Target-seq: single workflow for detection of genome integration site, DNA translocation and off-target events.
Designed donor DNA delivery through viral or nonviral systems to target loci in the host genome is a critical step for gene therapy. Adeno-associated virus and lentivirus are leading vehicles for in vivo and ex vivo delivery of therapeutic genes due to their high delivery and editing efficiency. Nonviral editing tools, such as CRISPR/Cas9, are getting more attention for gene modification. However, there are safety concerns; for example, tumorigenesis due to off-target effects and DNA rearrangement. Analysis tools to detect and characterize on-target and off-target genome modification post editing in the host genome are pivotal for evaluating the success and safety of gene therapy. We developed Target-seq combined with different analysis tools to detect the genome integration site, DNA translocation and off-target events.


Feasibility of a telephone-based counseling program for informal caregivers of hospice patients.
Informal caregivers of hospice patients experience multiple stressors that can negatively impact physical, psychological, and emotional health. The Caregiver Life Line (CaLL) study was a one-group, intervention-only feasibility study of a telephone cognitive-behavioral stress-management intervention consisting of 10 to 12 weekly telephone counseling calls designed specifically for informal caregivers of home-based hospice patients. Caregivers, who were recruited via seven participating hospices, were assessed at baseline, 3, and 6 months. Psychosocial measures included depression, perceived stress, benefit finding, social support, and quality of life (QoL). Process evaluation was also conducted at 6 months. Of the 25 participants, 19 completed the 12-week intervention. The mean age was 60.3 years; 91% were female and 70% married. Forty-eight percent were caring for a spouse; 51% were caring for a parent or other family member. Participating caregivers and hospice staff reported high levels of satisfaction with the intervention. Although the small sample size and lack of a control group limit our ability to assess the efficacy of the intervention, psychosocial outcomes changed in the expected direction and participants reported high levels of self-efficacy regarding their ability to implement the adaptive coping skills. Interestingly, we observed a decrease in physical QoL across the three time points. This pilot study demonstrated that the CaLL intervention is feasible and acceptable to informal hospice caregivers. This report describes important lessons learned from the pilot study with the intention of informing future intervention studies with this population.


GA binding protein augments autophagy via transcriptional activation of BECN1-PIK3C3 complex genes.
Macroautophagy is a vesicular catabolic trafficking pathway that is thought to protect cells from diverse stressors and to promote longevity. Recent studies have revealed that transcription factors play important roles in the regulation of autophagy. In this study, we have identified GA binding protein (GABP) as a transcriptional regulator of the combinatorial expression of BECN1-PIK3C3 complex genes involved in autophagosome initiation. We performed bioinformatics analyses that demonstrated highly conserved putative GABP sites in genes that encode BECN1/Beclin 1, several BECN1 interacting proteins, and downstream autophagy proteins including the ATG12-ATG5-ATG16L1 complex. We demonstrate that GABP binds to the promoter regions of BECN1-PIK3C3 complex genes and activates their transcriptional activities. Knockdown of GABP reduced BECN1-PIK3C3 complex transcripts, BECN1-PIK3C3 complex protein levels and autophagy in cultured cells. Conversely, overexpression of GABP increased autophagy. Nutrient starvation increased GABP-dependent transcriptional activity of BECN1-PIK3C3 complex gene promoters and increased the recruitment of GABP to the BECN1 promoter. Our data reveal a novel function of GABP in the regulation of autophagy via transcriptional activation of the BECN1-PIK3C3 complex.


High affinity choline transporter immunoreactivity in rat ileum myenteric nerves.
Recently, an antibody against the choline transporter (CHT), an essential molecule involved in ACh uptake, was used to label cholinergic nerves in the central nervous system; however, the enteric nervous system (ENS) was not examined. The present study localised CHT immunoreactivity (CHT-IR) within the rat ileum ENS and determined whether it colocalised with immunoreactivity for markers of cholinergic, tachykinergic and nitrergic circuitry. Segments of rat ileum were fixed, prepared for sectioning or whole-mounts and incubated with anti-CHT antisera followed by a fluorescent secondary antibody. Samples were double-labelled with antibodies to nitric oxide synthase, substance P (SP), common choline acetyltransferase (cChAT) and vesicular acetylcholine transporter (VAChT). CHT-IR was present in varicosities of nerve fibres in the myenteric plexus and muscle layers of rat ileum. In the myenteric ganglia, CHT-IR was found in nerve fibres and the cytoplasm of some nerve cell bodies. In the myenteric ganglia, no CHT/cChAT-immunoreactive neurons were present. A small number of CHT/SP-immunoreactive neurons and CHT/SP-immunoreactive nerve fibres clustered around unlabelled neurons. CHT-IR colocalised with VAChT-IR in the myenteric plexus but only half of the CHT-immunoreactive myenteric nerve fibres were VAChT-immunoreactive and half of VAChT-immunoreactive fibres were CHT-immunoreactive. In the circular muscle, 75% of CHT-immunoreactive fibres were VAChT-immunoreactive. Thus, the anti-CHT antiserum labels neurons and nerve fibres in the rat ENS. It does not label cholinergic cChAT-immunoreactive neurons, although it does immunostain cholinergic VAChT-immunoreactive nerve fibres and a population of nerves that are not VAChT-immunoreactive.


Water Sorption, Water Solubility, and Rheological Properties of Resin-Based Dental Composites Incorporating Immobilizable Eugenol-Derivative Monomer.
The present study aimed to evaluate the properties of new dental formulations incorporating a new polymerizable-derivative of eugenol (EgGMA). The experimental composites were prepared (by weight) with 35% resin-based matrix (1:1, bisphenol A-glycidyl methacrylate/triethylene glycol dimethacrylate) and 65% reinforcing materials (4:3, hydroxyapatite/zirconium oxide). A portion of 0.0, 2.5, and 5.0% of the resins with respect to the total composite was replaced by EgGMA monomer to obtain TBEg0, TBEg2.5, and TBEg5, respectively. The complex viscosity (at 25 and 37 °C), degree of conversion (DC), and water sorption (WSP) and water solubility (WSL) (3 cycles of sorption-desorption process) were investigated. Data were statistically analyzed using one-way and Tukey post-hoc tests. The results revealed a viscosity reduction with shear-thinning behavior as the EgGMA amount and temperature increased. The average complex viscosities at a lower frequency (ω = 1.0 rad/s) and at 25 °C were 234.7 ± 13.4, 86.4 ± 16.5, and 57.3 ± 17.1 (kPa·s) for TBEg0, TBEg2.5, and TBEg5, respectively. The inclusion of EgGMA led to a lower DC and WSP but higher WSL, compared to those of the reference (TBEg0). However, no significant differences between TBEg2.5 and control were detected (p > 0.05). Therefore, the incorporation of EgGMA in a low quantity, e.g., up to 8.45 mol% of resins, within the matrix may enhance the composite's performance, including handling and solubility properties without any apparent effect on DC and water sorption, making it a promising monomeric biomaterial for various applications including restorative dentistry.


Hepatocellular neoplasms after intrahepatic transplantation of ovarian fragments into ovariectomized rats.
Intrahepatic transplantation of ovarian fragments in ovariectomized rats results in morphological abnormalities. The liver acini draining blood from ovarian grafts show alterations resembling chemically induced amphophilic hepatocellular preneoplasias. We investigated the long-term development of these estrogen-induced foci of altered hepatocytes (FAH). We divided 451 Lewis rats into one main group (MG) and 11 (7 female, 4 male) control groups and observed them for up to 30 months. MG animals were ovariectomized and received ovarian transplants into the right liver part. Different combinations of castration, transplantation of ovarian or testicular fragments, and administration of antiestrogenic toremifene were used in controls. In the MG, transplants showed signs of gonadotropic stimulation, and estrogen levels were strongly increased in the downstream liver acini. After 6 and 12 months, FAH developed in hepatocytes downstream of the transplants. After 18 months, 27% of the MG animals showed transformation of FAH into hepatocellular adenomas; this figure increased to 42% after 24 months (8/19), significantly outnumbering four spontaneous adenomas that developed between 18 and 30 months in 258 control animals. Hepatocellular carcinoma (HCC) appeared only in the MG. At 24 and 30 months, 18 HCCs developed; thus, 78% of MG animals showed at least one carcinoma. Administration of toremifene in ovariectomized and transplanted animals completely prevented hepatocarcinogenesis. Testicular grafts showed no influence on liver tissue. In conclusion, initially adaptive but preneoplastic alterations in hepatocytes downstream of intrahepatically transplanted ovarian fragments may transform into HCC, indicating a strong hepatocarcinogenic potential of high local levels of endogenous estrogens in the rat liver.


Balancing my disease: women's perspectives of living with inflammatory bowel disease.
The purpose of this research was to examine the lived experiences of women with inflammatory bowel disease, by specifically exploring their management of their illness after diagnosis. Inflammatory bowel disease is a chronic autoimmune disease that has no known cause or cure. It is composed of two types of conditions: Crohn's disease and Ulcerative Colitis which have similar signs and symptoms, but have distinct physiological properties. Currently, Canada has the highest reported prevalence and incidence rates of inflammatory bowel disease in the world (Molodecky et al. 2012). Despite these increasing rates, there is a lack of knowledge and understanding of the burdens associated with inflammatory bowel disease, in particular from the participant's perspective. Heuristic phenomenology was the theoretical orientation used for this study. Eight women (aged 30-50 years) with inflammatory bowel disease who resided in Southern Ontario were recruited. Each participant completed background questionnaires and a one-on-one semi-structured interview. Several salient themes were identified; however, this paper will solely focus on one theme, balancing my disease. To achieve balance, the women described changing dietary behaviours, adjusting daily routines and managing symptoms through medications. These modifications were described as being important in controlling inflammatory bowel disease symptomatology on a day-to-day basis. There are many triggers for inflammatory bowel disease symptomatology. As such, the means in which those with inflammatory bowel disease manage their conditions varies. Participants discussed using a combination of strategies to control their condition. This information is important for patients living with inflammatory bowel disease, their support people, and healthcare professionals to identify various key strategies to improve the quality of life and well-being of those affected by inflammatory bowel disease. Furthermore, this research provided a voice to women diagnosed with inflammatory bowel disease by allowing them to openly convey their experiences of living with inflammatory bowel disease.


