Anti-vitamin D, vitamin D in SLE: preliminary results.
The aim of this study was to detect antibodies to vitamin D in systemic lupus erythematosus (SLE) and other autoimmune diseases. The results may shed light to a novel aspect of vitamin D deficiency in autoimmune diseases. Sera from 171 patients with SLE, 56 with antiphospholipid syndrome (APS), and 18 with pemphigus vulgaris (PV) were studied employing an enzyme-linked immunosorbent assay for anti-vitamin D antibodies along with 94 healthy blood donors. In parallel, vitamin D concentrations in the serum were determined by a DiaSorin commercial kit (LIAISON 25 OH vitamin D). Antibody-positive and antibody-negative individuals were compared with respect to demographic variables, SLE disease activity index (SLEDAI) score, autoantibodies profile, and serum vitamin D levels. Anti-vitamin D antibodies were detected in 7 (4%) of 171 patients with SLE, in 2 (3.5%) of 56 of sera from patients with APS, and in 2 (11%) of 18 sera from patients with PV. Vitamin D levels were similar in both SLE groups with and without anti-vitamin D antibodies. Demographic features, organ involvement, SLEDAI score, and autoantibodies did not differ between the groups. Except for anti-dsDNA antibodies, in which anti-vitamin D antibodies were strongly associated with these antibodies in sera from SLE patients (P = 0.0004). Anti-vitamin D antibodies are observed in a subset of patients with SLE, APS, and PV, and are associated with anti-dsDNA antibodies in SLE. Further studies are required to explore the potential diagnostic and prognostic role of these novel antibodies in SLE.


Magnetohydrodynamic interference with the edge pedestal motional Stark effect diagnostic on DIII-D.
Accurate measurement of internal magnetic field direction using motional Stark effect (MSE) polarimetry in the edge pedestal is desired for nearly all tokamak scenario work. A newly installed 500 kHz 32-channel digitizer on the MSE diagnostic of DIII-D allows full spectral information of the polarimeter signal to be recovered for the first time. Fourier analysis of this data has revealed magnetohydrodynamic (MHD) fluctuations in the plasma edge pedestal at ρ ≥ 0.92. By correlating edge localized mode fluctuations seen on lock-in amplifier outputs with MSE spectrograms, it has been shown that edge pedestal tearing mode fluctuations cause interference with MSE second harmonic instrument frequencies. This interference results in unrecoverable errors in the real-time polarization angle measurement that are more than an order of magnitude larger than typical polarimeter uncertainties. These errors can cause as much as a 38% difference in local q. By using a redundant measure of the linear polarization found at the fourth harmonic photo-elastic modulator (PEM) frequency, MHD interference can be avoided. However, because of poorer signal-to-noise the fourth harmonic signal computed polarization angle shows no improvement over the MHD polluted second harmonics. MHD interference could be avoided in future edge pedestal tokamak polarimeters by utilizing PEMs with higher fundamental frequencies and a greater separation between their frequencies.


Electrical effects of histamine on monolayers formed in culture from enriched canine gastric chief cells.
To develop techniques for studying transport properties and secretory function of selected cell types in the gastric mucosa, separated fractions of dispersed canine fundic mucosal cells were placed in short-term culture to form epithelial monolayers. Cell fractions enriched in either chief, parietal, or mucous cells were prepared by using counterflow centrifugation and were plated on type I collagen. An epithelial monolayer formed by approximately equal to 36 hr. Immunofluorescence with an antipepsinogen I antibody revealed pepsinogen-containing granules in greater than 95% of the cells, regardless of whether the monolayers were formed from the mucous, chief, or parietal cell-enriched fractions. Upon achieving confluency, chief cell monolayers were mounted in Ussing chambers to study their electrical properties. Under basal conditions, monolayers (n = 6) had a spontaneous potential difference (PD) (+/- SEM) of 26 +/- 4 mV (apical surface negative), a short-circuit current (Isc) (+/- SEM) of 16 +/- 2 microA/cm2, and a transepithelial resistance (R) (+/- SEM) of 1,480 +/- 210 omega X cm2. Histamine increased the short-circuit current, an effect blocked by an H2-receptor antagonist. Seventy percent of the spontaneous PD was amiloride sensitive, suggesting sodium absorption accounted for a major component of the PD. These preparative techniques yield highly enriched chief cell monolayers, which maintain morphological and functional cellular differentiation for greater than 48 hr in culture, thus allowing study of oriented functions of a selected cell type. The present studies indicate that an H2 receptor enhances electrogenic ion transport in chief cell monolayers, indicating that histamine can act on fundic mucosal cells other than just parietal cells.


Physical Activity to Reduce Systemic Inflammation Associated With Chronic Pain and Obesity: A Narrative Review.
The increasing prevalence of chronic pain and obesity has significant health and cost implications for economies in the developed and developing world. Evidence suggests that there is a positive correlation between obesity and chronic pain and the link between them is thought to be systemic inflammation. The aim of this narrative review was to explore the physiological links between chronic musculoskeletal pain and obesity and to consider the potential role of regular physical activity in providing a means of managing obesity-related chronic pain. Systemic inflammation, mechanical overload, and autonomic dysfunction are associated with increased prevalence and severity of chronic pain in individuals with obesity. It has been proposed, therefore, that interventions that target systemic inflammation could help to reduce chronic pain in obese individuals. Reduction in abdominal fat has been shown to alleviate pain and reduce the systemic markers of inflammation that contribute to chronic pain. Interventions that include exercise prescription have been shown to reduce both abdominal fat and systemic inflammation. Furthermore, exercise is also known to reduce pain perception and improve mental health and quality of life that also improves pain outcomes. However, adherence to formal exercise prescription is poor and therefore exercise programmes should be tailored to the interests, needs, and abilities of individuals to reduce attrition.


Cervical ripening as an outpatient procedure in the pandemic - minimizing the inpatient days and lowering the socioeconomic costs.
With an increasing incidence of labor induction the socioeconomic costs are increasing and the burden on hospital capacities is rising. In addition, the worldwide SARS-CoV-2 pandemic asks for improvements in patient care during pregnancy and delivery while decreasing the patient-staff contact. Here, we are retrospectively analyzing and comparing a mechanical ripening device that is utilized as an outpatient procedure to misoprostol and dinoprostone as inpatient induction methods in a low risk cohort. This is a retrospective comparative analysis of obstetric data on patients who presented for cervical ripening and labor induction. Ninety-six patients received a mechanical ripening agent as an outpatient procedure. As a control group, we used 99 patients with oral misoprostol (PGE1) and 42 patients with vaginal dinoprostone (PGE2) for cervical ripening in an inpatient setting. Data from 2016 until 2020 were analysed. Baseline characteristics showed no significant differences. Delivery modes were similar in all groups. The time period from patient admission to onset of labor was significantly shorter in the outpatient group (p<0.001): 10.9 h/0.5 days (±13.6/0.6) for osmotic dilator vs. 17.9 h/0.7 days (±13.1/0.5) for oral misoprostol vs. 21.8 h/0.8 days (±15.9/0.7) for vaginal dinoprostone. With 20.4 h/0.8 days (±14.3/0.6) the osmotic dilator group displayed significantly the shortest inpatient stay from admission to delivery (p=0.027). The patient subgroup of misoprostol had 25.7 h/1.1 days (±14.9/0.6) of inpatient stay from admission to delivery and the patient group of dinoprostone 27.5 h/1.1 days (±16.0/0.7). There were fewer hospital days in the outpatient group: 84.9 h/3.5 days vs. 88.9 h/3.7 days vs. 93.6 h/3.9 days (outpatient osmotic dilator vs. inpatient misoprostol and dinoprostone, respectively). New approaches are required to decrease individual contacts between patients and staff while maintaining a high quality patient care in obstetrics. This analysis reveals that outpatient mechanical cervical ripening can be as safe and effective as inpatient cervical ripening with PGE1/PGE2, while lowering patient-staff contact and total hospital stays and therefore decreasing the socioeconomic costs.


Mineralocorticoid receptors are indispensable for nongenomic modulation of hippocampal glutamate transmission by corticosterone.
The adrenal hormone corticosterone transcriptionally regulates responsive genes in the rodent hippocampus through nuclear mineralocorticoid and glucocorticoid receptors. Via this genomic pathway the hormone alters properties of hippocampal cells slowly and for a prolonged period. Here we report that corticosterone also rapidly and reversibly changes hippocampal signaling. Stress levels of the hormone enhance the frequency of miniature excitatory postsynaptic potentials in CA1 pyramidal neurons and reduce paired-pulse facilitation, pointing to a hormone-dependent enhancement of glutamate-release probability. The rapid effect by corticosterone is accomplished through a nongenomic pathway involving membrane-located receptors. Unexpectedly, the rapid effect critically depends on the classical mineralocorticoid receptor, as evidenced by the effectiveness of agonists, antagonists, and brain-specific inactivation of the mineralocorticoid but not the glucocorticoid receptor gene. Rapid actions by corticosterone would allow the brain to change its function within minutes after stress-induced elevations of corticosteroid levels, in addition to responding later through gene-mediated signaling pathways.


Cerebral melanoma metastases: a critical review on diagnostic methods and therapeutic options.
Malignant melanoma represents the third most common cause for cerebral metastases after breast and lung cancer. Central nervous system (CNS) metastases occur in 10 to 40% of patients with melanoma. Most of the symptoms of CNS melanoma metastases are unspecific and depend on localization of the lesion. All patients with new neurological signs and a previous primary melanoma lesion must be investigated. Although primary diagnosis may rely on computed tomography scan, magnetic resonance images are usually used in order to study more precisely the characteristics of the lesions in and to embase the surgical plan. Other possible complementary exams are: positron emission tomography, iofetamine cintilography, immunohistochemistry of liquor, monoclonal antibody immunocytology, optical coherence tomography, and transcriptase-polymerase chain reaction. Treatment procedures are indicated based on patient clinical status, presence of unique or multiple lesions, and family agreement. Often surgery, radiosurgery, whole brain radiotherapy, and chemotherapy are combined in order to obtain longer remissions and optimal symptom relieve. Corticoids may be also useful in those cases that present with remarkable peritumoral edema and important mass effect. Despite of the advance in therapeutic options, prognosis for patients with melanoma brain metastases remains poor with a median survival time of six months after diagnosis.


