive surgery instead of transnasal endoscopic repair.


Role of vinculin in regulating focal adhesion turnover.
Although vinculin (-/-) mouse embryo fibroblasts assemble focal adhesions (FAs), they spread more slowly, less extensively, and close a wound more rapidly than vinculin (+/+) cells. To investigate the structure and dynamics of FAs in these cells, we used real-time interference reflection microscopy (IRM) thus avoiding the need to express exogenous GFP-tagged FA proteins which may be misregulated. This showed that the FAs were smaller, less abundant and turned over more rapidly in vinculin null compared to wild-type cells. Expression of vinculin rescued the spreading defect and resulted in larger and more stable FAs. Phosphatidylinositol 4,5-bisphosphate (PIP2) is thought to play a role in vinculin activation by relieving an intramolecular association between the vinculin head (Vh) and tail (Vt) that masks the ligand binding sites in Vh and Vt. To investigate the role of the vinculin/PIP2 interaction in FA dynamics, we used a vinculin mutant lacking the C-terminal arm (residues 1053-1066) and referred to as the deltaC mutation. This mutation reduced PIP2 binding to a Vt deltaC polypeptide by >90% compared to wild type without affecting binding to Vh or F-actin. Interestingly, cells expressing the vinculin deltaC mutant assembled remarkably stable FAs. The results suggest that vinculin inhibits cell migration by stabilising FAs, and that binding of inositol phospholipids to Vt plays an important role in FA turnover.


Electrospun amorphous solid dispersions of poorly water-soluble drugs: A review.
The development of oral dosage forms for poorly water-soluble active pharmaceutical ingredients (APIs) is a persistent challenge. A range of methods has been explored to address this issue, and amorphous solid dispersions (ASDs) have received increasing attention. ASDs are typically prepared by starting with a liquid precursor (a solution or melt) and applying energy for solidification. Many techniques can be used, with the emergence of electrospinning as a potent option in recent years. This method uses electrical energy to induce changes from liquid to solid. Through the direct applications of electrical energy, electrospinning can generate nanofiber-based ASDs from drug-loaded solutions, melts and melt-solutions. The technique can also be combined with other approaches using the application of mechanical, thermal or other energy sources. Electrospinning has numerous advantages over other approaches to produce ASDs. These advantages include extremely rapid drying speeds, ease of implentation, compatibility with a wide range of active ingredients (including those which are thermally labile), and the generation of products with large surface areas and high porosity. Furthermore, this technique exhibits the potential to create so-called 'fifth-generation' ASDs with nanostructured architectures, such as core/shell or Janus systems and their combinations. These advanced systems can improve dissolution behaviour and provide programmable drug release profiles. Additionally, the fiber components and their spatial distributions can be precisely controlled. Electrospun fiber-based ASDs can maintain an incorporated active ingredient in the amorphous physical form for prolonged periods of time because of their homogeneous drug distribution within the polymer matrix (typically they comprise solid solutions), and ability to inhibit molecular motion. These ASDs can be utilised to generate oral dosage forms for poorly water-soluble drugs, resulting in linear or multiple-phase release of one or more APIs. Electrospun ASDs can also be exploited as templates for manipulating molecular self-assembly, offering a bridge between ASDs and other types of dosage forms. This review addresses the development, advantages and pharmaceutical applications of electrospinning for producing polymeric ASDs. Material preparation and analysis procedures are considered. The mechanisms through which performance has been improved are also discussed.


