Herbivory eliminates fitness costs of mutualism exploiters.
A common empirical observation in mutualistic interactions is the persistence of variation in partner quality and, in particular, the persistence of exploitative phenotypes. For mutualisms between hosts and symbionts, most mutualism theory assumes that exploiters always impose fitness costs on their host. We exposed legume hosts to mutualistic (nitrogen-fixing) and exploitative (non-nitrogen-fixing) symbiotic rhizobia in field conditions, and manipulated the presence or absence of insect herbivory to determine if the costly fitness effects of exploitative rhizobia are context-dependent. Exploitative rhizobia predictably reduced host fitness when herbivores were excluded. However, insects caused greater damage on hosts associating with mutualistic rhizobia, as a consequence of feeding preferences related to leaf nitrogen content, resulting in the elimination of fitness costs imposed on hosts by exploitative rhizobia. Our experiment shows that herbivory is potentially an important factor in influencing the evolutionary dynamic between legumes and rhizobia. Partner choice and host sanctioning are theoretically predicted to stabilize mutualisms by reducing the frequency of exploitative symbionts. We argue that herbivore pressure may actually weaken selection on choice and sanction mechanisms, thus providing one explanation of why host-based discrimination mechanisms may not be completely effective in eliminating nonbeneficial partners.


Downregulation of HMGA1 Mediates Autophagy and Inhibits Migration and Invasion in Bladder Cancer via miRNA-221/TP53INP1/p-ERK Axis.
MicroRNAs (miRNAs) have been implicated in regulating the development and metastasis of human cancers. MiR-221 is reported to be an oncogene in multiple cancers, including bladder cancer (BC). Deregulation of autophagy is associated with multiple human malignant cancers. Whether and how miR-221 regulates autophagy and how miR-221 has been regulated in BC are poorly understood. This study explored the potential functions and mechanisms of miR-221 in the autophagy and tumorigenesis of BC. We showed that the downregulation of miR-221 induces autophagy via increasing TP53INP1 (tumor protein p53 inducible nuclear protein 1) and inhibits migration and invasion of BC cells through suppressing activation of extracellular signal-regulated kinase (ERK). Furthermore, the expression of miR-221 is regulated by high-mobility group AT-hook 1 (HMGA1) which is overexpressed in BC. And both miR-221 and HMGA1 are correlated with poor patient survival in BC. Finally, the downregulation of HMGA1 suppressed the proliferative, migrative, and invasive property of BC by inducing toxic autophagy via miR-221/TP53INP1/p-ERK axis. Collectively, our findings demonstrate that the downregulation of miR-221 and HMGA1 mediates autophagy in BC, and both of them are valuable therapeutic targets for BC.


Endocrine therapy of breast cancer.
Breast cancer remains one of the first leading causes of death in women, and currently endocrine treatment is of major therapeutic value in patients with estrogen-receptor positive tumors. Selective estrogen-receptor modulators (SERMs), such as tamoxifen and raloxifene, aromatase inhibitors, and GnRH agonists are the drugs of choice. Tamoxifen, a partial nonsteroidal estrogen agonist, is a type II competitive inhibitor of estradiol at its receptor, and the prototype of SERMs. Aromatase inhibitors significantly lower serum estradiol concentration in postmenopausal patients, having no detectable effects on adrenocortical steroids formation, while GnRH agonists suppress ovarian function, inducing a menopause-like condition in premenopausal women. Endocrine therapy has generally a relatively low morbidity, leading to a significant reduction of mortality for breast cancer. The aim of chemoprevention is to interfere early with the process of carcinogenesis, reducing the risk of cancer development. As preventive agents, raloxifene and tamoxifene are equivalent, while raloxifene has more potent antiresorptive effects in postmenopausal osteoporosis. Endocrine treatment is usually considered a standard choice for patients with estrogen-receptor positive cancers and non-life-threatening advanced disease, or for older patients unfit for aggressive chemotherapy regimens. Several therapeutic protocols used in patients with breast cancer are associated with bone loss, which may lead to an increased risk of fracture. Bisphosphonates are the drugs of choice to treat such a drug-induced bone disease. The aim of this review is to outline current understanding on endocrine therapy of breast cancer.


[Development of a professional social responsibility questionnaire for physicians].
Professional Social Responsibility is now imperative for the practice of medicine. However, there are no instruments to assess it among physicians. To construct and evaluate the factorial structure and reliability of a questionnaire designed to measure socially responsible behavior in physicians. The Questionnaire on Medical Socially Responsible Behavior, consisting of 34 items, was constructed. It was applied to 284 physicians and medical students. After eliminating respondents who omitted questions, a valid sample of 214 individuals aged 23 to 67 years (51.4% males) was obtained. We assessed the factorial structure, reliability, discriminative ability of the items and correlation between factors. Exploratory factorial analysis, conducted using the principal axis method, identified the presence of three factors and considered 30 items. The reliability of the factors, assessed using Cronbach's alpha, ranged from 0.73 to 0.89. Only one item had a low correlation of 0.3. Correlations between the three factors were direct and high. The developed questionnaire presents a definite factorial structure, with internally consistent and correlated factors and with adequate psychometric properties.


Viro-immunological response of drug-naive HIV-1-infected patients starting a first-line regimen with viraemia >500,000 copies/ml in clinical practice.
Virological success (VS) and immunological reconstitution (IR) of antiretroviral-naive HIV-1-infected patients with pre-therapy viral load (VL) >500,000 copies/ml was assessed after 12 months of treatment according to initial drug-class regimens. An observational multicentre retrospective study was performed. VS was defined as the first VL <50 copies/ml from treatment start. IR was defined as an increase of at least 150 CD4+ T-lymphocytes from treatment start. Survival analysis was used to estimate the probability and predictors of VS and IR by 12 months of therapy. 428 HIV-1-infected patients were analysed. Patients were grouped according to the different first-line drug-classes used: a non-nucleoside reverse transcriptase inhibitor (NNRTI) plus two nucleoside reverse transcriptase inhibitors (NRTIs; NNRTI-group; n=105 [24.5%]); a protease inhibitor (PI) plus two NRTIs (PI-group; n=260 [60.8%]); a four-drug regimen containing a PI-regimen plus an integrase inhibitor (PI+INI-group; n=63 [14.7%]). Patients in the PI-group showed the lowest probability of VS (PI-group: 72.4%; NNRTI-group: 75.5%; PI+INI-group: 81.0%; P<0.0001). By Cox regression, patients in PI+INI and NNRTI-groups showed a higher adjusted hazard ratio (95% CI) of VS compared to those in the PI-group (PI+INI-group: 1.48 [1.08, 2.03]; P=0.014; NNRTI-group: 1.37 [1.06-1.78]; P=0.015). The probability of IR was 76.2%, and was similar among groups. Patients with AIDS showed a lower adjusted hazard ratio (95% CI) of IR compared to non-AIDS presenters (0.70 [0.54, 0.90]; P=0.005). In this multicentre retrospective study, patients with viraemia >500,000 copies/ml who start a first-line regimen containing PI+INI or NNRTI yield a better VS compared to those receiving a PI-based regimen.


Normal laparoscopic anatomy of the bovine abdomen.
Three laparoscopic procedures were performed on each of 6 adult Jersey cows in the first trimester of gestation to describe normal laparoscopic anatomy of the bovine abdomen. Also, a technique for laparoscopy of the cranioventral portion of the abdomen was described. Right paralumbar fossa, left paralumbar fossa, and cranioventral midline laparoscopy were performed 72 hours apart of each cow. Physical examination findings, CBC, serum biochemical analysis, and peritoneal fluid analysis before and 72 hours after the first surgery were used to assess the effects of the procedures on the cows. Exploratory celiotomy was performed 2 weeks after the last laparoscopy. The cows were then reexamined 6 weeks after the last procedure. The t-test for paired data was used for statistical analysis; the level of significance was P < 0.05. Laparoscopy was performed without complication in all cows. Adverse effects of laparoscopy, individually or serially, were not observed. Significant differences were not found between CBC, serum biochemical, and peritoneal fluid variables taken before and 72 hours after surgery.


BRCA1-associated growth arrest is RB-dependent.
BRCA1 is a susceptibility gene for breast and ovarian cancer with growth-inhibitory activity for which the mechanism of action remains unclear. When introduced into cells, BRCA1 inhibits growth of some but not all cell lines. In an attempt to uncover the mechanism of growth suppression by BRCA1, we examined a panel of cell lines for their ability to reduce colony outgrowth in response to BRCA1 overexpression. Of all variables tested, only those cells with wild-type pRb were sensitive to BRCA1-induced growth suppression. In cells with an intact rb gene, inactivation of pRb by HPV E7 abrogates the growth arrest imposed by BRCA1. In accordance with these observations, we found that BRCA1 could not suppress BrdUrd uptake in primary fibroblasts from rb-/- mice and exhibited an intermediate ability to inhibit DNA synthesis in rb+/- as compared with rb+/+ cells. We further found that the BRCA1 protein complexes with the hypophosphorylated form of pRb. This binding is localized to amino acids 304-394 of BRCA1 protein and requires the ABC domain of pRb. In-frame deletion of BRCA1 fragment involved in interaction with pRb completely abolished the growth-suppressive property of BRCA1. Although it has been reported that BRCA1 interacts with p53, we find the p53 status did not affect the ability of BRCA1 to suppress colony formation. Our data suggest that the growth suppressor function of BRCA1 depends, at least in part, on Rb.


Vena cava defect repair using a polytetrafluoroethylene graft after a radical nephrectomy and vena cava resection: A case report.
The gold standard treatment for large renal masses is a radical nephrectomy and the removal of tumor thrombi from the large vessels. Here, we discussed the repair of a vena cava defect using a polytetrafluoroethylene (PTFE) graft after a radical nephrectomy and vena cava resection. A 69-year-old male patient presented to our clinic with right-sided pain and 10 kg of weight loss over the previous 3 months. The computed tomography showed that the right kidney was 23 x 13 cm in size, with a 7 x 6 x 7 cm contrast-enhanced mass at the renal ilum level. The patient underwent a radical nephrectomy, and the vena cava defect was repaired using a PTFE graft. There was also tumor infiltration in the proximal third of the left renal vein. The renal vein defect was also repaired using a PTFE graft, and the end of the graft was sutured to the vena cava graft at a right angle. The histopathological examination showed a Fuhrman grade 4 renal cell carcinoma (RCC) with focal sarcomatoid differentiation areas. The management of patients with RCCs and inferior vena cava (IVC) tumor thrombi should be planned with an experienced team, including a cardiovascular surgeon and liver transplantation team. In these patients, the comorbidities, life expectancy, and imaging methods should be considered for treatment planning in experienced centers. The tumor stage, probability of invasion, and patient's performance status should also be determined using magnetic resonance imaging during the preoperative period. Finally, the needs for a graft or tubular patch, sternotomy, and chemotherapeutic agents after the nephrectomy should be discussed using a multidisciplinary approach.