Adhesional small bowel obstruction following anti-reflux surgery in children--comparison of 232 laparoscopic and open fundoplications.
Adhesions are a major cause of morbidity following abdominal surgery. There is debate whether laparoscopic abdominal surgery results in a lower incidence of adhesional small bowel obstruction (ASBO) compared with the equivalent open procedure. The aim of this study was to study the incidence of ASBO following laparoscopic and open anti-reflux surgery. The case notes of all patients undergoing fundoplication (open or laparoscopic) between November 1997 and November 2005 were reviewed retrospectively. Laparotomy due to ASBO was chosen as the endpoint as it was a clearly identifiable and incontrovertible endpoint. The incidence of ASBO following each operation was compared using Fisher's exact test (p<0.05 was considered significant). A total of 232 patients were analysed. There were no ASBO following 170 laparoscopic fundoplications. There were 5 episodes (8.1%) of ASBO in 3 patients (4.8%) following 62 open procedures. Two of the three patients who developed ASBO after open fundoplication had undergone concomitant Ladd's procedure. There was no significant difference in the rate of ASBO following laparoscopic and open fundoplication alone. ASBO is a serious complication following any abdominal surgery. The data from this study indicate that the risk of ASBO following laparoscopic fundoplication is negligible. Although there is a suggestion that the risk of ASBO is lower following laparoscopic fundoplication compared with open fundoplication, there was no significant difference between the two groups as Ladd's procedure was performed in 2 of the open cases.


Breast Cancer Classification from Ultrasound Images Using Probability-Based Optimal Deep Learning Feature Fusion.
After lung cancer, breast cancer is the second leading cause of death in women. If breast cancer is detected early, mortality rates in women can be reduced. Because manual breast cancer diagnosis takes a long time, an automated system is required for early cancer detection. This paper proposes a new framework for breast cancer classification from ultrasound images that employs deep learning and the fusion of the best selected features. The proposed framework is divided into five major steps: (i) data augmentation is performed to increase the size of the original dataset for better learning of Convolutional Neural Network (CNN) models; (ii) a pre-trained DarkNet-53 model is considered and the output layer is modified based on the augmented dataset classes; (iii) the modified model is trained using transfer learning and features are extracted from the global average pooling layer; (iv) the best features are selected using two improved optimization algorithms known as reformed differential evaluation (RDE) and reformed gray wolf (RGW); and (v) the best selected features are fused using a new probability-based serial approach and classified using machine learning algorithms. The experiment was conducted on an augmented Breast Ultrasound Images (BUSI) dataset, and the best accuracy was 99.1%. When compared with recent techniques, the proposed framework outperforms them.


[The effects of transcorneal administration of prostaglandin E2 on rabbit eyes].
The pharmacologic effects of prostaglandin E2 (PGE2) on the anterior ocular segment were investigated. PGE2 was administered with a glass cylinder attached to the cornea in concentrations ranging from 0.05 to 500 micrograms/ml. The intraocular inflammatory reaction was assessed according to the changes in the anterior chamber flare, ocular tension, and components of the aqueous humor. We also performed experiments designed to inhibit the reactions induced by PGE2 using anti-inflammatory agents. The PGE2 elicited a single peak reaction in the anterior chamber flare, which recovered after several hours, and a transient elevation in the ocular tension in the early stage after PGE2 administration. Quantitative analysis of these two reactions revealed that both were dependent on the dose of PGE2 administered. The concentrations of ascorbic acid and glutathione, particularly that of the reduced form of the latter, were transiently reduced in the aqueous humor. The anterior chamber flares were suppressed by agents that inhibit arachdonic acid metabolism. These findings suggest that the metabolism of arachdonic acid becomes newly activated as a result of stimulation by PGE2 of extrinsic origin.


Thymoma with osseous metaplasia.
A 69-year-old woman with a 15-year history of abnormal chest shadow was referred to our hospital. An enhanced chest CT scan of the anterior mediastinum revealed a mass containing a high-density area. The preoperative radiologic diagnosis was thymoma. Operation was performed in October 2000. Histologically, multiple ossified areas were observed within the tumor. Intratumoral ossification has never been reported in thymoma. Therefore, we report the first case of thymoma associated with multiple foci of osseous metaplasia.


Proteomic analysis of the secretome of human umbilical vein endothelial cells using a combination of free-flow electrophoresis and nanoflow LC-MS/MS.
Human umbilical vein endothelial cells are the most widely used in vitro model for endothelial cells. Their secreted proteins, however, have not been comprehensively analysed so far. In this study, we accomplished to map the secretome of human umbilical vein endothelial cells by combining free-flow electrophoresis with nanoflow LC-MS/MS. This comprehensive analysis provides a basis for future comparative studies of protein secretion by endothelial cells in response to cardiovascular risk factors and is available on our website http://www.vascular-proteomics.com.


Liquid chromatographic/electrospray ionization tandem mass spectrometric study of polyphenolic composition of four cultivars of Fragaria vesca L. berries and their comparative evaluation.
High-performance liquid chromatography coupled with ion spray mass spectrometry in the tandem mode with both negative and positive ionization was used for investigating a variety of polyphenolic compounds in four genotypes of Fragaria vesca berries. About 60 phenolic compounds belonging to the compound classes of phenolic acids, ellagitannins, ellagic acid derivatives, flavonols, monomeric and oligomeric flavanols, dihydrochalcones and anthocyanins were reported, providing for the first time a quite complete picture of polyphenolic composition of F. vesca berries. Some of the polyphenols herein investigated, such as a tris-galloyl-hexahydroxydiphenoyl-hexose, two castalagin/vescalagin-like isomers and peonidin-malonylglucoside, were described for the first time. Principal component analysis applied on original HPLC-MS/MS data, acquired in multiple reaction monitoring mode, successfully discriminated the four investigated cultivars on the basis of their polyphenolic composition, highlighting the fundamental role of mass spectrometry for food characterization.


Anionic halide···alcohol clusters in the solid state.
The cationic (1,3,5-triazapentadiene)Pt(II) complexes [1](Cl)2, [2](Cl)2, [3](Br)2, and [4](Cl)2, were crystallized from ROH-containing systems (R = Me, Et) providing alcohol solvates studied by X-ray diffraction. In the crystal structures of [1-4][(Hal)2(ROH)2] (R = Me, Et), the Hal(-) ion interacts with two or three cations [1-4](2+) by means of two or three or four contacts thus uniting stacked arrays of complexes into the layers. The solvated MeOH or EtOH molecules occupy vacant space, giving contacts with [1-4](2+), and connects to the Hal(-) ion through a hydrogen bridge via the H(1O)O(1S) H atom forming, by means of the Hal(-)···HOR (Hal = Cl, Br) contact, the halide-alcohol cluster. Properties of the Cl(-)···HO(Me) H-bond in [1][(Cl)2(MeOH)2] were analyzed using theoretical DFT methods.


Tobacco and Cannabis Use During and After Pregnancy in California.
As the social and legal acceptance of cannabis use grows, health professionals must understand and mitigate the impact of cannabis use in the perinatal period. Here we compare the prevalence of tobacco and cannabis use during and after pregnancy in California, a state that recently legalized cannabis use. Measures of tobacco and cannabis use during and after pregnancy were obtained from California's Maternal and Infant Health Assessment, an annual population-based survey of California resident women with a live birth. To allow analysis of county-level variation, we pooled data from the 35 counties with the largest numbers of births from 2017 to 2019. Cannabis use was more than twice as common as cigarette smoking among pregnant women (4.9% vs. 2.1%) in California. This difference was even more pronounced in some counties; for example, in Los Angeles, cannabis use was four times more prevalent than cigarette use. Either during or soon after birth, 7.3% of women in California reported cannabis use. Of those who smoked tobacco cigarettes prior to pregnancy, 73% quit before their third trimester of pregnancy, though 33.0% of these women reported a post-partum relapse in cigarette use. States that have legalized cannabis must attend to the increasing prevalence of perinatal cannabis use, as well as concurrent use with tobacco and other substances. Efforts to support cannabis cessation should draw from successful public health approaches in tobacco control.


[Gastroesophageal reflux disease and malignancy].
Severe gastroesophageal reflux disease (GERD) predisposes to the development of Barrett's esophagus defined by the replacement of the squamous epithelium of the distal esophagus by a columnar epithelium. Barrett's esophagus is a precancerous condition which has a risk of malignant transformation to adenocarcinoma. Barrett's adenocarcinoma remains a relatively rare condition. Its incidence among patients followed up for Barrett's esophagus is approximately 0.5% patients per year. Besides GERD, obesity, male gender and age are the main risk factors for the development of Barrett's mucosa and esophageal adenocarcinoma. Men above 50 presenting with ancient and frequent GERD symptoms could benefit from an upper-GI endoscopy to detect Barrett's esophagus. Though still controversial, surveillance of Barrett's mucosa could allow detection of high grade dysplasia and esophageal adenocarcinoma at an early stage, enabling a curative treatment. Patients who cannot undergo surgery may be treated using endoscopic techniques such as endoscopic mucosal resection.


Associations of Problematic Internet Use, Weight-Related Self-Stigma, and Nomophobia with Physical Activity: Findings from Mainland China, Taiwan, and Malaysia.
Insufficient physical activity is a common problem for university students because they may engage in sedentary lifestyle owing to excessive time spent on their smartphones and social media use. This may result in problematic internet use (PIU) and nomophobia (fear of not having a mobile phone). Moreover, prior evidence shows that weight-related self-stigma is an important factor contributing to low physical activity. Therefore, the present study examined the associations between PIU, nomophobia, and physical activity among university students across mainland China, Taiwan, and Malaysia. Participants (3135 mainland Chinese, 600 Taiwanese, and 622 Malaysian) completed the Bergen Social Media Addiction Scale (BSMAS), Smartphone Application-Based Addiction Scale (SABAS), Nomophobia Questionnaire (NMPQ), Weight Self-Stigma Questionnaire (WSSQ), and International Physical Activity Questionnaire Short Form (IPAQ-SF). The measurement invariance of the assessed questionnaires was supported across the three regions. The present findings analyzed using partial least squares structural equation modeling showed that (i) greater nomophobia was associated with higher levels of physical activity, (ii) greater weight-related self-stigma was associated with higher levels of physical activity, and (iii) greater nomophobia was associated with greater weight-related self-stigma. Although the present findings suggest the possibility that experiencing some level of nomophobia or weight-related self-stigma appears to help improve physical activity, it is not recommended that these be encouraged, but reducing PIU should be targeted as a means to improve physical activity.