A promising subgroup identification method based on a genetic algorithm for censored survival data.
Modern precision medicine requires drug development to account for patients' heterogeneity, as only a subgroup of the patient population is likely to benefit from the targeted therapy. In this paper, we propose a novel method for subgroup identification based on a genetic algorithm. The proposed method can detect promising subgroups defined by predictive biomarkers in which the treatment effects are much higher than the population average. The main idea is to search for the subgroup with the greatest predictive ability in the entire subgroup space via a genetic algorithm. We design a real-valued representation of subgroups that evolves according to a genetic algorithm and derive an objective function that properly evaluates the predictive ability of the subgroups. Compared with model- or tree-based subgroup identification methods, the distinctive search strategy of this new approach offers an improved capability to explore subgroups defined by multiple predictive biomarkers. By embedding a resampling scheme, the multiplicity and complexity issues inherent in subgroup identification methods can be addressed flexibly. We evaluate the performance of the proposed method in comparison with two other methods using simulation studies and a real-world example. The results show that the proposed method exhibits good properties in terms of multiplicity and complexity control, and the subgroups identified are much more accurate. Although we focus on the implementation of censored survival data, this method could easily be extended for the realization of continuous and categorical endpoints.


Accuracy of affordable instruments for hearing screening in adults and the elderly.
To evaluate the accuracy of affordable instruments for hearing screening of adults and the elderly. This study was carried out with users of a Hearing Health Service of the Unified Health System. All were screened with the MoBASA smartphone application, the Telehealth audiometer (TH) and the electronic version of the Hearing Handicap Inventory for the Elderly - screening version - eHHIE-S. The examiners were blinded to the results of the screening tests and pure tone audiometry (PTA). Hearing impairment was considered for those with a PTA quadritonal mean greater than 40 dB in the best ear. Sensitivity, specificity and positive and negative predictive values (PPV and NPV, respectively) were calculated. The Kappa index was used as an agreement indicator between the PTA and the screening results. The sample consisted of 80 individuals between 18 and 94 years old (55.18 ± 20.21). In the PTA test, 21 individuals (26.25%) had typical hearing and 59 (73.75%) hearing loss. In the hearing screening tests, sensitivity, specificity, PPV and NPV values greater than 75% were observed with the MoBASA as well as in terms of sensitivity and NPV of the TH and the eHHIE-S. The TH and the eHHIE-S specificity and PPV were less than 75%. The Kappa index indicated a substantial agreement (0.6) between the PTA and the MoBASA screening results. The TH and the eHHIE-S showed regular agreement (0.3). MoBASA proved to be an accurate method for hearing screening of adults and the elderly with disabling hearing loss.


Integrase Strand Transfer Inhibitor Resistance Mutations in Antiretroviral Therapy-Naive and Treatment-Experienced HIV Patients in South Korea.
Updated guidelines for the treatment of antiretroviral therapy (ART)-negative patients with HIV recommend integrase strand transfer inhibitor (INSTI)-based regimens. HIV patients are tested for resistance to antiretrovirals, and the reported prevalence of transmitted INSTI resistance remains rare worldwide. However, no data related to INSTI resistance in Korean HIV patients have been reported. We aimed to determine the prevalence of INSTI resistance-related mutations in South Korea. We subjected both ART-naive (n = 58) and ART-experienced Korean HIV patients (n = 41) to genotypic resistance analysis and determined a prevalence of INSTI major resistance mutations of 3.4% (n = 2) among ART-naive patients and 22.0% (n = 9) among ART-experienced patients. In the former group, both major INSTI resistance cases involved the nonpolymorphic E92Q mutation in the integrase strand. Our findings suggest that INSTI resistance testing should be included in the standard resistance screening protocols for Korean HIV patients.


Symbiotic effectiveness of indigenous soybean bradyrhizobia as related to serological, morphological, rhizobitoxine, and hydrogenase phenotypes.
A Collection of 360 isolates of Bradyrhizobium japonicum was developed from soybean (Glycine max [L.] Merrill) nodules taken from 18 locations in Delaware. The isolates were characterized serologically with an enzyme-linked immunosorbent assay, morphologically by colony type on yeast extract-mannitol agar, and for production of rhizobitoxine symptoms with soybean plants. These analyses revealed 12 and 3 groups based on serology and morphology, respectively. The more common identifiable isolates were in serogroups 94, 6, 122, and 76. Nearly 33% of the isolates were rated nonreactive with all of the antisera tested. Overall, 18% of the isolates produced rhizobitoxine symptoms, and these were associated with five known serogroups (31, 46, 76, 94, and 130) and the nonreactive grouping, but with only one colony type. A subsample of 92 isolates was rated for N(2)-fixing ability in the greenhouse and for hydrogenase phenotype in the laboratory. The nitrogen content of plant shoots was strongly and comparably related to both the serological and morphological groupings. Rhizobitoxine and hydrogenase phenotypes were relatively poor predictors of symbiotic effectiveness. Among the serologically reactive isolates, those in serogroups 38-115, 122, and 110 fixed the most N(2), whereas one colony type (that containing isolates producing rhizobitoxine) was clearly inferior to the remaining two morphological groupings. Isolates displaying hydrogenase activity (approximately 15% of the isolates tested) correlated with three serologically reactive groupings (serogroups 110 and 122 and a 122/123 cross-reactive group) and two colony types, none of which coincided with groupings containing bradyrhizobia rated positive for rhizobitoxine production.


Involvement of Smads in TGFbeta1-induced furin (fur) transcription.
Furin is recognized as being one of the main convertases of the cellular constitutive secretion pathway but the mechanisms regulating its expression are still unknown. We have previously demonstrated that TGFbeta1 up-regulates its own converting enzyme, furin, creating a novel activation/regulation cycle of potential importance in a variety of physiological and pathophysiological conditions. The fur (fes upstream region) gene is regulated via three alternative promoters; P1, P1A, and P1B. To gain insight into the molecular mechanism(s) underlying this up-regulation, we performed transient cell transfections with P1, P1A, and P1B promoter luciferase constructs. Transfection experiments in HepG2 cells revealed that fur P1 promoter is the strongest and the most sensitive to TGFbeta1 stimulation (5 ng/ml) (3.2-fold vs. 2.4-fold for P1A and 2.1-fold for P1B). Cotransfection with either a dominant negative mutant form of Smad2 [Smad2(3SA)] or a known Smad inhibitor [Smad7] inhibit constitutive and TGFbeta1-induced luciferase activity indicating the participation of endogenous Smads. Increased levels of TGFbeta1-induced transcriptional activation of the P1 promoter by overexpression of Smad2 and/or Smad4 is greatly reduced in the presence of Smad2(3SA) and completely inhibited by Smad7, suggesting the participation of endogenous Smad2/Smad4 complexes. Furthermore, the fork-head activin signal transducer (FAST-1), known to interact with Smad2/Smad4 complexes, is a potent stimulator of TGFbeta1-induced transactivation of the fur P1 promoter. Five prime-deletion analysis of this promoter identified the proximal region (between positions -8734 and -7925), as the nucleotide stretch that carries most of the transcriptional activation of fur P1 promoter by Smad2. Overall, the present data demonstrate that Smad2 and Smad4 possibly in complex with FAST-1 or other DNA binding partners participate in the constitutive and inducible transactivation of the fur P1 promoter. This represents the first detailed study of the transcriptional regulation of the fur gene.


Risk factors for low urinary citrate in calcium nephrolithiasis: low vegetable fibre intake and low urine volume to be added to the list.
Risk factors for low urinary citrate excretion were assessed in 34 consecutive male recurrent idiopathic calcium stone formers (RCSF) who collected two 24-h urines while on free-choice diet. Overt hypocitraturia (hypo-cit) was defined as UCit x V < 1.70 mmol/day, and 'low' citraturia (low-cit) as UCit x V between 1.70 and 2.11 mmol/day. Twenty-three RCSF had normocitraturia (normo-cit), six low-cit and five hypo-cit. UCit x V positively correlated with urine volume (VOLUME, r = 0.44, P = 0.009), vegetable fibre intake (fibers, r = 0.46, P = 0.009) and GI-alkali absorption (alkali, r = 0.47, P = 0.006), and volume, fibres and alkali tended to be lower among RCSF with low-/hypo-cit. A 3-day NH4Cl loading test (0.95 mEq/kg BW daily in 3 doses) was performed in RCSF as well as in 14 age-matched healthy male controls (C). On a plot of urine pH versus serum bicarbonate, 10 of 11 RCSF with low-/hypo-cit, but only six of 23 with normo-cit (P = 0.0004) fell off the normal range, indicating incomplete RTA. Two or more risk factors simultaneously occurred in only four of 23 RCSF with normo-cit, but in eight of 11 with low-/hypo-cit (P = 0.002). In conclusion, incomplete RTA is the most prevalent risk factor for low-/hypo-cit in RCSF, and decreases in vegetable fibres and urine volume emerge as two new risk factors for low urinary CIT.


Angular induction as a function of contact and target orientation.
The Poggendorff effect is seen as misalignment of two obliques, or misprojection of one, when the obliques are placed outside a set of parallel lines. To understand better the mechanisms behind this effect, the orientation of the lines which are normally parallel was systematically manipulated. The results indicate that projection bias is affected by the orientation of either line, is at a minimum where the line is orthogonal to the oblique, and is maximal at small angles. This is in line with classic theories which attribute the illusion to misperception of angular size. However, such explanations presuppose that in order to be effective the induction line must be proximal to the oblique so that an angle can be formed. Results are reported which show that the angle formed by the oblique and a line placed at a distance from the oblique, serving as the target of the projection, follows an angular rule of effectiveness similar to what is seen when the line is placed directly in contact with the oblique. The underlying process is described as 'angular induction'.


Production of Monoclonal Antibodies Specific to FimA of Porphyromonas gingivalis and Their Inhibitory Activity on Bacterial Binding.
The FimA of Porphyromonas gingivalis is a crucial pathogenic component of the bacteria and has been implicated as a target for vaccine development against the periodontal diseases. In this study, the purified fimbriae (FimA subunit polymers) protein was used for immunization in their native form and B hybridoma clones producing antibodies specific to FimA were established. The monoclonal antibodies prepared from selected two clones, designated #123 (IgG2b/ kappa) and #265 (IgG1/kappa), displayed different patterns of binding activity against the cognate antigen. Both antibodies reacted with conformational epitopes expressed by partially dissociated oligomers, but not with monomer as elucidated by Western blot analysis. Ascites fluid containing the monoclonal antibodies showed the inhibitory activity against P. gingivalis to saliva-coated hydroxyapatite beads, an in vitro model for the pellicle-coated tooth surface. These results suggest that the monoclonal antibodies could be used as vaccine material against the periodontal diseases through passive immunization.