Complete budding and asymmetric division of primitive model cells to produce daughter vesicles with different interior and membrane compositions.
Asymmetric cell division is common in biology and plays critical roles in differentiation and development. Unicellular organisms are often used as model systems for understanding the origins and consequences of asymmetry during cell division. Although basic as compared to mammalian cells, these are already quite complex. We report complete budding and asymmetric fission of very simple nonliving model cells to produce daughter vesicles that are chemically distinct in both interior and membrane compositions. Our model cells are based on giant lipid vesicles (GVs, 10-30 μm) encapsulating a polyethylene glycol (PEG)/dextran aqueous two-phase system (ATPS) as a crowded and compartmentalized cytoplasm mimic. Ternary lipid compositions were used to provide coexisting micrometer-scale liquid disordered (L(d)) and liquid ordered (L(o)) domains in the membranes. ATPS-containing vesicles formed buds when sucrose was added externally to provide increased osmotic pressure, such that they became not only morphologically asymmetric but also asymmetric in both their interior and their membrane compositions. Further increases in osmolality drove formation of two chemically distinct daughter vesicles, which were in some cases connected by a lipid nanotube (complete budding), and in others were not (fission). In all cases, separation occurred at the aqueous-aqueous phase boundary, such that one daughter vesicle contained the PEG-rich aqueous phase and the other contained the dextran-rich aqueous phase. PEGylated lipids localized in the L(o) domain resulted in this membrane domain preferentially coating the PEG-rich bud prior to division, and subsequently the PEG-rich daughter vesicle. Varying the mole ratio of lipids resulted in excess surface area of L(o) or L(d) membrane domains such that, upon division, this excess portion was inherited by one of the daughter vesicles. In some cases, a second "generation" of aqueous phase separation and budding could be induced in these daughter vesicles. Asymmetric fission of a simple self-assembled model cell, with production of daughter vesicles that harbored different protein concentrations and lipid compositions, is an example of the seemingly complex behavior possible for simple molecular assemblies. These compartmentalized and asymmetrically dividing ATPS-containing GVs could serve as a test bed for investigating possible roles for spatial and organizational cues in asymmetric cell division and inheritance.


In vitro skin irritation: facts and future. State of the art review of mechanisms and models.
The skin is the main target tissue for exogenous noxes, protecting us from harmful environmental hazards, UV-irradiation and endogenous water loss. It is composed of three layers, whereas the outermost epidermis is a squamous epithelium that mainly consists of keratinocytes. These cells execute a terminal differentiation, which finally results in the assembly of the stratum corneum. This layer, consisting of cornified keratinocytes, is an effective barrier against a vast number of substances. Apart of this, keratinocytes play crucial roles in the immune surveillance and the initiation, modulation and regulation of inflammation in the epidermis. Regarding cutaneous inflammatory reactions, skin irritation is one of the most common adverse effect in humans. For reasons of human safety assessment new chemicals are still evaluated for irritant potentials by application to animals followed by visible changes such as erythema and oedema. Testing for skin irritation in animals potentially cause them pain and discomfort. Furthermore, the results are not always predictive for those found in humans. In order to replace animal testing and to improve the prediction of irritants, the cosmetic and toiletry industry, in Europe represented by Colipa, develops and uses several alternative in vitro test systems. In this respect, the use of in vitro reconstructed organotypic skin equivalents are mostly favored, because of their increasingly close resemblance to human skin. Due to ethical and scientific questions and on account of the 7th amendment of the European Council Directive 76/768/EEC, the authors see the requirement to drive the development of alternative tests for irritants. Therefore, this article centres on cosmetic ingredients and provides the readership an overview of the state of art of cellular mechanisms of skin irritation and summarizes the results of the commonly used skin equivalents to evaluate irritation in vitro.


Diagnosis on death certificates of sudden death due to ischemic heart disease.
This study was undertaken to evaluate the quality of diagnoses of sudden death (SD) caused by ischemic heart disease (IHD) on death certificates. A random sample of 10% (100 cases) was drawn from all such deaths which occurred in 1970 among Oklahoma City residents. The medical records of each case were reviewed and the quality of the diagnosis was rated, by the use of predetermined standard criteria, as confirmed and valid or unconfirmed and invalid. Among 100 IHD deaths, SD constitute 45%. Of these SDs, 18% were unobserved, without information as to the manifestations of the attack or the interval from onset of symptoms to death. According to criteria of the American Heart Association and WHO Expert Committee, the diagnosis in this 18% was unconfirmed and invalid. Thus there is insufficient quantitative evidence to justify the use of SD, as found on death certificates, as an indication of frequency of ischemic heart disease.


High Throughput Assay to Examine Egg-Laying Preferences of Individual Drosophila melanogaster.
Recently, egg-laying preference of Drosophila has emerged as a genetically tractable model to study the neural basis of simple decision-making processes. When selecting sites to deposit their eggs, female flies are capable of ranking the relative attractiveness of their options and choosing the "greater of two goods." However, most egg-laying preference assays are not practical if one wants to take a systematic genetic screening approach to search for the circuit basis underlying this simple decision-making process, as they are population-based and laborious to set up. To increase the throughput of studying of egg-laying preferences of single females, we developed custom chambers that each can simultaneously assay egg-laying preferences of up to thirty individual flies as well as a protocol that ensures each female has a high egg-laying rate (so that their preference is readily discernable and more convincing). Our approach is simple to execute and produces very consistent results. Additionally, these chambers can be equipped with different attachments to allow video recording the egg-laying animals and to deliver light for optogenetics studies. This article provides the blueprints for fabricating these chambers and the procedure for preparing the flies to be assayed in these chambers.


