MicroRNAs define distinct human neuroblastoma cell phenotypes and regulate their differentiation and tumorigenicity.
Neuroblastoma (NB) is the most common extracranial solid tumor in children. NB tumors and derived cell lines are phenotypically heterogeneous. Cell lines are classified by phenotype, each having distinct differentiation and tumorigenic properties. The neuroblastic phenotype is tumorigenic, has neuronal features and includes stem cells (I-cells) and neuronal cells (N-cells). The non-neuronal phenotype (S-cell) comprises cells that are non-tumorigenic with features of glial/smooth muscle precursor cells. This study identified miRNAs associated with each distinct cell phenotypes and investigated their role in regulating associated differentiation and tumorigenic properties. A miRNA microarray was performed on the three cell phenotypes and expression verified by qRT-PCR. miRNAs specific for certain cell phenotypes were modulated using miRNA inhibitors or stable transfection. Neuronal differentiation was induced by RA; non-neuronal differentiation by BrdU. Changes in tumorigenicity were assayed by soft agar colony forming ability. N-myc binding to miR-375 promoter was assayed by chromatin-immunoprecipitation. Unsupervised hierarchical clustering of miRNA microarray data segregated neuroblastic and non-neuronal cell lines and showed that specific miRNAs define each phenotype. qRT-PCR validation confirmed that increased levels of miR-21, miR-221 and miR-335 are associated with the non-neuronal phenotype, whereas increased levels of miR-124 and miR-375 are exclusive to neuroblastic cells. Downregulation of miR-335 in non-neuronal cells modulates expression levels of HAND1 and JAG1, known modulators of neuronal differentiation. Overexpression of miR-124 in stem cells induces terminal neuronal differentiation with reduced malignancy. Expression of miR-375 is exclusive for N-myc-expressing neuroblastic cells and is regulated by N-myc. Moreover, miR-375 downregulates expression of the neuronal-specific RNA binding protein HuD. Thus, miRNAs define distinct NB cell phenotypes. Increased levels of miR-21, miR-221 and miR-335 characterize the non-neuronal, non-malignant phenotype and miR-335 maintains the non-neuronal features possibly by blocking neuronal differentiation. miR-124 induces terminal neuronal differentiation with reduction in malignancy. Data suggest N-myc inhibits neuronal differentiation of neuroblastic cells possibly by upregulating miR-375 which, in turn, suppresses HuD. As tumor differentiation state is highly predictive of patient survival, the involvement of these miRNAs with NB differentiation and tumorigenic state could be exploited in the development of novel therapeutic strategies for this enigmatic childhood cancer.


Surviving the wait: defining support while awaiting breast cancer surgery.
This paper is a report of a descriptive study of the common meanings, shared experiences and practices of social support of women within the days between breast cancer diagnosis and treatment initiation. Support needs, types of social support and support outcomes during and after breast cancer treatment have been explored worldwide. However, to promote women's psychological wellbeing it is essential to understand how women define support in the highly stressful period initially following diagnosis. Secondary analysis of narrative texts using interpretive phenomenology from 18 women in the Midwestern United States newly diagnosed with breast cancer who were interviewed in 2005 for a study of women's pretreatment thought processes. 'Surviving the wait for surgery by balancing support needs to maintain a hopeful outlook' was the overarching pattern linking six other related themes: (1) controlling access to information for self and to others, (2) knowing which supportive network members to access, (3) controlling anxiety through distraction to maintain hope while waiting, (4) being in good hands and comfortable with decision (provider support), (5) protecting others through concealment and being strong to maintain hope and (6) accepting care from others vs. maintaining a nurturing role. Implications for nurses working with women in the days following breast cancer diagnosis include assessing women's definitions and availability of support; respecting varied needs for informational support; providing a supportive clinical environment; educating clinicians, family and friends regarding unsupportive responses within the cultural context and validating women's control and balancing of support needs.


Mitochondria as a target for neuroprotection: role of methylene blue and photobiomodulation.
Mitochondrial dysfunction plays a central role in the formation of neuroinflammation and oxidative stress, which are important factors contributing to the development of brain disease. Ample evidence suggests mitochondria are a promising target for neuroprotection. Recently, methods targeting mitochondria have been considered as potential approaches for treatment of brain disease through the inhibition of inflammation and oxidative injury. This review will discuss two widely studied approaches for the improvement of brain mitochondrial respiration, methylene blue (MB) and photobiomodulation (PBM). MB is a widely studied drug with potential beneficial effects in animal models of brain disease, as well as limited human studies. Similarly, PBM is a non-invasive treatment that promotes energy production and reduces both oxidative stress and inflammation, and has garnered increasing attention in recent years. MB and PBM have similar beneficial effects on mitochondrial function, oxidative damage, inflammation, and subsequent behavioral symptoms. However, the mechanisms underlying the energy enhancing, antioxidant, and anti-inflammatory effects of MB and PBM differ. This review will focus on mitochondrial dysfunction in several different brain diseases and the pathological improvements following MB and PBM treatment.


Cerebral Embolic Protection Devices During Transcatheter Aortic Valve Implantation: Systematic Review and Meta-Analysis.
Silent ischemic embolic lesions are common after transcatheter aortic valve implantation (TAVI). The use of embolic protection devices (EPD) may reduce the occurrence of these embolic lesions. Thus, a quantitative overview and credibility assessment of the literature was necessary to draw a robust message about EPD. Therefore, the aim of this meta-analysis was to study whether the use of EPD reduces silent ischemic and clinically evident cerebrovascular events associated with TAVI. We conducted a comprehensive search to identify studies that evaluated patients undergoing TAVI with or without EPD. Random-effects meta-analyses were performed to estimate the effect of EPD compared with no-EPD during TAVI using aggregate data. Sixteen studies involving 1170 patients (865/305 with/without EPD) fulfilled the inclusion criteria. The EPD delivery success rate was reported in all studies and was achieved in 94.5% of patients. Meta-analyses evaluating EPD versus without EPD strategies could not confirm or exclude any differences in terms of clinically evident stroke (relative risk, 0.70; 95% confidence interval [CI], 0.38-1.29; P=0.26) or 30-day mortality (relative risk, 0.58; 95% CI, 0.20-1.64; P=0.30). There were no significant differences in new-single, multiple, or total number of lesions. The use of EPD was associated with a significantly smaller ischemic volume per lesion (standardized mean difference, -0.52; 95% CI, -0.85 to -0.20; P=0.002) and smaller total volume of lesions (standardized mean difference, -0.23; 95% CI, -0.42 to -0.03; P=0.02). Subgroup analysis by type of valve showed an overall trend toward significant reduction in new lesions per patient using EPD (standardized mean difference, -0.41; 95% CI, -0.82 to 0.00; P=0.05), driven by self-expanding devices. The use of EPD during TAVI may be associated with smaller volume of silent ischemic lesions and smaller total volume of silent ischemic lesions. However, EPD may not reduce the number of new-single, multiple, or total number of lesions. There was only very low quality of evidence showing no significant differences between patients undergoing TAVI with or without EPD with respect to clinically evident stroke and mortality.


Prevalence and mobility of integrative and conjugative elements within a Streptomyces natural population.
Horizontal Gene Transfer (HGT) is a powerful force generating genomic diversity in bacterial populations. HGT in Streptomyces is in large part driven by conjugation thanks to plasmids, Integrative and Conjugative elements (ICEs) and Actinomycete ICEs (AICEs). To investigate the impact of ICE and AICE conjugation on Streptomyces genome evolution, we used in silico and experimental approaches on a set of 11 very closely related strains isolated from a millimeter scale rhizosphere population. Through bioinformatic searches of canonical conjugation proteins, we showed that AICEs are the most frequent integrative conjugative elements, with the central chromosome region being a hotspot for integrative element insertion. Strains exhibited great variation in AICE composition consistent with frequent HGT and/or gene loss. We found that single insertion sites can be home to different elements in different strains (accretion) and conversely, elements belonging to the same family can be found at different insertion sites. A wide variety of cargo genes was present in the AICEs with the potential to mediate strain-specific adaptation (e.g., DNA metabolism and resistance genes to antibiotic and phages). However, a large proportion of AICE cargo genes showed hallmarks of pseudogenization, consistent with deleterious effects of cargo genes on fitness. Pock assays enabled the direct visualization of conjugal AICE transfer and demonstrated the transfer of AICEs between some, but not all, of the isolates. Multiple AICEs were shown to be able to transfer during a single mating event. Although we did not obtain experimental evidence for transfer of the sole chromosomal ICE in this population, genotoxic stress mediated its excision from the chromosome, suggesting its functionality. Our results indicate that AICE-mediated HGT in Streptomyces populations is highly dynamic, with likely impact on strain fitness and the ability to adapt to environmental change.