Molecular evolution of cadherin-related neuronal receptor/protocadherin(alpha) (CNR/Pcdh(alpha)) gene cluster in Mus musculus subspecies.
The mouse cadherin-related neuronal receptor/protocadherin (CNR/Pcdh) gene clusters are located on chromosome 18. We sequenced single-nucleotide polymorphisms (SNPs) of the CNR/Pcdh(alpha)-coding region among 12 wild-derived and four laboratory strains; these included the four major subspecies groups of Mus musculus: domesticus, musculus, castaneus, and bactrianus. We detected 883 coding SNPs (cSNPs) in the CNR/Pcdh(alpha) variable exons and three in the constant exons. Among all the cSNPs, 586 synonymous (silent) and 297 nonsynonymous (amino acid exchanged) substitutions were found; therefore, the K(a)/K(s) ratio (nonsynonymous substitutions per synonymous substitution) was 0.51. The synonymous cSNPs were relatively concentrated in the first and fifth extracellular cadherin domain-encoding regions (ECs) of CNR/Pcdh(alpha). These regions have high nucleotide homology among the CNR/Pcdh(alpha) paralogs, suggesting that gene conversion events in synonymous and homologous regions of the CNR/Pcdh(alpha) cluster are related to the generation of cSNPs. A phylogenetic analysis revealed gene conversion events in the EC1 and EC5 regions. Assuming that the common sequences between rat and mouse are ancestral, the GC content of the third codon position has increased in the EC1 and EC5 regions, although biased substitutions from GC to AT were detected in all the codon positions. In addition, nonsynonymous substitutions were extremely high (11 of 13, K(a)/K(s) ratio 5.5) in the laboratory mouse strains. The artificial environment of laboratory mice may allow positive selection for nonsynonymous amino acid variations in CNR/Pcdh(alpha) during inbreeding. In this study, we analyzed the direction of cSNP generation, and concluded that subspecies-specific nucleotide substitutions and region-restricted gene conversion events may have contributed to the generation of genetic variations in the CNR/Pcdh genes within and between species.


Combinatorial expression of Lef1, Lhx2, Lhx5, Lhx9, Lmo3, Lmo4, and Prox1 helps to identify comparable subdivisions in the developing hippocampal formation of mouse and chicken.
We carried out a study of the expression patterns of seven developmental regulatory genes (Lef1, Lhx2, Lhx9, Lhx5, Lmo3, Lmo4, and Prox1), in combination with topological position, to identify the medial pallial derivatives, define its major subdivisions, and compare them between mouse and chicken. In both species, the medial pallium is defined as a pallial sector adjacent to the cortical hem and roof plate/choroid tela, showing moderate to strong ventricular zone expression of Lef1, Lhx2, and Lhx9, but not Lhx5. Based on this, the hippocampal formation (indusium griseum, dentate gyrus, Ammon's horn fields, and subiculum), the medial entorhinal cortex, and part of the amygdalo-hippocampal transition area of mouse appeared to derive from the medial pallium. In the chicken, based on the same position and gene expression profile, we propose that the hippocampus (including the V-shaped area), the parahippocampal area (including its caudolateral part), the entorhinal cortex, and the amygdalo-hippocampal transition area are medial pallial derivatives. Moreover, the combinatorial expression of Lef1, Prox1, Lmo4, and Lmo3 allowed the identification of dentate gyrus/CA3-like, CA1/subicular-like, and medial entorhinal-like comparable sectors in mouse and chicken, and point to the existence of mostly conserved molecular networks involved in hippocampal complex development. Notably, while the mouse medial entorhinal cortex derives from the medial pallium (similarly to the hippocampal formation, both being involved in spatial navigation and spatial memory), the lateral entorhinal cortex (involved in processing non-spatial, contextual information) appears to derive from a distinct dorsolateral caudal pallial sector.


Enhanced retinal responses in Huntington's disease patients.
Huntington's disease (HD) is a fatal progressive neurodegenerative disease characterized by chorea, cognitive impairment and psychiatric symptoms. Retinal examination of HD patients as well as in HD animal models have shown evidence of retinal dysfunction. However, a detailed retinal study employing clinically available measurement tools has not been reported to date in HD. The goal of this study was to assess retinal responses measured by electroretinogram (ERG) between HD patients and controls and evaluate any correlation between ERG measurements and stage of disease. Eighteen patients and 10 controls with inclusion criteria of ages 18-70 years (average age HD subjects: 52.1 yrs and control subjects: 51.9 yrs) were recruited for the study. Subjects with previous history of retinal or ophthalmologic disease were excluded. Retinal function was examined by full-field ERG in both eyes of each subject. Amplitudes and latencies to increasing flash intensities in both light- and dark-adaptation were measured in all subjects. Statistical analyses employed generalized estimating equations, which account for repeated measures per subject. We analyzed the b-wave amplitudes of ERG response in all flash intensities and with 30 Hz flicker stimulation. We found statistically significant increased amplitudes in HD patients compared to controls at light-adapted (photopic) 24.2 and 60.9 cd.sec/m2 intensities, dark-adapted (scotopic, red flash) 0.22 cd.sec/m2 intensity, and a trend toward significance at light-adapted 30 Hz flicker. Furthermore, we found a significant increase in light-adapted ERG response from female compared to male HD patients, but no significant difference between gender amongst controls. We also noted a positive association between number of CAG repeats and ERG response at the smallest light adapted intensity (3.1 cd.sec/m2). ERG studies revealed significantly altered retinal responses at multiple flash intensities in subjects with an HD expansion allele compared to controls. Significant differences were observed with either light-adapted tests or the dark-adapted red flash which suggests that the enhanced responses in HD patients is specific to the cone photoreceptor pathway.


Precise Control of Two-Dimensional Hexagonal Platelets via Scalable, One-Pot Assembly Pathways Using Block Copolymers with Crystalline Side Chains.
Crystallization-driven self-assembly (CDSA) of block copolymers (BCPs) in selective solvents provides a promising route for direct access to two-dimensional (2D) platelet micelles with excellent uniformity, although significant limitations also exist for this robust approach, such as tedious, multistep procedures, and low yield of assembled materials. Herein, we report a facile strategy for massively preparing 2D, highly symmetric hexagonal platelets with precise control over their dimensions based on BCPs with crystalline side chains. Mechanistic studies unveiled that the formation of hexagonal platelets was subjected to a hierarchical self-assembly process, involving an initial stage of formation of kinetically trapped spheres upon cooling driven by solvophobic interactions, and a second stage of fusion of such spheres to the 2D nuclei to initiate the lateral growth of hexagonal platelets via sequential particle attachments driven by thermodynamically ordered reorganization of the BCP upon aging. Moreover, the size of the developed 2D hexagonal platelets could be finely regulated by altering the copolymer concentration over a broad concentration range, enabling scale-up to a total solids concentration of at least 6% w/w. Our work reveals a new mechanism to create uniform 2D core-shell nanoparticles dictated by crystallization and particle fusion, while it also provides an alternative facile strategy for the design of soft materials with precise control of their dimensions, as well as for the scalability of the derived nanostructures.


Effects of pseudowollastonite (CaSiO3) bioceramic on in vitro activity of human mesenchymal stem cells.
We report the effects of two pseudowollastonite (beta-CaSiO(3)) substrates on the attachment, viability, proliferation and osteogenic differentiation of human mesenchymal stem cells (hMSCs), and provide detailed mechanistic links of surface texture, soluble factors and culture media to cell activities. Cell attachment and viability were lower for psWf (fine-grained, roughness 0.74 microm) than for psWc (coarse-grained, roughness 1.25 microm) surface, and were ascribed to the greater specific area of the finer psWf particles resulting in higher release rate of Si, which is cytotoxic at high levels. Interestingly, proliferation was greater on psWf. Osteogenic differentiation occurred on both surfaces, indicated by calcium phosphate bone nodule formation and by osteocalcin, osteopontin and core-binding factor alpha-1 gene expression. Gene levels were lower on psWf than on psWc at day 8 in growth medium, explained by differences in Ca and/or Si concentrations between the two surfaces. Similar gene expression on both surfaces at day 16 in both growth and osteogenic induction media was attributed to pro-osteogenic effects of Ca and P at specific concentrations and complementary Ca and P levels on the two surfaces. In summary, soluble factors from substrates may be more important for osteogenic differentiation in growth medium than small surface roughness variations within a factor of 2. Optimum concentration ranges exist for individual soluble factors to balance cell toxicity/growth versus osteogenic differentiation, and soluble factors together have complex, cooperative or opposing, effects on a given cell activity.


Coupled transcriptome and proteome analysis of human lymphotropic tumor viruses: insights on the detection and discovery of viral genes.
Kaposi's sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV) are related human tumor viruses that cause primary effusion lymphomas (PEL) and Burkitt's lymphomas (BL), respectively. Viral genes expressed in naturally-infected cancer cells contribute to disease pathogenesis; knowing which viral genes are expressed is critical in understanding how these viruses cause cancer. To evaluate the expression of viral genes, we used high-resolution separation and mass spectrometry coupled with custom tiling arrays to align the viral proteomes and transcriptomes of three PEL and two BL cell lines under latent and lytic culture conditions. The majority of viral genes were efficiently detected at the transcript and/or protein level on manipulating the viral life cycle. Overall the correlation of expressed viral proteins and transcripts was highly complementary in both validating and providing orthogonal data with latent/lytic viral gene expression. Our approach also identified novel viral genes in both KSHV and EBV, and extends viral genome annotation. Several previously uncharacterized genes were validated at both transcript and protein levels. This systems biology approach coupling proteome and transcriptome measurements provides a comprehensive view of viral gene expression that could not have been attained using each methodology independently. Detection of viral proteins in combination with viral transcripts is a potentially powerful method for establishing virus-disease relationships.


Temporal corneal phacoemulsification in patients with filtered glaucoma.
To evaluate the effect of temporal clear corneal phacoemulsification on intraocular pressure (IOP) in eyes that underwent prior trabeculectomy. Retrospective case-control study. Forty consecutive patients who underwent temporal clear corneal phacoemulsification subsequent to trabeculectomy (trabeculectomy-phacoemulsification group) were identified, and 40 control patients who underwent trabeculectomy alone (trabeculectomy group) were matched to the case patients for length of follow-up, age, IOP, number of antiglaucoma medications, number of 5-fluorouracil injections, race, sex, and diagnosis. Intraocular pressure before vs 1 year after phacoemulsification in the trabeculectomy-phacoemulsification group compared with IOP in the trabeculectomy group and survival analysis of IOP control after trabeculectomy in the 2 groups. In the trabeculectomy-phacoemulsification group, IOP 1 year after phacoemulsification was not significantly different from the prephacoemulsification IOP value (P = .65). Kaplan-Meier survival analysis showed that the rates of IOP control 3, 6, and 9 years after trabeculectomy in the trabeculectomy-phacoemulsification group were 80%, 66%, and 44%, respectively; in the trabeculectomy group, these were 79%, 69%, and 55%, respectively. These survival curves were not statistically different (P = .55). Cataract surgery by temporal clear corneal phacoemulsification in eyes with filtering blebs after trabeculectomy does not adversely affect long-term IOP control.