A Biodegradable Hemostatic Gelatin/Polycaprolactone Composite for Surgical Hemostasis.
Massive bleeding is the leading cause of battlefield-related deaths and the second leading cause of deaths in civilian trauma centers. One of the challenges of managing severe wounds is the need to promote hemostasis as quickly as possible, which can be achieved by using hemostatic dressings. In this study, we fabricated 2 kinds of gelatin/polycaprolactone composites with 2 ratios of gelatin/polycaprolactone, 1:1 and 2:1 (GP11 and GP21, respectively). Scanning electron microscopy revealed that the GP11 composite exhibited rougher and more porous structure than the GP21 composite did. Furthermore, both composites showed similar biocompatibility as that of tissue culture polystyrene. Moreover, both GP composites tended to show a gradual decrease in contact angle to zero within 40 minutes. The in vitro blood plasma coagulation assay revealed that the prothrombin time was significantly longer for the GP composites than it was for the Quikclot composite, whereas the activated partial thromboplastin time of the GP11 composite was significantly shorter than that of the gauze. Furthermore, the GP11 had the largest platelet adsorption of all the composites. The in vivo coagulation test showed an obvious shortening of the bleeding time with the Quikclot and GP21 compared with gauze sample. In conclusion, the GP composites showed superior biocompatibility and hemostasis to the gauze and comparable effects with the Qickclot composite. Therefore, the GP composites have the potential for development as biodegradable surgical hemostatic agents.


Phenotypic analysis of HIV-1 genotypic drug-resistant isolates from China, using a single-cycle system.
Drug resistance in HIV-1 is one of the main causes of failure of antiretroviral therapy. Phenotypic detection of drug-resistant HIV-1 can provide guidance in selecting the optimal treatment regimen. Traditional phenotype assays are labor intensive and time consuming. Thus, a rapid and convenient phenotype assay with a single cycle of replication was developed and used in this study. Two restriction endonuclease sites, ApaI and AgeI, were inserted into the plasmid pSG3Δenv(,) using site-directed mutagenesis. The reverse transcriptase and protease genes of HIV-1 were amplified from patients and cloned into the modified pSG3Δenv. Sixteen original recombinant pseudoviruses were generated. The phenotypic susceptibility of these 16 recombinant pseudoviruses to 12 antiretroviral drugs was determined using a luciferase reporter system, and the phenotype and genotype results were compared. A modified phenotype assay with a single-cycle system was established, and its reproducibility and feasibility were validated. Approximately 89% of the phenotype results were in agreement with the genotype results; this slight disagreement may have been due to complex and multiple resistance mutations. The phenotype results showed that individual pseudoviruses with four thymidine analog mutations (TAMs).[M41L, T67N, L210W, and T215Y] in combination with various other mutations had different levels of resistance to nucleoside reverse transcriptase inhibitors (NRTIs). Mutations E44A, T69D, and V118I influenced the pattern of resistance of TAMs. The level of resistance to non-NRTIs (NNRTIs) was also variable when different NNRTI-resistance mutations were combined. The single-cycle pseudovirus phenotypic susceptibility detection system reflects HIV-1 drug resistance, especially for complex resistance mutants, and could be used to screen new antiretroviral candidates.


First report of Phytopythium vexans causing root rot on Macadamia integrifolia in China.
Soil-borne plant-pathogenic Phytopythium spp. can cause root rot and damping off on important plant species, resulting in serious economic loss. A survey in October 2021 identified soil-borne diseases occurring on Macadamia integrifolia in Yunnan Province, China. Microbes were isolated from necrotic roots of 23 trees with root rot symptoms by growing on cornmeal-based oomycete-selective 3P (Haas 1964) and P5APR (Jeffers and Martin, 1986) media at 24ºC in the dark for 7 days. Of the 56 single-hyphal isolates obtained, 18 were morphologically similar to Phytopythium vexans (van der Plaats-Niterink 1981; de Cock et al. 2015). Isolates LC04 and LC051 were selected for molecular analyses. The internal transcribed spacer (ITS) region and the cytochrome c oxidase subunit II (CoxII) gene were PCR-amplified using universal primers ITS1/ITS4 (White et al. 1990) and oomycete-specific primers Cox2-F/Cox2-RC4 (Choi et al. 2015), respectively. The PCR products were sequenced with the amplification primers and sequences were lodged in Genbank (Accession no. OM346742, OM415989 for ITS, OM453644, OM453643 for CoxII for isolates LC04 and LC051, respectively). The top BLAST hit in the Genbank nr database for all four sequences was Phytopythium vexans (>99% identity). A maximum-likelihood phylogenetic tree was constructed with analogous concatenated ITS and CoxII sequences from either type or voucher specimens of 13 Phytopythium species in the same phylogenetic clade as P. vexans (Table 1; Bala et. al 2010). Isolates LC04 and LC051 grouped most closely to P. vexans, with LC051 basal and sister to LC04 and P. vexans voucher specimen CBS119.80 with 100% support (Fig. 1). Millet seed inoculated with agar pieces colonized by P. vexans LC04 and LC51 was used to fulfill Koch's postulates (Li et al. 2015) in a completely randomized experimental design. Four 6-month-old M. integrifolia var. Keaau (660) seedlings were transplanted into pasteurized commercial potting mix containing 0.5% (w/w) inoculum. Plants were grown in free draining pots and watered once a day. At 14 days post-inoculation, roots were discolored compared to control plants inoculated with millet seed mixed with agar plugs lacking P. vexans (Fig. 2). By 30 days post-inoculation, infected roots were discolored with obvious decay and reduction in root system size. Control plants were symptomless. P. vexans was successfully re-isolated from two lesioned roots from each plant. The infection experiment was done twice, demonstrating that P. vexans LC04 and LC51 caused root disease on M. integrifolia. P. vexans causes root rot, damping-off, crown rot, stem rot or patch canker on economically important trees in many parts of the world, including seven plant species in China (Farr and Rossman 2022). This is the first report of pathogenic P. vexans on M. integrifolia in China. Reports of pathogenic P. vexans on multiple hosts in several parts of the world suggest it should be considered a quarantine risk and included in risk mitigation or pest management plans that include other species of Phytopythium, or species of Pythium or Phytophthora, to which P. vexans has many similarities (de Cock et al. 2015).


Overcoming Self-Incompatibility in Diploid Potato Using CRISPR-Cas9.
Potato breeding can be redirected to a diploid inbred/F1 hybrid variety breeding strategy if self-compatibility can be introduced into diploid germplasm. However, the majority of diploid potato clones (Solanum spp.) possess gametophytic self-incompatibility that is primarily controlled by a single multiallelic locus called the S-locus which is composed of tightly linked genes, S-RNase (S-locus RNase) and multiple SLFs (S-locus F-box proteins), which are expressed in the style and pollen, respectively. Using S-RNase genes known to function in the Solanaceae gametophytic SI mechanism, we identified S-RNase alleles with flower-specific expression in two diploid self-incompatible potato lines using genome resequencing data. Consistent with the location of the S-locus in potato, we genetically mapped the S-RNase gene using a segregating population to a region of low recombination within the pericentromere of chromosome 1. To generate self-compatible diploid potato lines, a dual single-guide RNA (sgRNA) strategy was used to target conserved exonic regions of the S-RNase gene and generate targeted knockouts (KOs) using a Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (Cas9) approach. Self-compatibility was achieved in nine S-RNase KO T0 lines which contained bi-allelic and homozygous deletions/insertions in both genotypes, transmitting self compatibility to T1 progeny. This study demonstrates an efficient approach to achieve stable, consistent self-compatibility through S-RNase KO for use in diploid potato breeding approaches.


Total synthesis of (+)-macquarimicin A.
The first total synthesis of (+)-macquarimicin A (1), a novel inhibitor of neutral sphingomyelinase (N-SMase) with antiinflammatory activity, has been accomplished. The present work determined the absolute configuration of (+)-1 and revised the C(2)-C(3) geometry to be Z. The synthesis features a transannular Diels-Alder reaction, which constructed the tetracyclic framework stereoselectively, and a convergent and efficient synthetic pathway, which afforded (+)-macquarimicin A (1) in 27 steps (longest linear sequence) with 9.9% overall yield.


Gut Microbiota and Development of Vibrio cholerae-Specific Long-Term Memory B Cells in Adults after Whole-Cell Killed Oral Cholera Vaccine.
Cholera is a diarrheal disease caused by Vibrio cholerae that continues to be a major public health concern in populations without access to safe water. IgG- and IgA-secreting memory B cells (MBC) targeting the V. cholerae O-specific polysaccharide (OSP) correlate with protection from infection in persons exposed to V. cholerae and may be a major determinant of long-term protection against cholera. Shanchol, a widely used oral cholera vaccine (OCV), stimulates OSP MBC responses in only some people after vaccination, and the gut microbiota is a possible determinant of variable immune responses observed after OCV. Using 16S rRNA sequencing of feces from the time of vaccination, we compared the gut microbiota among adults with and without MBC responses to OCV. Gut microbial diversity measures were not associated with MBC isotype or OSP-specific responses, but individuals with a higher abundance of Clostridiales and lower abundance of Enterobacterales were more likely to develop an MBC response. We applied protein-normalized fecal supernatants of high and low MBC responders to THP-1-derived human macrophages to investigate the effect of microbial factors at the time of vaccination. Feces from individuals with higher MBC responses induced significantly different IL-1β and IL-6 levels than individuals with lower responses, indicating that the gut microbiota at the time of vaccination may "prime" the mucosal immune response to vaccine antigens. Our results suggest the gut microbiota could impact immune responses to OCVs, and further study of microbial metabolites as potential vaccine adjuvants is warranted.


Hypoplastic basement membrane of the lens anlage in the inheritable lens aplastic mouse (lap mouse).
Adult homozygous lap mice show various eye abnormalities such as aphakia, retinal disorganization, and dysplasia of the cornea and anterior chamber. In the fetal eye of a homozygous lap mouse, the lens placode appears to develop normally. However, the lens vesicle develops abnormally to form a mass of cells without a cavity, and the mass vanishes soon afterward. Apoptotic cell death is associated with the disappearance of the lens anlage. We examined the basement membranes of the lens anlage of this mutant by immunohistochemical methods under light microscopy using antibodies against basement membrane components of the lens anlage, type IV collagen, fibronectin, laminin, heparan sulfate proteoglycan, and entactin and by transmission electron microscopy. Immunohistochemistry showed the distribution and intensity of antibody binding to the lens anlage to be almost the same for each these antibodies regardless of the stage of gestation or whether the anlagen were from normal BALB/c or lap mice. Thus, positive continuous reactions were observed around the exterior region of the lens anlage from day 10 of gestation for type IV collagen, fibronectin, laminin, heparan sulfate proteoglycan antibodies, and at least from day 11of gestation for entactin antibody. The basement membrane lamina densa of both normal and lap mice was shown by electron microscopy to be discontinuous at days 10 and 10.5 of gestation. However, by day 11 the lamina densa was continuous in the lens anlagen of normal mice but still discontinuous in the lap mice. By day 12 of gestation, the lamina densa had thickened markedly in normal mice, whereas in lap mice it remained discontinuous and its thinness indicated hypoplasia. These results indicate that, while all basement components examined are produced and deposited in the normal region of the lens anlage in the lap mouse, the basement membrane is, for some reason, imperfectly formed. The time at which hypoplasia of the basement membrane was observed in this mutant coincided with the stage during which apoptosis in the lens anlage occurred. This result may indicate a possibility of the relationship between the basement membrane and apoptosis in this mutant.