MR Imaging of Rectal Cancer: Radiomics Analysis to Assess Treatment Response after Neoadjuvant Therapy.
Purpose To investigate the value of T2-weighted-based radiomics compared with qualitative assessment at T2-weighted imaging and diffusion-weighted (DW) imaging for diagnosis of clinical complete response in patients with rectal cancer after neoadjuvant chemotherapy-radiation therapy (CRT). Materials and Methods This retrospective study included 114 patients with rectal cancer who underwent magnetic resonance (MR) imaging after CRT between March 2012 and February 2016. Median age among women (47 of 114, 41%) was 55.9 years (interquartile range, 45.4-66.7 years) and median age among men (67 of 114, 59%) was 55 years (interquartile range, 48-67 years). Surgical histopathologic analysis was the reference standard for pathologic complete response (pCR). For qualitative assessment, two radiologists reached a consensus. For radiomics, one radiologist segmented the volume of interest on high-spatial-resolution T2-weighted images. A random forest classifier was trained to separate the patients by their outcomes after balancing the number of patients in each response category by using the synthetic minority oversampling technique. Statistical analysis was performed by using the Wilcoxon rank-sum test, McNemar test, and Benjamini-Hochberg method. Results Twenty-one of 114 patients (18%) achieved pCR. The radiomic classifier demonstrated an area under the curve of 0.93 (95% confidence interval [CI]: 0.87, 0.96), sensitivity of 100% (95% CI: 0.84, 1), specificity of 91% (95% CI: 0.84, 0.96), positive predictive value of 72% (95% CI: 0.53, 0.87), and negative predictive value of 100% (95% CI: 0.96, 1). The diagnostic performance of radiomics was significantly higher than was qualitative assessment at T2-weighted imaging or DW imaging alone (P < .02). The specificity and positive predictive values were significantly higher in radiomics than were at combined T2-weighted and DW imaging (P < .0001). Conclusion T2-weighted-based radiomics showed better classification performance compared with qualitative assessment at T2-weighted and DW imaging for diagnosing pCR in patients with locally advanced rectal cancer after CRT. © RSNA, 2018 Online supplemental material is available for this article.


Azobenzene photoswitching without ultraviolet light.
Most azobenzene-based photoswitches use UV light for photoisomerization. This can limit their application in biological systems, where UV light can trigger unwanted responses, including cellular apoptosis. We have found that substitution of all four ortho positions with methoxy groups in an amidoazobenzene derivative leads to a substantial (~35 nm) red shift of the n-π* band of the trans isomer, separating it from the cis n-π* transition. This red shift makes trans-to-cis photoswitching possible using green light (530-560 nm). The cis state is thermally stable with a half-life of ~2.4 days in the dark in aqueous solution. Reverse (cis-to-trans) photoswitching can be accomplished with blue light (460 nm), so bidirectional photoswitching between thermally stable isomers is possible without using UV light at all.


The SMRT corepressor is regulated by a MEK-1 kinase pathway: inhibition of corepressor function is associated with SMRT phosphorylation and nuclear export.
The SMRT (silencing mediator of retinoic acid and thyroid hormone receptor) corepressor participates in the repression of target gene expression by a variety of transcription factors, including the nuclear hormone receptors, promyelocytic leukemia zinc finger protein, and B-cell leukemia protein 6. The ability of SMRT to associate with these transcription factors and thereby to mediate repression is strongly inhibited by activation of tyrosine kinase signaling pathways, such as that represented by the epidermal growth factor receptor. We report here that SMRT function is potently inhibited by a mitogen-activated protein kinase (MAPK) kinase kinase (MAPKKK) cascade that operates downstream of this growth factor receptor. Intriguingly, the SMRT protein is a substrate for phosphorylation by protein kinases operating at multiple levels in this MAPKKK pathway, including the MAPKs, MAPK-extracellular signal-regulated kinase 1 (MEK-1), and MEK-1 kinase (MEKK-1). Phosphorylation of SMRT by MEKK-1 and, to a lesser extent, MEK-1 inhibits the ability of SMRT to physically tether to its transcription factor partners. Notably, activation of MEKK-1 or MEK-1 signaling in transfected cells also leads to a redistribution of the SMRT protein from a nuclear compartment to a more perinuclear or cytoplasmic compartment. We suggest that SMRT-mediated repression is regulated by the MAPKKK cascade and that changes both in the affinity of SMRT for its transcription factors and in the subcellular distribution of SMRT contribute to the loss of SMRT function that is observed in response to kinase signal transduction.


Phosphatase inhibition potentiates IL-6 production by mast cells in response to FcepsilonRI-mediated activation: involvement of p38 MAPK.
Mast cells are crucial effector cells in the immune response through mediator secretion and release of cytokines. A coordinated balance between protein kinases and phosphatases plays an essential role in the regulation of mast cell mediator secretion. We have previously shown that treatment of mast cells with okadaic acid (OA), a protein phosphatase 2A (PP2A) inhibitor, results in a dose-dependent increase in interleukin (IL)-6 production. We show here for the first time a synergism between OA and immunoglobulin E (IgE)-mediated IL-6 secretion by murine bone marrow-derived mast cells (BMMC). Selective p38 mitogen-activated protein kinase (p38 MAPK) inhibition reduces OA and IgE-mediated IL-6 production. Regulation of p38 MAPK by PP2A was demonstrated, as OA treatment caused a dose-dependent increase in p38 MAPK phosphorylation. Antigen-mediated activation of murine mast cells also resulted in an increase in p38 MAPK phosphorylation, which was potentiated by cotreatment of the cells with OA. Lastly, in two mast cell lines (human mast cell-1 5C6 and murine MC/9) and primary-cultured murine BMMC, we show by coimmunoprecipitation an interaction between p38 MAPK and PP2A. These data support a role for PP2A through interaction with p38 MAPK in the regulation of IgE-dependent mast cell activation.


Resistance to caspase-8 and -9 fragments in a malignant pleural mesothelioma cell line with acquired cisplatin-resistance.
Apoptotic cysteine-aspartate proteases (caspases) are essential for the progression and execution of apoptosis, and detection of caspase fragmentation or activity is often used as markers of apoptosis. Cisplatin (cis-diamminedichloroplatinum (II)) is a chemotherapeutic drug that is clinically used for the treatment of solid tumours. We compared a cisplatin-resistant pleural malignant mesothelioma cell line (P31res1.2) with its parental cell line (P31) regarding the consequences of in vitro acquired cisplatin-resistance on basal and cisplatin-induced (equitoxic and equiapoptotic cisplatin concentrations) caspase-3, -8 and -9 fragmentation and proteolytic activity. Acquisition of cisplatin-resistance resulted in basal fragmentation of caspase-8 and -9 without a concomitant increase in proteolytic activity, and there was an increased basal caspase-3/7 activity. Similarly, cisplatin-resistant non-small-cell lung cancer cells, H1299res, had increased caspase-3 and -9 content compared with the parental H1299 cells. In P31 cells, cisplatin exposure resulted in caspase-9-mediated caspase-3/7 activation, but in P31res1.2 cells the cisplatin-induced caspase-3/7 activation occurred before caspase-8 or -9 activation. We therefore concluded that in vitro acquisition of cisplatin-resistance rendered P31res1.2 cells resistant to caspase-8 and caspase-9 fragments and that cisplatin-induced, initiator-caspase independent caspase-3/7 activation was necessary to overcome this resistance. Finally, the results demonstrated that detection of cleaved caspase fragments alone might be insufficient as a marker of caspase activity and ensuing apoptosis induction.


Serologic testing with ANCA, ASCA, and anti-OmpC in children and young adults with Crohn's disease and ulcerative colitis: diagnostic value and correlation with disease phenotype.
Serologic testing is increasingly being utilized to evaluate children with suspected inflammatory bowel disease (IBD). The aim of this paper was to evaluate the sensitivity and specificity of a currently available panel involving four antibodies: deoxyribonuclease (DNase)-sensitive perinuclear antineutrophil cytoplasmic antibody (DNase-sensitive pANCA), IgA and IgG antibodies to Saccharomyces cerevisiae (IgA and IgG ASCA), and antibody to Escherichia coli outer membrane porin (anti-OmpC). We also wished to determine whether antibody levels correlated with disease activity, and whether a specific antibody pattern correlated with location and outcome of disease in children. We studied sera from 81 children with Crohn's disease (CD), 54 with ulcerative colitis (UC), and 63 controls. Clinical data, disease activity, and disease diagnosis were gathered at the time of serum sampling, and charts were re-reviewed at time of the study to determine long-term outcome. Enzyme-linked immunosorbent assay was utilized to determine titers of antibodies to ASCA, DNase-sensitive pANCA, and anti-OmpC; the presence of perinuclear staining for ANCA was confirmed by immunofluorescence. We identified ASCA antibodies in 44% of CD patients, 0% of UC patients, and 1 control patient. DNase-sensitive pANCA antibodies were found in 70% of patients with UC, 18% of CD patients (predominantly Crohn's colitis), and 3% of controls. Anti-OmpC as an isolated assay had low sensitivity for both CD (24%) and UC (11%), and displayed a 5% false-positive rate. However, anti-OmpC did identify a small number of IBD patients not detected by the other assays. If any one or more of the four antibodies was positive, the overall sensitivity of the four antibody panel was 65% for CD and 76% for UC, with a specificity of 94%. Patients who were ASCA-positive were more likely to have disease of the ileum or ileum and right colon than patients who were ASCA-negative (58%vs 18%, p < 0.001). Patients with ASCA-positive were also more likely to require ileocecal resection (36%vs 13%, p < 0.05). A currently available commercial antibody panel has good sensitivity and excellent specificity for CD and UC. The ASCA antibodies, while highly specific for CD, identify predominantly the subset of children with disease of the ileum and ascending colon who may be at increased risk of surgery.