Peritoneal Seeding Is More Common in Gastric Cancer Patients with FGFR2 Amplification or High Tumor Mutation Burden.
Fibroblast growth factor receptor-2 (FGFR2) gene alterations have been identified in solid tumors. FGFR2 amplification is found in 2−9% of gastric carcinomas. We hypothesized that FGFR2 could be associated with peritoneal seeding and studied 360 advanced gastric carcinoma patients; 222 (61.7%) were male, 246 (73.7%) had poorly differentiated histology, and 175 (48.6%) presented with peritoneal seeding. High tumor mutation burden (TMB) was observed in 44 (12.2%) patients, high microsatellite instability (MSI) was observed in 12 (3.33%) patients, ERBB2 amplification was observed in 44 (12.2%) patients, EBV positivity was observed in 10 (10/278; 3.6%) patients, and PD-L1 positivity was observed in 186 (186/264; 70.5%) cases. We found FGFR2 amplification in 26 (7.2%) patients, of which 12 (46.2%) were female and 22 (84.6%) had poorly differentiated histology. In these 26 cases, the copy number of FGFR2 amplification ranged from 3.7 to 274. Eighteen of them showed seeding, and this association was statistically significant (18/26, 69.2%; 157/334, 47%; p = 0.023). In addition, high TMB was significantly associated with seeding (p = 0.028; OR = 1.83). Poorly differentiated histology was significantly associated with seeding (p = 0.04) but not with FGFR2 amplification (p > 0.1). Seeding was frequent in gastric carcinoma patients with FGFR2 amplification, in patients with high TMB, or in those who were female. The subgroup of patients with FGFR2 amplification could be potential candidates for targeted therapeutic agents.


Low sero-prevalence of hepatitis delta antibodies in HIV/ hepatitis B co-infected patients attending an urban HIV clinic in Uganda.
Co-infection with hepatitis B (HBV) and hepatitis D (HDV) is common among human immunodeficiency virus (HIV) infected individuals in developing countries and it aggressively accelerates progression of liver disease to cirrhosis and other complications. There is scarcity of data on HDV in sub-Saharan Africa .We investigated the sero-prevalence and factors associated with HDV antibody among HIV/HBV co-infected patients attending a large urban HIV clinic in Uganda. We screened 189 HIV/HBV co-infected individuals for anti-HDV immunoglobulin G (IgG) and performed logistic regression to determine the associated factors. Socio-demographic, clinical data, immunological status, and liver fibrosis (as determined by the Aspartate transaminase to platelet ratio index and transient elastography) were included. Participants were predominately young and of sound immunologic status (median age 40 years, median CD4 440 cells/µl). 98% were on ART regimens containing anti-HBV active medications (95.2% were on TDF/3TC while 4.8% on 3TC containing regimen). Median duration on ART was 36 months (IQR 22-72). Anti-HDV was detected in 6/198, 3.2% (95% CI 1.14-6.92%), associated with male gender and a duration of more than 5 years since HIV diagnosis. The sero-prevalence of HDV antibodies among the HIV/HBV co-infected patients is low in a Ugandan urban cohort.


[Expression of human annexin V in different fetal tissues].
To investigate the expression of human annexin-V (HA-V) in relation to HBV infection in different fetal tissues. Immunohistochemistry was employed to detect the expression and distribution of HA-V in the liver, kidney, ovary, heart, fallopian tube, spleen, and thymus gland of human fetus. HA-V expression was detected in different tissues including the ovary, liver, intrahepatic bile duct, heart, kidney, lymphocytic cells in the thymus gland, epithelial cells of the fallopian, and cortical and medullary cells of the spleen. HA-V was distributed mainly in the cytoplasm of the cells. The liver tissues exhibited greater gray scale for HA-V expression than in the other tissues (P<0.05) and no significant difference was observed in the other tissues than the liver (P>0.05) in image analysis with Photoshop 7.0. HA-V is an inherent protein in fetal tissues with possible relation to HBV infection of different tissues as a HBV receptor. Greater amount of HA-V in the liver may account for the vulnerability of the liver to HBV infection.


Lobar torsion after video-assisted thoracoscopic lobectomy: 2 case reports.
Lung torsion is a rare but potentially lethal complication after pulmonary resection. We report two cases of right middle lobar torsion after video-assisted thoracoscopic (VATS) right upper lobectomy in our unit. The two cases were diagnosed definitely 7 and 14 days after VATS surgery by computed tomography (CT) scan and bronchoscopy, respectively. Rethoracotomy and right middle lobectomy were performed in both patients. VATS lobectomy contributes to the development of lobar torsion so this complication should be considered. Carefully checking anatomical structures before thoracotomic closure and correctly suturing the middle lobe to the consecutive lobe are effective ways to prevent this complication.


Enzymatic cross-linking of beta-lactoglobulin: conformational properties using FTIR spectroscopy.
In this study, we use FTIR spectroscopy to probe the conformational changes of beta-lactoglobulin (beta-LG)-the main constituent of whey proteins-as subjected to enzymatic cross-linking by transglutaminase. We investigate both the amide I region (1600-1700 cm(-1)) and the C-H stretching region (2800-3100 cm(-1)). In the amide I region, spectra of denatured conformations of beta-LG, known to be necessary for cross-linking, differ according to the denaturation procedure, i.e., chemical or thermal treatment. Denaturation by chemical denaturants, dithiothreitol (DTT) or beta-mercaptoethanol, show no effect on the alpha-helix, while shifting the monomer dimer equilibrium toward higher monomer concentration. On the other hand, denaturing by thermal treatment dissociates the beta-sheets in the native structure, leading to new intermolecular beta-sheets being formed. Preheated then enzyme cross-linked beta-LG molecules show very similar spectra in the amide I region to the molecules with no cross-linking, indicating minimal effects of the cross-links on the carbonyl stretching mode. However, chemically denatured (using beta-mercaptoethanol) then enzyme cross-linked beta-LG molecules show noticeable diminution in the alpha-helix band and formation of strong hydrogen-bonded intermolecular beta-sheets. In the C-H stretching region, preheated then enzyme cross-linked beta-LG molecules exhibit a different degree of exposure of aliphatic amino acids due to the enzyme action. The same behavior is observed for DTT-treated then enzyme cross-linked beta-LG molecules. Generally, the changes in the C-H stretching region clearly indicate that hydrophobic interactions are altered upon enzymatic cross-linking.


[Establishment of an undifferential orchitis model in rats and changes of spermatogenic epithelium].
To explore the changes of rat testicular spermatogenic epithelium stimulated by bacterial lipopolysaccharide (LPS) in vivo. Twenty Wistar rats were divided into two groups: control group and experimental group. The control group was treated with pyrogen-free saline (1 ml/kg) and the experimental group was injected ip with saline containing LPS (1 mg/kg) once every two days. Two groups were operated after ten days in order to investigate the testicular pathological changes by HE staining and the expression of proliferating cell nuclear antigen( PCNA), alpha-catenin in spermatogenic epithelium by immunohistochemistry assay. The testes of the experimental group showed inflammatory changes. The positive expression of PCNA in seminiferous epithelium was significantly lower than that of control group. The number of positive cells in every seminiferous, in which only spermatogonia were stained in experimental group were 59 +/- 5 and it showed significant decrease compared with the control (P < 0.01). Furthermore, the percentage of such seminiferous tubules was 0.673 +/- 0.054 and increased apparently (P < 0.01). The expression of alpha-catenin in testicular tissue of the experimental group declined (P < 0.01), and cellular positive granular light density was 0.150 +/- 0.014. The ability of spermatogonium proliferation and the function of conglutination of cells under inflammatory condition of the testes declined, which may be one of the etiologies of male infertility.


Attention Measures of Accuracy, Variability, and Fatigue Detect Early Response to Donepezil in Alzheimer's Disease: A Randomized, Double-blind, Placebo-Controlled Pilot Trial.
Donepezil is widely used to treat Alzheimer's disease (AD), but detecting early response remains challenging for clinicians. Acetylcholine is known to directly modulate attention, particularly under high cognitive conditions, but no studies to date test whether measures of attention under high load can detect early effects of donepezil. We hypothesized that load-dependent attention tasks are sensitive to short-term treatment effects of donepezil, while global and other domain-specific cognitive measures are not. This longitudinal, randomized, double-blind, placebo-controlled pilot trial (ClinicalTrials.gov Identifier: NCT03073876) evaluated 23 participants newly diagnosed with AD initiating de novo donepezil treatment (5 mg). After baseline assessment, participants were randomized into Drug (n = 12) or Placebo (n = 11) groups, and retested after approximately 6 weeks. Cognitive assessment included: (a) attention tasks (Foreperiod Effect, Attentional Blink, and Covert Orienting tasks) measuring processing speed, top-down accuracy, orienting, intra-individual variability, and fatigue; (b) global measures (Alzheimer's Disease Assessment Scale-Cognitive Subscale, Mini-Mental Status Examination, Dementia Rating Scale); and (c) domain-specific measures (memory, language, visuospatial, and executive function). The Drug but not the Placebo group showed benefits of treatment at high-load measures by preserving top-down accuracy, improving intra-individual variability, and averting fatigue. In contrast, other global or cognitive domain-specific measures could not detect treatment effects over the same treatment interval. The pilot-study suggests that attention measures targeting accuracy, variability, and fatigue under high-load conditions could be sensitive to short-term cholinergic treatment. Given the central role of acetylcholine in attentional function, load-dependent attentional measures may be valuable cognitive markers of early treatment response.


Detecting amplicons of loop-mediated isothermal amplification.
Loop-mediated isothermal amplification (LAMP) assays are used to detect diverse pathogens. Initially, LAMP amplicons were detected using electrophoresis; later, real-time monitoring based on turbidity was developed to overcome the problem of contamination with environmental DNA. Recently, real-time monitoring of fluorescence signals using a quenching primer and probe has improved the reliability of amplification signals. Here, methods of detecting LAMP amplicons are reviewed.


TumGrowth: An open-access web tool for the statistical analysis of tumor growth curves.
The analysis of tumor growth curves is standard practice in experimental oncology including tumor immunology. In experimental oncology, cancer cells are inoculated into rodents (mostly mice) and their growth is monitored by measuring tumor diameter, surface or volume over time as a function of distinct treatments. Then, different groups of tumors/treatments are compared among each other for their evolution and possible responses to treatment. The R package TumGrowth has been created as a software tool allowing to carry out a series of statistical comparisons across or between groups of tumor growth curves obtained in a standard laboratory, for experimenters with limited knowledge in statistics. TumGrowth is freely available online at https://kroemerlab.shinyapps.io/TumGrowth/ and can be downloaded into any computer. It offers an exhaustive panoply of tools to visualize and analyze complex data sets including longitudinal, cross-sectional and time-to-endpoint measurements.


Amino Acid Sensing in Metabolic Homeostasis and Health.
Sensing and responding to changes in nutrient levels, including those of glucose, lipids, and amino acids, by the body is necessary for survival. Accordingly, perturbations in nutrient sensing are tightly linked with human pathologies, particularly metabolic diseases such as obesity, type 2 diabetes mellitus, and other complications of metabolic syndromes. The conventional view is that amino acids are fundamental elements for protein and peptide synthesis, while recent studies have revealed that amino acids are also important bioactive molecules that play key roles in signaling pathways and metabolic regulation. Different pathways that sense intracellular and extracellular levels of amino acids are integrated and coordinated at the organismal level, and, together, these pathways maintain whole metabolic homeostasis. In this review, we discuss the studies describing how important sensing signals respond to amino acid availability and how these sensing mechanisms modulate metabolic processes, including energy, glucose, and lipid metabolism. We further discuss whether dysregulation of amino acid sensing signals can be targeted to promote metabolic disorders, and discuss how to translate these mechanisms to treat human diseases. This review will help to enhance our overall understanding of the correlation between amino acid sensing and metabolic homeostasis, which have important implications for human health.