Triple-slit nanoaperture for transmission enhancement of a cavity-aperture.
A cavity-aperture has a problem of low transmission efficiency due to its nano-sized aperture despite its potential for plasmonic color filters. In this study, a triple-slit aperture is proposed as the nanoaperture in the center of the cavity-aperture to improve the transmittance. It provides one centered nanoslit and two symmetric wedge structures to each of three cavities corresponding to incident polarization, and induces the strong confinement and transmission of electric fields due to plasmonic resonances at the two types of nanostructures. The transmittance of the triple-slit aperture is theoretically five times and experimentally two times higher than that of a circular aperture. Furthermore, expansive studies on polarization-insensitive nanoapertures with six-fold rotational symmetry will contribute to the development of plasmonic color filters and imaging devices.


Microbial specialization by prions.
Microbial prions facilitate a variety of phenotypic switches. Recently-developed tools that can directly interrogate, in the living cell, the aggregation state of a protein have enabled a wider range of experiments for prion-mediated behaviors. With such tools, the roles of the yeast prion [SWI+] in migration and mating were studied. Although [SWI+] cells were consistently less fit than their [swi-] counterparts under traditional laboratory conditions, in these new phenotypic paradigms [SWI+] cells demonstrated a distinct advantage. [SWI+] cells dispersed over a larger area under conditions resembling rainfall and outcrossed more frequently. We postulate that many behaviors in microorganisms may be modulated by stochastic prion switching. In diverse and changing natural environments, prion switching at low frequency may promote greater fitness of the population by specializing a small number of individuals with altered responses to their environments.


Copper-Catalyzed Three-Component Difunctionalization of Aromatic Alkenes with 2-Amino-1,4-naphthoquinones and α-Bromocarboxylates.
A copper-catalyzed three-component difunctionalization of aromatic alkenes to access 1,4-naphthoquinone derivatives with diverse structures is described. Experiments show that the difunctionalization reaction is accompanied by ester exchange reaction with the solvent. In this method, α-bromocarboxylates are used as radical precursors and 2-amino-1,4-naphthoquinones as radical trapping reagents. The substrate scope is broad because various aromatic alkenes, 2-amino-1,4-naphthoquinones, and α-bromocarboxylates are employed in the reaction, and corresponding products are obtained in moderate to good yields.


Medial approach in elbow arthroscopy.
The author undertook a cadaveric dissection study to confirm the hypothesis that starting with the anterior medial portal in elbow arthroscopy is safer than starting with the anterior radial portal. In six cadaveric elbows, the capsule was distended with saline. Both anterior medial and anterior radial approaches were made with the elbow flexed to 90 degrees. Four and one-half millimeter arthroscopic sheaths were inserted and obturators were then left in place while the saline was drained; expanding polyurethane foam was used to distend the capsule. We allowed the polyurethane foam to harden and then dissected all elbows, with special attention given to exposure of the radial and medial nerve and the brachial artery. The hardened foam allowed for continued capsular distension during these dissections and recreated normal distances from instrument portals to neurovascular bundles. The minimum distance from the path of the arthroscopic sheath to the large neurovascular structures was then measured. The distance from the medial portal to the nearest neurovascular structure (median nerve, brachial artery) averaged 23 mm. The distance from the radial portal to the nearest neurovascular structure (radial nerve) averaged 3 mm. The ulnar nerve averaged a 25 mm clearance from the medial portal. Even when the medial portal was made by an incorrect method, the minimum clearance to the median nerve averaged 11 mm. The most frequently recommended current standard technique for elbow arthroscopy involves beginning with an anterior radial portal. However, the findings in this study suggest that an anterior medial portal is a superior starting point.(ABSTRACT TRUNCATED AT 250 WORDS)


Narrative review: the role of leptin in human physiology: emerging clinical applications.
Leptin is a hormone secreted by adipose tissue in direct proportion to amount of body fat. The circulating leptin levels serve as a gauge of energy stores, thereby directing the regulation of energy homeostasis, neuroendocrine function, and metabolism. Persons with congenital deficiency are obese, and treatment with leptin results in dramatic weight loss through decreased food intake and possible increased energy expenditure. However, most obese persons are resistant to the weight-reducing effects of leptin. Recent studies suggest that leptin is physiologically more important as an indicator of energy deficiency, rather than energy excess, and may mediate adaptation by driving increased food intake and directing neuroendocrine function to converse energy, such as inducing hypothalamic hypogonadism to prevent fertilization. Current studies investigate the role of leptin in weight-loss management because persons who have recently lost weight have relative leptin deficiency that may drive them to regain weight. Leptin deficiency is also evident in patients with diet- or exercise-induced hypothalamic amenorrhea and lipoatrophy. Replacement of leptin in physiologic doses restores ovulatory menstruation in women with hypothalamic amenorrhea and improves metabolic dysfunction in patients with lipoatrophy, including lipoatrophy associated with HIV or highly active antiretroviral therapy. The applications of leptin continue to grow and will hopefully soon be used therapeutically.


Intra- and Interrater Reliability of Short-Term Measurement of Heart Rate Variability on Rest in Individuals Post-COVID-19.
Individuals affected by COVID-19 have an alteration in autonomic balance, associated with impaired cardiac parasympathetic modulation and, consequently, a decrease in heart rate variability (HRV). This study examines the inter- and intrarater reliability of HRV) parameters derived from short-term recordings in individuals post-COVID. Sixty-nine participants of both genders post-COVID were included. The RR interval, the time elapsed between two successive R-waves of the QRS signal on the electrocardiogram (RRi), were recorded during a 10 min period in a supine position using a portable heart rate monitor (Polar® V800 model). The data were transferred into Kubios® HRV standard analysis software and analyzed within the stable sessions containing 256 sequential RRi. The intraclass correlation coefficient (ICC) ranged from 0.920 to 1.000 according to the intrarater analysis by Researcher 01 and 0.959 to 0.999 according to the intrarater by Researcher 02. The interrater ICC ranged from 0.912 to 0.998. The coefficient of variation was up to 9.23 for Researcher 01 intrarater analysis, 6.96 for Researcher 02 intrarater analysis and 8.83 for interrater analysis. The measurement of HRV in post-COVID-19 individuals is reliable and presents a small amount of error inherent to the method, supporting its use in the clinical environment and in scientific research.


Thin transparent W-doped indium-zinc oxide (WIZO) layer on glass.
Annealing effect on structural and electrical properties of W-doped IZO (WIZO) films for thin film transistors (TFT) was studied under different process conditions. Thin WIZO films were deposited on glass substrates by RF magnetron co-sputtering technique using indium zinc oxide (10 wt.% ZnO-doped In2O3) and WO3 targets in room temperature. The post annealing temperature was executed from 200 degrees C to 500 degrees C under various O2/Ar ratios. We could not find any big difference from the surface observation of as grown films while it was found that the carrier density and sheet resistance of WIZO films were controlled by O2/Ar ratio and post annealing temperature. Furthermore, the crystallinity of WIZO film was changed as annealing temperature increased, resulting in amorphous structure at the annealing temperature of 200 degrees C, while clear In2O3 peak was observed for the annealed over 300 degrees C. The transmittance of as-grown films over 89% in visible range was obtained. As an active channel layer for TFT, it was found that the variation of resistivity, carrier density and mobility concentration of WIZO film decreased by annealing process.


Double- and Multi-Femtosecond Pulses Produced by Birefringent Crystals for the Generation of 2D Laser-Induced Structures on a Stainless Steel Surface.
Laser-induced textures have been proven to be excellent solutions for modifying wetting, friction, biocompatibility, and optical properties of solids. The possibility to generate 2D-submicron morphologies by laser processing has been demonstrated recently. Employing double-pulse irradiation, it is possible to control the induced structures and to fabricate novel and more complex 2D-textures. Nevertheless, double-pulse irradiation often implies the use of sophisticated setups for modifying the pulse polarization and temporal profile. Here, we show the generation of homogeneous 2D-LIPSS (laser-induced periodic surface structures) over large areas utilizing a simple array of birefringent crystals. Linearly and circularly polarized pulses were applied, and the optimum process window was defined for both. The results are compared to previous studies, which include a delay line, and the reproducibility between the two techniques is validated. As a result of a systematic study of the process parameters, the obtained morphology was found to depend both on the interplay between fluence and inter-pulse delay, as well as on the number of incident pulses. The obtained structures were characterized via SEM (scanning electron microscopy) and atomic force microscopy. We believe that our results represent a novel approach to surface structuring, primed for introduction in an industrial environment.


Jejunal diverticulosis.
Jejunal diverticulosis is an uncommon condition, but it is often associated with a number of symptoms including postprandial abdominal pain, distension, flatulence, borborygmus, nausea, vomiting, diarrhea and constipation and with complications such as vomiting blood or coffee grounds material, melena and shock. The authors report such a condition in a 71-year-old man. Resection is the treatment of choice for relieving symptoms, and for preventing or treating complications.


Bilateral glaucomatocyclitic crisis in a patient with Holmes Adie syndrome.
A patient with pre-existing bilateral tonic pupils presented with simultaneous bilateral glaucomatocyclitic crisis. Deep tendon reflexes were absent although they were documented to be present 6 years ago. A possibility of a progressive autonomic dysfunction in both these conditions is discussed.