Health effects of exposure to organic dust in workers of a modern hatchery.
The aim of the presented study was to determine the health status of workers occupationally exposed to moderate amounts of organic dust, employed in a modern hatchery with an efficient ventilation system. A group of 32 hatchery workers was examined. As a reference group, 50 urban dwellers not exposed to any kind of organic dust were examined. All people were interviewed for the presence of work-related symptoms and subjected to physical and spirometric examinations. Blood sera were examined for the presence of precipitins against 13 antigens associated with organic dust, and for the presence of total and chicken-specific No significant differences were found between the spirometric values in the group of hatchery workers and the reference group. Positive precipitin reactions were noted mostly with the antigens of Gram-negative bacteria associated with organic dust. The frequencies of positive reactions to antigens of Escherichia coli and Acinetobacter baumannii in hatchery workers were significantly greater compared to the reference group (p<0.05). Precipitin reactions to Gram-positive non-branching bacteria, actinomycetes, fungi and birdactions to antigens of Escherichia coli and Acinetobacter baumannii in hatchery workers were significantly greater compared to the reference group (p<0.05). Precipitin reactions to Gram-positive non-branching bacteria, actinomycetes, fungi and bird protein, were rare or absent. The mean concentration of total IgE in sera of hatchery workers was nearly 3 times greater compared to the reference group, and the difference proved to be statistically significant (p<0.05). No specific IgE antibodies against chicken feathers were detected in the blood of hatchery workers and referents. In conclusion, the examined hatchery workers showed a moderate frequency of work-related symptoms, no decline in lung function and low reactivity to most microbial and bird protein allergens. These results suggest that the effects of exposure to organic dust in workers of modern hatcheries with an efficient ventilation system are less compared to the workers of poultry farms, such as broiler or egg laying houses.


A restricted role for TYK2 catalytic activity in human cytokine responses revealed by novel TYK2-selective inhibitors.
TYK2 is a JAK family protein tyrosine kinase activated in response to multiple cytokines, including type I IFNs, IL-6, IL-10, IL-12, and IL-23. Extensive studies of mice that lack TYK2 expression indicate that the IFN-α, IL-12, and IL-23 pathways, but not the IL-6 or IL-10 pathways, are compromised. In contrast, there have been few studies of the role of TYK2 in primary human cells. A genetic mutation at the tyk2 locus that results in a lack of TYK2 protein in a single human patient has been linked to defects in the IFN-α, IL-6, IL-10, IL-12, and IL-23 pathways, suggesting a broad role for TYK2 protein in human cytokine responses. In this article, we have used a panel of novel potent TYK2 small-molecule inhibitors with varying degrees of selectivity against other JAK kinases to address the requirement for TYK2 catalytic activity in cytokine pathways in primary human cells. Our results indicate that the biological processes that require TYK2 catalytic function in humans are restricted to the IL-12 and IL-23 pathways, and suggest that inhibition of TYK2 catalytic activity may be an efficacious approach for the treatment of select autoimmune diseases without broad immunosuppression.


Acute Heart Failure Is a Malignant Process: But We Can Induce Remission.
Acute heart failure is a common and increasingly prevalent condition, affecting >10 million people annually. For those patients who survive to discharge, early readmissions and death rates are >30% everywhere on the planet, making it a malignant condition. Beyond these adverse outcomes, it represents one of the largest drivers of health care costs globally. Studies in the past 2 years have demonstrated that we can induce remissions in this malignant process if therapy is instituted rapidly, at the first acute heart failure episode, using full doses of all available effective medications. Multiple studies have demonstrated that this goal can be achieved safely and effectively. Now the urgent call is for all stakeholders, patients, physicians, payers, politicians, and the public at large to come together to address the gaps in implementation and enable health care providers to induce durable remissions in patients with acute heart failure.


TERC Variants Associated with Short Leukocyte Telomeres: Implication of Higher Early Life Leukocyte Telomere Attrition as Assessed by the Blood-and-Muscle Model.
Short leukocyte telomere length (LTL) is associated with atherosclerotic cardiovascular disease (ASCVD). Mendelian randomisation studies, using single nucleotide polymorphisms (SNPs) associated with short LTL, infer a causal role of LTL in ASCVD. Recent results, using the blood-and-muscle model, indicate that higher early life LTL attrition, as estimated by the ratio between LTL and skeletal muscle telomere length (MTL), rather than short LTL at conception, as estimated by MTL, should be responsible of the ASCVD-LTL connection. We combined LTL and MTL measurements and SNPs profiling in 402 individuals to determine if 15 SNPs classically described as associated with short LTL at adult age were rather responsible for higher LTL attrition during early life than for shorter LTL at birth. Two of these SNPs (rs12696304 and rs10936599) were associated with LTL in our cohort (p = 0.027 and p = 0.025, respectively). These SNPs, both located on the TERC gene, were associated with the LTL/MTL ratio (p = 0.007 and p = 0.037, respectively), but not with MTL (p = 0.78 and p = 0.32 respectively). These results suggest that SNPs located on genes coding for telomere maintenance proteins may contribute to a higher LTL attrition during the highly replicative first years of life and have an impact later on the development of ASCVD.


Role of human aquaporin 5 in colorectal carcinogenesis.
While overexpression of several aquaporins (AQPs) has been reported in different types of human cancer, the role of AQPs in carcinogenesis has not been clearly defined. Here, by immunochemistry, we have found expression of AQP5 protein in 62.8% (59/94) of resected colon cancer tissue samples as well as association of AQP5 with liver metastasis. We then demonstrated that overexpression of human AQP5 (hAQP5) induces cell proliferation in colon cancer cells. Overexpression of wild-type hAQP5 increased proliferation and phosphorylation of extracellular signal-regulated kinase-1/2 in HCT116 colon cancer cells whereas these phenomena in hAQP5 mutants (N185D and S156A) were diminished, indicating that both membrane association and serine/threonine phosphorylation of AQP5 are required for proper function. Interestingly, overexpression of AQP1 and AQP3 showed no differences in extracellular signal-regulated kinase-1/2 phosphorylation, suggesting that AQP5, unlike AQP1, may be involved in signal transduction. Moreover, hAQP5-overexpressing cells showed an increase in retinoblastoma protein phosphorylation through the formation of a nuclear complex with cyclin D1 and CDK4. Small interfering RNA analysis confirmed that hAQP5 activates the Ras signaling pathway. These data not only describe the induction of hAQP5 expression during colorectal carcinogenesis but also provide a molecular mechanism for colon cancer development through the interaction of hAQP5 with the Ras/extracellular signal-regulated kinase/retinoblastoma protein signaling pathway, identifying hAQP5 as a novel therapeutic target.


Measuring recreational visitation at U.S. National Parks with crowd-sourced photographs.
Land managers rely on visitation data to inform policy and management decisions. However, visitation data is often costly and burdensome to obtain, and provides a limited depth of information. In this paper, we assess the validity of using crowd-sourced, online photographs to infer information about the habits and preferences of recreational visitors by comparing empirical data from the National Park Service to photograph data from the online platform Flickr for 38 National Parks in the western United States. Using multiple regression analysis, we find that the number of photos posted monthly in a park can reliably indicate the number of visitors to a park in a given month. Through additional statistical testing we also find that the home locations of photo-takers, provided voluntarily on an online profile, accurately show the home origins of park visitors. Together, these findings validate a new method for measuring recreational visitation, opening an opportunity for land managers worldwide to track and understand visitation by augmenting current data collection methods with crowd-sourced, online data that is easy and inexpensive to obtain. In addition, it enables future research on how visitation rates change with changes in access, management or infrastructure, weather events, or ecosystem health, and facilitates valuation research, such as travel cost studies.


[Effect of weight reduction on the clinical and hormonal condition of obese anovulatory women].
Obesity is associated to a variety of endocrine abnormalities that might lead to chronic anovulation in women. Weight loss may improve the hormonal profile and then restore ovulation in some of them. The aim of the current study was to evaluate the effect of a weight loss program on the clinical and hormonal characteristics of anovulatory obese women attending our reproductive clinic. We studied a group of 30 anovulatory obese patients between 18 and 35 years old without any thyroid disease. Before and after a weight loss of at least 5% of initial body weight we analyzed LH, FSH, estradiol, prolactine, testosterone, dehydroepiandrosterone sulfate, oral glucose tolerance test and progesterone, weight, BMI, waist/hip ratio and total body fat percentage. The mean weight loss was 9.5 +/- 4.3 kg. which represents a weight loss of 10.96% from initial body weight, with 26 patients (86.6%) resuming spontaneous ovulation. The women's mean plasma testosterone, LH, estradiol and DHEA-S decreased significantly and there was significantly increased on progesterone. At the beginning a total of 12 patients showed an impaired oral glucose tolerance test and after weight loss 9 of them improved it. The results shown in this study demonstrate that even a small weight reduction and a decrease in total body fat percentage improve the hormonal profile and restore ovulation in anovulatory obese women. Thus weight loss should be considered before starting with ovulation induction therapy.