[Onychomycosis frequency in psoriatic patients in a tertiary care hospital].
The changes in psoriatic nails can closely resemble an onychomycosis. Therefore, the fungal infection may be underdiagnosed. It was investigated the frequency of mycosis in fingernails and toenails in 150 patients with psoriasis in a dermatology department. The clinical data suggestive of onychomycosis were investigated. Nail scales were obtained and cultured on Sabouraud dextrose agar with and without antibiotic. A direct examination with KOH was also performed. Out of 150 patients, 67 (45 %) had healthy nails; 42 (28 %) presented onychomycosis and 41 (27 %) showed nail changes without infection. Fingernail changes were more associated with psoriatic onychopathy (82.5 %), unlike toenail changes that were more frequently caused by fungal infection (26.4 % vs. 9.45 % in psoriasis). Out of 20 positive cultures, 22 fungi were isolated, of which 11 belonged to Candida spp. (50 %). As risk factor to develope an onychomycosis, only the psoriasis evolution time showed a significant difference (p = 0.033). In patients with psoriasis, fingernail disorders are mainly due to the own disease, while toenail disorders changes can be associated with onychomycosis. The main etiological agents were yeasts from the genus Candida. The only factor associated with a higher incidence of onychomycosis in these patients was a long lasting psoriasis.


Mucosal delivery of RNA vaccines by Newcastle disease virus vectors.
The rapid evolution of SARS-CoV-2 since its pandemic outbreak has underscored the need for improved SARS-CoV-2 vaccines that efficiently reduce not only hospitalizations and deaths, but also infections and transmission. This might be achieved by a new generation of intranasally administered SARS-CoV-2 vaccines to stimulate protective mucosal immunity. Among all different approaches, preclinical and clinical information using Newcastle Disease Virus (NDV)-vectors expressing S of SARS-CoV2 as a COVID-19 vaccine show the potential of this vaccine platform as an affordable, highly immunogenic, safe strategy to intranasally vaccinate humans against SARS-CoV-2 and other infectious diseases. These vaccine vectors consist on the use of a harmless avian negative strand RNA virus to deliver intranasally a self-replicating RNA expressing the vaccine antigen in the cells of the respiratory mucosa. The vector also incorporates the antigen in the virus particle used for RNA delivery, thus combining the properties of nanoparticle-based and RNA-based vaccines. Other advantages of NDV-based vectors include the worldwide availability of manufacturing facilities for their production and their stability at non-freezing temperatures. While phase 3 clinical studies to evaluate efficacy are still pending, phase 1 and 2 clinical studies have demonstrated the safety and immunogenicity of NDV-S vaccines against SARS-CoV-2.


Aggregating and prostanoid-releasing effects of platelet-activating factor and leukotrienes on human polymorphonuclear leukocytes and platelets.
Aggregating and prostanoid-releasing properties of the inflammatory mediators, platelet-activating factor (PAF) and leukotrienes B4, C4 and D4 were studied in human polymorphonuclear leukocytes (PMNL) and in human platelet-rich plasma. Leukotriene B4 (LTB4) and PAF both induced a reversible aggregation of human PMNL with concomitant stimulation of PGE2 formation, whereas LTC4 had no effect on human PMNL. Arachidonic acid (AA) caused an irreversible aggregation of PMNL which was accompanied by formation of both PGE2 and TXB2. Inhibition of TXB2 synthesis by indomethacin or by OKY-1581, a thromboxane synthetase inhibitor, had no effect on the PMNL aggregation induced by LTB4, PAF or AA. Leukotrienes B4, C4 and D4 caused neither aggregation nor TXB2 release in human platelet-rich plasma. PAF, on the other hand, induced a dose-dependent, reversible platelet aggregation which was not accompanied by TXB2 formation nor inhibited by OKY-1581. The present study indicates that in addition to inducing PMNL aggregation, LTB4 is capable of releasing arachidonate metabolites from human PMNL but not from human platelets. Also the responses of PMNL and platelets to PAF seem to differ as the PAF-induced PMNL aggregation was accompanied by increased prostanoid formation whereas the PAF-induced reversible platelet aggregation was obviously independent from arachidonate metabolism.


Transcriptome-based discovery of pathways and genes related to reproduction of the black tiger shrimp (Penaeus monodon).
The black tiger shrimp (Penaeus monodon) is an aquatic animal with considerable economic importance. Poor reproductive maturation in captivity impedes sustainable aquaculture production of this species. This study aims to provide transcriptomic information on reproductive organs using 454 pyrosequencing technology. The transcriptome analysis of ovaries and testes revealed 41,136 transcripts with 20,192 contigs. We found novel sets of transcripts completing several important reproductive pathways such as gonadotropin-releasing hormone (GnRH) signaling and progesterone-mediated oocyte maturation. In addition, we found transcripts encoding for receptors crucial for initiation of the maturation process, such as GnRH receptor (GnRHR), voltage-dependent calcium channel L type alpha-1C (CACNA1C) and epidermal growth factor receptor (EGFR). Moreover, we found a putative novel vigillin encoding for an estrogen-induced polysome-associated protein, which has not been reported in penaeid shrimp. These results suggest that the regulatory mechanism of the pathways important to reproductive maturation might be similar to those in the vertebrate. The obtained data will consequently accelerate the study of reproductive biology of this important species to ensure a sustainable shrimp farming industry.


Overexpression of AtDREB1D transcription factor improves drought tolerance in soybean.
Drought is one of the major abiotic stresses that affect productivity in soybean (Glycine max L.) Several genes induced by drought stress include functional genes and regulatory transcription factors. The Arabidopsis thaliana DREB1D transcription factor driven by the constitutive and ABA-inducible promoters was introduced into soybean through Agrobacterium tumefaciens-mediated gene transfer. Several transgenic lines were generated and molecular analysis was performed to confirm transgene integration. Transgenic plants with an ABA-inducible promoter showed a 1.5- to two-fold increase of transgene expression under severe stress conditions. Under well-watered conditions, transgenic plants with constitutive and ABA-inducible promoters showed reduced total leaf area and shoot biomass compared to non-transgenic plants. No significant differences in root length or root biomass were observed between transgenic and non-transgenic plants under non-stress conditions. When subjected to gradual water deficit, transgenic plants maintained higher relative water content because the transgenic lines used water more slowly as a result of reduced total leaf area. This caused them to wilt slower than non-transgenic plants. Transgenic plants showed differential drought tolerance responses with a significantly higher survival rate compared to non-transgenic plants when subjected to comparable severe water-deficit conditions. Moreover, the transgenic plants also showed improved drought tolerance by maintaining 17-24 % greater leaf cell membrane stability compared to non-transgenic plants. The results demonstrate the feasibility of engineering soybean for enhanced drought tolerance by expressing stress-responsive genes.


Using intergenerational photovoice to understand family strengths among Native American children and their caregivers.
The purpose of the current study was to examine Native American children and caregivers' perspectives of family and cultural strengths using photovoice and to identify lessons learned from the first-ever implementation of intergenerational photovoice with Native Americans. Participants were Native American, low-income caregivers (n = 6) and their children (n = 12) between the ages of 10 and 15 who participated in six photovoice sessions. The themes that emerged from photos and group discussion included myriad challenges faced by Native American families including exposure to community violence, substance abuse, and criminal offending and incarceration. Themes also emerged that highlighted the strengths of Native families that were used to overcome identified challenges, including religion/spirituality, engagement in traditional cultural practices (e.g., prayer, song, dance), healthy activities (e.g., running, meditation). These data provided foundational information that is currently being used, along with other data, to develop a culturally grounded, strengths-focused, family-based program (Tiwahe Wicagwicayapi [Strengthening/Growing Families in Lakota]) to prevent adverse childhood experiences. We also discuss the challenges of intergenerational photovoice and lessons learned to inform future intergenerational photovoice projects.


Fosfomycin, antimicrobial activity in vitro and in vivo.
Antibacterial activities of fosfomycin were investigated both in vitro and in vivo for the purpose of comparative evaluation on its fundamental properties with other antimicrobial agents. The MIC was determined with nutrient agar (Difco) inoculated with one loopful of 1,000-fold dilution (about 10(6) cells/ml) of bacterial suspension cultured overnight in nutrient broth. This substance showed antibacterial activity to most gram-positive and gram-negative bacteria, being strongest to Enterobacteriaceae with a peak of the MIC at 1.56 mug/ml in Salmonella. It was also active against P. aeruginosa with a peak of the MIC at 6.25 mug/ml in its sensitivity distribution. Intravenous and subcutaneous fosfomycin Na salt and oral fosfomycin Ca salt were given to 5-week-old ddN strain male mice challenged with clinical isolates of P. aeruginosa, E. coli and P. mirabilis. Therapeutic effect was observed in all these test organisms. In P. aeruginosa, it was more effective than carbenicillin.


["In vitro" activity of gentamicin, sisomicin and tobramycin against "pseudomonas aeruginosa" strains from hospital sources (author's transl)].
The in vitro activity of two amynoglicosidic antibiotics, tobramycin and sisomicin, against 215 strains of Pseudomonas aeruginosa isolated from clinical specimens during the years 1972-1975, was evaluated in comparison with that of gentamicin. Tobramycin showed a greater activity than sisomicin and gentamicin. At the concentration of 5 mu/ml tobramycin resulted inhibitory on 94% of strains and sisomicin and gentamicin on 78.6% and 49.8% of strains respectively. The susceptibility of Pseudomonas aeruginosa strains to gentamicin decreased from 1972 to 1975, with an increasing of resistant strains of 23%. In an evaluation of possible cross-resistance of 14 strains of Pseudomonas aeruginosa highly resistant (MIC greater than or equal to 25 gamma/ml) to gentamicin, 13 showed a high degree of resistance also to sisomicin; on the other hand 7 strains highly resistant to gentamicin was sensitive to tobramycin at concentration less than or equal to 10 gamma/ml.


Glucose 6-phosphate formation by L-type pentose phosphate pathway reactions of rat liver in vitro: further evidence.
An investigation of the mechanism in vitro of the non-oxidative segment of the pentose phosphate pathway using [5-14C]ribose 5-phosphate as a prediction labelling substrate with rat liver enzyme preparation is reported. Glucose 6-phosphate formed during the initial 0.5 h of reaction was heavily labelled in C-1 and thus is consistent with the prediction of the liver (L)-type pentose phosphate pathway (theoretically C-1/C-6 = 0.5). The reaction sequences of the fat (F-) type pentose phosphate pathway exclusively confine 14C to C-6 of glucose 6-phosphate. The presence of L-type reactions was further affirmed by the formation of D-arabinose 5-phosphate and D-glycero-D-ido-octulose phosphate which were identified and measured during the initial 2 h of incubation using gas liquid chromatography. After 3 h the concentration of 14C in C-1 of glucose 6-phosphate gradually declined and by 17 h of incubation the ratio of 14C in C-1/C-6 was 0.1. Possible reasons for the late changes in 14C-isotope distributions towards a pattern consistent with a contribution of F-type pentose phosphate pathway are given.