Validation of ELDO approaches for retrospective assessment of cumulative eye lens doses of interventional cardiologists-results from DoReMi project.
The first validation results of the two approaches developed in the ELDO project for retrospective assessment of eye lens doses for interventional cardiologists (ICs) are presented in this paper. The first approach (a) is based on both the readings from the routine whole body dosimeter worn above the lead apron and procedure-dependent conversion coefficients, while the second approach (b) is based on detailed information related to the occupational exposure history of the ICs declared in a questionnaire and eye lens dose records obtained from the relevant literature. The latter approach makes use of various published eye lens doses per procedure as well as the appropriate correction factors which account for the use of radiation protective tools designed to protect the eye lens. To validate both methodologies, comprehensive measurements were performed in several Polish clinics among recruited physicians. Two dosimeters measuring whole body and eye lens doses were worn by every physician for at least two months. The estimated cumulative eye lens doses, calculated from both approaches, were then compared against the measured eye lens dose value for every physician separately. Both approaches results in comparable estimates of eye lens doses and tend to overestimate rather than underestimate the eye lens doses. The measured and estimated doses do not differ, on average, by a factor higher than 2.0 in 85% and 62% of the cases used to validate approach (a) and (b), respectively. In specific cases, however, the estimated doses differ from the measured ones by as much as a factor of 2.7 and 5.1 for method (a) and (b), respectively. As such, the two approaches can be considered accurate when retrospectively estimating the eye lens doses for ICs and will be of great benefit for ongoing epidemiological studies.


Blastomonas aquatica sp. nov., a bacteriochlorophyll-containing bacterium isolated from lake water.
Yellow or orange-to-brown pigmented, ovoid or rod-shaped, Gram-negative staining, aerobic strains PE 4-5(T) and N5-10 m-1 were isolated from brackish water in Lake Peng Co and fresh to brackish water in Lake Namtso on the Tibetan Plateau, China. Bacteriochlorophyll a was produced by the isolates. The predominant cellular fatty acids were C16 : 1, C17 : 1 and C18 : 1 unsaturated fatty acids, C17 : 1ω6c (55.3%), C17 : 1ω8c (13.0%) and C18 : 1ω7c (10.4%) for PE 4-5(T) and C18 : 1ω7c (54.7%) and C16 : 1ω7c (18.0%) for N5-10 m-1. The polar lipid profiles of strains PE 4-5(T) and N5-10 m-1 were composed of diphosphatidylglycerol, phosphatidylcholine (not detected in N5-10 m-1), phosphatidyldimethylethanolamine, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol, sphingoglycolipid and an unknown phospholipid. The predominant respiratory quinone was ubiquinone Q10 and the DNA G+C content was 66.0 mol% for both strains. The16S rRNA gene sequence of strain PE 4-5(T) shared 99.0% similarity with that of N5-10 m-1, and 97.56% similarity with those of Blastomonas natatoria LMG 17322(T) and Blastomonas ursincola DSM 9006(T), respectively. The DNA-DNA hybridization relatedness between strains PE 4-5(T) and N5-10 m-1 was 79.0 ± 1.0%, but below 70% with the type strains in the genus Blastomonas . Based on the variability of phylogenetic and phenotypic characteristics, the isolates should be classified as representatives of a novel species of the genus Blastomonas; the name Blastomonas aquatica sp. nov. is proposed. The type strain is PE 4-5(T) ( =JCM 30179(T) =CGMCC 1.12851(T)).


Imaging large arrays of supported lipid bilayers with a macroscope.
Herein, the authors present fluorescence resonance energy transfer (FRET) and two-dimensional protein saturation data acquired from spatially addressed arrays of solid supported lipid bilayers (SLBs). The SLB arrays were imaged with an epifluorescencetotal internal reflection macroscope. The macroscope allowed 1x imaging and had a relatively high numerical aperture (0.4). Such powerful light gathering and large field of view capabilities make it possible to simultaneously image dozens of addressed SLBs. Three experiments have been performed. First, a 9x7 array of supported lipid bilayer was fabricated and imaged in which each bilayer element was individually addressed. Second, a FRET assay was developed between Texas Red-DHPE (1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine) and NBD-PE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-n-(7-nitro-2-1,3-benzoxadiazol-4-yl)). The concentration of dye could be varied at each address and the value of the Forster radius (7.3+/-0.6 nm) was easily abstracted. Third, a ligandreceptor recognition assay was designed to show the two-dimensional number density of proteins which can be bound at saturation. It was found for the streptavidinbiotin pair that the protein saturated at the interface above 3 mol % biotin concentration. This corresponded to a two-dimensional footprint of 40 nm(2) for the streptavidin molecule. These results clearly open the door to using individually addressed bilayers for obtaining large amounts of biophysical data at the supported bilayeraqueous interface. Such abilities will be crucial to obtaining sufficient data for determining the interfacial mechanisms for a variety of membraneprotein interactions.


Site-directed mutagenesis of a thermostable alpha-amylase from Bacillus stearothermophilus: putative role of three conserved residues.
The relationship between structure, activity, and stability of the thermostable Bacillus stearothermophilus alpha-amylase was studied by site-directed mutagenesis of the three most conserved residues. Mutation of His-238 to Asp involved in Ca2+ and substrate binding reduced the specific activity and thermal stability, but did not affect the pH and temperature optima. Replacement of Asp-331 by Glu in the active site caused almost total inactivation. Interestingly, in prolonged incubation this mutant enzyme showed an altered end-product profile by liberating only maltose and maltotriose. Conservative mutation of the conserved Arg-232 by Lys, for which no function has yet been proposed, resulted in lowered specific activity: around 12% of the parental enzyme. This mutant enzyme had a wider pH range but about the same temperature optimum and thermal stability as the wild-type enzyme. Results obtained with different mutants were interpreted by computer aided molecular modeling.


Somatic mutation and functional polymorphism of a novel regulatory element in the HGF gene promoter causes its aberrant expression in human breast cancer.
The HGF gene is transcriptionally silenced in normal differentiated breast epithelial cells, but its repression fails to occur in mammary carcinoma tissues and cell lines. The molecular mechanisms underpinning aberrant HGF expression in breast cancer cells are unknown. Here we report the discovery of a DNA element located 750 bp upstream from the transcription start site in the human HGF promoter that acts as a transcriptional repressor and is a target of deletion mutagenesis in human breast cancer cells and tissues. This HGF promoter element consists of a mononucleotide repeat of 30 deoxyadenosines (30As), which we have termed "deoxyadenosine tract element" (DATE). Functional studies revealed that truncation mutations within DATE have profound local and global effects on the HGF promoter region by modulating chromatin structure and DNA-protein interactions, leading to constitutive activation of the HGF promoter in human breast carcinoma cell lines. We found that 51% of African Americans and 15% of individuals of mixed European descent with breast cancer harbor a truncated DATE variant (25As or fewer) in their breast tumors and that the truncated allele is associated with cancer incidence and aberrant HGF expression. Notably, breast cancer patients with the truncated DATE variant are substantially younger than those with a wild-type genotype. We also suggest that DATE may be used as a potential genetic marker to identify individuals with a higher risk of developing breast cancer.


Electron paramagnetic resonance (EPR) dosimetry using lithium formate in radiotherapy: comparison with thermoluminescence (TL) dosimetry using lithium fluoride rods.
Solid-state radiation dosimetry by electron paramagnetic resonance (EPR) spectroscopy and thermoluminescence (TL) was utilized for the determination of absorbed doses in the range of 0.5-2.5 Gy. The dosimeter materials used were lithium formate and lithium fluoride (TLD-100 rods) for EPR dosimetry and TL dosimetry, respectively. 60Co gamma-rays and 4, 6, 10 and 15 MV x-rays were employed. The main objectives were to compare the variation in dosimeter reading of the respective dosimetry systems and to determine the photon energy dependence of the two dosimeter materials. The EPR dosimeter sensitivity was constant over the dose range in question, while the TL sensitivity increased by more than 5% from 0.5 to 2.5 Gy, thus displaying a supralinear dose response. The average relative standard deviation in the dosimeter reading per dose was 3.0% and 1.2% for the EPR and TL procedures, respectively. For EPR dosimeters, the relative standard deviation declined significantly from 4.3% to 1.1% over the dose range in question. The dose-to-water energy response for the megavoltage x-ray beams relative to 60Co gamma-rays was in the range of 0.990-0.979 and 0.984-0.962 for lithium formate and lithium fluoride, respectively. The results show that EPR dosimetry with lithium formate provides dose estimates with a precision comparable to that of TL dosimetry (using lithium fluoride) for doses above 2 Gy, and that lithium formate is slightly less dependent on megavoltage photon beam energy than lithium fluoride.


A Novel Concept for Partial Liver Transplantation in Nonresectable Colorectal Liver Metastases: The RAPID Concept.
Selected patients with nonresectable colorectal liver metastases benefit from liver transplantation and have acceptable 5-year survival rates. However, allocating full-sized grafts to this group of patients is difficult due to the scarcity of grafts. This could be improved by utilizing small partial grafts, which mandates effective strategies to overcome the problems regarding insufficient functional liver mass. We have developed a protocol incorporating previously reported experiences from living donor transplantation and recent developments in liver surgery, facilitating transplantation of very small liver grafts. At the time of transplantation, segments 1 to 3 are resected in the recipient and orthotopically replaced by a segment 2 to 3 allograft. Portal inflow is modulated by redirecting the portal flow to the graft with concomitant focus on keeping the portal vein pressure below 20 mm Hg. A second-stage hepatectomy is performed as soon as the graft has regenerated to a sufficient volume. A graft weighing 330 g was transplanted to a 50-year-old man weighing 92 kg, and the portal vein to the right remnant liver was closed. The volume of the liver graft was doubled 2 weeks after the first procedure, and it increased further after the second procedure, with extended right hepatectomy performed at day 23 after transplantation. There were no signs of liver failure or small-for-size syndrome. The current protocol and ongoing study could represent a possible strategy to increase the availability of liver transplantation to patients with nonresectable liver tumors such as hepatocellular carcinoma and colorectal liver metastases.