Determinants of Successful Exclusive Breastfeeding for Saudi Mothers: Social Acceptance Is a Unique Predictor.
Recent guidelines motivate health care professionals to promote exclusive breastfeeding (EBF). The reported rate of EBF is low in Saudi Arabia. This study aimed to explore the determinants of successful exclusive breastfeeding for Saudi mothers. A cross-sectional, survey-based study was conducted in family medicine clinics. The dependent variable was the actual practice of EBF. Independent variables were the mothers' demographic information, comfortableness with breastfeeding in public, knowledge and attitudes about breastfeeding, previous experience of successful breastfeeding, and a previous feeding plan. Statistical analysis was carried out using bivariate analysis and multinomial logistic regression. Out of the 322 respondents, only 28% practiced exclusive breastfeeding for the first six months. Perceived insufficient milk (p = 0.011) was associated with a lower EBF rate. Mothers' degrees of comfort with breastfeeding in front of their relatives (p = 0.024) and in front of friends (p = 0.028) were significantly associated with their infants' actual feeding practices for the first six months of their infants' lives. Mothers reported that the absence of a suitable place for breastfeeding caused them to stop breastfeeding (p = 0.043) and was associated with their infant's actual feeding practices for the first six months of their infant's lives. An antenatal breastfeeding intention was considered a significant predictor of EBF; OR: 7.31 (95% CI: 2.24-23.84). Mothers who do not stop breastfeeding when they get sick have a 5.054 times higher chance of continuing EBF (95% CI: 1.037-24.627) than the formula-only feeding group. Thus, social acceptance is a unique predictor for their success in exclusive breastfeeding. Mothers have good intentions and a desire to breastfeed. Therefore, they must be guided through their pregnancy and postpartum period to overcome breastfeeding issues.


MAL, but not MAL2, expression promotes the formation of cholesterol-dependent membrane domains that recruit apical proteins.
Our recent studies have been aimed at understanding the mechanisms regulating apical protein sorting in polarized epithelial cells. In particular, we have been investigating how lipid rafts serve to sort apical proteins in the biosynthetic pathway. The recent findings that lipid domains are too small or transient to host apically destined cargo have led to newer versions of the hypothesis that invoke proteins required for lipid domain coalescence and stabilization. MAL (myelin and lymphocyte protein) and its highly conserved family member, MAL2, have emerged as possible regulators of this process in the direct and indirect apical trafficking pathways respectively. To test this possibility, we took a biochemical approach. We determined that MAL, but not MAL2, self-associates, forms higher-order cholesterol-dependent complexes with apical proteins and promotes the formation of detergent-resistant membranes that recruit apical proteins. Such biochemical properties are consistent with a role for MAL in raft coalescence and stabilization. These findings also support a model whereby hydrophobic mismatch between the long membrane-spanning helices of MAL and the short-acyl-chain phospholipids in the Golgi drive formation of lipid domains rich in raft components that are characterized by a thicker hydrophobic core to alleviate mismatch.


Myopia and peripheral ocular aberrations.
We measured wave aberrations over the central 42 degrees x 32 degrees visual field for a 5-mm pupil for groups of 10 emmetropic (mean spherical equivalent = 0.11 +/- 0.50 D) and 9 myopic (MSE = -3.67 +/- 1.91 D) young adults. Relative peripheral refractive errors over the measured field were generally myopic in both groups. Mean values of C(4)(0) were almost constant across the measured field and were more positive in emmetropes (+0.023 +/- 0.043 microm) than in myopes (-0.007 +/- 0.045 microm). Coma varied more rapidly with field angle in myopes: modeling suggested that this difference reflected the differences in mean anterior corneal shape and axial length in the two groups. In general, however, overall levels of RMS aberration differed only modestly between the two groups, implying that it is unlikely that high levels of aberration contribute to myopia development.


Super-Resolution Microscopy Unveils Dynamic Heterogeneities in Nanoparticle Protein Corona.
The adsorption of serum proteins, leading to the formation of a biomolecular corona, is a key determinant of the biological identity of nanoparticles in vivo. Therefore, gaining knowledge on the formation, composition, and temporal evolution of the corona is of utmost importance for the development of nanoparticle-based therapies. Here, it is shown that the use of super-resolution optical microscopy enables the imaging of the protein corona on mesoporous silica nanoparticles with single protein sensitivity. Particle-by-particle quantification reveals a significant heterogeneity in protein absorption under native conditions. Moreover, the diversity of the corona evolves over time depending on the surface chemistry and degradability of the particles. This paper investigates the consequences of protein adsorption for specific cell targeting by antibody-functionalized nanoparticles providing a detailed understanding of corona-activity relations. The methodology is widely applicable to a variety of nanostructures and complements the existing ensemble approaches for protein corona study.


The effect of the genomic GC content bias of prokaryotic organisms on the secondary structures of their proteins.
One of the main characteristics of prokaryotic genomes is the ratio in which guanine-cytosine bases are used in their DNA sequences. This is known as the genomic GC content and varies widely, from values below 20% to values greater than 74%. It has been demonstrated that the genomic GC content varies in accordance with the phylogenetic distribution of organisms and influences the amino acid composition of their corresponding proteomes. This bias is particularly important for amino acids that are coded by GC content-rich codons such as alanine, glycine, and proline, as well as amino acids that are coded by AT-rich codons, such as lysine, asparagine, and isoleucine. In our study, we extend these results by considering the effect of the genomic GC content on the secondary structure of proteins. On a set of 192 representative prokaryotic genomes and proteome sequences, we identified through a bioinformatic study that the composition of the secondary structures of the proteomes varies in relation to the genomic GC content; random coils increase as the genomic GC content increases, while alpha-helices and beta-sheets present an inverse relationship. In addition, we found that the tendency of an amino acid to form part of a secondary structure of proteins is not ubiquitous, as previously expected, but varies according to the genomic GC content. Finally, we discovered that for some specific groups of orthologous proteins, the GC content of genes biases the composition of secondary structures of the proteins for which they code.


External endometriosis in 801 operated patients.
Details of medical history, symptomatology, operative findings and certain other special features are reported for patients with external endometriosis treated operatively during the period 1969-76. The incidence of endometriosis in this survey was 19% of all gynecological laparotomies. The mean age of the patients at the time of the operation was 36.7 years. The most common sites of endometriosis were the retrocervix and the ovaries. Of the patients, 559 (70%) had complained of dysmenorrhea; the pain generally began before the bleeding. Also typical was the exacerbation of the pain with increasing age. Twenty-seven per cent of the patients also complained of infertility. Surgery was performed because of acute abdominal symptoms in 6% of cases. The incidence of endometriosis in postmenopausal women was 2.5% and that in young women, 1%. Endometriosis recurred in 15% of the cases during the follow-up period. Gestagen therapy after the operation had no effect on the recurrence rate.


[The vomeronasal organ (VNO) of newborn guinea pigs is functionally sound].
The present investigation demonstrates that the vascularization of the vomeronasal organ in the newborn guinea pig is capable of generating a pumping mechanism for suction of stimuli from the nasal cavity into the lumen of the organ.


Components of action representations evoked when identifying manipulable objects.
We examined the influence of holding planned hand actions in working memory on the time taken to visually identify objects with handles. Features of the hand actions and position of the object's handle were congruent or incongruent on two dimensions: alignment (left vs. right) and orientation (horizontal vs. vertical). When an object was depicted in an upright view, subjects were slower to name it when its handle was congruent with the planned hand actions on one dimension but incongruent on the other, relative to when the object handle and actions were congruent on both or neither dimension. This pattern is consistent with many other experiments demonstrating that a cost occurs when there is partial feature overlap between a planned action and a perceived target. An opposite pattern of results was obtained when the depicted object appeared in a 90° rotated view (e.g., a beer mug on its side), suggesting that the functional goal associated with the object (e.g., drinking from an upright beer mug) was taken into account during object perception and that this knowledge superseded the influence of the action afforded by the depicted view of the object. These results have implications for the relationship between object perception and action representations, and for the mechanisms that support the identification of rotated objects.


Volitional Generation of Reproducible, Efficient Temporal Patterns.
One of the extraordinary characteristics of the biological brain is the low energy expense it requires to implement a variety of biological functions and intelligence as compared to the modern artificial intelligence (AI). Spike-based energy-efficient temporal codes have long been suggested as a contributor for the brain to run on low energy expense. Despite this code having been largely reported in the sensory cortex, whether this code can be implemented in other brain areas to serve broader functions and how it evolves throughout learning have remained unaddressed. In this study, we designed a novel brain-machine interface (BMI) paradigm. Two macaques could volitionally generate reproducible energy-efficient temporal patterns in the primary motor cortex (M1) by learning the BMI paradigm. Moreover, most neurons that were not directly assigned to control the BMI did not boost their excitability, and they demonstrated an overall energy-efficient manner in performing the task. Over the course of learning, we found that the firing rates and temporal precision of selected neurons co-evolved to generate the energy-efficient temporal patterns, suggesting that a cohesive rather than dissociable processing underlies the refinement of energy-efficient temporal patterns.


Cloning and Functional Analysis of Histones H3 and H4 in Nuclear Shaping during Spermatogenesis of the Chinese Mitten Crab, Eriocheir sinensis.
During spermatogenesis in most animals, the basic proteins associated with DNA are continuously changing and somatic-typed histones are partly replaced by sperm-specific histones, which are then successively replaced by transition proteins and protamines. With the replacement of sperm nuclear basic proteins, nuclei progressively undergo chromatin condensation. The Chinese Mitten Crab (Eriocheir sinensis) is also known as the hairy crab or river crab (phylum Arthropoda, subphylum Crustacea, order Decapoda, and family Grapsidae). The spermatozoa of this species are aflagellate, and each has a spherical acrosome surrounded by a cup-shaped nucleus, peculiar to brachyurans. An interesting characteristic of the E. sinensis sperm nucleus is its lack of electron-dense chromatin. However, its formation is not clear. In this study, sequences encoding histones H3 and H4 were cloned by polymerase chain reaction amplification. Western blotting indicated that H3 and H4 existed in the sperm nuclei. Immunofluorescence and ultrastructural immunocytochemistry demonstrated that histones H3 and H4 were both present in the nuclei of spermatogonia, spermatocytes, spermatids and mature spermatozoa. The nuclear labeling density of histone H4 decreased in sperm nuclei, while histone H3 labeling was not changed significantly. Quantitative real-time PCR showed that the mRNA expression levels of histones H3 and H4 were higher at mitotic and meiotic stages than in later spermiogenesis. Our study demonstrates that the mature sperm nuclei of E. sinensis contain histones H3 and H4. This is the first report that the mature sperm nucleus of E. sinensis contains histones H3 and H4. This finding extends the study of sperm histones of E. sinensis and provides some basic data for exploring how decapod crustaceans form uncondensed sperm chromatin.