Antioxidant activity of knotwood extractives and phenolic compounds of selected tree species.
The antioxidant potency and the radical scavenging capacity of superoxide and peroxyl radicals were assessed for 13 hydrophilic knotwood extracts of commercially important wood species, or fractions thereof, as well as for five pure wood-derived lignans and the flavonoid taxifolin. The chemical composition of the knotwood extracts was determined by gas chromatography combined with mass spectrometry. Most of the investigated wood species were rich in hydrophilic extractives (10-20% of the dry wood) with one or a few compounds dominating in each extract. All extracts had a high antioxidative potency and/or radical scavenging capacity as compared to the well-known antioxidants Trolox and butylated hydroxyanisole. The pure wood-derived lignans and taxifolin also had a high antioxidative potency and/or radical scavenging capacity. However, the antioxidant potency and/or radical scavenging capacity of several of the hydrophilic knotwood extracts were higher than that of the dominating compounds in pure form.


Immune responses to inactivated vaccine in people naturally infected with hantaviruses.
An inactivated Hantaan virus vaccine for hemorrhagic fever with renal syndrome (HFRS) was given by injection to 15 people who were naturally infected with either Hantaan or Seoul viruses. Immunofluorescent antibody (IFA), reversed passive hemagglutination inhibition (RPHI), hemagglutination inhibition (HI), and neutralization antibody (NA) assays were used to measure the antibody titers of the vaccinated people before and after three doses of vaccine. The results indicated that IFA and RPHI antibody titers were boosted significantly (P < 0.05) after the vaccination. Either Hantaan or Seoul virus could induce two-way cross-reactive neutralization antibody responses in humans. After HTNV vaccine immunization, the NA titers of people with natural infection increased significantly (P < 0.05) to both Hantaan and Seoul viruses, while the relative dominance between these two type responses was still similar to that of natural infection. It is worthwhile to studying the procedure further to inoculate two different virus vaccines for improving the cross-protective effect.


Effects of variations in renal hemodynamics on the time course of renin secretion rate.
The effects of variations in renal hemodynamics on the time course of renin secretion were studied in dogs anesthetized with pentobarbital-chloralose. Hemodynamic changes were induced either locally in kidneys perfused in situ via an extracorporeal circuit (with or without a pump system) or systemically by hemorrhage or nitroprusside infusion. In the autoperfused kidney the reduction of renal perfusion pressure to approximately one-half of the arterial pressure by inflow occlusion caused an increase in renal conductance (renal vasodilation) and an increase in renin secretion rate (RSR). In the pump-perfused kidney, a step increase in renal blood flow (RBF) caused renal vasoconstriction and a decrease in RSR; a step decrease in RBF caused renal vasodilation and an increase in RSR. Following step changes in RBF, the time constant of the alterations of renal conductance was 56.5 s, and the time constant of the RSR responses was 80.1 s. The total time required to reach a steady state for RSR lagged behind that for renal conductance by approximately 5 min. These differences reflect the time needed for the kidney to release renin in response to changes in renal vascular caliber. The results suggest that renin release occurs in response to the autoregulatory dilation of the renal arterioles. When systemic hypotension was induced by nitroprusside infusion, RSR also increased together with the renal conductance. Following hemorrhage, however, RSR increased despite a decrease in renal conductance, reflecting the role of neurohumoral factors in causing renin release in this case. The comparison of renin secretion following different types of hemodynamic alterations serves to elucidate the mechanisms of renin secretion.


Material properties of bone from the femoral neck and calcar femorale of patients with osteoporosis or osteoarthritis.
Patients with osteoarthritis (OA) rarely fracture the neck of the femur whereas this is common in patients with osteoporosis (OP). The reasons for this are not clear. In this study, cores of trabecular bone and thin slices of bone from the calcar were obtained from the femoral neck of patients with OA or OP following hip arthroplasty and a normal group post mortem. The mechanical properties, densities and material composition were measured. The aim was to determine whether differences in these properties could explain why osteoarthritic patients did not suffer from fractured neck of femur. No difference was found in the density or stiffness of the calcar between the groups, though there appeared to be a small increase in mineralization in the OA bone compared with the OP. However, there was a 72% increase in the volume of trabecular bone in the OA group compared with a loss of about 20% in the OP group. This increased apparent density of the OA trabecular bone resulted in a greater stiffness, yield strength and energy absorbed to yield, whereas the same properties of OP bone were not significantly lower than normal. These marked changes in the OA bone could result in a redistribution of stresses due to loads caused by falling, thereby absorbing the energy of impact and preventing the formation of a fracture surface.


Removal of UV photoproducts from an adenovirus-encoded reporter gene following infection of unirradiated and UV-irradiated human fibroblasts.
Ad5HCMVsp1lacZ is a recombinant nonreplicating adenovirus containing the lacZ gene under the control of the human cytomegalovirus immediate early promoter. Previous reports show that lacZ expression for UV-irradiated Ad5HCMVsp1lacZ is greater in nucleotide excision repair (NER) proficient compared to NER deficient human fibroblasts from patients with xerodermapigmentosum (XP) and Cockaye's syndrome (CS) and that pre-UV-treatment of normal fibroblasts results in an enhanced expression of the lacZ gene from UV-irradiated Ad5HCMVsp1lacZ. We have used a quantitative PCR technique to examine whether UV photoproducts are actually removed from the lacZ gene following infection of human fibroblasts with UV-irradiated Ad5HCMVsp1lacZ. Primers flanking a 2.6-kb region of the lacZ reporter gene were added to equal amounts of DNA extracted from Ad5HCMVsp1lacZ infected cells and amplified by PCR using radiolabelled nucleotides as substrates. Results show a significant removal of UV photoproducts in normal human fibroblasts, but a reduced removal in NER deficient XP and CS cells and an enhanced removal in pre-UV-treated normal fibroblasts.


Episodic loadings of phosphorus influence growth and composition of benthic algae communities in artificial stream mesocosms.
Phosphorus (P) is an essential macronutrient for algal communities, but in excess can exacerbate stream eutrophication. However, P loadings to streams vary temporally from continuous to episodic as a result of inputs from point and non-point sources, respectively. P loading pattern can thus alter the temporal availability of P and may influence effects of P enrichment on algal communities. We assessed how P loading pattern influences algal biomass and composition by conducting a 29-day P enrichment experiment in nine artificial streams exposed to either: (1) continuous P enrichment; (2) episodic P enrichment, or; (3) no P enrichment. P enrichment increased algal biomass accrual, but peak biomass did not differ between continuously and episodically enriched treatments. Maximum absolute growth rates were also comparable between P enriched treatments. However, episodic P additions sustained elevated rates of biomass accrual, whereas absolute growth rates in the continuously enriched communities declined towards the end of the experiment. P enrichment resulted in comparable increases in relative abundance of chlorophytes and decreased proportions of bacillariophytes and charophytes in algal communities for continuously and episodically enriched treatments. However, composition of bacillariophyte (diatom) assemblages differed significantly among all P enrichment treatments in accordance with species autecological attributes for P. Our results demonstrate that episodic and continuous P enrichment may augment algal biomass similarly. Yet, P loading pattern regulated the composition of algal communities. Thus, remedial management strategies for the control of nuisance algae production may require focus on the predominant source of P to streams. Finally, species specific responses of diatom assemblages to P enrichment and associated loading patterns suggests this taxonomic group may have potential as diagnostic indicators for identifying the presence of key nutrient sources associated with eutrophication of stream ecosystems.


Insect cell expression and purification of recombinant SARS-COV-2 spike proteins that demonstrate ACE2 binding.
The COVID-19 pandemic caused by SARS-CoV-2 infection has led to socio-economic shutdowns and the loss of over 5 million lives worldwide. There is a need for the identification of therapeutic targets to treat COVID-19. SARS-CoV-2 spike is a target of interest for the development of therapeutic targets. We developed a robust SARS-CoV-2 S spike expression and purification protocol from insect cells and studied four recombinant SARS-CoV-2 spike protein constructs based on the original SARS-CoV-2 sequence using a baculovirus expression system: a spike protein receptor-binding domain that includes the SD1 domain (RBD) coupled to a fluorescent tag (S-RBD-eGFP), spike ectodomain coupled to a fluorescent tag (S-Ecto-eGFP), spike ectodomain with six proline mutations and a foldon domain (S-Ecto-HexaPro(+F)), and spike ectodomain with six proline mutations without the foldon domain (S-Ecto-HexaPro(-F)). We tested the yield of purified protein expressed from the insect cell lines Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tni) and compared it to previous research using mammalian cell lines to determine changes in protein yield. We demonstrated quick and inexpensive production of functional glycosylated spike protein of high purity capable of recognizing and binding to the angiotensin converting enzyme 2 (ACE2) receptor. To further confirm functionality, we demonstrate binding of eGFP fused construct of the spike ectodomain (S-Ecto-eGFP) to surface ACE2 receptors on lung epithelial cells by flow cytometry analysis and show that it can be decreased by means of receptor manipulation (blockade or downregulation).