Clinical screening of Nocardia in sputum smears based on neural networks.
Nocardia is clinically rare but highly pathogenic in clinical practice. Due to the lack of Nocardia screening methods, Nocardia is often missed in diagnosis, leading to worsening the condition. Therefore, this paper proposes a Nocardia screening method based on neural networks, aiming at quick Nocardia detection in sputum specimens with low costs and thereby reducing the missed diagnosis rate. Firstly, sputum specimens were collected from patients who were infected with Nocardia, and a part of the specimens were mixed with new sputum specimens from patients without Nocardia infection to enhance the data diversity. Secondly, the specimens were converted into smears with Gram staining. Images were captured under a microscope and subsequently annotated by experts, creating two datasets. Thirdly, each dataset was divided into three subsets: the training set, the validation set and the test set. The training and validation sets were used for training networks, while the test set was used for evaluating the effeteness of the trained networks. Finally, a neural network model was trained on this dataset, with an image of Gram-stained sputum smear as input, this model determines the presence and locations of Nocardia instances within the image. After training, the detection network was evaluated on two datasets, resulting in classification accuracies of 97.3% and 98.3%, respectively. This network can identify Nocardia instances in about 24 milliseconds per image on a personal computer. The detection metrics of mAP50 on both datasets were 0.780 and 0.841, respectively. The Nocardia screening method can accurately and efficiently determine whether Nocardia exists in the images of Gram-stained sputum smears. Additionally, it can precisely locate the Nocardia instances, assisting doctors in confirming the presence of Nocardia.


Receptors for the anaphylatoxin C5a (CD88) on human mesangial cells.
In these studies, we determined whether there are receptors for the anaphylatoxin C5a (C5aR, CD88) on human mesangial cells (HMC). To prepare Abs to C5aR, we first synthesized an immunogenic peptide spanning residues 8-32 of the molecule, and this peptide was used to immunize rabbits. Anti-C5aR antiserum, but not preimmune serum, stained fixed and unfixed HMC in culture. By Western blotting anti-C5aR, Abs identified a 49.6-kDa protein in HMC. By reverse-transcription PCR, a cDNA product of 558 bp was amplified corresponding to the expected size of C5aR cDNA. A cDNA of the same size was amplified simultaneously from human PBL. Restriction mapping of the products amplified from HMC and from PBL gave restriction fragments of the same size. Incubation of HMC with increasing doses of C5a caused a progressive increase in the levels of the transcription factors activator protein-1 (AP-1) and cAMP response element binding protein (CREB), but C5a had no effect on the level of nuclear factor-kappaB (NF-kappaB). The effects of C5a on AP-1 were concentration and time dependent and peaked after 60 min. In contrast, the C5a metabolite C5adesArg had no significant effect on AP-1 levels. Preincubation of HMC with rabbit anti-C5aR antiserum inhibited partially the effect of C5a on AP-1. However, anti-C5aR Abs alone had no appreciable effects on AP-1. C5a caused a significant up-regulation of mRNA for the early response genes c-jun and c-fos on HMC. These results provide evidence for the presence of C5aR in adult HMC in culture and indicate that, after binding to C5aR, the anaphylatoxin C5a causes significant up-regulation of certain transcription factors and early response genes.


Anoikis-associated signatures predict prognosis and immune response in bladder cancer.
Objective: Anoikis is a type of programmed cell death that occurs in normal epithelial and endothelial cells. However, the specific role of anoikis regulators (ANRs) in bladder cancer (BLCA) remains unknown. Therefore, the objective of this study was to find subgroups that could identify different levels of anoikis resistance in BLCA and construct an anoikis scoring system to assess prognosis. Method: By obtaining ANRs from public datasets, subgroups of BLCA with varying degrees of anoikis resistance were identified, and risk was determined. Result: ANRs affects the occurrence and prognosis of BLCA and can be predicted by establishing risk models. Conclusion: The anoikis scoring system and anoikis-associated risk profiles may help develop more effective and personalized treatment strategies for BLCA patients.