Chondrosarcoma of the radius with distant metastasis in a dog.
A 9-year-old castrated male Doberman Pinscher was admitted for evaluation of lameness of the left forelimb. Radiography and examination of biopsy specimens revealed a moderately differentiated chondrosarcoma of the proximal portion of the radius. The dog was treated by local excision of the neoplasm, which involved resection of the radial head and proximal portion of the radius. Despite the large size of the dog and the weight-bearing forces exerted on the cubital joint, major problems with ambulation did not develop after surgery. Functional use of the limb returned slowly; however, substantial limb use was observed despite the development of mild degenerative changes of the joint and migration of the humeroulnar articulation. Six months after surgery, metastasis of a widely disseminated, poorly differentiated chondrosarcoma to the subcutaneous tissues and thoracic and abdominal cavities was diagnosed. Local redevelopment of the chondrosarcoma in the area of the cubital joint was not detected. Resection of the radial head and proximal portion of the radius may be considered a viable, alternative, limb-sparing technique. The biologically aggressive nature of this chondrosarcoma of the appendicular skeleton indicated that additional information was needed before a reliable prognosis could be established for this dog with this tumor type. Reports of low rates of metastasis have been based on insufficient numbers of dogs to adequately or accurately determine the long-term prognosis of dogs with chondrosarcoma of the appendicular skeleton.


Considerations concerning the use of counting active personal dosimeters in pulsed fields of ionising radiation.
Active personal electronic dosimeters (APDs) exhibit limitations in pulsed radiation fields, which cannot be overcome without the use of new detection technology. As an interim solution, this paper proposes a method by which some conventional dosimeters can be operated in a way such that, based on the basic knowledge about the pulsed radiation field, any dosimetric failure of the dosimeter is signalised by the instrument itself. This method is not applicable to all combinations of APD and pulsed radiation field. The necessary requirements for the APD and for the parameters of the pulsed radiation field are given in the paper. Up to now, all such requirements for APDs have not been tested or verified in a type test. The suitability of the method is verified for the use of one APD used in two clinical pulsed fields.


Effect of antisense oligonucleotide against mouse dentine matrix protein 1 on mineralization ability and calcium ions metabolism in odontoblast-like cell line MDPC-23.
To study the mineralization ability and the dynamic changes of intracellular and extracellular concentrations of calcium ions in the odontoblast-like cell line MDPC-23 affected by antisense oligonucleotide (AS-ODN) against mouse dentine matrix protein 1 (DMP1). The expression of DMP1 in MDPC-23 cells was detected by an immunohistochemical method and its blocking outcome by the Western blot method. The alkaline phosphatase (ALP) activity, size and number of mineralized nodules, and the intracellular free ([Ca2+]if), total ([Ca2+]it) and the extracellular ([Ca2+]e) calcium ion concentrations in MDPC-23 cells in the experimental group affected with AS-ODN were compared with those in the control group (paired-samples t-test). Dentine matrix protein 1 was stably expressed in a stable way in MDPC-23 cells; the expression was only just detectable at 12 h and became negative after 24 h affected by AS-ODN. Compared with the control groups, ALP activity of MDPC-23 cells in the AS-ODN group was decreased (P < 0.05), and both the number and size of mineralized nodules were smaller than those in the control group. [Ca2+]if in the AS-ODN group increased and then decreased after 24 h. [Ca2+]it dropped substantially to the lowest point at 24 h (P < 0.01). [Ca2+]e increased before treatment for 24 h and then dropped, however, it was still higher than that of the control group. Antisense oligonucleotide against DMP1 could decrease mineralization ability and affect the intracellular and extracellular concentrations of calcium ions in MDPC-23 cells. This would indicate that DMP1 regulates the metabolism and transportation of calcium ions in odontoblasts, and thus boosts dentine mineralization.


Characterization of the stomatin domain involved in homo-oligomerization and lipid raft association.
The cytoplasmically oriented monotopic integral membrane protein stomatin forms high-order oligomers and associates with lipid rafts. To characterize the domains that are involved in oligomerization and detergent-resistant membrane (DRM) association, we expressed truncation and point mutants of stomatin and analyzed their size and buoyancy by ultracentrifugation methods. A small C-terminal region of stomatin that is largely hydrophobic, Ser-Thr-Ile-Val-Phe-Pro-Leu-Pro-Ile (residues 264-272), proved to be crucial for oligomerization, whereas the N-terminal domain (residues 1-20) and the last 12 C-terminal amino acids (residues 276-287) were not essential. The introduction of alanine substitutions in the region 264-272 resulted in the appearance of monomers. Remarkably, only three of these residues, Ile-Val-Phe (residues 266-268), were found to be indispensable for the DRM association. Interestingly, the exchange of Pro-269 and to some extent the residues 270-272, which are essential for oligomerization, did not affect the DRM association of stomatin. This suggests that the formation of oligomers is not necessary for the association of stomatin with DRMs. Internal deletions near the membrane anchoring domain resulted in the formation of intermediate size oligomers suggesting a conformational interdependence of large parts of the C-terminal region. Fluorescence recovery after photobleaching analysis of the tagged, monomeric, non-DRM mutant ST-(1-262)-green fluorescent protein and wild type stomatin StomGFP showed a significantly higher lateral mobility of the truncation mutant in the plasma membrane suggesting a membrane interaction of the respective C-terminal region also in vivo.


Focal adhesion kinase signaling and the aggressive melanoma phenotype.
Focal adhesion kinase (FAK) mediates myriad cellular functions and has been found to be overexpressed in numerous human cancers. We recently explored the role of FAK in promoting the aggressive phenotype of melanoma cells, characterized by increased invasion, migration, and vasculogenic mimicry (VM) potential. We found FAK to be phosphorylated on its key tyrosine residues (397 and 576) in aggressive melanoma cells cultured on a three-dimensional type 1 collagen matrix in vitro, as well as in radial and vertical growth phase melanomas in situ. Furthermore, expressing FAK-related non-kinase (FRNK) in melanoma cells directly resulted in the inhibition of the aggressive phenotype, as demonstrated by decreased invasion, migration and VM potential, in part by blocking an Erk1/2 mediated signaling pathway. Additional data indicated that increased FAK activity may promote cellular proliferation and anchorage independent growth of aggressive melanoma. Together these observations implicate FAK as a promoter of the aggressive melanoma phenotype, thereby identifying a rational target for therapeutic intervention of malignant melanoma.


Multiple-length-scale patterning of magnetic nanoparticles by stamp assisted deposition.
Control of the size and spatial distribution of materials at multiple length scales is one of the most compelling issues in nanotechnology research. We report a multiple-length-scale patterning of pure magnetic particles as well as biocompatible magnetic particles based on a printing technique named micro-injection molding in capillaries. The magnetic particles were prepared by a technique of co-precipitation of ferric and ferrous salts in an alkali medium. We demonstrate that the morphology and the size of the patterning nanoparticles can be controlled by simply controlling the concentration of the solution. Our method exploits the self-organization of the nanoparticles in a solution confined between a stamp and the surfaces of a substrate, exploiting confinement and competing interactions between the adsorbate and the substrate. Our approach represents a remarkable example of an integrated top-down/bottom-up process.


In the wake of progress--ethical problems of renal failure treated by dialysis.
The ability of dialysis in the 1960s to prolong the lives of patients with terminal renal failure created a running and changing debate in bioethics. The first issue was the selection of suitable individuals to be treated in the limited facilities available. Progress in technology was ahead of any medical or ethical consensus. The costs of treatment soon exceeded the will or means of health funders to pay, creating a system of covert rationing and fiscal fudges with which physicians reluctantly colluded. Evidence, epidemiology and public opinion eventually prevailed and the UK now provides a satisfactory, if imperfect, programme of renal replacement therapy (RRT) for most patients with end-stage renal failure who can benefit. The ethical challenges in developed countries are now different and relate to end of life decisions. In poorer countries the issues are starker as treatment has to be rationed or provided only for those who can afford it.


Dihydrotestosterone Regulates Hair Growth Through the Wnt/β-Catenin Pathway in C57BL/6 Mice and In Vitro Organ Culture.
Dihydrotestosterone (DHT) is the most potent androgen that regulates hair cycling. Hair cycling involves cross-talk between the androgen and Wnt/β-catenin pathways. However, how DHT regulates hair follicle (HF) growth through the Wnt/β-catenin pathway has not been well investigated. This study aimed to investigate the roles of DHT in hair growth in vivo and in vitro. Human scalp HFs were treated with different concentrations of DHT (10-5, 10-6, 10-7, 10-8, and 10-9 mol/L) for 10 days. The effects of DHT on hair shaft elongation, the proliferation of hair matrix cells, and the levels of β-catenin, GSK-3β, and phosphorylated GSK-3β (ser9) were evaluated in the cultured HFs. The effects of DHT were further investigated in C57BL/6 mice. Moreover, the growth of cultured human HFs was observed after interfering with the β-catenin pathway through inhibitors or activators in the presence or absence of DHT. We found that different concentrations of DHT had different effects on human HFs in vitro and C57BL/6 mice. At 10-6 mol/L, DHT inhibited HF growth and β-catenin/p-GSK-3β expression, whereas 10-7 mol/L DHT induced HF growth and β-catenin/p-GSK-3β expression. In addition, a β-catenin inhibitor (21H7) inhibited HF growth in vitro, while a β-catenin activator (IM12) promoted HF growth in vitro and antagonized the inhibition of HFs by high levels of DHT. These results suggest that DHT plays a pivotal role in region-specific hair growth, which may be related to the Wnt/β-catenin pathway.