Mechanisms of DNA damage repair in adult stem cells and implications for cancer formation.
Maintenance of genomic integrity in tissue-specific stem cells is critical for tissue homeostasis and the prevention of deleterious diseases such as cancer. Stem cells are subject to DNA damage induced by endogenous replication mishaps or exposure to exogenous agents. The type of DNA lesion and the cell cycle stage will invoke different DNA repair mechanisms depending on the intrinsic DNA repair machinery of a cell. Inappropriate DNA repair in stem cells can lead to cell death, or to the formation and accumulation of genetic alterations that can be transmitted to daughter cells and so is linked to cancer formation. DNA mutational signatures that are associated with DNA repair deficiencies or exposure to carcinogenic agents have been described in cancer. Here we review the most recent findings on DNA repair pathways activated in epithelial tissue stem and progenitor cells and their implications for cancer mutational signatures. We discuss how deep knowledge of early molecular events leading to carcinogenesis provides insights into DNA repair mechanisms operating in tumours and how these could be exploited therapeutically.


A black beam borne by an incandescent field self-traps in a photopolymerizing medium.
We report that a self-trapped black optical beam that is spatially and temporally incoherent forms spontaneously in a nascent photopolymerization system. The black beam inscribes a permanent cylindrical channel, which prevents the propagation of visible light even under passive conditions (in the absence of polymerization). The finding opens a powerful new mechanism to manipulate light signals from incoherent sources such as LEDs through selective suppression of light propagation. This contrasts with approaches employed by photonic crystals and optical waveguides, which concentrate and guide light intensity within spatially localized regions. The self-trapped black beam forms when a broad incandescent beam bearing a negligible depression was launched into a photopolymerizable medium. Because of refractive index changes caused by polymerization, the depression narrows, deepens, and continually rejects the visible spectrum of light until it stabilizes as a black beam that propagates over long distances (≫ effective Rayleigh range) without significant divergence. As refractive index changes due to polymerization are irreversible, the cylindrical region occupied by the self-trapped black beam is inscribed as a black channel waveguide in the medium.


Global threat of COVID 19 and evacuation of the citizens of different countries
Beginning from China on December 2019, COVID-19 epidemic has spreaded all over the world in a short period of time and has been a pandemic. In challenge with this pandemic quarantine technique has been used widely after tens of years. In the course of the pandemic, many countries evacuated their citizens from affected regions and combined the evacuation with quarantine process. Some examples of these countries who evacuated their citizens are Germany, Italy, Spain, and USA. In further times, during the course of pandemic, according to spread, other countries evacuated their citizens from these countries too. Despite being the origin of the pandemic, in later times Wuhan was also a place where people were evacuated to. Evacuation and quarantine have caused social and psychological impacts on people and some of them took place in mainstream media. In this review article, evacuation and quarantine processes as well as the society's reactions to these, have been compiled.


Effectiveness of Glaucoma Counseling on Rates of Follow-up and Glaucoma Knowledge in a South Indian Population.
To evaluate the impact of traditional counseling and patient-centered counseling, either alone or with recorded audio counseling reinforcement, on glaucoma knowledge and clinical follow-up. Prospective randomized controlled trial. Newly diagnosed adult glaucoma patients were randomized to 1 of 3 categories of glaucoma counseling: traditional counseling, patient-centered counseling, or patient-centered counseling with audio counseling reinforcement. Demographic and clinical information from each subject was ascertained, and all subjects completed the Glaucoma Knowledge Assessment before and after counseling sessions at the time of diagnosis and at 1-month follow-up. Patients were instructed to return to clinic for routine follow-up at 1, 3, 6, 9, and 12 months after enrollment. A multivariate logistic regression model was used to determine factors associated with appropriate clinical follow-up. Overall, only 13.5% of subjects had appropriate clinical follow-up at 1 year, defined as attending at least 3 follow-up visits during that interval, and there was no significant difference between counseling groups. The mean glaucoma knowledge assessment score (GKAS) improved by 77.6% with the initial counseling intervention (P < .0001), decreased by 17.4% within a 1-month period following initial counseling, and improved by 22.8% (P < .001) after the second counseling intervention. Monthly household income over 2500 rupees, GKAS greater than 5 after initial counseling, and undergoing any ocular surgical procedure were all independent predictors of appropriate follow-up. While all 3 counseling methods resulted in transient improvement of patient knowledge regarding glaucomatous disease, follow-up rates were poor for all groups. Poor retention of glaucoma knowledge may impact the likelihood of patient follow-up, and reinforcement with repeated counseling may be beneficial with regard to both disease knowledge and follow-up.


Review of Early Postoperative Metrics for Children Undergoing Resection of Congenital Pulmonary Airway Malformations and Report of Pleuropulmonary Blastoma at a Single Institution.
The purpose of this study is to describe a single institution's 11-year experience treating children with congenital pulmonary airway malformations (CPAMs) and pleuropulmonary blastoma (PPB). An institutional database was sampled for all patients aged 0 to 18 years from January 1, 2005, to December 31, 2015. Patients with a pathologic diagnosis of CPAM or PPB during this period were reviewed. A total of 51 patients with a pathologic diagnosis of CPAM (n = 45; 88.2%) or PPB (n = 6; 11.8%) underwent surgical resection. Among patients treated for PPB, one death occurred approximately 13 months after diagnosis. Although four patients with PPB (four out of six; 66.7%) had radiographic indicators highly suggestive of malignancy prior to surgery, two had a preoperative diagnosis of CPAM (two out of six; 33.3%). Twenty-four patients (24 out of 45; 53.3%) with CPAM underwent resection after developing symptoms and 21 (21 out of 45; 46.7%) were symptomatic at the time of surgery. Mann-Whitney's tests revealed a statistically significant difference in postoperative length of stay (median: 6 vs. 3 days; p < 0.001) and days with thoracostomy tube in place (median 3 vs. 2 days; p = 0.003) for symptomatic versus asymptomatic patients, respectively. CPAM patients appear to recover faster from surgery, if performed before the onset of symptoms. There may be a benefit to waiting until at least 3 months of age to complete resection in the asymptomatic patient. A low threshold for resection should be maintained in patients where delineating CPAM from PPB is difficult.


Telehealth examination of the lumbar spine.
Telehealth evaluations of musculoskeletal conditions have increased due to the stay-at-home policies enacted during the COVID-19 pandemic. Back pain is one of the most common complaints in primary care. While telehealth may never supplant in-person evaluation of back pain, it is imperative in a changing world to learn to perform this evaluation via telephone or video. Virtual visits rely on history-taking and patient self-reported descriptions of pain elicited from self-palpation or specific movements while on the telephone with the clinician. Video examinations provide a unique way of evaluating the lower back compared to telephone because of the ability to visualize the actions of the patient. To create an evaluation pathway for examination of the lumbar spine via telehealth. Our group has created a step-by-step evaluation pathway to help physicians direct their patients through typical lumbar examination elements, including inspection, palpation, range of motion, and strength, special, and functional testing. We have developed a table of questions and instructions and a glossary of images of each maneuver to facilitate lumbar spine examination via telemedicine. This paper provides a guide for extracting clinically relevant information while performing telemedicine examinations of the lumbar spine.


High prevalence of papillomavirus-associated penile intraepithelial neoplasia in sexual partners of women with cervical intraepithelial neoplasia.
To determine whether neoplastic cervical lesions in women are associated with papillomavirus infections in their sexual partners, we used a colposcope to examine male sexual partners of women with cervical flat condyloma (294 cases) or cervical intraepithelial neoplasia (186 cases), before and after 5 percent acetic acid was applied to the penis and the anogenital area. Condylomata acuminata, papules, and macules were observed in 309 of the 480 men (64.4 percent). In 204 of them (42.5 percent), macules or slightly elevated papules were detected only after application of acetic acid. Condylomata acuminata or lesions showing histologic features of condyloma were found in 121 partners (41.2 percent) of women with condyloma, but in only 10 partners (5.4 percent) of women with cervical intraepithelial neoplasia. Penile lesions showing histologic features of intraepithelial neoplasia were found in 61 partners (32.8 percent) of women with cervical intraepithelial neoplasia, but in only 4 partners (1.4 percent) of women with flat condyloma. Thirty-six (60 percent) of the 60 macules or papules tested contained papillomavirus DNA sequences. Human papillomavirus types 16 and 33 were almost exclusively found in penile intraepithelial neoplasia. Type 6, type 11, and the recently recognized type 42 were found in lesions showing features of condyloma or minimal histologic changes. As yet uncharacterized papillomaviruses were found in 15 percent of the specimens. These data support the concept that cervical carcinomas and precancerous lesions in women may be associated with genital papillomavirus infection in their male sexual partners.