Low-molecular-weight heparin dosage in newborn foals.
Heparin is used in humans as prophylaxis of hypercoagulable states and disseminated intravascular coagulation (DIC). However, babies need a higher heparin dose than do adults. Septic neonate foals are at high risk of hypercoagulable state and DIC, and there is limited objective information about heparin dose for equine neonates. To assess whether neonate foals require higher dosages of low-molecular-weight heparin (LMWH) than adults. Eighteen healthy and 11 septic neonate foals. Experimental and clinical studies. Firstly, healthy foals were randomly distributed in 2 groups, 1 receiving 50 IU/kg SC of dalteparin and the 2nd group receiving 100 IU/kg SC of dalteparin, once daily for 3 days. Blood samples were collected before and 3, 6, 27, and 51 hours after the 1st LMWH administration. Plasma antifactor-Xa activity was measured, together with hemostatic and hematologic parameters used to assess the risk of bleeding. Subsequently, septic foals were treated blindly either with placebo (saline) or 100 IU/kg of dalteparin for 3 days. Plasma antifactor-Xa activity and other hemostatic parameters were determined before and after treatment. Plasma antifactor-Xa activity in healthy foals was below prophylactic activity when using the adult dosage (50 IU/kg), whereas prophylactic activities were achieved when using the double dosage (100 IU/kg). No hemorrhagic events and erythrocyte-related complications were observed with either dosage. In the clinical study, only 4/6 septic foals had plasma antifactor-Xa activity adequate for prophylaxis. Equine neonates require higher dosages of LMWH compared with adults to reach prophylactic heparinemia.


[Long-term results of comprehensive treatment for cervical cancer with poor prognosis].
The data on a randomized treatment of 100 patients with locally advanced cervical carcinoma are discussed. Half of the patients received complex therapy while the rest--radiation alone (control). The modality also included immunotherapy with epithalamin 100 mg, intramuscularly, neoadjuvant intraarterial polychemotherapy with 5-fluorouracil 2 g/m2 and cisplatin 100 mg/m2, concurrent radiotherapy with an interval and 4-6 courses of adjuvant polychemotherapy. As a consequence, short-term results improved by 18.8% in cases tumor size under T3; end-results--by 14%. Improvement in survival was accounted for by causing tumor process to slow down considerably. The adjuvant polychemotherapy component was thought to be responsible for a 25.4% increase in one-year survival and 12.5%--in 3-year survival.


Ultrastructural aspects of activation and contraction of vascular smooth muscle.
Ultrastructural studies of potential intracellular calcium storage sites and of the organization of contractile proteins in vascular smooth muscle are reviewed. The sarcoplasmic reticulum (SR) is a system of closed tubules present in every smooth muscle examined. The volume of the SR varies in different smooth muscles (from approximately 2.0 to 7.5% of cytoplasmic volume) and correlates with the ability of a given smooth muscle to contract in calcium-free media. The SR accumulates the divalent cation strontium and forms couplings with the surface membrane. Mitochondria are other potential sites of calcium accumulation in vascular smooth muscle, as indicated by the respiration supported accumulation of calcium (with a Km of approximately 17 muM) by isolated mitochondria and by the energy dependent accumulation of barium by mitochondria in situ. The presence of barium in mitochondrial granules in tissues contracted with barium and the presence of strontium and calcium in appropriately incubated preparations have been verified with electron probe microanalysis. Calcium has also been demonstrated in mitochondrial granules in frozen dried cardiac muscle sections and in cultured vascular smooth muscle cells. This technique appears suitable for eventual quantitation of mitochondrial calcium content in vascular and other smooth muscles. Thin (actin, 50-80 A), thick (myosin, approximately 155 A), and intermediate (approximately 100 A) filaments are present in suitably fixed vascular smooth muscle. In rabbit portal anterior mesenteric vein intermediate high voltage stereo electron microscopy shows the myosin filaments to be tapered and approximately 2.2 mum long: significantly longer than the myosin filaments in vertebrate striated muscle. Actin filaments insert on dense bodies. The ultrastructural findings are compatible with physiological evidence suggesting the contributions of intracellular organelles to the regulation of cytoplasmic free calcium levels and the operation of a sliding filament mechanism of contraction in vertebrate smooth muscle.