Chitin Heterodisaccharide, Released from Chitin by Chitinase and Chitin Oligosaccharide Deacetylase, Enhances the Chitin-Metabolizing Ability of Vibrio parahaemolyticus.
Vibrio parahaemolyticus RIMD2210633 secretes both chitinase and chitin oligosaccharide deacetylase and produces β-N-acetyl-d-glucosaminyl-(1,4)-d-glucosamine (GlcNAc-GlcN) from chitin. Previously, we reported that GlcNAc-GlcN induces chitinase production by several strains of Vibrio harboring chitin oligosaccharide deacetylase genes (T. Hirano, K. Kadokura, T. Ikegami, Y. Shigeta, et al., Glycobiology 19:1046-1053, 2009). The metabolism of chitin by Vibrio was speculated on the basis of the findings of previous studies, and the role of chitin oligosaccharide produced from chitin has been well studied. However, the role of GlcNAc-GlcN in the Vibrio chitin degradation system, with the exception of the above-mentioned function as an inducer of chitinase production, remains unclear. N,N'-Diacetylchitobiose, a homodisaccharide produced from chitin, is known to induce the expression of genes encoding several proteins involved in chitin metabolism in Vibrio strains (K. L. Meibom, X. B. Li, A. Nielsen, C. Wu, et al., Proc Natl Acad Sci U S A 101:2524-2529, 2004). We therefore hypothesized that GlcNAc-GlcN also affects the expression of enzymes involved in chitin metabolism in the same manner. In this study, we examined the induction of protein expression by several sugars released from chitin using peptide mass fingerprinting and confirmed the expression of genes encoding enzymes involved in chitin metabolism using real-time quantitative PCR analysis. We then confirmed that GlcNAc-GlcN induces the expression of genes encoding many soluble enzymes involved in chitin degradation in Vibrio parahaemolyticus Here, we demonstrate that GlcNAc-GlcN enhances the chitin-metabolizing ability of V. parahaemolyticusIMPORTANCE We demonstrate that β-N-acetyl-d-glucosaminyl-(1,4)-d-glucosamine (GlcNAc-GlcN) enhances the chitin-metabolizing ability of V. parahaemolyticus Members of the genus Vibrio are chitin-degrading bacteria, and some species of this genus are associated with diseases affecting fish and animals, including humans (F. L. Thompson, T. Iida, and J. Swings, Microbiol Mol Biol Rev 68:403-431, 2004; M. Y. Ina-Salwany, N. Al-Saari, A. Mohamad, F.-A. Mursidi, et al., J Aquat Anim Health 31:3-22, 2019). Studies on Vibrio are considered important, as they may facilitate the development of solutions related to health, food, and aquaculture problems attributed to this genus. This report enhances the current understanding of chitin degradation by Vibrio bacteria.


Total synthesis and initial biological evaluation of new B-ring-modified bryostatin analogs.
[Structure: see text] The total synthesis and preliminary biological evaluation of the first bryostatin analogs (bryologs) to incorporate B-ring substitution are reported. Asymmetric syntheses of two new polyketide "spacer" domains are described, one exploiting the pseudosymmetry of the C1-C13 region. These fragments are convergently joined to the "recognition" domain through a remarkably versatile macrotransacetalization process. The resulting new analogs exhibit potent nanomolar or picomolar affinity to protein kinase C (PKC), comparable to or better than that found for bryostatin.


[Psychosomatic medicine: old resources and new technologies].
Two primary research directions closely coexist in psychosomatic medicine. One is the most traditional, associated with an assessment of the psychological aspects of the connection, interconnection and mutual impact of mental and somatic pathology. The second, based on the rapid development of biological medicine in the last decade, studies causal associations and looks for shared mechanisms. In our review, we consider the previous main stages in the psychosomatic medicine and the prospective approaches to its further study. Evaluation of the etiopathogenesis of the entire set of mental and somatic symptoms in their interaction and dynamics can help identify individual subpopulations of patients with shared pathobiochemical and neurophysiological disorders. The recent interpretation of the biopsychosocial model is mainly related to the etiology and pathogenesis of mental disorders and also provides a good perspective for research on these issues. Today, there are sufficient opportunities to study all three domains of the model. Productive study of the biological, personal and social domains is also possible on the base of evidence-based design using modern research technologies.


Determination of pK(a) values of diastereomers of phosphinic pseudopeptides by CZE.
A CE method was used for the determination of acidity constants (pK(a)) of a series of ten phosphinic pseudopeptides, which varied in number and type of ionogenic groups. Effective electrophoretic mobilities were measured in the 1.8-12.0 pH range in the BGEs of constant ionic strength of 25 mM. Effective electrophoretic mobilities, corrected to standard temperature of 25 degrees C, were subjected to non-linear regression analysis and the obtained apparent pK(a) values were recalculated to thermodynamic pK(a)'s by extrapolation to zero ionic strength according to the extended Debye-Hückel model. The pK(a) values of the phosphinic acid group fell typically in the 1.5-2.25 interval, C-terminal carboxylic groups in the 2.94-3.50 interval, carboxylic groups of the lateral chain of glutamate and aspartate in the 4.68-4.97 interval, imidazolyl moiety of histidine in the 6.55-8.32 interval, N-terminal amino groups in the 7.65-8.28 interval and epsilon-amino group of the lateral chain of lysine in the 10.46-10.61 interval. Further, separation of diastereomers of the phosphinic pseudopeptides was investigated in achiral BGEs. Evaluation of the resolution of the diastereomers as a function of pH of the BGE revealed that most suitable pH region for separation of the diastereomers is around the pK(a) values of the central phosphinic acid group of the pseudopeptides. Successful separation of some diastereomers was, however, achieved in the neutral and alkaline BGEs as well.


Implementation of bioavailability in standard setting and risk assessment: suggestions based on a workshop with emphasis on metals.
Bioavailability is increasingly recognised as the key issue linking increased levels of toxicants with actually occurring adverse effects in ecosystems, whilst taking the modifying effects of the abiotic components of the environment into account. Various factors may affect bioavailability in the field, and often these factors are time- and space-dependent. This is one of the main reasons why legislators have been reluctant in implementing bioavailability in risk assessment procedures. Over the last few years, however, considerable scientific progress has been made with regard to better understanding of chemical and ecological mechanisms responsible for rendering chemicals available for uptake and toxicity. As a consequence, legislators face the challenge to anticipate the scientific progress and to implement bioavailability in legislation. This paper discusses the possibilities of implementing various methodologies within a maximum period of time of three years.


Infolding of the ligamentum flavum: a cause of spinal cord compression after reduction of cervical facet injuries.
Controversy exists regarding management of cervical facet injuries. Previous literature has focused on associated disc herniations reported to cause neurologic injury upon reduction. Although rupture of the ligamentum flavum has been noted with these injuries, its clinical significance has not been examined. In this case report, we present two patients in whom neurologic deterioration occurred due to infolding of the torn ligamentum flavum with spinal cord compression after reduction of cervical facet subluxations. Both had large flaps of ligamentum flavum arising from the caudal lamina which infolded upon reduction and became trapped between the spinal cord and cephalad lamina. Both patients regained normal motor function after removal of the pathologically infolded ligamentum. Neither patient had a disc herniation, hypotensive/anemic/hypoxic event, or epidural hematoma that could have otherwise been causative of the neurologic deficit. Pathologic infolding of ligamentum flavum, in addition to extruded disc herniations, should be recognized as another potential cause for spinal cord compression with reduction of cervical facet injuries. In particular, if there is a long flap of flavum arising from the caudal lamina poised to become entrapped in the spinal canal with reduction and the patient has a congenitally narrow canal, the surgeon should consider removal of the ligamentum flavum prior to reduction.


Detection of altered extracellular matrix in surface layers of unstable carotid plaque: an optical spectroscopy, birefringence and microarray genetic analysis.
Erosion and rupture of surface layers in atherosclerotic plaque can cause heart attack and stroke; however, changes in luminal surface composition are incompletely defined. Laser-induced fluorescence spectroscopy (LIFS), with limited tissue penetration, was used to investigate the surface of unstable carotid plaque and correlated with microscopy, birefringence and gene expression. Arterial matrix collagens I, III and elastin were assessed in unstable plaques (n = 25) and reference left internal mammary arteries (LIMA, n = 10). LIFS in addition to selective histological staining with picrosirius red, Movat pentachrome and immunostaining revealed decreased elastin and increased collagen I and III (P < 0.05) in carotid plaque when compared with LIMA. Within plaque, collagen I was elevated in the internal carotid region versus the common carotid region. Polarized light microscopy detected layers of aligned collagen and associated mechanical rigidity of the fibrous cap. Microarray analysis of three carotid and three LIMA specimens confirmed up-regulation of collagen I, III and IV, lysyl oxidase and MMP-12. In conclusion, LIFS analysis coupled with microscopy revealed marked regional differences in collagen I, III and elastin in surface layers of carotid plaque; indicative of plaque instability. Birefringence measurements demonstrated mechanical rigidity and weakening of the fibrous cap with complementary changes in ECM gene expression.


Neurogenic claudication and root claudication treated with calcitonin. A double-blind trial.
Forty-two patients with either neurogenic claudication or unilateral root claudication were analyzed in a double-blind comparison of salmon calcitonin (SCT) and placebo, receiving either 100 IU SCT or 1 ml saline four times a week for 8 weeks. Five of 20 SCT and one of 22 placebo patients were classified as responders. There was no statistically significant difference between the treatment groups in the proportion of responders. Seven of eighteen of the placebo group who later received salmon calcitonin improved their walking distance. The authors have not established that this is an organic response.


Hormonal regulation during secretion of alpha-fetoprotein in hepatoma cells grown in synthetic medium.
The variant cell line of H4-II-E-C3 cells derived from the Reuber H-35 hepatoma cells has been established using protein- and lipid-free synthetic medium. This H4-II-E-C3-V line can synthesize and secrete considerable amounts of alpha-fetoprotein (AFP) and albumin. The addition of 5 X 10(-7) M dexamethasone to the medium stimulated the excretion of AFP without increasing total AFP synthesis, whereas 8.7 X 10(-8) M insulin inhibited the excretion of AFP without a significant inhibition of intracellular AFP synthesis. However, neither dexamethasone nor insulin altered either the cellular or secreted levels of albumin. Cells were pulse labeled with [35S]methionine and then chased after addition of excess unlabeled methionine. AFP appeared in the medium after 10 min, and 50% of the protein was secreted after 110 min. The rate of secretion of AFP was much slower than that of albumin, 50% of which was secreted after 25 min. Dexamethasone, 5 X 10(-7) M, caused a marked enhancement in the rate of AFP secretion, with 50% released after 75 min. Insulin, 8.7 X 10(-8) M, by contrast, caused a marked delay in AFP secretion with only 20% released after 180 min and then a plateau was approached. Since the intracellular AFP was excreted 55% after 180 min the remaining 25% of newly made AFP was suggested to be degraded during secretion. The kinetics of movement of AFP during secretion and endoglycosidase H treatment of intracellular and secreted AFP suggested that insulin impeded the transport of AFP from the rough endoplasmic reticulum to the Golgi apparatus.


Not to be and then to be: visual representation of ignored unfamiliar faces.
Negative priming, the increase in response time and/or errors to targets previously encountered as distractors, is explained by inhibitory mechanisms that block the access of distractor representations to response systems. The processing of unfamiliar human faces was investigated using negative priming. Observers viewed a row of faces to decide whether 2 target faces were the same or different. Response latencies were longer when 1 or both targets had appeared as distractors on the immediately preceding trial--evidence that never-before seen faces are represented and require inhibition. Response latencies were shorter when face targets had appeared as distractors, either corrupted with high-frequency noise or contrast inverted--evidence that representations are facilitated. Finally, response latencies remained unaltered when face targets had appeared as upside-down distractors--evidence that not all distractor representations afford response priming. The visual system indeed represents ignored unfamiliar faces, but blocks these representations only if they vie with targets for the control of action.