Evaluation of CD44 variant expression in oral, head and neck squamous cell carcinomas using a triple approach and its clinical significance.
Cancer stem cells possess the qualities of self-renewal, tumorigenesis and the ability to recapitulate a heterogeneous tumor. Our group was the first to isolate head and neck squamous cell carcinoma (HNSCC) stem cells using the cell surface marker CD44. CD44 is a trans-membrane glycoprotein with a multitude of key-functions that regulate cancer cell proliferation and metastasis. The variety of CD44 functions is due to tissue-specific patterns of glycosylation of the extracellular portion, and to the multiple protein isoforms (CD44 variants, CD44v) generated by alternative splicing. This study investigates the expression pattern of CD44 variants in HNSCC. Ten cell lines from the most common HNSCC locations and representative of various clinical outcomes were assayed by quantitative realtime PCR, flow cytometry and immunofluorescence comparatively with normal oral keratinocytes. The CD44 v4 and v6 were exclusively abundant in HNSCC while the isoform v1,2 was expressed in normal oral keratinocytes. Of interest, the highest level of CD44v6 expression was detected in advanced metastatic HNSCC, suggesting a link between CD44v6 expression and HNSCC metastasis, while the highest CD44v4 was detected in a stage IV HNSCC refractory to chemotherapy which developed recurrence. Oral-derived HNSCC expressed the highest CD44v4 and v6, and levels corresponded with staging, showing also an increasing tendency with recurrence and metastasis. CD44v were detected predominantly in smaller cells (a characteristic that has been associated with stem cell properties) or cells with mesenchymal morphology (a characteristic that has been associated with the migratory and invasive potential of epithelial tumor cells), suggesting that CD44v differential expression in HNSCC may be representative of the morphological changes inherent during tumor progression towards a more aggressive potential, and thus contributing to the individual tumor biology. The mechanism of CD44 variant involvement in HNSCC progression and metastasis is under investigation.


Inhibitory effect of Thymus vulgaris and Origanum vulgare essential oils on virulence factors of phytopathogenic Pseudomonas syringae strains.
Pseudomonas syringae is a phytopathogenic bacterium that causes lesions in leaves during the colonisation process. The damage is associated with production of many virulence factors, such as biofilm and phytotoxins. The essential oils of Thymus vulgaris (thyme) and Origanum vulgare (oregano) have been demonstrated to inhibit P. syringae. The aim of this study was to investigate the effects of T. vulgaris and O. vulgare essential oils on production of virulence factors of phytopathogenic P. syringae strains, including anti-biofilm and anti-toxins activities. The broth microdilution method was used for determination of MIC and biofilm inhibition assays. Coronatine, syringomycin and tabtoxin were pheno- and genotypically evaluated. Both oils showed good inhibitory activity against P. syringae, with MIC values from 1.43 to 11.5 mg·ml-1 for thyme and 5.8 to 11.6 mg·ml-1 for oregano. Biofilm formation, production of coronatine, syringomycin and tabtoxin were inhibited by thyme and oregano essential oil in most strains. The results presented here are promising, demonstrating the bactericidal activity and reduction of virulence factor production after treatment with thyme and oregano oil, providing insight into how they exert their antibacterial activity. These natural products could be considered in the future for the control of diseases caused by P. syringae.


Adenovirus E1A requires synthesis of a cellular protein to establish a stable transcription complex in injected Xenopus laevis oocytes.
The Escherichia coli-expressed adenovirus E1A 13S mRNA product injected into Xenopus oocytes was active, as assessed by its ability to stimulate the transcription of an injected gene which is normally responsive to E1A in mammalian cells. In the presence of the protein synthesis inhibitors pactamycin or cycloheximide, E1A was correctly posttranslationally modified (phosphorylated) and transported to the nucleus; but it failed to stimulate the transcription of an injected gene containing the human heat shock protein 70 promoter. The basal (unstimulated) level of transcription of the gene was unaffected by these inhibitors. If oocytes were cultured in the presence of cycloheximide after E1A stimulated transcription, however, the high level of transcription was maintained for several hours without new protein synthesis. Results of competition studies with the same promoter (the heat shock protein 70 promoter) linked to two marked genes demonstrated that once the induction of transcription by E1A took place, the stimulated levels of transcription were maintained, even when they were challenged with excess competitor DNA. Results of these studies suggest that E1A requires the synthesis of a cellular protein to form a stable transcription complex.


Multi-spin echo spatial encoding provides three-fold improvement of temperature precision during intermolecular zero quantum thermometry.
Intermolecular multiple quantum coherences (iMQCs) are a source of MR contrast with applications including temperature imaging, anisotropy mapping, and brown fat imaging. Because all applications are limited by signal-to-noise ratio (SNR), we developed a pulse sequence that detects intermolecular zero quantum coherences with improved SNR. A previously developed pulse sequence that detects iMQCs, HOMOGENIZED with off resonance transfer (HOT), was modified with a multi-spin echo spatial encoding scheme (MSE-HOT). MSE-HOT uses a series of refocusing pulses to generate a stack of images that are averaged in postprocessing for higher SNR. MSE-HOT performance was quantified by measuring its temperature accuracy and precision during hyperthermia of ex vivo red bone marrow samples. MSE-HOT yielded a three-fold improvement in temperature precision relative to previous pulse sequences. Sources of improved precision were 1) echo averaging and 2) suppression of J-coupling in the methylene protons of fat. MSE-HOT measured temperature change with an accuracy of 0.6°C. MSE-HOT improved the temperature accuracy and precision of HOT to a level that is sufficient for hyperthermia of bone marrow.


Characterization of CsTSI in the Biosynthesis of Theanine in Tea Plants (Camellia sinensis).
Theanine is a unique major amino acid in tea plants responsible for umami taste and mental health benefits of tea. However, theanine biosynthesis and physiological role in tea plants are not fully understood. Here, we demonstrate that tea plant theanine synthetase is encoded by a glutamine synthetase gene CsTSI. The expression pattern of CsTSI is closely correlated with theanine and glutamine levels in various tissues. CsTSI transcripts were accumulated in root tip epidermal cells, pericycle and procambial cells, where CsTSI presents as a cytosolic protein. Ectopic expression of the gene in Arabidopsis led to greater glutamine and theanine production than controls when fed with ethylamine (EA). RNAi knockdown or overexpression of CsTSI in tea plant hairy roots reduced or enhanced theanine and glutamine contents, respectively, compared with controls. The CsTSI recombinant enzymes used glutamate as an acceptor and ammonium or EA as a donor to synthesize glutamine and theanine, respectively. CsTSI expression in tea roots responded to nitrogen supply and deprivation and was correlated with theanine contents. This study provides fresh insights into the molecular basis for the biosynthesis of theanine, which may facilitate the breeding of high-theanine tea plants for improving the nutritional benefit of tea.


The physics of boundary conditions in reaction-diffusion problems.
The use of fully or partially absorbing boundary conditions for diffusion-based problems has become paradigmatic in physical chemistry and biochemistry to describe reactions occurring in solutions or in living media. However, as chemical states may indeed disappear, particles cannot, unless such degradation happens physically and should, thus, be accounted for explicitly. Here, we introduce a simple, yet general idea that allows one to derive the appropriate boundary conditions self-consistently from the chemical reaction scheme and the geometry of the physical reaction boundaries. As an illustration, we consider two paradigmatic examples, where the known results are recovered by taking specific physical limits. More generally, we demonstrate that our mathematical analysis delivers physical insight that cannot be accessed through standard treatments.


Reversible inactivation of alpha-ketoglutarate dehydrogenase in response to alterations in the mitochondrial glutathione status.
In a previous study, we found that treatment of rat heart mitochondria with H(2)O(2) resulted in a decline and subsequent recovery in the rate of state 3 NADH-linked respiration. These effects were shown to be mediated by reversible alterations in NAD(P)H utilization and in the activities of specific Krebs cycle enzymes alpha-ketoglutarate dehydrogenase (KGDH) and succinate dehydrogenase. The purpose of the current study was to examine potential mechanism(s) by which H(2)O(2) reversibly alters KGDH activity. We report here that inactivation is not simply due to direct interaction of H(2)O(2) with KGDH. In addition, incubation of mitochondria with deferroxamine, an iron chelator, or 1,3-dimethyl-2-thiourea, an oxygen radical scavenger, prior to addition of H(2)O(2) did not alter the rate or extent of inactivation. Thus, inactivation does not appear to involve a more potent oxygen radical formed upon metal-catalyzed oxidation. Inactive KGDH from H(2)O(2)-treated mitochondria was reactivated with dithiothreitol, implicating oxidation of a protein sulfhydryl(s). However, the thioredoxin system had no effect, indicating that enzyme inactivation is not due to the formation of intra- or intermolecular disulfide(s) or a sulfenic acid. Upon incubation of mitochondria with H(2)O(2), reduced GSH levels fell rapidly prior to enzyme inactivation but recovered at the same time as enzyme activity. Importantly, treatment of inactive KGDH with glutaredoxin facilitated the GSH-dependent recovery of KGDH activity. Glutaredoxin is characterized as a specific and efficient catalyst of protein deglutathionylation. Thus, the results of the current study indicate that KGDH activity appears to be modulated through enzymatic glutathionylation and deglutathionylation. These studies demonstrate a novel mechanism by which KGDH activity and mitochondrial function can be modulated by redox status.