Comparison of formula accuracy for intraocular lens power calculation based on measurements by a swept-source optical coherence tomography optical biometer.
To analyze the results of intraocular lens (IOL) power calculation using measurements by a swept-source optical coherence tomography (SS-OCT) optical biometer. IRCCS G.B. Bietti Foundation, Rome, Italy. Evaluation of a diagnostic test instrument. Preoperative measurements by the OA-2000 (Tomey Inc.) were taken in a consecutive series of patients undergoing cataract surgery with one IOL model (AcrySof SN60WF; Alcon Laboratories, Inc.). Measurements were entered into the following formulas: Barrett Universal II, Emmetropia Verifying Optical (EVO), Haigis, Hoffer Q, Holladay 1, Holladay 2, Holladay 2 with axial length adjustment, Kane, Olsen, Panacea, SRK/T, T2, and VRF. When refraction was measured at 1 month postoperatively, the mean arithmetic prediction error, the median absolute error (MedAE), and the percentage of eyes with a error of ±0.50 D or less were calculated after constant optimization. We enrolled 150 eyes. All formulas yielded excellent outcomes. The MedAE ranged between 0.200 D and 0.259 D, with a statistically significant difference among formulas (P = .0004). The lowest MedAE values were obtained with the Barrett, EVO, Kane, Olsenstandalone, Radial Basis Function (RBF), and T2 formulas. The percentage of eyes with a prediction error of ±0.50 D or less ranged between 80.00% and 90.67%, with a statistically significant difference (P < .0001). The Barrett, EVO, Holladay 2 with axial length adjustment, Kane, RBF, and T2 achieved the highest percentages (≥88%). Measurements provided by the SS-OCT optical biometer enable accurate IOL power calculation because all formulas yielded a prediction error within 0.50 D in at least 80% of eyes.


Polymorphism of the promoter region of C-509T of transforming growth factor-beta1 gene and ulcerative colitis.
Transforming growth factor-beta is a regulatory protein that plays a key role in inflammatory, fibrotic, and immunological events in the intestinal mucosa. Recently, attention has been focused on the role of transforming growth factor-beta in the etiopathogenesis of inflammatory bowel disease. Enhanced expression of transforming growth factor-beta mRNA in the lamina propria and a disordered expression pattern of transforming growth factor-beta1 receptors I and II in epithelial cells have been documented in the colonic mucosa of patients with ulcerative colitis and Crohn's disease. Based on these associations, we report in this study, the restriction fragment length polymorphism-polymerase chain reaction and allele frequencies of the transforming growth factor-beta gene polymorphisms in a population of Iranian patients with ulcerative colitis and healthy controls. We analyzed whether these two polymorphisms are related with the disease characteristics. One hundred fifty-seven (75 males and 82 females) unrelated patients with ulcerative colitis attending the Departments of Gastroenterology, Nemazi and Faghihi Hospitals, affiliated to Shiraz University of Medical Sciences, Shiraz, Iran were enrolled into this study. Ninety-four age- and sex-matched healthy volunteers with no history of chronic bloody diarrhea and abdominal pain (41 males and 53 females) served as the control group. The change at position -509 (C/T) of the transforming growth factor-beta1 gene was studied using restriction fragment length polymorphism-polymerase chain reaction in this study. The mean age of patients was 36.4 (range: 23 - 51) years. The genotype at position -509 (C/T) in 58 (37%) patients with ulcerative colitis, and 39 (41.5%) normal subjects, were homozygous as CC. In addition, 65 (41.5%) patients and 44 (47%) normal individuals were heterozygous as CT. Thirty-four (21.5%) of 157 patients, and 11 (11.5%) of 94 normal individuals, were homozygous as TT. There was no statistically significant difference between patients and normal female individuals in respect to genotype distribution and allele frequency at the said position (P = 0.138). No association could be found between transforming growth factor-beta1 -509 (C/T) promoter gene polymorphism and patients with ulcerative colitis.


Subclinical Neuropathy in Children With Type I Diabetes Mellitus: Tertiary Care Centre Experience.
Diabetic peripheral neuropathy is a common complication of diabetes mellitus (DM) type 1. However, it can occur without evidence of symptoms or clinical signs of neuropathy labeled as subclinical neuropathy, which neurophysiological studies can best detect. To evaluate the prevalence of subclinical neuropathy among children with DM type 1, determine the association with blood sugar control, and evaluate the pattern of nerve involvement in neurophysiological studies. This cross-sectional study evaluated 100 children with DM type 1, aged five to 15 years, at least one year after the diagnosis. Subclinical neuropathy was evaluated using nerve conduction study. Glycemic control was assessed using hemoglobin A1c (HbA1c). The mean age of subjects was 11.5 ± 0.25 years. The average age at the onset of the disease was 5.95 ± 0.25 years. There were 64 patients who had electrophysiological evidence of peripheral neuropathy. The most observed electrophysiological changes were distal latency abnormalities in the left and right peroneal nerves in 39 and 33 patients, respectively. Sensory nerve amplitude, peak latency, and conduction velocity were normal in all patients (100%). HbA1c level did not show a statistically significant association with the incidence of subclinical neuropathy. Subclinical neuropathy was prevalent in children with DM type 1. Poor glucose control was only associated with an increased odds ratio of subclinical neuropathy.


Effect of food on the absorption of vigabatrin.
1. This study, involving eight healthy volunteers, revealed that food intake does not have a clinically significant effect on vigabatrin kinetics. 2. Kinetics of vigabatrin in the fasting state showed low inter-individual variation. 3. The plasma concentration vs time profile followed a bi-exponential decline with a terminal half-life of 6.8 h and a relative bioavailability of 92% +/- 11%. 4. In the clinical setting, vigabatrin may be taken at times convenient for the patient.


Rural radiology services in Texas: an assessment in quality.
Health professionals share a common interest in improving the quality of healthcare provided to the notion's underserved citizens, many who live in rural areas. This article examines the quality of radiology services found in rural Texas hospitals as reported by radiology managers through online survey research in terms of staffing, radiologist availability, and quality control measures. A 22 question survey was developed and 72 of 106 possible responses were obtained for a 67.9% response rate representing 45% of the total population. Texas is a licensure state for radiologic technologists. Texas rural counties with fewer than 50 thousand citizens have only 9% of MRTs, 10% of LMRTs, and 12% of NCTs licensed to practice radiology imaging. Licensing all three levels of technologists through the some administrative body could result in more standard educational and training requirements, thereby increasing the quality of care given by these individuals. Patients seen at lower-volume rural facilities benefit from convenient scheduling and lower prices and are beginning to see faster reporting with the prevalence of teleradiology and voice recognition dictation.


Glial cells promote dendritic development in rat sympathetic neurons in vitro.
Many types of glial-neuronal interactions occur during the development of the nervous system. To determine how such interactions might affect the development of autonomic ganglia, we compared the morphology of embryonic rat sympathetic neurons grown in the absence and in the presence of ganglionic nonneuronal cells in serum-free medium. Dye injections, electron microscopy, and immunocytochemistry were used to distinguish axons from dendrites. In cultures without nonneuronal cells, most (greater than 80%) sympathetic neurons extended only a single axonal process, and this unipolar state persisted for at least 8 weeks. Coculture with ganglionic nonneuronal cells caused sympathetic neurons to become multipolar and to extend multiple (range 1-17) dendrites. Morphometric measurements made after 1 month of coculture indicated that the amount of dendritic growth that occurred in vitro (mean number of dendrites/cell = 7.5; total dendritic length = 1,050 micron) was similar to that normally occurring during a comparable period in situ. In contrast to its prominent effects on dendritic growth, coculture did not cause changes in the number of axons/neuron or in the uptake of neurotransmitter. Cultures with ganglionic nonneuronal cells were immunostained for antigens present on the surfaces of fibroblasts (Thy-1.1, fibronectin) and of glia of the peripheral nervous system (laminin). Fewer than 1% of the nonneuronal cells displayed immunoreactivity for fibroblastic antigens; in contrast, greater than or equal to 99% reacted with antibody to laminin. Moreover, reconstitution experiments revealed that purified populations of laminin-positive Schwann cells promoted dendritic growth. Fibroblasts and heart cells lacked this activity. These data indicate that glia selectively promote dendritic development in sympathetic neurons maintained in serum-free medium.


Polymer Brush-Induced Hollow Colloids via Diffusion-Controlled Silication.
We report on a new strategy to synthesize asymmetrical hollow colloidal particles by exploiting limited chemical diffusion that occurs at the periphery of a solvated polymer brush on the particle surface. The polymer brush-in our case poly(glycidyl methacrylate)-bears hydroxyl groups upon hydrolysis and is partially cross-linked under the Stöber condition of silication. Desolvation of the polymers creates a cavity. While elucidating this new mechanism, we demonstrate that particles with various types of cavities and tunable properties can be synthesized, including the ones bearing hemispherical and crescent shapes, as well as particles with wrinkled surfaces. Furthermore, we show that the hollow particles adopt preferred orientations because of their shape and composition attributes, which is further explored to facilitate the confined synthesis of nanocrystals.


Fuel selection during prolonged arm and leg exercise with 13C-glucose ingestion.
To compare fuel selection during prolonged arm (AE) and leg exercise (LE) with water or glucose ingestion. Ten subjects (VO2max: 4.77 +/- 0.20 and 3.36 +/- 0.15 L x min(-1) for LE and AE, respectively) completed 120 min of LE and AE at 50% of the mode-specific maximal power output (353 +/- 18 and 160 +/- 9 W, respectively) with ingestion of water (20 mL x kg(-1)) or 13C-glucose (2 g x kg(-1)). Substrate oxidation was measured using indirect respiratory calorimetry corrected for urea excretion and 13CO2 production at the mouth. The contribution of protein oxidation to the energy yield (%En) was higher during AE than LE (approximately 8% vs approximately 4%) because of the lower energy expenditure and was not significantly modified with glucose ingestion. With water ingestion, the %En from CHO oxidation was not significantly different during LE and AE (64 +/- 2% and 66 +/- 2%, respectively). Glucose ingestion significantly increased the %En from total CHO oxidation during AE (78 +/- 3%) but not during LE (71 +/- 2%). Exogenous glucose oxidation was not significantly different in AE and LE (56 +/- 4 and 65 +/- 3 g, respectively), but the %En from exogenous glucose was higher during AE than LE (30 +/- 1% and 24 +/- 1%) because of the lower energy expenditure. When glucose was ingested, the %En from endogenous CHO oxidation was significantly reduced during both AE (66 +/- 2% to 48 +/- 3%) and LE (64 +/- 2% to 47 +/- 3%) and was not significantly different in the two modes of exercise. The difference in fuel selection between AE and LE when water was ingested was modest with a slightly higher reliance on CHO oxidation during AE. The amount of exogenous glucose oxidized was lower but its %En was higher during AE because of the lower energy expenditure.