Exploring missed opportunities for influenza vaccination and influenza vaccine co-administration patterns among Italian older adults: a retrospective cohort study.
Missed opportunities constitute a main driver of suboptimal seasonal influenza vaccination (SIV) coverage in older adults. Vaccine co-administration is a way to reduce these missed opportunities. In this study, we quantified missed opportunities for SIV, identified some of their socio-structural correlates and documented SIV co-administration patterns. In this registry-based retrospective cohort study, we verified the SIV status of all subjects aged ≥65 years who received at least one dose of coronavirus disease 2019 (COVID-19), pneumococcal or herpes zoster vaccines during the 2022/23 influenza season. The frequency of concomitant same-day administration of SIV with other target vaccines was also assessed. Among 41 112, 5482 and 3432 older adults who received ≥1 dose of COVID-19, pneumococcal and herpes zoster vaccines, missed opportunities for SIV accounted for 23.3%, 5.0% and 13.2%, respectively. Younger, male and foreign-born individuals were generally more prone to missing SIV. The co-administration of SIV with other recommended vaccines was relatively low, being 11.0%, 53.1% and 17.1% in COVID-19, pneumococcal and herpes zoster cohorts, respectively. A sizeable proportion of older adults who received other recommended vaccines during the last influenza season did not receive SIV. This share of missed opportunities, which are subject to some social inequalities, may be addressed by increasing vaccine co-administration rates and implementing tailored health promotion interventions.


Multi-fragment site-directed mutagenic overlap extension polymerase chain reaction as a competitive alternative to the enzymatic assembly method.
Methods for introducing multiple site-directed mutations are important experimental tools in molecular biology. Research areas that use these methods include the investigation of various protein modifications in cellular processes, modifying proteins for efficient recombinant expression, and the stabilization of mRNAs to allow for increased protein expression. Introducing multiple site-directed mutations is also an important tool in the field of synthetic biology. There are two main methods used in the assembling of fragments generated by mutagenic primers: enzymatic assembly and overlap extension polymerase chain reaction (OE-PCR). In this article, we present an improved OE-PCR method that can be used for the generation of large DNA fragments (up to 7.4 kb) where at least 13 changes can be introduced using a genomic template. The improved method is faster (due to fewer reaction steps) and more accurate (due to fewer PCR cycles), meaning that it can effectively compete with the enzymatic assembly method. Data presented here show that the site-directed mutations can be introduced anywhere between 50 and 1800 bp from each other. The method is highly reliable and predicted to be applicable to most DNA engineering when the introduction of multiple changes in a DNA sequence is required.


Short- and long-term COX-2 inhibition reverses endothelial dysfunction in patients with hypertension.
Hypertension is associated with endothelial dysfunction that is attributable to oxidative stress and a proinflammatory state. Under these conditions, enhanced expression of cyclooxygenase-2 might lead to increased production of vasoconstrictor prostanoids and reactive oxygen species that reduce the bioavailability of endothelium-derived nitric oxide. To investigate the contribution of cyclooxygenase-2 activity to endothelial dysfunction in human hypertension, we evaluated brachial artery vasodilator function by ultrasound in 29 hypertensive patients before and after treatment with the selective cyclooxygenase-2 inhibitor celecoxib or placebo in a randomized, double-blind study. Brachial artery flow-mediated dilation improved from a baseline of 7.9+/-4.5% to 9.9+/-5.1% (P=0.005) 3 hours after the first dose and to 10.1+/-6.1% (P=0.006) after 1 week of treatment with celecoxib. In contrast, placebo treatment had no significant effect on flow-mediated dilation (8.1+/-4.4%, 8.3+/-3.5%, and 8.0+/-3.2%, respectively). Neither treatment altered nitroglycerin-mediated dilation, extent of reactive hyperemia, or baseline arterial diameter. Celecoxib treatment had no significant effect on the urinary concentrations of F2 isoprostane or thromboxane metabolites. However, urinary concentrations of the prostacyclin metabolite 2,3-dinor-6-ketoprostglandin F1alpha were significantly lower after 1 week of celecoxib treatment. Thus, cyclooxygenase-2 products contribute to endothelial dysfunction in hypertension, and treatment with a cyclooxygenase-2 inhibitor could have a beneficial effect in this setting. However, cyclooxygenase-2 inhibition also has an adverse effect on prostacyclin production that could promote thrombosis, and the net clinical consequences of improved endothelial function versus loss of prostacyclin merits further investigation.


Nurse manager turnover in New South Wales during the 1990s.
The principal aim of the research reported in this paper was to explore the nature and incidence of various environmental, personal, and work-related factors that are implicated in turnover and retention amongst nurse managers. The study sought to ascertain whether or not turnover patterns during the 1990s were similar for nurse managers working in different specialist areas and settings of health services within New South Wales, and to discover the main reasons why nurse managers change their jobs or stay in their jobs. This paper reports the main findings, and highlights how the factors implicated in nurse manager turnover and retention, whilst not significantly dissimilar across different cohorts of nurse managers, do not parallel those that are implicated in turnover amongst nurses generally. The research has found that the main reason nurse managers change their jobs is for career development, often within the same organisation or, otherwise, the same health sector or geographic area. Second to this is leaving due to dissatisfying aspects of the job and/or work environment. Some differences have been observed in the propensity to change jobs and in the relative importance of particular influences on job changes in certain practice areas and health sectors. Seven per cent of nurse managers would like to quit their present jobs. However, almost 70% are happy to stay either because the job suits their skills and qualifications, the job is personally satisfying/rewarding, they enjoy the company of their coworkers, or their hours of work suit their needs.


The Role of Artificial Intelligence in Surgery: What do General Surgery Residents Think?
Artificial intelligence (AI) holds significant potential in medical education and patient care, but its rapid emergence presents ethical and practical challenges. This study explored the perspectives of surgical residents on AI's role in medicine. We performed a cross-sectional study surveying general surgery residents at a university-affiliated teaching hospital about their views on AI in medicine and surgical training. The survey covered demographics, residents' understanding of AI, its integration into medical practice, and use of AI tools like ChatGPT. The survey design was inspired by a recent national survey and underwent pretesting before deployment. Of the 31 participants surveyed, 24% identified diagnostics as AI's top application, 12% favored its use in identifying anatomical structures in surgeries, and 20% endorsed AI integration into EMRs for predictive models. Attitudes toward AI varied based on its intended application: 77.41% expressed concern about AI making life decisions and 70.97% felt excited about its application for repetitive tasks. A significant 67.74% believed AI could enhance the understanding of medical knowledge. Perception of AI integration varied with AI familiarity (P = .01), with more knowledgeable respondents expressing more positivity. Moreover, familiarity influenced the perceived academic use of ChatGPT (P = .039) and attitudes toward AI in operating rooms (P = .032). Conclusion: This study provides insights into surgery residents' perceptions of AI in medical practice and training. These findings can inform future research, shape policy decisions, and guide AI development, promoting a harmonious collaboration between AI and surgeons to improve both training and patient care.


Two-year recall for people with no diabetic retinopathy: a multi-ethnic population-based retrospective cohort study using real-world data to quantify the effect.
The English Diabetic Eye Screening Programme (DESP) offers people living with diabetes (PLD) annual screening. Less frequent screening has been advocated among PLD without diabetic retinopathy (DR), but evidence for each ethnic group is limited. We examined the potential effect of biennial versus annual screening on the detection of sight-threatening diabetic retinopathy (STDR) and proliferative diabetic retinopathy (PDR) among PLD without DR from a large urban multi-ethnic English DESP. PLD in North-East London DESP (January 2012 to December 2021) with no DR on two prior consecutive screening visits with up to 8 years of follow-up were examined. Annual STDR and PDR incidence rates, overall and by ethnicity, were quantified. Delays in identification of STDR and PDR events had 2-year screening intervals been used were determined. Among 82 782 PLD (37% white, 36% South Asian, and 16% black people), there were 1788 incident STDR cases over mean (SD) 4.3 (2.4) years (STDR rate 0.51, 95% CI 0.47 to 0.55 per 100-person-years). STDR incidence rates per 100-person-years by ethnicity were 0.55 (95% CI 0.48 to 0.62) for South Asian, 0.34 (95% CI 0.29 to 0.40) for white, and 0.77 (95% CI 0.65 to 0.90) for black people. Biennial screening would have delayed diagnosis by 1 year for 56.3% (1007/1788) with STDR and 43.6% (45/103) with PDR. Standardised cumulative rates of delayed STDR per 100 000 persons for each ethnic group were 1904 (95% CI 1683 to 2154) for black people, 1276 (95% CI 1153 to 1412) for South Asian people, and 844 (95% CI 745 to 955) for white people. Biennial screening would have delayed detection of some STDR and PDR by 1 year, especially among those of black ethnic origin, leading to healthcare inequalities.


Detecting Childhood Pneumonia Using Handcrafted and Deep Learning Cough Sound Features and Multilayer Perceptron.
Pneumonia is one of the leading causes of morbidity and mortality in children. This is especially true in resource poor regions lacking diagnostic facilities, bringing about the need for rapid diagnostic tests for pneumonia. Cough is a common symptom of acute respiratory diseases, including pneumonia, and the sound of cough can be indicative of the pathological variations caused by respiratory infections. As such, in this paper we study objective cough sound evaluation for differentiating between pneumonia and other acute respiratory diseases. We use a dataset of 491 cough sounds from 173 children diagnosed either as having pneumonia or other acute respiratory diseases. We extract features which describe the temporal, spectral, and cepstral characteristics of the cough sound. These features are combined with feature embeddings from a pretrained deep learning network and used to train a multilayer perceptron for classification. The proposed method achieves a sensitivity and specificity of 84% and 73% respectively in differentiating between pneumonia and other acute respiratory diseases using cough sounds alone.


Helical propensity in an intrinsically disordered protein accelerates ligand binding.
Many intrinsically disordered proteins fold upon binding to other macromolecules. The secondary structure present in the well-ordered complex is often formed transiently in the unbound state. The consequence of such transient structure for the binding process is, however, not clear. The activation domain of the activator for thyroid hormone and retinoid receptors (ACTR) is intrinsically disordered and folds upon binding to the nuclear coactivator binding domain (NCBD) of the CREB binding protein. A number of mutants was designed that selectively perturbs the amount of secondary structure in unbound ACTR without interfering with the intermolecular interactions between ACTR and NCBD. Using NMR spectroscopy and fluorescence-monitored stopped-flow kinetic measurements we show that the secondary structure content in helix 1 of ACTR indeed influences the binding kinetics. The results thus support the notion of preformed secondary structure as an important determinant for molecular recognition in intrinsically disordered proteins.