Highly specific bacteriophage-associated polysaccharide hydrolases for Klebsiella aerogenes type 8.
Two phage-bound polysaccharide hydrolases specific for Klebsiella aerogenes type 8 exopolysaccharides were isolated. Each enzyme was specific for the polysaccharide produced by the host strain. One enzyme hydrolysed a pyruvylated and acetylated polymer, while the other only acted on the substrate lacking these substituents. Both enzymes were endogalactosidases releasing tetrasaccharides from their substrates which were only hydrolysed to a limited extent.


Mitochondrial genome-wide analysis of nuclear DNA methylation quantitative trait loci.
Mitochondria have a complex communication network with the surrounding cell and can alter nuclear DNA methylation (DNAm). Variation in the mitochondrial DNA (mtDNA) has also been linked to differential DNAm. Genome-wide association studies have identified numerous DNAm quantitative trait loci, but these studies have not examined the mitochondrial genome. Herein, we quantified nuclear DNAm from blood and conducted a mitochondrial genome-wide association study of DNAm, with an additional emphasis on sex- and prediabetes-specific heterogeneity. We used the Young Finns Study (n = 926) with sequenced mtDNA genotypes as a discovery sample and sought replication in the Ludwigshafen Risk and Cardiovascular Health study (n = 2317). We identified numerous significant associations in the discovery phase (P < 10-9), but they were not replicated when accounting for multiple testing. In total, 27 associations were nominally replicated with a P < 0.05. The replication analysis presented no evidence of sex- or prediabetes-specific heterogeneity. The 27 associations were included in a joint meta-analysis of the two cohorts, and 19 DNAm sites associated with mtDNA variants, while four other sites showed haplogroup associations. An expression quantitative trait methylation analysis was performed for the identified DNAm sites, pinpointing two statistically significant associations. This study provides evidence of a mitochondrial genetic control of nuclear DNAm with little evidence found for sex- and prediabetes-specific effects. The lack of a comparable mtDNA data set for replication is a limitation in our study and further studies are needed to validate our results.


A complexity-informed in-depth case study into the sustainability and impact of a culture of health: The TR14ers community youth dance group.
There are calls for researchers to study existing community assets and activities that appear to improve health and have achieved longevity. The TR14ers Community Dance Charity Limited is a community youth dance group that has been running since 2005 providing free weekly sessions for children and adolescents in an economically disadvantaged town in the UK. An in-depth case study employing qualitative, quantitative and participatory methods was undertaken with the TR14ers (current participants and those who have left, co-ordinators and families) over 6 months with the aim of understanding the sustainable processes and impact of the Group. The 12 complex systems' leverage points described by Meadows and the five domains of adolescent wellbeing developed by the United Nations H6+ Technical Working Group on Adolescent Health and Well-Being were used as frameworks to recognise the complexity of community assets like the TR14ers. The quantitative and qualitative data indicated that being part of the TR14ers contributed to multiple health and wellbeing outcomes. The positive experiences of being a TR14er led members to actively recruit others through word of mouth and public performances. Central to the TR14ers is a commitment to children's rights, which is communicated formally and informally throughout the membership informing how and what the Group does, leading to the structure and delivery of the Group evolving over time. Members sought to ensure the sustainability of the Group after they had left and were keen to mentor younger members to develop and become the leaders. Based on the insights from this case study we suggest that efforts to develop cultures of health, like the TR14ers, should focus on the core values of the activity or intervention that underpin what it does and how within the local context.


A Phase I, First-in-Human Study of AMG 780, an Angiopoietin-1 and -2 Inhibitor, in Patients with Advanced Solid Tumors.
To assess the toxicity, pharmacokinetics, tumor vascular response, tumor response, and pharmacodynamics of AMG 780, a mAb designed to inhibit the interaction between angiopoietin-1 and -2 and the Tie2 receptor. This was a phase I dose-escalation study of patients with advanced solid tumors refractory to standard treatment without previous antiangiogenic treatment. AMG 780 was administered by intravenous infusion every 2 weeks in doses from 0.1 to 30 mg/kg. The primary endpoints were incidences of dose-limiting toxicity (DLT) and adverse events (AE), and pharmacokinetics. Secondary endpoints included tumor response, changes in tumor volume and vascularity, and anti-AMG 780 antibody formation. Forty-five patients were enrolled across nine dose cohorts. Three patients had DLTs (0.6, 10, and 30 mg/kg), none of which prevented dose escalation. At 30 mg/kg, no MTD was reached. Pharmacokinetics of AMG 780 were dose proportional; median terminal elimination half-life was 8 to 13 days. No anti-AMG 780 antibodies were detected. At week 5, 6 of 16 evaluable patients had a >20% decrease in volume transfer constant (K(trans)), suggesting reduced capillary blood flow/permeability. The most frequent AEs were hypoalbuminemia (33%), peripheral edema (29%), decreased appetite (27%), and fatigue (27%). Among 35 evaluable patients, none had an objective response; 8 achieved stable disease. AMG 780 could be administered at doses up to 30 mg/kg every 2 weeks in patients with advanced solid tumors. AMG 780 treatment resulted in tumor vascular effects in some patients. AEs were in line with toxicity associated with antiangiopoietin treatment. Clin Cancer Res; 22(18); 4574-84. ©2016 AACR.


Malignant fibrous histiocytomas of the head and neck.
Fibrous histiocytomas are soft tissue neoplasms of histiocytic origin showing partial fibroblastic and histiocytic differentiation. The histopathology of these tumors and their nomenclature are discussed. A case of primary intraosseous malignant fibrous histiocytoma of the mandible is presented and added to the previously reported nine cases. One hundred and ten cases of fibrous histiocytoma are reviewed, a rapidly increasing number of reported cases in the deep tissues of the head and neck region. No definite reliable histologic criteria have been found to predict the true malignant lesions. Malignant fibrous histiocytomas show the most aggressive behavior in the head and neck region with a metastatic rate of 30-42%. Aggressive surgery remains the treatment of choice with a five year survival rate of approximately 60%, chemotherapy and radiotherapy being used as adjunctive treatment.


Hepatocellular carcinoma: A global view.
Hepatocellular carcinoma (HCC) is a global health problem, although developing countries are disproportionally affected: over 80% of HCCs occur in such regions. About three-quarters of HCCs are attributed to chronic HBV and HCV infections. In areas endemic for HCV and HBV, viral transmission occurs at an early age, and infected individuals develop HCC in mid-adulthood. As these are their most productive years of life, HCC accounts for a substantial burden on the health-care system and drain of productive capacity in the low-income and middle-income countries most affected by HCV and HBV infections. Environments with disparate resource levels require different strategies for the optimal management of HCC. In high-resource environments, guidelines from the American Association for the Study of Liver Diseases or European Association for the Study of the Liver should be applied. In intermediate-resource or low-resource environments, the fundamental focus should be on primary prevention of HCC, through universal HBV vaccination, taking appropriate precautions and antiviral treatments. In intermediate-resource and low-resource environments, the infrastructure and capacity for abdominal ultrasonography, percutaneous ethanol injection, radiofrequency ablation and surgical resection should be established. Programs to provide targeted therapy at low cost, similar to the approach used for HIV therapy in the developing world, should be pursued.


Microsurgical Management of the Middle Cerebral Artery Bifurcation Aneurysms: An Anatomic Feasibility Study.
The proper head positioning decreases the surgical complications by enabling a better surgical maneuverability. Middle cerebral artery (MCA) bifurcation aneurysms have been classified by Dashti et al. [Surg Neurol. 2007 May;67(5):441-56] as the intertruncal, inferior, lateral, insular, and complex types based on dome projection. Our aim was to identify the optimum head positions and to explain the anatomic variables, which may affect the surgical strategy of MCA bifurcation aneurysms. The lateral supraorbital approach bilaterally was performed in the 4 cadaveric heads. All steps of the dissection were recorded using digital camera. The distal Sylvian fissure (SF) dissection may be preferred for insular type and the proximal SF dissection may be preferred for all other types. Fifteen degrees head rotation was found as the most suitable position for the intertruncal, lateral type and subtype of complex aneurysms related with superior trunk. Thirty degrees head rotation was found the most suitable position for the inferior type, insular type, and subtype of complex aneurysms related with inferior trunk. The head positioning in middle cerebral bifurcation aneurysms surgery is a critical step. It should be tailored according to the projection and its relationship with the parent vessels of the middle cerebral bifurcation.