Multimodal In Vivo Tracking of Chimeric Antigen Receptor T Cells in Preclinical Glioblastoma Models.
Iron oxide nanoparticles have been used to track the accumulation of chimeric antigen receptor (CAR) T cells with magnetic resonance imaging (MRI). However, the only nanoparticle available for clinical applications to date, ferumoxytol, has caused rare but severe anaphylactic reactions. MegaPro nanoparticles (MegaPro-NPs) provide an improved safety profile. We evaluated whether MegaPro-NPs can be applied for in vivo tracking of CAR T cells in a mouse model of glioblastoma multiforme. We labeled tumor-targeted CD70CAR (8R-70CAR) T cells and non-tumor-targeted controls with MegaPro-NPs, followed by inductively coupled plasma optical emission spectroscopy, Prussian blue staining, and cell viability assays. Next, we treated 42 NRG mice bearing U87-MG/eGFP-fLuc glioblastoma multiforme xenografts with MegaPro-NP-labeled/unlabeled CAR T cells or labeled untargeted T cells and performed serial MRI, magnetic particle imaging, and histology studies. The Kruskal-Wallis test was conducted to evaluate overall group differences, and the Mann-Whitney U test was applied to compare the pairs of groups. MegaPro-NP-labeled CAR T cells demonstrated significantly increased iron uptake compared with unlabeled controls ( P < 0.01). Cell viability, activation, and exhaustion markers were not significantly different between the 2 groups ( P > 0.05). In vivo, tumor T2* relaxation times were significantly lower after treatment with MegaPro-NP-labeled CAR T cells compared with untargeted T cells ( P < 0.01). There is no significant difference in tumor growth inhibition between mice injected with labeled and unlabeled CAR T cells. MegaPro-NPs can be used for in vivo tracking of CAR T cells. Because MegaPro-NPs recently completed phase II clinical trial investigation as an MRI contrast agent, MegaPro-NP is expected to be applied to track CAR T cells in cancer immunotherapy trials in the near future.


Comparison of total laparoscopic hysterectomy and abdominal hysterectomy.
The aim of this prospective study is to evaluate and compare to the outcomes of total laparoscopic hysterectomy (TLH) and total abdominal hysterectomy (TAH) who performed in our clinic. We performed surgical procedures at Necmettin Erbakan University Faculty of Medicine, Department of Obstetrics and Gynecology between January 2013 and April 2014. Forty patients who underwent TLH (group 1) compared to 40 patients who underwent TAH (group 2). The mean age of the cases, body mass index (BMI), duration of operation, the amount of blood loss, rates of complications and post operative hospital stay were compared for two groups. There were no statistically significant differences between the two groups regarding age, body mass index (BMI), specimen weight, pre-operative hemoglobin (Hb) value and rates of the complications. The mean post-operative Hb value was significantly higher in group 1 than group 2 (11.5±0.8 gr/dl vs. 10.8±1.7, p=0.02). The mean time of operation was significantly longer in group 1 than in group 2 (105.4±22.9 minutes vs. 74±18, p<0.001). The mean duration of hospital stay was statistically shorter in group 1 compared to the group 2 (2.48±0.6 days vs. 4.88±1.2, p<0.001). Total laparoscopic hysterectomy is safe and feasible method for gynecological diseases. TLH may offer specific benefits for properly selected patients. Its advantages are lower peri-operative morbidity, improvement of quality of life, shorter hospital stay and faster return to activity.


Effect of Interfacial Structure on Mechanical Properties of Graphene Reinforced Al2O3-WC Matrix Ceramic Composite.
The interfacial structures and interfacial bonding characteristics between graphene and matrix in graphene-reinforced Al2O3-WC matrix ceramic composite prepared by two-step hot pressing sintering were systematically investigated. Three interfacial structures including graphene-Al2O3, graphene-Al2OC and graphene-WC were determined in the Al2O3-WC-TiC-graphene composite by TEM. The interfacial adhesion energy and interfacial shear strength were calculated by first principles, and it has been found that the interfacial adhesion energy and interfacial shear strength of the graphene-Al2OC interface (0.287 eV/nm2, 59.32 MPa) were far lower than those of graphene-Al2O3 (0.967 eV/nm2, 395.77 MPa) and graphene-WC (0.781 eV/nm2, 229.84 MPa) interfaces. Thus, the composite with the strong and weak hybrid interfaces was successfully obtained, which was further confirmed by the microstructural analysis. This interfacial structure could induce strengthening mechanisms such as load transfer, grain refinement, etc., and toughening mechanisms such as crack bridging, graphene pull-out, etc., which effectively improved mechanical properties.


An Enhanced Region Proposal Network for object detection using deep learning method.
Faster Region-based Convolutional Network (Faster R-CNN) is a state-of-the-art object detection method. However, the object detection effect of Faster R-CNN is not good based on the Region Proposal Network (RPN). Inspired by RPN of Faster R-CNN, we propose a novel proposal generation method called Enhanced Region Proposal Network (ERPN). Four improvements are presented in ERPN. Firstly, our proposed deconvolutional feature pyramid network (DFPN) is introduced to improve the quality of region proposals. Secondly, novel anchor boxes are designed with interspersed scales and adaptive aspect ratios. Thereafter, the capability of object localization is increased. Thirdly, a particle swarm optimization (PSO) based support vector machine (SVM), termed PSO-SVM, is developed to distinguish the positive and negative anchor boxes. Fourthly, the classification part of multi-task loss function in RPN is improved. Consequently, the effect of classification loss is strengthened. In this study, our proposed ERPN is compared with five object detection methods on both PASCAL VOC and COCO data sets. For the VGG-16 model, our ERPN obtains 78.6% mAP on VOC 2007 data set, 74.4% mAP on VOC 2012 data set and 31.7% on COCO data set. The performance of ERPN is the best among the comparison object detection methods. Furthermore, the detection speed of ERPN is 5.8 fps. Additionally, ERPN obtains good effect on small object detection.


Species-specific activation of TLR4 by hypoacylated endotoxins governed by residues 82 and 122 of MD-2.
The Toll-like receptor 4/MD-2 receptor complex recognizes endotoxin, a Gram-negative bacterial cell envelope component. Recognition of the most potent hexaacylated form of endotoxin is mediated by the sixth acyl chain that protrudes from the MD-2 hydrophobic pocket and bridges TLR4/MD-2 to the neighboring TLR4 ectodomain, driving receptor dimerization via hydrophobic interactions. In hypoacylated endotoxins all acyl chains could be accommodated within the binding pocket of the human hMD-2. Nevertheless, tetra- and pentaacylated endotoxins activate the TLR4/MD-2 receptor of several species. We observed that amino acid residues 82 and 122, located at the entrance to the endotoxin binding site of MD-2, have major influence on the species-specific endotoxin recognition. We show that substitution of hMD-2 residue V82 with an amino acid residue with a bulkier hydrophobic side chain enables activation of TLR4/MD-2 by pentaacylated and tetraacylated endotoxins. Interaction of the lipid A phosphate group with the amino acid residue 122 of MD-2 facilitates the appropriate positioning of the hypoacylated endotoxin. Moreover, mouse TLR4 contributes to the agonistic effect of pentaacylated msbB endotoxin. We propose a molecular model that explains how the molecular differences between the murine or equine MD-2, which both have sufficiently large hydrophobic pockets to accommodate all five or four acyl chains, influence the positioning of endotoxin so that one of the acyl chains remains outside the pocket and enables hydrophobic interactions with TLR4, leading to receptor activation.


Influence of a hybrid form of cardiac rehabilitation on exercise tolerance in coronary artery disease patients with and without diabetes.
The effectiveness of stationary and ambulatory cardiac rehabilitation of patients with coronary artery disease (CAD) and diabetes has been proven by some authors, but data concerning the effects of hybrid forms of cardiac rehabilitation (HCR) in this population are lacking. A home-based telerehabilitation is a promising form of secondary prevention of cardiovascular diseases in this group of patients. The objective of the study was to compare the effects of HCR in CAD patients with and without diabetes mellitus (DM). The secondary endpoint was the assessment of CAD risk factors like low exercise capacity and obesity, in both groups of patients. This was a retrospective study, which comprised 125 patients with CAD aged 57.31 ± 5.61 years referred for HCR. They were assigned to Group D (with diabetes; n = 37) or Group C (without diabetes; n = 88). HCR was carried out as a comprehensive procedure that included all core components of cardiac rehabilitation according to guidelines. Before and after HCR all patients underwent a symptom-limited exercise test performed according to the Bruce protocol on a treadmill. Before HCR the maximal workload was higher in Group C than in Group D (8.13 ± 2.82 METs vs. 6.77 ± 1.88 METs; p = 0.023), but after HCR the difference was not significant. In both groups an increase in the maximal workload after HCR was observed (Group D: before HCR 6.81 ± 1.91 METs, after HCR 8.30 ± 2.04 METs; p < 0.001; Group C: before HCR 8.31 ± 2.71 METs, after HCR 9.13 ± 2.87 METs; p = 0.001). Resting heart rate, double product, and heart rate recovery 1 (HRR1) declined in both groups. No significant differences in changes in exercise test parameters between both groups' parameters were found. HCR was effective in patients with DM. The adherence was high. Patients with DM had higher rates of obesity and significantly lower exercise tolerance than patients without DM. Patients from both groups gained similar benefit from HCR in terms of physical capacity, resting heart rate, and heart rate recovery.


Expression of class II molecules on intestinal epithelial cells in humans. Differences between normal and inflammatory bowel disease.
Expression of class II antigens on human intestinal epithelial cells was assessed using a sensitive avidinbiotin-peroxidase technique. HLA-DR was present predominantly in the normal small bowel with diminished but evident expression in the colon. HLA-DP staining was less prominent, and HLA-DQ was absent. In inflammatory bowel disease the expression of both HLA-DR and HLA-DP was increased, but that for HLA-DQ remained absent, suggesting an inherent defect in the ability of intestinal epithelial cells to express HLA-DQ. In related experiments, an interferon gamma-treated malignant epithelial cell line T84 also failed to stain for HLA-DQ and HLA-DP despite the presence of HLA-DR. Isolated RNAs for all three subclasses of HLA-D were detectable by slot-blot analysis, suggesting that the lack of HLA-DQ expression relates to posttranscriptional defects in intestinal epithelium. These and other differences with conventional class II antigen-positive accessory cells (macrophages/B cells) may help to explain the unique properties of intestinal epithelial cells as antigen-presenting cells.