Anthrapyrazoles, a new class of intercalating agents with high-level, broad spectrum activity against murine tumors.
A series of 5-[(aminoalkyl)amino]-substituted anthra[1,9-cd] pyrazol-6(2H)ones (anthrapyrazoles) were synthesized. These compounds, which differ from the anthracenediones in that an additional pyrazole ring has been fused to the anthracene system in place of one carbonyl group, were evaluated in vivo for their anticancer activity in eight different mouse tumor systems. Compounds were selected for testing primarily on the basis of their high levels of activity P388 leukemia and occasionally for structural considerations. Sixty-seven % of the 21 analogues studied were curative in the National Cancer Institute P388 screen. Many of the compounds tested were highly active against each of the tumors of the National Cancer Institute panel. Thus 82, 73, 45, and 80% of the compounds tested were curative for L1210 leukemia, B16 melanoma, M5076 sarcoma, and the MX-1 mammary xenograft, respectively. Several of the compounds studied were curative against every tumor of the above panel. Because of the high activity of the anthrapyrazole series as a class in the National Cancer Institute tumor panel, additional testing was necessary to allow selection of clinical candidates. Twenty-one anthrapyrazoles were tested against mammary adenocarcinoma 16C, colon adenocarcinoma 11a, and the Ridgway osteogenic sarcoma. Four compounds, PD 113,309 (Cl-937), PD 113,785 (Cl-941), PD 111,815 (Cl-942), and PD 115,593, were judged superior to the rest on the basis of the expanded panel testing. The preclinical data to date suggest that these anthrapyrazoles are similar to doxorubicin in both degree and spectrum of activity. Each of these anthrapyrazoles were significantly more active than were the other synthetic intercalating agents, the anthracenediones mitoxantrone and ametantrone, against the tumors of the expanded panel. On the basis of their high level of broad spectrum activity in preclinical systems, ease of formulation, possible lack of cross-resistance with doxorubicin, and potential lack of cardiotoxicity, Cl-937, Cl-941, and Cl-942 have been selected for further preclinical evaluation and possible clinical development.


Synthesis of delta(E)Phe-containing tripeptide via photoisomerization and its conformation in solution.
A new synthetic route to (E)-beta-phenyl-alpha,beta-dehydroalanine (delta(E)Phe)-containing peptide was presented via photochemical isomerization of the corresponding (Z)-beta-phenyl-alpha,beta-dehydroalanine (delta(Z)Phe)-containing peptide. By applying this method to Boc-Ala-delta(Z)Phe-Val-OMe (Z-I: Boc, t-butoxycarbonyl; OMe, methoxy), Boc-Ala-delta(E)Phe-Val-OMe (E-I) was obtained. The identification of peptide E-I was evidenced by 1H-nmr, 13C-nmr, and uv absorption spectroscopy, elemental analysis, and hydrogenation. The conformation of peptide E-I in CDCl3 was investigated by 1H-nmr spectroscopy (solvent dependence of NH chemical shift and difference nuclear Overhauser effect). Interestingly, peptide E-I differed from peptide Z-I in the hydrogen-bonding mode. Namely, for peptide Z-I, only Val NH participates in intramolecular hydrogen bonding, which leads to a type II beta-turn conformation supported by hydrogen bonding between CO(Boc) and NH(Val). On the other hand, for peptide E-I, two NHs, delta(E)Phe NH and Val NH, participate in intramolecular hydrogen bonding. In both peptides, a remarkable NOE (approximately 11-13%) was observed for Ala C(alpha) H-deltaPhe NH pair. Based on the nmr data and conformational energy calculation, it should be concluded that peptide E-I takes two consecutive gamma-turn conformations supported by hydrogen bonding between CO(Boc) and NH(delta(E)Phe), and between CO(Ala) and NH(Val) as its plausible conformation.


Urinary incontinence, mental health and loneliness among community-dwelling older adults in Ireland.
Urinary incontinence (UI) is associated with worse health among older adults. Little is known however, about its relation with loneliness or the role of mental health in this association. This study examined these factors among older adults in Ireland. Data were analyzed from 6903 community-dwelling adults aged ≥ 50 collected in the first wave of The Irish Longitudinal Study on Ageing (TILDA) in 2009-11. Information was obtained on the self-reported occurrence (yes/no) and severity (frequency/activity limitations) of UI in the past 12 months. Loneliness was measured using the UCLA Loneliness Scale short form. Information was also obtained on depression (CES-D), anxiety (HADS-A) and other sociodemographic variables. Logistic regression analysis was used to examine the association between variables. In a model adjusted for all potential confounders except mental disorders, compared to no UI, any UI was associated with significantly higher odds for loneliness (odds ratio: 1.51). When depression was included in the analysis, the association was attenuated and became non-significant while the inclusion of anxiety had a much smaller effect. Similarly, although frequency of UI and activity limitations due to UI were both significantly associated with loneliness prior to adjustment for mental disorders, neither association remained significant after adjustment for both depression and anxiety. UI is associated with higher odds for loneliness among older community-dwelling adults but this association is largely explained by comorbid mental health problems, in particular, depression.


Appendiceal Intussusception: A Diagnostic Challenge.
Appendiceal intussusception is a rare condition in children characterized by an invagination of the appendix into the cecum to various degrees. The treatment is appendectomy; however since symptoms are not specific, clinical diagnosis is challenging and frequently only intraoperative. We present a series of five patients with appendiceal intussusception and discuss features that may direct the pediatric surgeon to achieve early recognition and provide optimal treatment. The database of a tertiary medical center was retrospectively reviewed for all patients treated for appendiceal intussusception during the period from January 1995 to January 2016. Data collected by chart review included demographics, clinical characteristics, imaging studies, surgical technique, and outcome. The findings were analyzed by descriptive statistics. This series included five patients (two females and three males) with ages ranging between 27 and 42 months (mean: 35.2). Patients presented with intermittent abdominal pain (IAP, all five patients), alternate vomiting (three of five patients), alternate diarrhea (two of five patients), fever (two of five patients), and rectal bleeding (one of five patients). The average length of symptoms was 22.6 days. Eighteen diagnostic studies were performed, including abdominal ultrasound for all patients, barium enema for three patients with secondary ileocolic intussusception, and abdominal computed tomography (CT) for one patient. The average number of studies per patient was 3.6. In surgery, the appendiceal intussusception was found to be complete in four patients, whereas it was partial in the remaining patient. In all patients, appendectomy was performed with resection of a small rim of cecal wall due to marked congestion and edema in an attempt to decrease recurrence. The mainstay of clinical presentation is intermittent abdominal pain while patients may be completely asymptomatic between attacks. Appendiceal intussusception may act as a leading point to ileocolic intussusception and is frequently concealed by it. The treatment is appendectomy. Both pediatric surgeons and radiologists should be aware of this occurrence to provide adequate management and avoid complications.


Functional Outcomes Are Similar After Early and Late Arthroscopic One-Tunnel Transosseous Repair of Triangular Fibrocartilage Complex Foveal Tears.
To compare outcomes at different time periods following arthroscopic triangular fibrocartilage complex (TFCC) transosseous foveal repair within 6 months, between 6 and 12 months, and more than 12 months from injury. Consecutive patients treated with arthroscopic TFCC foveal repair using the uniform one-tunnel transosseous suture technique by a surgeon from 2014 to 2017 were retrospectively reviewed. The patients were assigned to 1 of 3 groups according to time between injury and surgery. Pain visual analog scale (VAS); grip strength; modified Mayo wrist score (MMWS); Quick disabilities of the arm, shoulder, and hand (QuickDASH) score; and distal radioulnar joint stability were assessed at minimum 2 years postoperatively, along with minimal clinically important difference, and overall patient satisfaction. This study cohort consisted of 80 patients: group A (<6 months, n = 38), group B (6-12 months, n = 20), and group C (>12 months, n = 22). No differences were found among groups in VAS, grip strength, and MMWS and QuickDASH. Overall, patients exhibited significant functional improvement at 2 years (VAS: 3-0, P < .001; grip strength: 77.1%-95.6%, P < .001; MMWS: 65-90, P < .001, QuickDASH: 20.5-4.5, P < .001). Median changes in outcome variables and the proportion of patients achieving minimal clinically important difference for the QuickDASH were similar among groups. Seventy-eight patients (97%) achieved distal radioulnar joint stability, and 70 patients (87%) were satisfied with treatment. Although this current study has insufficient statistical power, the available data suggest that patients with a TFCC foveal tear who underwent arthroscopic transosseous repair surgery more than 12 months after injury could expect to experience similar functional improvement compared with patients who underwent surgery within 6 months or between 6 and 12 months following injury. Level III, retrospective comparative study.


Prevalence of Antibiotic Resistome in Ready-to-Eat Salad.
Ready-to-eat salad harbors microorganisms that may carry various antibiotic resistance genes (ARGs). However, few studies have focused on the prevalence of ARGs on salad, thus underestimating the risk of ARGs transferring from salad to consumers. In this small-scale study, high-throughput quantitative PCR was used to explore the presence, prevalence and abundance of ARGs associated with serving salad sourced from two restaurant types, fast-food chain and independent casual dining. A total of 156 unique ARGs and nine mobile genetic elements (MGEs) were detected on the salad items assessed. The abundance of ARGs and MGEs were significantly higher in independent casual dining than fast-food chain restaurants. Absolute copies of ARGs in salad were 1.34 × 107 to 2.71 × 108 and 1.90 × 108 to 4.87 × 108 copies per g salad in fast-food and casual dining restaurants, respectively. Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes were the dominant bacterial phyla detected from salad samples. Pseudomonas, Acinetobacter, Exiguobacterium, Weissella, Enterobacter, Leuconostoc, Pantoea, Serratia, Erwinia, and Ewingella were the 10 most dominant bacterial genera found in salad samples. A significant positive correlation between ARGs and MGEs was detected. These results integrate knowledge about the ARGs in ready-to-eat salad and highlight the potential impact of ARGs transfer to consumers.


The role of protein characteristics in the formation and fluorescence of Au nanoclusters.
Protein-encapsulated gold nanoclusters have shown many advantages over other gold nanocluster systems, including green synthesis, biocompatibility, high water solubility, and the ease of further conjugation. In this article, we systematically investigated the effects of the protein size and amino acid content on the formation and fluorescent properties of gold nanoclusters using four model proteins (bovine serum albumin, lysozyme, trypsin, and pepsin). We discovered that the balance of amine and tyrosine/tryptophan containing residues was critical for the nanocluster formation. Protein templates with low cysteine contents caused blue shifts in the fluorescent emissions and difference in fluorescent lifetimes of the gold nanoclusters. Furthermore, the protein size was found to be a critical factor for the photostability and long-term stability of gold nanoclusters. The size of the protein also affected the Au nanocluster behaviour after immobilization.