[Tobacco use in Brazilian school adolescents: association with mental health and family context].
To study the association between aspects of mental health and the family context with tobacco experimentation and consumption among Brazilian schoolchildren. A cross-sectional study using data from the National Survey of Schoolchildren's Health in Brazil conducted on 109,104 schoolchildren from the ninth year of elementary school, day scholars in public and private schools in all Brazilian state capitals and the Federal District. Descriptive analysis of variables, bivariate and multivariate analyses were performed with logistic regression estimates, adjusted for socioeconomic variables, obtaining the odds ratio (OR) and 95% confidence interval (95%CI). Among the participants, 52.2% were female. Twenty point nine percent of the students reported having experimented with cigarettes at least once. Adolescents who experienced more often feelings of loneliness (OR: 2.07; 95%CI: 1.98-2.16), difficulty sleeping (OR: 2.37; 95%CI: 2.52-2.48) and lower social interaction (OR: 1.27; 95%CI: 1.26-1.32), were more likely to smoke, regardless of socioeconomic status. The schoolchildren of parents or caregivers (OR: 2.39; 95%CI: 2.29-2.49) who did not know what their children did in their free time (OR: 1.52; 95%CI: 1.46-1.59) were associated with more frequent cigarette smoking. An association was seen between aspects of mental health and the family context with the experimentation and consumption of tobacco in adolescents enrolled in the study in Brazil. These associations are independent of school type, sex and skin colour. It is important that programmes for the prevention of tobacco use in adolescence emphasise these findings.


Fine particle receptor modeling in the atmosphere of Mexico City.
Source apportionment analyses were carried out by means of receptor modeling techniques to determine the contribution of major fine particulate matter (PM2.5) sources found at six sites in Mexico City. Thirty-six source profiles were determined within Mexico City to establish the fingerprints of particulate matter sources. Additionally, the profiles under the same source category were averaged using cluster analysis and the fingerprints of 10 sources were included. Before application of the chemical mass balance (CMB), several tests were carried out to determine the best combination of source profiles and species used for the fitting. CMB results showed significant spatial variations in source contributions among the six sites that are influenced by local soil types and land use. On average, 24-hr PM2.5 concentrations were dominated by mobile source emissions (45%), followed by secondary inorganic aerosols (16%) and geological material (17%). Industrial emissions representing oil combustion and incineration contributed less than 5%, and their contribution was higher at the industrial areas of Tlalnepantla (11%) and Xalostoc (8%). Other sources such as cooking, biomass burning, and oil fuel combustion were identified at lower levels. A second receptor model (principal component analysis, [PCA]) was subsequently applied to three of the monitoring sites for comparison purposes. Although differences were obtained between source contributions, results evidence the advantages of the combined use of different receptor modeling techniques for source apportionment, given the complementary nature of their results. Further research is needed in this direction to reach a better agreement between the estimated source contributions to the particulate matter mass.


Reference values for selected ophthalmic diagnostic tests of the ferret (Mustela putorius furo).
To perform selected ophthalmic diagnostic tests in healthy ferrets with the aim of establishing normal physiological reference values for this species. A total of 15 healthy, unrelated ferrets were used to test most of the parameters in this investigation. Eight of the 15 ferrets were used for central corneal thickness evaluation. Ages varied from 1.5 to 6 years of age. Selected diagnostic ocular tests were performed including Schirmer tear test, tonometry using an applanation tonometer (Tonopen), central corneal thickness using an ultrasonic pachymeter (Sonomed, Micropach, Model 200P +) and culture of the normal conjunctival bacterial flora. Staphylococcus sp. and Corynebacterium sp. were isolated from healthy conjunctival and eyelid margins, suggesting they are normal constituents of the conjunctival flora of the ferret. Results for selected ocular diagnostic tests investigated here for the ferret eye were as follows: intraocular pressure: 14.50 +/- 3.27 mmHg; Schirmer tear test: 5.31 +/- 1.32 mm/min; central corneal thickness: 0.337 +/- 0.020 mm. No statistically significant differences between ages or genders were found for any of the results. The reference data for the ocular tests obtained in this investigation will help veterinary ophthalmologists to more accurately diagnose ocular diseases in the ferret. Knowledge of these reference values will be particularly useful to diagnose discrete or unusual pathological changes of the ferret eye.


A medication-estimated health status measure for predicting primary care visits: the Long-Term Therapeutic Groups Index.
Managed care is one of the means advocated for health care reforms. The Malaysian government has proposed managed care for its citizens. In the Malaysian private health care sector, managed care is practised on a small scale with crude risk adjustment. The main determinant of an individual's health service utilization is their health status (HS). HS is used as a risk adjuster for capitation payment. Prescribed medications represent a useful source for HS estimation. We aimed to develop and validate a medication-based HS estimate and to incorporate it in the Andersen model of health service utilization. This is a preparatory step in studying the feasibility of developing a model for risk assessment in the Malaysian context. Data were collected retrospectively from an academic year from computerized databases in University Sains Malaysia (USM) about users of USM primary care services. A user is a USM health scheme beneficiary who made at least one visit in the academic year to USM-assigned primary care providers. Socio-demographic variables, enrolment period, medications prescribed and number of visits were also collected. Chronic illness medications and some non-chronic illness medications were used to calculate the Long-Term Therapeutic Groups Index (LTTGI) which is an estimate of the HS of users. Using a random 50% of users, weighted least square methods were used to develop a model that predicts a user's number of visits. The other 50% were used for validation. Socio-demographic variables explained 15% of variability in number of primary care visits among users. Adding the LTTGI improved the explanatory power of the model to 36% (P < 0.001). A similar contribution of the LTTGI was noted in the validation. The Long-Term Therapeutic Groups Index was successfully developed. Variability in number of primary care visits can be predicted by LTTGI-based models.


1H magnetic resonance in the study of hepatic encephalopathy in humans.
1H magnetic resonance (1H MR) studies of the brain in patients with liver diseases have shown several abnormalities that may be relevant for the pathogenesis of hepatic encephalopathy. 1H magnetic resonance imaging shows a typical pallidal hyperintensity on T1-weighted images. This abnormality appears to be secondary to the accumulation of manganese in basal ganglia because of portal-systemic shunting. No direct correlation between the magnitude of pallidal hyperintensity and the grade of hepatic encephalopathy has been found, but some studies have related pallidal hyperintensity to parkinsonism. 1H magnetic resonance spectroscopy shows relative to creatine an increase in glutamine/glutamate (Glx) signal and a decrease of choline containing compounds (Cho) and myo-inositol. Abnormalities in the Glx signal have been interpreted as an increase in brain glutamine secondary to the metabolism of ammonia in astrocytes. Disturbances of Cho and myo-inositol have been interpreted as a compensatory response to the increase in intracellular osmolality caused by the accumulation of glutamine in astrocytes. In addition, magnetization transfer imaging shows signs compatible with low-grade cerebral edema. Altogether, 1H MR studies suggest the accumulation of manganese and the development of osmotic abnormalities in the brain of patients with cirrhosis. These abnormalities appear to participate in some of the neurological manifestations of hepatic encephalopathy.


Truncated dystrophin ameliorates the dystrophic phenotype of mdx mice by reducing sarcolipin-mediated SERCA inhibition.
Duchenne muscular dystrophy (DMD) and the less severe Becker muscular dystrophy (BMD) are due to mutations in the DMD gene. Previous reports show that in-frame deletion of exons 45-55 produces an internally shorted, but functional, dystrophin protein resulting in a very mild BMD phenotype. In order to elucidate the molecular mechanism leading to this phenotype, we generated exon 45-55 deleted dystrophin transgenic/mdx (Tg/mdx) mice. Muscular function of Tg/mdx mice was restored close to that of wild type (WT) mice but the localization of the neuronal type of nitric oxide synthase was changed from the sarcolemma to the cytosol. This led to hyper-nitrosylation of the ryanodine receptor 1 causing increased Ca2+ release from the sarcoplasmic reticulum. On the other hand, Ca2+ reuptake by the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA) was restored to the level of WT mice, suggesting that the Ca2+ dysregulation had been compensated by SERCA activation. In line with this, expression of sarcolipin (SLN), a SERCA-inhibitory peptide, was upregulated in mdx mice, but strongly reduced in Tg/mdx mice. Furthermore, knockdown of SLN ameliorated the cytosolic Ca2+ homeostasis and the dystrophic phenotype in mdx mice. These findings suggest that SLN may be a novel target for DMD therapy.


Specific inhibition of HCN channels slows rhythm differently in atria, ventricle and outflow tract and stabilizes conduction in the anoxic-reoxygenated embryonic heart model.
The hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are expressed in pacemaker cells very early during cardiogenesis. This work aimed at determining to what extent these channels are implicated in the electromechanical disturbances induced by a transient oxygen lack which may occur in utero. Spontaneously beating hearts or isolated ventricles and outflow tracts dissected from 4-day-old chick embryos were exposed to a selective inhibitor of HCN channels (ivabradine 0.1-10microM) to establish a dose-response relationship. The effects of ivabradine on electrocardiogram, excitation-contraction coupling and contractility of hearts submitted to anoxia (30min) and reoxygenation (60min) were also determined. The distribution of the predominant channel isoform, HCN4, was established in atria, ventricle and outflow tract by immunoblotting. Intrinsic beating rate of atria, ventricle and outflow tract was 164+/-22 (n=10), 78+/-24 (n=8) and 40+/-12bpm (n=23, mean+/-SD), respectively. In the whole heart, ivabradine (0.3microM) slowed the firing rate of atria by 16% and stabilized PR interval. These effects persisted throughout anoxia-reoxygenation, whereas the variations of QT duration, excitation-contraction coupling and contractility, as well as the types and duration of arrhythmias were not altered. Ivabradine (10microM) reduced the intrinsic rate of atria and isolated ventricle by 27% and 52%, respectively, whereas it abolished activity of the isolated outflow tract. Protein expression of HCN4 channels was higher in atria and ventricle than in the outflow tract. Thus, HCN channels are specifically distributed and control finely atrial, ventricular and outflow tract pacemakers as well as conduction in the embryonic heart under normoxia and throughout anoxia-reoxygenation.