Study on the Relationship between Emulsion Properties and Interfacial Rheology of Sugar Beet Pectin Modified by Different Enzymes.
The contribution of rheological properties and viscoelasticity of the interfacial adsorbed layer to the emulsification mechanism of enzymatic modified sugar beet pectin (SBP) was studied. The component content of each enzymatic modified pectin was lower than that of untreated SBP. Protein and ferulic acid decreased from 5.52% and 1.08% to 0.54% and 0.13%, respectively, resulting in a decrease in thermal stability, apparent viscosity, and molecular weight (Mw). The dynamic interfacial rheological properties showed that the interfacial pressure and modulus (E) decreased significantly with the decrease of functional groups (especially proteins), which also led to the bimodal distribution of particle size. These results indicated that the superior emulsification property of SBP is mainly determined by proteins, followed by ferulic acid, and the existence of other functional groups also promotes the emulsification property of SBP.


A Fibrinogen Alpha Fragment Mitigates Chemotherapy-Induced MLL Rearrangements.
Rearrangements in the Mixed Lineage Leukemia breakpoint cluster region (MLLbcr) are frequently involved in therapy-induced leukemia, a severe side effect of anti-cancer therapies. Previous work unraveled Endonuclease G as the critical nuclease causing initial breakage in the MLLbcr in response to different types of chemotherapeutic treatment. To identify peptides protecting against therapy-induced leukemia, we screened a hemofiltrate-derived peptide library by use of an enhanced green fluorescent protein (EGFP)-based chromosomal reporter of MLLbcr rearrangements. Chromatographic purification of one active fraction and subsequent mass spectrometry allowed to isolate a C-terminal 27-mer of fibrinogen α encompassing amino acids 603 to 629. The chemically synthesized peptide, termed Fα27, inhibited MLLbcr rearrangements in immortalized hematopoietic cells following treatment with the cytostatics etoposide or doxorubicin. We also provide evidence for protection of primary human hematopoietic stem and progenitor cells from therapy-induced MLLbcr breakage. Of note, fibrinogen has been described to activate toll-like receptor 4 (TLR4). Dissecting the Fα27 mode-of action revealed association of the peptide with TLR4 in an antagonistic fashion affecting downstream NFκB signaling and pro-inflammatory cytokine production. In conclusion, we identified a hemofiltrate-derived peptide inhibitor of the genome destabilizing events causing secondary leukemia in patients undergoing chemotherapy.


Analysis of fluorescently labeled substance P analogs: binding, imaging and receptor activation.
BACKGROUND: Substance P (SP) is a peptide neurotransmitter found in central and peripheral nerves. SP is involved in the control of smooth muscle, inflammation and nociception. The amino acid sequence of SP is Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2. Five different forms of fluorescently labeled SP have recently been synthesized, in which Alexa 488, BODIPY Fl, fluorescein, Oregon Green 488 or tetramethylrhodamine has been covalently linked to SP at Lys3. Here, these novel analogs are characterized as to their ligand binding, receptor activation and fluorescence labeling properties. RESULTS: Competition binding studies, using radiolabeled [125I] SP, revealed that all of the labeled forms of SP, except for Alexa 488-SP, effectively competed with radiolabeled SP for binding at the rat SP receptor. With the exception of Alexa 488-SP, all of the SP analogs produced Ca++ elevations and fluorescence labeling of the SP receptor expressed in Chinese hamster ovary cells. In SP-responsive neurons, BODIPY Fl-SP and Oregon Green 488-SP were as effective as unlabeled SP in producing a reduction of the M-type K+ current. Fluorescein-SP produced variable results, while tetramethylrhodamine-SP was less potent and Alexa 488-SP was less effective on intact neurons. CONCLUSIONS: The above results show that fluorescent labeling of SP altered the biological activity and the binding properties of the parent peptide. Oregon Green 488 and BODIPY FL-SP are the most useful fluorophores for labeling SP without affecting its biological activity. Given these results, these probes can now be utilized in further investigations of the mechanisms of SPR function, including receptor localization, internalization and recycling.


Maintenance of genetic integrity in frozen and freeze-dried mouse spermatozoa.
Chromosome stability was maintained in mouse spermatozoa after freeze-drying or freezing without cryoprotection in a simple Tris.HCl buffer containing EGTA (50 mM) and NaCl (50 mM). The ability of spermatozoa to activate oocytes spontaneously was not destroyed by freeze-drying or freezing without cryoprotection in this solution. Embryos derived after injecting oocytes with sperm heads from rehydrated freeze-dried and from thawed spermatozoa developed normally. Provided the DNA integrity of the sperm nucleus is maintained, embryos can be generated by the intracytoplasmic sperm injection technique (ICSI) from severely damaged spermatozoa that are no longer capable of normal physiological activity. This procedure was effective for preserving spermatozoa from strains (C57BL/6J, 129/SvJ, and BALB/c) in which the fertility of spermatozoa frozen conventionally is extremely poor. The technique provides an effective means of storing mouse spermatozoa from many different inbred, mutant, and transgenic strains for biomedical research.


Evidence-based care for depression in Maine: dissemination of the Kaiser Permanente Nurse Telecare Program.
This paper describes the program model, implementation and preliminary results from a dissemination of a nurse case management program for treating depression in primary care. The program design was modeled after the Kaiser Permanente Nurse TeleCare program, which in a randomized clinical trial had previously demonstrated significant improvement in depression outcomes and patient satisfaction over usual care. As illustrated in this pilot by patient outcomes measured using the Hamilton Depression Rating Scale, the SF-12 Mental Health Composite Score, and the Work Role, Household and Leisure Time Functioning, the authors believe that it is possible to implement successful interventions in smaller primary care practices in community-based settings.


Transition to independent accommodation for adults with schizophrenia.
The aim of this exploratory study was to investigate the process of transition to independent accommodation for people with schizophrenia. The study employed a grounded theory approach. Multiple semi-structured and responsive e-mails and interviews were conducted with a total of seven participants currently living in independent accommodation. Through constant-comparative analysis of interview and e-mail data, three central processes were identified. The three processes are developing a sense of control; establishing a relationship between illness and place; and attaining a sense of belonging. By attending to each of these three processes, mental health professionals may better assist people with schizophrenia to make a successful transition to independent accommodation, and in turn, to living well in the community.


Exogenous Application of Zinc to Mitigate the Salt Stress in Vigna radiata (L.) Wilczek-Evaluation of Physiological and Biochemical Processes.
Salt stress adversely affects the growth and productivity of crops. However, reports suggest that the application of various micronutrients could help the plant to cope with this stress. Hence, the objective of the study was to examine the effect of exogenous application of Zinc (Zn) on salt tolerance in Vigna radiata (L.) Wilczek (mungbean). Mungbean is considered to be an economically important crop and possess a strategic position in Southeast Asian countries for sustainable crop production. It is rich in quality proteins, minerals and vitamins. Three weeks old grown seedlings were subjected to NaCl (150 mM and 200 mM) alone or with Zn (250 µM). After 21 days of treatment, plants were harvested for investigating morphological, physiological and biochemical changes. We found that the Zn application mitigates the negative effect upon plant growth to a variable extent. This may be attributed to the increased shoot and root length, improved chlorophyll and carotenoid contents, enhanced total soluble sugar (TSS), total soluble protein (TSP) and proline accumulation, decreased H2O2 content and increased enzymatic antioxidant activities. Zn's application improved the performance of the enzymes such as phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) of the secondary metabolism, which resulted in the improvement of total phenol and flavonoids. The antioxidant activities such as 1,1diphenyl 2-picryl hydrazine (DPPH) and ferrous reducing antioxidant power assay (FRAP) of the plants also showed improved results in their salt only treatments. Furthermore, hydrogen peroxide (H2O2) and superoxide radical (SOD) scavenging activity were also improved upon the application of 250 µM zinc. Thus, Zn application in low doses offers promising potential for recovering plants suffering from salinity stress. In conclusion, we assume that zinc application improved salt tolerance in mungbean through the improvement of various physiological and photochemical processes which could prove to be useful in nutrient mediated management for crop improvement.


Iron stress in open-ocean cyanobacteria (Synechococcus, Trichodesmium, and Crocosphaera spp.): identification of the IdiA protein.
Cyanobacteria are prominent constituents of the marine biosphere that account for a significant percentage of oceanic primary productivity. In an effort to resolve how open-ocean cyanobacteria persist in regions where the Fe concentration is thought to be limiting their productivity, we performed a number of Fe stress experiments on axenic cultures of marine Synechococcus spp., Crocosphaera sp., and Trichodesmium sp. Through this work, we determined that all of these marine cyanobacteria mount adaptive responses to Fe stress, which resulted in the induction and/or repression of several proteins. We have identified one of the Fe stress-induced proteins as an IdiA homologue. Genomic observations and laboratory data presented herein from open-ocean Synechococcus spp. are consistent with IdiA having a role in cellular Fe scavenging. Our data indicate that IdiA may make an excellent marker for Fe stress in open-ocean cyanobacterial field populations. By determining how these microorganisms respond to Fe stress, we will gain insight into how and when this important trace element can limit their growth in situ. This knowledge will greatly increase our understanding of how marine Fe cycling impacts oceanic processes, such as carbon and nitrogen fixation.


Multifunctional Flame-Retardant, Thermal Insulation, and Antimicrobial Wood-Based Composites.
Wood has been used in a variety of applications in our daily lives and military industry. Nevertheless, its flammability causes potential fire risks and hazards. Improving the flame retardancy of wood is a challenging task. Herein, a phytic acid-based flame retardant (referred to as AMPA) was synthesized based on supramolecular reactions between melamine and p-amino-benzene sulfonic acid followed by a reaction with phytic acid using deionized water as the solvent. A composite wood was prepared by removing lignin to tailor the unique mesoporous structure of the material, followed by coating AMPA on the surfaces of wood microchannels. The limiting oxygen index of wood has been improved to 52.5% with the addition of 5.6 wt % AMPA. The peak heat release rate for the prepared composite wood was reduced by 81% compared to that for delignified wood, which demonstrates the excellent flame-retardant performance of the prepared composite wood. Furthermore, AMPA and mesoporous structures endow antimicrobial and thermal insulation functions. Hence, this work provides a feasible method for preparing flame-retardant wood-based materials for diversified applications.