[General anaesthesia with sevoflurane in dental surgery with use of LMA-Flexible and LMA-ProSeal].
To compare the laryngeal mask airways (LMA), LMA - Flexible (LMA-F) and LMA-ProSeal (PLMA) with respect to airway management in dental surgery. 35 patients, with the pathology of stomatological area (cysts of the lower jaw; diseases maxillary sinus) were investigated during the operation. Anesthesia was induced with propofol, fentanyl and atracurium besylate. Anesthesia was maintained with sevoflurane and neuromuscular blockade with mechanical ventilation. The operation lasted 30-40 min. For airway management was used LMA-F in 20 patients, in 15 patients was made PLMA. There were no statistically significant differences between LMA-F/PLMA groups for SpO(2), ET CO(2) or airway pressure during anesthesia. As for the gas exchange no difference was noticed in both groups (p<0.05). Undesirable laryngo-phryngeal reflexes were not noticed in total myoplegia. The reaction of cardiovascular system while installating LMA-F and PLMA was absent. As for the gas exchange no difference was noticed in both groups (p<0.05). A correctly placed LMA-F or PLMA during general anaesthesia with sevoflurane is effective for airway management in surgery.


Bowel preparation for pediatric colonoscopy procedures.
Colonoscopy is a routinely performed procedure in children and adolescents. Proper visualization of the intestinal mucosa, completion of the procedure, including examination of the terminal ileum, detection of pathological lesions, and therapeutic maneuvers, all are highly dependent on the quality of the bowel preparation. A significant proportion of patients, in some reports up to one third, are inadequately prepared for the examination, which leads to extended procedure time, incomplete examination, or need for repeat procedure. Aside from efficacy and safety, the most important aspects of colon preparation in pediatrics are ease of administration, palatability, dietary restriction, and minimization of disruption of daily routine. An ideal preparation does not exist and a wide variety of regimens are being used. Several of these have been investigated in pediatric clinical trials. This article reviews the published literature with an emphasis on the most commonly used agents, their mechanism of action, efficacy and ease of use, and safety.


Use of GLP-1 agonists in high risk patients prior to bariatric surgery: a cohort study.
Body mass index (BMI) > 50 kg/m2 is associated with relatively increased morbidity and mortality with bariatric surgery (BS). There is reluctance to consider these patients operative candidates without preoperative weight loss. Glucagon-like peptide-1 (GLP-1) agonists have demonstrated effective weight loss in the post-BS setting. This study aims to determine the safety and efficacy of GLP-1 agonists in the pre-habilitation of patients with BMI > 50 kg/m2. This is a retrospective review of bariatric surgery patients with BMI > 50 kg/m2 from a single bariatric center. Patients were compared by preoperative GLP-1 therapy status. All patients received medical, surgical, psychiatric, and nutritional evaluation and counseling. Preoperative BMI, change in weight from program intake until surgery, time to surgery, and perioperative complications were evaluated. 31 patients were included in the analysis. 18 (58%) received a GLP-1 agonist preoperatively. GLP-1 agonist use was associated with a 5.5 ± 3.2-point reduction in BMI compared to 2.9 ± 2.4 amongst controls (p = 0.026). There was no difference in the mean length of time in the bariatric program prior to surgery between groups (p = 0.332). There were no reported complications related to GLP-1 use in the preoperative setting and no difference in perioperative complications between groups (p = 0.245). GLP-1 agonist use in patients with a BMI > 50 kg/m2 results in significantly more weight loss prior to bariatric surgery, without increased time to surgery or complication rate. Further study is required to evaluate the long-term impact of preoperative GLP-1 agonist use prior to bariatric surgery. This therapy may improve perioperative and long-term outcomes in the very high-risk BMI population.


Diagnosing testicular function using 31P magnetic resonance spectroscopy: a current review.
Patients with low sperm counts combined with normal concentrations of gonadotrophins, and in whom physical examination and post-ejaculatory urine analysis are normal, present a diagnostic dilemma. This situation can be caused by testicular failure or by ductal obstruction, which have very different clinical prognoses. Ductal obstruction might be correctable by microsurgical vasovaso/vasoepididymostomy, whereas this approach is of no use in primary testicular failure. A possible diagnostic step for these patients is a testicular biopsy to differentiate between hypospermatogenesis and a normal gonad. However, to date testicular biopsy is seldom performed because of its invasive character. An alternative accurate, non-invasive method to assess testicular function could be very helpful in the evaluation of idiopathic azoospermia or idiopathic oligozoospermia. During the past decade, magnetic resonance (MR) spectroscopy has been developed from a scientific tool into a non-invasive clinical diagnostic tool and has also been used to study testicular function. Recent studies have shown that 31P-MR spectroscopy, based upon differences in the ratio of peaks of phosphomonoester to beta-adenosinetriphosphate, is a non-invasive technique able to differentiate between groups of patients with testicular failure and ductal obstruction, and it correlates reasonably well with the averaged mean Johnsen score of testicular biopsy. The role for a non-invasive technique in the diagnosis of male infertility, such as 31P-MR spectroscopy, can be manifold. It serves not only as an alternative for biopsy but can also be used to assess obstruction as the cause of infertility in patients with subnormal sperm counts, and to predict the chances of pregnancy in patients planned for vasovasostomy to correct a prior vasectomy. However, the main limitation to MR spectroscopy becoming a universal clinical diagnostic technique is the limited availability of 1.5 Tesla MR scanners.


THRX-198321 is a bifunctional muscarinic receptor antagonist and beta2-adrenoceptor agonist (MABA) that binds in a bimodal and multivalent manner.
Biphenyl-2-yl-carbamic acid 1-{9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)-ethylamino]-nonyl}-piperidin-4-yl ester (THRX-198321) is a single molecule composed of a muscarinic acetylcholine receptor (mAChR) antagonist moiety, represented by the fragment MA, linked by a C9 polymethylene chain to a β(2)-adrenoceptor (β(2)AR) agonist moiety, represented by the fragment 8-hydroxy-5-((R)-1-hydroxy-2-methylamino-ethyl)-1H-quinolin-2-one (BA). THRX-198321 exhibited high affinity for mAChR (M(2) pK(I,App) = 10.57 ± 0.09; M(3) pK(I,App) = 10.07 ± 0.11) and β(2)AR (pK(I,App) = 9.54 ± 0.15), with potent mAChR antagonist (M(2) pK(I,Fn) = 9.69 ± 0.23; M(3) pK(I,Fn) = 10.05 ± 0.17) and β(2)AR agonist (pEC(50) = 9.25 ± 0.02) activities. Consistent with multivalent interactions, THRX-198321 binding affinity was >300-fold higher at mAChR and 29-fold higher at β(2)AR relative to its monovalent fragments biphenyl carbamic acid piperidinyl ester (MA) and BA, respectively. THRX-198321 was a competitive antagonist at mAChR (M(2) pK(B) = 9.98 ± 0.13; M(3) pK(B) = 10.31 ± 0.89), whereas THRX-198321 agonist activity at β(2)AR was competitively inhibited by propranolol. Interactions of THRX-198321 with an allosteric site on mAChR and a novel extracellular allosteric site on β(2)AR, respectively, were detected by measuring THRX-198321-evoked changes in the dissociation rates for the orthosteric radioligands, [N-methyl-(3)H]scopolamine methyl chloride (M(2) pEC(50,diss) = 6.73 ± 0.10; M(3) pEC(50,diss) = 5.02 ± 0.14) and [4,6-propyl-(3)H]dihydroalprenolol (β(2)AR pEC(50,diss) = 3.82 ± 0.25). The carbostyril-linker fragment (BA-L) binds to the allosteric site of mAChR (M(2) pEC(50,diss) = 5.06 ± 0.03; M(3) pEC(50,diss) = 4.15 ± 0.25), whereas the MA fragment binds to the allosteric site of β(2)AR (pEC(50,diss) = 3.60 ± 0.18). Collectively, these observations suggest that THRX-198321 exhibits a multivalent bimodal orientation in the orthosteric and allosteric binding pockets of mAChR and β(2)AR, a phenomenon that may be unique to this class of molecule.


The obesity paradox in elderly obese patients undergoing coronary artery bypass surgery.
We sought to determine whether the protective role of the 'obesity paradox' was present among elderly obese patients undergoing coronary artery bypass grafting (CABG) by median sternotomy. We retrospectively analyzed 1909 consecutive patients who underwent heart surgery between January 2006 and June 2009, and identified 396 patients who were ≥ 70 years of age and had isolated CABG. Subjects were divided into three groups according to their body mass index (BMI): obese (BMI ≥ 30 kg/m(2)), overweight (BMI 25-29.99 kg/m(2)) and normal (BMI 18.5-24.99 kg/m(2)). Of the 396 patients, 94 were obese, 167 were overweight, and 135 had a normal BMI. The composite of in-hospital complications and hospital mortality did not differ between the groups. Re-exploration for bleeding was required in none of the obese patients, which was statistically significant (P=0.05) compared to the other groups; otherwise there was no statistical difference for all other complications, including total length hospital stay and length of stay in the intensive care unit. Despite being labeled as higher risk candidates preoperatively, obese elderly patients undergoing CABG did not demonstrate an increased risk of postsurgical complications. We conclude that the 'obesity paradox' is present in this population, and they should not be excluded from receiving the benefits of CABG.


Silk fibroin nanoparticles constitute a vector for controlled release of resveratrol in an experimental model of inflammatory bowel disease in rats.
We aimed to evaluate the intestinal anti-inflammatory properties of silk fibroin nanoparticles, around 100 nm in size, when loaded with the stilbene compound resveratrol, in an experimental model of rat colitis. Nanoparticles were loaded with resveratrol by adsorption. The biological effects of the resveratrol-loaded nanoparticles were tested both in vitro, in a cell culture of RAW 264.7 cells (mouse macrophages), and in vivo, in the trinitrobenzenesulfonic acid model of rat colitis, when administered intracolonically. The resveratrol liberation in 1× phosphate-buffered saline (PBS; pH 7.4) was characterized by fast liberation, reaching the solubility limit in 3 hours, which was maintained over a period of 80 hours. The in vitro assays revealed immunomodulatory properties exerted by these resveratrol-loaded nanoparticles since they promoted macrophage activity in basal conditions and inhibited this activity when stimulated with lipopolysaccharide. The in vivo experiments showed that after evaluation of the macroscopic symptoms, inflammatory markers, and intestinal barrier function, the fibroin nanoparticles loaded with resveratrol had a better effect than the single treatments, being similar to that produced by the glucocorticoid dexamethasone. Silk fibroin nanoparticles constitute an attractive strategy for the controlled release of resveratrol, showing immunomodulatory properties and intestinal anti-inflammatory effects.


The reliability of MCLs with pure tones and damped wave trains.
Most Comfortable Loudness (MCL) was determined for each of 10 normal-hearing young adults using pulsed pure-tone trains and damped wave trains (DWTs) each at 2.5/sec, at frequencies of .25 and 2 kc/s; 4 MCLs were determined for each of the 4 signal type/frequency combinations. In a bracketing procedure employing steps of 20, 10, and finally 5 db, Ss were permitted to respond only "loud" or "soft" to each short train. Group mean test-retest differences within each stimulus type/frequency combination were only 1-2.5 db for all possible test-to-contiguous-test comparisons, i.e., better than with most other MCL procedures except the descending series in the Method of Limits. However, individual Ss differed by from 15-35 db within the 16 distributions (2 stimulus types X 2 frequencies X 4 replications). No statistical or practical differences were found between pure tones or DWTs, or across frequency. It was suggested that the range of individual differences renders any group mean MCL too imprecise a measure for selecting and prescribing a hearing aid, or for determining the effect that aid may have on an individual's performance.


Assessment of early treatment response on MRI in multiple myeloma: Comparative study of whole-body diffusion-weighted and lumbar spinal MRI.
To compare remission status at completion of chemotherapy for multiple myeloma (MM) with changes in total diffusion volume (tDV) calculated from whole-body diffusion-weighted imaging (WB-DWI) and fat fraction (FF) of lumbar bone marrow (BM) by modified Dixon Quant (mDixon Quant) soon after induction of chemotherapy, and to assess the predictive value of MRI. Fifty patients (mean age, 66.9 ± 10.5 years) with symptomatic myeloma were examined before and after two cycles of chemotherapy. From WB-DWI data, tDV was obtained with the threshold for positive BM involvement. Mean FF was calculated from lumbar BM using the mDixon Quant sequence. At the completion of chemotherapy, patients were categorized into a CR/very good PR (VGPR) group (n = 15; mean age, 67.6 ± 10.3 years) and a PR, SD or PD group (n = 35; mean age, 69.1 ± 8.6 years). ROC curves were plotted to assess performance in predicting achievement of CR/VGPR. At second examination, serum M protein, β2-microglobulin, and tDV were significantly decreased and hemoglobin, mean ADC, and FF were significantly increased in the CR/VGPR group and serum M protein was significantly increased in the PR/SD/PD group. The general linear model demonstrated that percentage changes in FF and M protein contributed significantly to achieving CR/VGPR (P = 0.02, P = 0.04, respectively). AUCs of ROC curves were 0.964 for FF and 0.847 for M protein. Early change in FF of lumbar BM and serum M protein soon after induction of chemotherapy contributed significantly to prediction of CR/VGPR.


Primary pyogenic ventriculitis caused by Neisseria meningitidis: case report and review of the literature.
Background. Pyogenic ventriculitis is a well-known complication of meningitis, brain abscesses and intraventricular drains. Primary pyogenic ventriculitis is a rare entity and few cases have been described so far. We report the first case of primary pyogenic ventriculitis in an adult caused by Neisseria meningitidis and present an overview of all reported adult primary pyogenic ventriculitis cases in the English literature. Methods. A PubMed search was performed using the terms ependymitis, ventricular empyema, pyocephalus and ventriculitis. Filter was set for adults and English. Articles in which pyogenic ventriculitis was a complication of well-known risk factors were excluded. A total of five cases of primary pyogenic ventriculitis were identified. Results. There were seven adult patients. Only one patient showed signs of meningeal irritation. Four patients had positive blood cultures with Escherichia coli (one patient), methicillin-resistant Staphylococcus aureus (one patient), one patient was bacteraemic with Enterococcus faecalis, Escherichia coli and Peptostreptococcus spp., and N. meningitidis (our patient). In four patients cerebrospinal fluid was sent for culture, which yielded methicillin-sensitive Staphylococcus aureus (one patient), Peptostreptococcus spp. (one patient), Streptococcus intermedius (one patient, identified via 16S PCR) and Listeria monocytogenes (one patient). Cerebrospinal fluid cell count was determined in four patients and showed pleocytosis in all four cases. Ventricular drainage was performed in four patients. Five patients survived. Discussion. We report the first case of pyogenic ventriculitis caused by N. meningitidis. Primary pyogenic ventriculitis is a rare entity with various clinical presentations caused by various bacterial species. Treatment consists of adequate antimicrobial therapy, and ventricular drainage may be necessary.


Interparticle Reactions: An Emerging Direction in Nanomaterials Chemistry.
Nanoparticles exhibit a rich variety in terms of structure, composition, and properties. However, reactions between them remain largely unexplored. In this Account, we discuss an emerging aspect of nanomaterials chemistry, namely, interparticle reactions in solution phase, similar to reactions between molecules, involving atomically precise noble metal clusters. A brief historical account of the developments, starting from the bare, gas phase clusters, which led to the synthesis of atomically precise monolayer protected clusters in solution, is presented first. Then a reaction between two thiolate-protected, atomically precise noble metal clusters, [Au25(PET)18]- and [Ag44(FTP)30]4- (PET = 2-phenylethanethiol, FTP = 4-fluorothiophenol), is presented wherein these clusters spontaneously exchange metal atoms, ligands, and metal-ligand fragments between them under ambient conditions. The number of exchanged species could be controlled by varying the initial compositions of the reactant clusters. Next, a reaction of [Au25(PET)18]- with its structural analogue [Ag25(DMBT)18]- (DMBT = 2,4-dimethylbenzenethiol) is presented, which shows that atom-exchange reactions happen with structures conserved. We detected a transient dianionic adduct, [Ag25Au25(DMBT)18(PET)18]2-, formed between the two clusters indicating that this adduct could be a possible intermediate of the reaction. A reaction involving a dithiolate-protected cluster, [Ag29(BDT)12]3- (BDT = 1,3-benzenedithiol), is also presented wherein metal atom exchange alone occurs, but with no ligand and fragment exchanges. These examples demonstrate that the nature of the metal-thiolate interface, that is, its bonding network and dynamics, play crucial roles in dictating the type of exchange processes and overall rates. We also discuss a recently proposed structural model of these clusters, namely, the Borromean ring model, to understand the dynamics of the metal-ligand interfaces and to address the site specificity and selectivity in these reactions. In the subsequent sections, reactions involving atomically precise noble metal clusters and one- and two-dimensional nanosystems are presented. We show that highly protected, stable clusters such as [Au25(PET)18]- undergo chemical transformation on graphenic surfaces to form a bigger cluster, Au135(PET)57. Finally, we present the transformation of tellurium nanowires (Te NWs) to Ag-Te-Ag dumbbell nanostructures through a reaction with an atomically precise silver cluster, Ag32(SG)19 (SG = glutathione thiolate). The starting materials and the products were characterized using high resolution electrospray ionization mass spectrometry, matrix assisted laser desorption ionization mass spectrometry, UV/vis absorption, luminescence spectroscopies, etc. We have analyzed principally mass spectrometric data to understand these reactions. In summary, we present the emergence of a new branch of chemistry involving the reactions of atomically precise cluster systems, which are prototypical nanoparticles. We demonstrate that such interparticle chemistry is not limited to metal clusters; it occurs across zero-, one-, and two-dimensional nanosystems leading to specific transformations. We conclude this Account with a discussion of the limitations in understanding of these reactions and future directions in this area of nanomaterials chemistry.


Hypoxia/Ischemia-Induced Rod Microglia Phenotype in CA1 Hippocampal Slices.
The complexity of microglia phenotypes and their related functions compels the continuous study of microglia in diseases animal models. We demonstrated that oxygen-glucose deprivation (OGD) induced rapid, time- and space-dependent phenotypic microglia modifications in CA1 stratum pyramidalis (SP) and stratum radiatum (SR) of rat organotypic hippocampal slices as well as the degeneration of pyramidal neurons, especially in the outer layer of SP. Twenty-four h following OGD, many rod microglia formed trains of elongated cells spanning from the SR throughout the CA1, reaching the SP outer layer where they acquired a round-shaped amoeboid phagocytic head and phagocytosed most of the pyknotic, damaged neurons. NIR-laser treatment, known to preserve neuronal viability after OGD, prevented rod microglia formation. In CA3 SP, pyramidal neurons were less damaged, no rod microglia were found. Thirty-six h after OGD, neuronal damage was more pronounced in SP outer and inner layers of CA1, rod microglia cells were no longer detectable, and most microglia were amoeboid/phagocytic. Damaged neurons, more numerous 36 h after OGD, were phagocytosed by amoeboid microglia in both inner and outer layers of CA1. In response to OGD, microglia can acquire different morphofunctional phenotypes which depend on the time after the insult and on the subregion where microglia are located.


Culture and the care of children with chronic conditions: their physicians' views.
Little is known about physicians' perceptions of the influence of culture on the health care of children with chronic and disabling conditions. To identify physicians' perceptions of the impact of the family's ethnocultural background on the health care of school-aged children with chronic conditions and recommendations for improving care. Qualitative study in 2 midwestern metropolitan areas. General community. Convenience sample of 52 physicians nominated by 60 African American, Hispanic, and European American families of school-aged children with chronic conditions. In-depth interviews were conducted with the physicians. Content analytic techniques were used to analyze the data. In 44% of the responses, the physicians reported that ethnocultural background did not influence the care the child received, noting that comparable care was provided to all of their patients. In 14% of the responses, the effect was unknown. The overall effect was negative in 26% of the responses and positive in 16%. Physicians' recommendations focused on 4 topics: improving the training and education of health care professionals and families; ensuring good communication between the child, family, and health care professionals; supporting families; and improving the access and provision of services for children from diverse cultural backgrounds. Although the majority of participants reported that ethnocultural background did not affect the care the child received from the health care system, physicians' recommendations reflected awareness of the influence of culture on the care of children with chronic conditions and the need for further training on this issue.


[Immunohistochemical study of hypertrophic pyloric stenosis].
In spite of the clinical experience about infantil Hypertrophic Pyloric Stenosis (IHPS), its etiopathology remains unknown. Recent studies have been focussed in immunohistochemistry techniques for valuing the neuronal development in the pyloric muscle. We took biopsy from 10 babies diagnosed of IHPS and 10 babies with similar age, died because of other causes. Immunohistochemical study was performed using monoclonal antibodies: S100 protein, GFAP, enolasa and neurofilaments NF. The results were showed semi-quantitativement as strong (++), moderate (+) and absent (-). Immunotinction of the myenteric plexus (ganglion cells and satellite) and nervous fibers of the muscle layer, were done. We observed a poor immunoreactivity in the muscle layer (longitudinal and circular) in the 60-70% of specimens of pyloric muscle in babies with IHPS. GFAP were absent in the 80% in the myenteric plexus. This poor innervation may be related to the etiopathogenesis of pyloric stenosis and hypertrophy.