[A case of tuberculosis with multiple lung nodules, abdominal lymphadenopathy, and splenomegaly].
Abdominal tuberculous lymphadenitis is very rare. We report a case of pulmonary tuberculosis showing marked abdominal lymphadenopathy and splenomegaly. A 95-year-old man was admitted to our hospital because of abnormal chest X-ray and body weight loss in last 6 months. He had low grade fever with no abdominal pain. He did not have past history of tuberculosis. Laboratory examination showed mild renal dysfunction and mild glucose intolerance. Soluble interleukin 2 recepter was highly elevated (3800 U/ml). Tumor markers, such as carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA), and progastrin-releasing peptide (Pro GRP) were all within normal limit. Chest X-ray showed multiple nodules in bilateral lung fields. Chest computed tomography showed multiple nodules in bilateral lungs, especially in upper part of lungs, right hilar lymphadenopathy and upper mediastinal lymphadenopathy. Abdominal and pelvic enhanced computed tomography showed marked abdominal lymphadenopathy and splenomegaly (67 x 49 mm). Abdominal lymph nodes were hepatoduodenal (50 x 50 mm), splenic hilar (40 x 25 mm), upper paraaortic (30 x 60 mm), and small superior mesenteric (10 x 10 mm) lymph nodes. FDG-PET showed accumulation in the nodules of right lung field, right hilar lymph nodes, upper mediastinal lymph nodes, and abdominal lymph nodes. Bronchial lavage fluid (BAL) smear for acid-fast bacilli was positive, polymerase chain reaction for Mycobacterium tuberculosis was positive and acid-fast bacilli was cultured. Transbronchial lung biopsy specimen demonstrated non-specific intraalveolar organization and alveolitis. The patient was diagnosed as pulmonary tuberculosis, but about abdominal lymphadenopathy and splenomegaly we had to differentiate malignant lymphoma, and for definite diagnosis, laparotomy was necessary. But considering his age and general condition, we followed up carefully with anti-tuberculosis therapy. Pulmonary tuberculosis, abdominal lymphadenopathy and splenomegaly all showed marked improvement 4 months after starting anti-tuberculosis therapy with isoniazid, rifampicin, and ethambutol, so we clinically diagnosed abdominal tuberculous lymphadenitis and splenic tuberculosis.


Immunochemiluminometric assay for measurement of thyrotropin in various thyroid disorders.
The measurement of serum TSH concentration by the new ICMA had good precision and was more efficacious than RIA, IRMA and IEMA in distinguishing hyperthyroid patients from euthyroid subjects. Moreover, the measurement of serum TSH by ICMA was less interfered by high concentrations of hCG in sera of patients with trophoblastic disease. However, the assay needs special equipment and the cost is currently higher than other available methods.


Polarization singularities hidden in a deep subwavelength confined electromagnetic field with angular momentum.
Topologies associated with polarization point and line singularities can provide tools for controlling light propagation. By using the Stokes parameter, we demonstrate the emergence of polarization singularities hidden in deep subwavelength confined electromagnetic fields with angular momentum. We show that when the incoming orbital angular momentum is appropriately chosen, highly confined electromagnetic fields with super-diffraction-limited spatial dimensions can be obtained. At the same time, a conversion of orbital to spin angular momentum occurs, leading to a non-trivial topology. Our method provides a platform for developing topological photonics and studying the behavior of polarization singularities under strong focusing.


P-wave duration in lead aVR and the risk of atrial fibrillation in hypertension.
Hypertension entails atrial remodeling that affect P-wave (PW) duration on electrocardiogram (ECG). PW indices (e.g., variance, dispersion, and terminal force) are associated with a higher risk for atrial fibrillation (AF), but their calculation requires multiple measurements of PW duration, limiting their use in clinical practice. We evaluated whether PW duration in specific ECG leads may identify patients with increased susceptibility to AF in a population of hypertensive patients. In a case-control study, AF and control subjects were matched for age, sex, and left atrial (LA) dimensions. PW duration was measured from digitally stored ECGs. Logistic regression was used to assess the association of PW duration and indices with AF. We enrolled 44 hypertensive AF patients (16 paroxysmal and 28 persistent) and 44 hypertensive controls. AF and control subjects were matched for sex (males, n = 27), age (67 ± 8 years), LA diameter (40 ± 5 mm), and were comparable for left ventricular mass (45 ± 11 g/m(2.7) vs 48 ± 12 g/m(2.7) , P = 0.19), ejection fraction (58 ± 7% in both groups), and prevalence of mild valvular heart disease (7% vs 5%; P = 0.64). PW duration in lead aVR was significantly higher in AF patients as compared with controls (115 ± 18 ms vs 101 ± 14 ms; P < 0.0001) and was the best independent predictor of AF in multivariable logistic regression (PW ≥ 100 ms: RR = 3.7; 95% CI: 1.3-10.3; P = 0.02). Simple measurement of PW duration in lead aVR allows effective identification of AF patients in a population of hypertensives. Confirmation of this finding in a larger population would provide a simple and effective risk marker of AF in hypertensive patients.


Synthesis of 2'-modified oligodeoxynucleotides via on-column conjugation.
Oligodeoxynucleotides modified at the 2'-position of 2'-amino-2'-deoxyuridine or uridine were prepared in high yield and purity using phosphoramidites 2 and 3, respectively. Oligodeoxynucleotide conjugates were prepared on the solid-phase synthesis support following selective unmasking of the nucleophile incorporated in these phosphoramidites. Synthesis of oligodeoxynucleotides modified at the 2'-position of an internal nucleotide provides molecules that are complementary to those previously prepared via a similar approach using C5-substituted pyrimidines. The efficiency of functionalization of the 2'-O-alkylamino-uridine derived from 3 in a protected oligodeoxynucleotide was less susceptible to steric hindrance than the 2'-amino-2'-deoxyuridine in the same polymeric substrate. However, the greater reactivity of the 2'-O-alkylamine containing nucleotide gave rise to undesired acetamide formation resulting from nucleophilic attack on the 5'-terminal acetate in capped failure sequences. This problem was overcome by using 2,2,2-trimethylacetyl anhydride as a capping agent during the automated synthesis cycles. Finally, the efficiency of the photochemical unmasking of the support bound alkylamine on a 1 mumole scale was improved by using two 20 min photolysis cycles, coupled with removing reaction byproducts between cycles.


Age exerts a continuous effect in the outcomes of Asian breast cancer patients treated with breast-conserving therapy.
Asians are diagnosed with breast cancer at a younger age than Caucasians are. We studied the effect of age on locoregional recurrence and the survival of Asian breast cancer patients treated with breast-conserving therapy. Medical records of 2492 patients treated with breast-conserving therapy between 1989 and 2012 were reviewed. The Kaplan-Meier method was used to estimate locoregional recurrence, breast cancer-free survival, and breast cancer-specific survival rates. These rates were then compared using log-rank tests. Outcomes and age were modeled by Cox proportional hazards. Fractional polynomials were then used to test for non-linear relationships between age and outcomes. Patients ≤ 40 years old were more likely to have locoregional recurrence than were older patients (Hazard ratio [HR] = 2.32, P < 0.001). Locoregional recurrence rates decreased year-on-year by 4% for patients with luminal-type breast cancers, compared with 8% for those with triple-negative cancers. Similarly, breast cancer-free survival rates increased year-on-year by 4% versus 8% for luminal-type and triple-negative cancers, respectively. Breast cancer-specific survival rates increased with age by 5% year-on-year. Both breast cancer-free survival and breast cancer-specific survival rates in patients with luminal cancers exhibited a non-linear ("L-shaped") relationship-where decreasing age at presentation was associated with escalating risks of relapse and death. The influence of age on overall survival was confounded by competing non-cancer deaths in older women, resulting in a "U-shaped" relationship. Young Asian breast cancer patients have a continuous year-on-year increase in rates of disease relapse and cancer deaths compared with older patients with no apparent threshold.


[Interpretation for the group standards in technical specification of long-term follow-up for end point in large population-based cohort study].
Long-term follow-up for end point is an extremely important and arduous task in large population-based cohort studies, which is also the key to the success of large cohort studies. Thus, the fundamental question of the achievements above is how to construct a large population- based cohort in a standardized way. The Chinese Preventive Medicine Association coordinated experienced researchers from Zhejiang Provincial Center for Disease Control and Prevention and other professional institutes to write up the group standard entitled Technical specification of long-term follow-up for end point in large population-based cohort study (T/CPMA 002-2019). The standard is drafted with principles of emphasizing the scientific, normative, applicability, and feasible nature. This group standard recommended the follow-up target population, time, content, methods, quality control, and indicators assessment. The standard aims to guide the large population-based cohorts that have been or intended to be established in China, including national cohorts, regional population cohorts, and special population cohorts, hence, to improve domestic scientific research level and the international influence, and to support decision-making and practice of disease prevention and control.


COVID-19 Pandemic and Remote Education Contributes to Improved Nutritional Behaviors and Increased Screen Time in a Polish Population-Based Sample of Primary School Adolescents: Diet and Activity of Youth during COVID-19 (DAY-19) Study.
The Coronavirus-19 disease (COVID-19) pandemic has influenced the nutrition of individuals, including the diet followed, food availability, and food security. However, thus far, only a few studies have been published regarding the diet and activity of children and adolescents. The aim of the present study was to analyze the influence of the COVID-19 pandemic and remote education in this period on the diet and physical activity in a Polish population-based sample of primary school adolescents. In June 2020, the Diet and Activity of Youth during COVID-19 (DAY-19) Study was conducted on a population recruited based on stratified random sampling from all regions (schools sampled from counties, and counties from voivodeships). The sample consisted of a total of 1334 adolescents aged 10-16 years. The study assessed the diet and physical activity of the participants using a validated questionnaire which included questions about the period of remote education and the period before the COVID-19 pandemic. The participants were asked about the following: consumption of fruit, vegetables, soft drinks, water, French fries, and fast food; eating meals in front of the television; and the number of days they are physically active and the number of hours they spend watching television. The obtained data were analyzed by stratifying the respondents by the gender, age, size of the city and total COVID-19 morbidity in the voivodeship. It was observed that, during the pandemic and the resultant remote education, the proportion of respondents who declared the recommended intake of fruits and vegetables had increased compared to that before the pandemic-a higher proportion consumed at least three portions of fruit per day (19.0% before pandemic vs. 27.4% during pandemic; p < 0.0001), as well as three and four or more portions of vegetables per day (11.9% vs. 14.5% and 7.5% vs. 11.1%; p = 0.0004). At the same time, the proportion of respondents consuming at least three cups of water per day had increased (41.1% vs. 47.9%; p = 0.0020), whereas the proportion of respondents who never or rarely eat their meals in front of the television had decreased (35.6% vs. 28.9%; p < 0.0001), and the proportion watching television for more than 2 h a day had increased (78.3% vs. 88.4%; p < 0.0001). Based on the results, it may be concluded that, during the period of remote education due to the COVID-19 pandemic, the dietary behaviors of the studied population of Polish adolescents were more beneficial, which included a higher intake of fruit, vegetables, and water, compared to before the pandemic. In spite of the increasing screen time, including eating in front of the television, there was no reduction in the number of days the respondents were physically active.