Parenchymal expression of CD40 exacerbates adenovirus-induced hepatitis in mice.
The healthy adult human liver expresses low levels of major histocompatibility complex class II (MHC II) and undetectable levels of immune costimulatory molecules. However, high levels of MHC II, CD40, and B7 family molecules are expressed in the activated Kupffer cells and hepatocytes of patients with viral hepatitis. The precise role of these molecules in viral clearance and immune-mediated liver injury is not well understood. We hypothesized that parenchymal CD40 expression enhances T cell recruitment and effector functions, which may facilitate viral clearance and alleviate liver injury. To test this hypothesis, we generated novel liver-specific, conditional CD40 transgenic mice, and we challenged them intravenously with a recombinant replication-deficient adenovirus carrying Cre recombinase (AdCre). Wild-type mice infected with AdCre developed a relatively mild course of viral hepatitis and recovered spontaneously. CD40 expression in the livers of transgenic animals, however, resulted in CD80 and CD86 expression. The dysregulation of population dynamics and effector functions of intrahepatic lymphocytes (IHLs) resulted in severe lymphocytic infiltration, apoptosis, necroinflammation, and serum alanine aminotransferase elevations in a dose-dependent fashion. To our surprise, an early expansion and subsequent contraction of IHLs (especially CD8(+) and natural killer cells), accompanied by increased granzyme B and interferon-γ production, did not lead to faster viral clearance in CD40 transgenic mice. Our results demonstrate that hepatic CD40 expression does not accelerate adenoviral clearance but rather exacerbates liver injury. This study unveils a previously unknown deleterious effect of hepatic CD40 on adenovirus-induced liver inflammation.


Cumulative effects of therapies on disability in relapsing multiple sclerosis.
Long-term effectiveness of treatment remains a key question in multiple sclerosis (MS) and the cumulative effects of past treatment have not been investigated so far. Explore the relationship between treatment exposure and disability risk in patients with relapsing-remitting multiple sclerosis (RRMS). A total of 2285 adult patients from the French nationwide cohort were included. Outcomes were irreversible EDSS4, and conversion to secondary progression of multiple sclerosis (SPMS). Associations between treatments and risk of disability were assessed using a novel weighted cumulative exposure model, assuming a 3-year lag to account for reverse causality. This flexible approach accounts for past exposure in a multivariate Cox proportional hazards model by computing a weight function. At baseline, mean ± standard deviation age of patients was 33.4 ± 8.9 years and 75.0% were women. A 15-year continuous treatment starting 20 years ago was associated with a decrease in risk of 26% for irreversible EDSS4, and 34% for SPMS compared to a 5-year treatment starting 10 years ago. The risk of disability decreased with increasing duration of exposure to disease-modifying treatment (DMT). Long-term use of treatments in RRMS has a stronger beneficial cumulative impact than only early uses and delays the occurrence of moderate disability and conversion to SPMS.


A PASS 3 evaluation of community residences in Wales.
PASS 3 was used to evaluate 14 residential services for people with mental retardation. Residents had a broad range of abilities. Size of residences ranged from one to seven residents. PASS scores were generally associated with both resident's ability level and smaller size of living unit. Overall, the residences were shown to be well-located and reasonably home-like. However, residence personnel tended to lack organized means and competencies to promote resident development and experience. Administrative practices were also weak. Attention must be given to factors other than residence size, location, building characteristics, and staffing parameters when services are commissioned and when service contracts are specified and reviewed.


Preliminary safety and efficacy results of laser immunotherapy for the treatment of metastatic breast cancer patients.
We report our preliminary results of a pilot clinical trial of late-stage breast cancer patients treated by laser immunotherapy (LIT), a local intervention using an 805 nm laser for non-invasive irradiation, indocyanine green for selective thermal effect, and immunoadjuvant (glycated chitosan) for immunological stimulation. Ten breast cancer patients were enrolled in this study; all patients were considered to be out of other available treatment options. Toxicity was individually evaluated through physical exams and laboratory tests. Adverse reactions only occurred in the area of treatment due to photothermal injury and local administration of immunoadjuvant. No grade 3 or 4 side effects were observed. Treatment efficacy of LIT was also evaluated by physical examination and tomography. In 8 patients available for evaluation, the objective response rate was 62.5% and the clinical beneficial response rate was 75%. While the study is still ongoing, the initial outcomes of this clinical trial show that LIT is well tolerated and is promising in the treatment of metastatic breast cancer.


Risk factors for dengue outbreaks in Odisha, India: A case-control study.
Environmental and climatic risk factors of dengue outbreak has been studied in detail. However, the socio-epidemiological association with the disease is least explored. The study aims to identify the social and ecological factors associated with emerging dengue in Odisha, India. A population-based case-control study (age and sex matched at the ratio of 1:1) was conducted in six districts of the state in 2017. A structured validated questionnaire was used to collect information for each consenting participant. An ecological household survey was done using a checklist during the month of July-September. Along with the descriptive statistics, conditional logistic regression model was used to calculate the adjusted odds ratio using STATA. Of 380 cases, nearly 55% were male and the median age was 33years. The adjusted odds of having dengue was nearly three times higher among the people having occupation which demands long travel, presence of breeding sites (1.7; 95% CI 1.2-2.6), presence of swampy area near home (1.5; 95% CI 1.1-2.1) and having travel history close to the index date (1.6; 95% CI 1.1-2.4). People staying in thatched houses had three times higher risk of the disease, however, households keeping the swampy areas clean had 50% less risk for the disease (0.5; 95% CI 0.31-0.67). Nearly 22.2% of cases had a travel history during the index date. Of them, 36% had diagnosis before the travel, whereas, 64% developed the disease after the returning from the travel. Household factors such as occupation and ecological condition of households play important roles in dengue outbreaks in Odisha. However, our study suggests travel/commuting are also essential factors to be considered during disease prevention planning.


Intramammary infections in the sow during the peripartum period.
The incidence and ype of aerobic bacteria causing intramammary infection (IMI) in two groups of peripartum sows were determined. The incidence of IMI in 24 sows near farrowing was 30.2 percent; the incidence in 16 machine-milked sows was 81.0 percent. Numerous mixed IMI were detected. The bacterium causing most IMI was Staphylococcus epidermidis, followed by beta hemolytic streptococci, aslpha hemolytic streptococci, Escherichia coli, and diphtheroids. The beta hemolytic streptococci and E. coli cultures were studied in detail. The relationship of bacterial IMI to agalactiae toxemia is discussed.


High-throughput functional affinity purification of mannose binding proteins from Oryza sativa.
We have used affinity chromatography in combination with mass spectrometry to isolate, identify, and assign a preliminary functional annotation to a large number of both known and novel proteins from rice. Rice (Oryza sativa) leaf, root, and seed tissue extracts were fractionated by column affinity chromatography using alpha-D-mannose as the ligand. Bound fractions were eluted and subjected to one-dimensional electrophoresis, followed by high-performance liquid chromatography-tandem mass spectrometric analysis of separated proteins. This multiplexed technology resulted in the isolation and identification of 136 distinct mannose binding proteins from rice. A comparative analysis demonstrates very little overlap of identified proteins between the respective tissues, and confirms the correctly compartmentalized presence of a significant number of proteins from largely tissue-specific biochemical pathways. Over 30% of the identified proteins with a previously annotated function are directly involved in sugar metabolism, including several highly expressed known rice lectins. Direct comparison of the peptide sequences identified in this study to those peptides identified in the most comprehensive survey of the rice proteome to date indicates that our current data represents a significant enrichment of proteins unique to this dataset. Nearly 15% of the identified proteins, identified on the basis of exact peptide matching to sequences in the rice genomic database, represent proteins without a previously known functional annotation, indicating the potential of this combined chromatographic approach to assign a preliminary function to novel proteins in a high-throughput fashion.


Anti-influenza activity of Alchemilla mollis extract: possible virucidal activity against influenza virus particles.
Influenza virus infection is a major public health problem that leads to significant morbidity and mortality. The emergence of resistance to the currently available anti-influenza agents has necessitated the development of new drugs with novel targets. Studying known ethno-medicinal plants is a promising approach for the discovery of new antiviral compounds. Alchemilla mollis is used in traditional medicine in Europe for different indications, including minimizing the symptoms of a sore throat. In this study, we found that A. mollis extract has anti-influenza activity, and investigated the mechanism underlying its inhibition of influenza virus replication. Plaque assays demonstrated that treatment of cells with A. mollis extract prior to infection did not inhibit influenza virus infection. However, plaque formation was markedly reduced when infected cells were overlaid with an agarose gel containing A. mollis extract. In addition, exposure of the virus to A. mollis extract prior to infection and treatment of cells during virus infection significantly suppressed plaque formation. Influenza virus-induced hemagglutination of chicken red blood cells was inhibited by A. mollis extract treatment. The inhibitory effect was observed against influenza A virus subtypes H1N1, H3N2, and H5N2. These findings suggest that A. mollis extract has virucidal or neutralizing activity against influenza virus particles. Furthermore, inhibitory effect of zanamivir synergistically increased after combination with A. mollis extract. Our results suggest that A. mollis extract has the potential to be developed as an antiinfluenza agent.


Choosing treatment options for patients with relapsed/refractory multiple myeloma.
Multiple myeloma (MM) is a clonal plasma cell disorder that is still incurable using conventional treatments. Over the last decade, advances in front-line therapy have led to an increase in survival, but there are still some doubts in the case of relapsed/refractory disease. We searched the PubMed database for articles on treatment options for patients with relapsed/refractory MM published between 1996 and 2013. These treatments included hematopoietic cell transplantation (HCT), rechallenges using previous chemotherapy regimens, and trials of new regimens. The introduction of new agents such as the immunomodulatory drugs (IMIDs) thalidomide and lenalidomide, and the first-in-its-class proteasome inhibitor bortezomib, has greatly improved clinical outcomes in patients with relapsed/refractory MM, but not all patients respond and those that do may eventually relapse or become refractory to treatment. The challenge is therefore to select the optimal treatment for each patient by balancing efficacy and toxicity. To do this, it is necessary to consider disease-related factors, such as the quality and duration of responses to previous therapies, and the aggressiveness of the relapse, and patient-related factors such as age, comorbidities, performance status, pre-existing toxicities and cytogenetic patterns. The message from the trials reviewed in this article is that the new agents may be used to re-treat relapsed/refractory disease, and that the sequencing of their administration should be modulated on the basis of the various disease and patient-related factors. Moreover, our understanding of the pharmacology and molecular action of the new drugs will contribute to the possibility of developing tailored treatment.