A staging scheme for the development of the moth midge Clogmia albipunctata.
Model organisms, such as Drosophila melanogaster, allow us to address a wide range of biological questions with experimental rigour. However, studies in model species need to be complemented by comparative studies if we are to fully understand the functional properties and evolutionary history of developmental processes. The establishment of new model organisms is crucial for this purpose. One of the first essential steps to establish a species as an experimental model is to carefully describe its life cycle and development. The resulting staging scheme serves as a framework for molecular studies, and allows us to homologise developmental processes between species. In this paper, we have characterised the life cycle and development of an emerging non-drosophilid dipteran model system: the moth midge Clogmia albipunctata. In particular, we focus on early embryogenesis (cleavage and blastoderm cycles before gastrulation), on formation and retraction of extraembryonic tissues, and on formation of the germ line. Considering the large evolutionary distance between the two species (approximately 250 million years), we find that the development of C. albipunctata is remarkably conserved compared to D. melanogaster. On the other hand, we detect significant differences in morphology and timing affecting the development of extraembryonic tissues and the germ line. Moreover, C. albipunctata shows several heterochronic shifts, and lacks head involution and associated processes during late stages of development.


Setting the bar: Standards for ecosystem services.
Progress in ecosystem service science has been rapid, and there is now a healthy appetite among key public and private sector decision makers for this science. However, changing policy and management is a long-term project, one that raises a number of specific practical challenges. One impediment to broad adoption of ecosystem service information is the lack of standards that define terminology, acceptable data and methods, and reporting requirements. Ecosystem service standards should be tailored to specific use contexts, such as national income and wealth accounts, corporate sustainability reporting, land-use planning, and environmental impact assessments. Many standard-setting organizations already exist, and the research community will make the most headway toward rapid uptake of ecosystem service science by working directly with these organizations. Progress has been made in aligning with existing organizations in areas such as product certification and sustainability reporting, but a major challenge remains in mainstreaming ecosystem service information into core public and private use contexts, such as agricultural and energy subsidy design, national income accounts, and corporate accounts.


Direct analysis in real time coupled with quadrupole-Orbitrap high-resolution mass spectrometry for rapid analysis of pyrethroid preservatives in wooden food contact materials.
A methodology is presented for the determination of four pyrethroid (PYR) preservatives in wooden food contact materials (FCMs) using direct analysis in real time (DART) coupled with quadrupole-Orbitrap high-resolution mass spectrometry (Q-Orbitrap HRMS). The sampling mode and critical parameters of the DART-Q-Orbitrap HRMS protocol were systematically investigated. Good linearity was achieved for the four analytes with correlation coefficients all greater than 0.99. The limits of detection (LODs) and limits of quantitation (LOQs) of the method were in the range of 0.04-0.20 mg kg-1 and 0.10-0.50 mg kg-1, respectively. The mean recoveries ranged from 72.1% to 82.7% with relative standard deviations (RSDs) from 5.2% to 11.8% at three spiked levels. The developed method was proved to be suitable for rapid screening of PYRs in complex wooden FCM samples to ensure product safety and consumer health.


Oxidized thioredoxin 1 places a leash on NLRP1 inflammasome activity.
The biology of the NACHT domain and leucine-rich repeat (NLR) and pyrin domain-containing 1 (NLRP1) inflammasome has perplexed researchers since this inflammasome was first described about two decades ago. The identification of oxidized thioredoxin 1 (TRX1) as a suppressor of NLRP1 recently linked cellular redox homeostasis to NLRP1 inflammasome signaling. Now, Zhang et al. present a molecular structure of TRX1-bound NLRP1 with unprecedented detail. This structure gives key insight into regulatory mechanisms governing NLRP1 activation and offers enormous potential for structure-based anti-inflammatory drug design.


Pregnane X receptor as the "sensor and effector" in regulating epigenome.
The pregnane X receptor (PXR, NR1I2) is a ligand-activated nuclear receptor which plays an essential role in organism's metabolic detoxification system by sensing the presence of xenobiotics and triggering detoxification responses. In addition to its role in xenobiotic metabolism, PXR has pleiotropic functions in regulating immune/inflammatory responses, cell proliferation, bile acid/cholesterol metabolism, glucose and lipid metabolism, steroid/endocrine homeostasis, and bone metabolism. Recent research suggests that the PXR is required for maintaining healthy commensalism between microbiota and gut. Interestingly, the metabolites such as indole derivatives from commensal microbes serve as the ligands for the PXR in intestinal epithelium forming an intricate mutualistic interaction between host and microbiota. PXR-regulated gene responses are controlled at epigenetic level by chromatin modifications, DNA methylation and noncoding RNA. Developmental alterations of the epigenome by exposure to the xenobiotics or diseases may produce persistent changes in PXR-regulated physiological responses. These new areas of research promise to vastly increase our understanding of PXR-regulated responses. In this review we highlight recent results on the epigenetic mechanisms for the PXR-regulated gene expression and discuss the physiological significance of these findings.


Asthma, Chronic Obstructive Pulmonary Disease, and Subsequent Risk for Incident Rheumatoid Arthritis Among Women: A Prospective Cohort Study.
Inflamed airways are hypothesized to contribute to rheumatoid arthritis (RA) pathogenesis due to RA-related autoantibody production, and smoking is the strongest environmental RA risk factor. However, the role of chronic airway diseases in RA development is unclear. We undertook this study to investigate whether asthma and chronic obstructive pulmonary disease (COPD) were each associated with RA. We performed a prospective cohort study of 205,153 women in the Nurses' Health Study (NHS, 1988-2014) and NHSII (1991-2015). Exposures were self-reported physician-diagnosed asthma or COPD confirmed by validated supplemental questionnaires. The primary outcome was incident RA confirmed by medical record review by 2 rheumatologists. Covariates (including smoking pack-years/status) were assessed via biennial questionnaires. Multivariable hazard ratios (HRs) and 95% confidence intervals (CIs) for RA were estimated using Cox regression. We identified 15,148 women with confirmed asthma, 3,573 women with confirmed COPD, and 1,060 incident RA cases during 4,384,471 person-years (median 24.0 years/participant) of follow-up in the NHS and NHSII. Asthma was associated with increased RA risk (HR 1.53 [95% CI 1.24-1.88]) compared to no asthma/COPD after adjustment for covariates, including smoking pack-years/status. Asthma remained associated with increased RA risk when analyzing only never-smokers (HR 1.53 [95% CI 1.14-2.05]). COPD was also associated with increased RA risk (HR 1.89 [95% CI 1.31-2.75]). The association of COPD with RA was most pronounced in the subgroup of ever-smokers age >55 years (HR 2.20 [95% CI 1.38-3.51]). Asthma and COPD were each associated with increased risk of incident RA, independent of smoking status/intensity and other potential confounders. These results provide support for the hypothesis that chronic airway inflammation may be crucial in RA pathogenesis.


A genetic association between microcephaly and lymphedema.
We discuss a family in which microcephaly and lymphedema are co-segregating as an apparently autosomal or X-linked dominant trait. A review of each malformation is presented with reference to the known genetic patterns of each. This combination of microcephaly and lymphedema may be a unique syndrome, previously undescribed because of subtleties of expression in affected individuals.


Fate and redistribution of perfluoroalkyl acids through AFFF-impacted groundwater.
Leaching of perfluoroalkyl acids (PFAAs) from a local point source, a fire-fighting training area, has led to extensive contamination of a groundwater aquifer which has spread underneath part of a nearby town, Oakey, situated in the State of Queensland, Australia. Groundwater is extracted by residents from privately owned wells for daily activities such as watering livestock and garden beds. The concentration of 10 PFAAs in environmental and biological samples (water, soil, grass, chicken egg yolk, serum of horses, cattle and sheep), as well as human serum was investigated to determine the extent of contamination in the town and discuss fate and redistribution of PFAAs. Perfluorooctane sulfonate (PFOS) was the dominant PFAA in all matrices investigated, followed by perfluorohexane sulfonate (PFHxS). PFOS concentrations measured in water ranged between <0.17-14μg/L, concentrations of PFHxS measured between <0.07-6μg/L. PFAAs were detected in backyards (soil, grass), livestock and chicken egg yolk. Significant differences (p<0.01) in PFOS and PFHxS concentrations in two groups of cattle were found, one held within the contamination plume, the other in the vicinity but outside of the contamination plume. In human serum PFOS concentrations ranged from 38 to 381μg/L, while PFHxS ranged from 39 to 214μg/L. Highest PFOS concentrations measured in human serum were >30-fold higher compared to the general Australian population. Through use of contaminated groundwater secondary sources of PFAA contamination are created on private property, leading to further redistribution of contamination and creation of additional human exposure pathways.


Nausea prophylaxis using transdermal scopolamine in the setting of patient-controlled analgesia.
We evaluated the effectiveness of transdermal scopolamine in patients receiving morphine via patient-controlled intravenous analgesia following intra-abdominal gynecologic surgery. Soon after arrival in the post-anesthesia recovery unit (time 0), patients were randomized either to receive or not receive a postauricular transdermal scopolamine patch. Nausea and vomiting were scored on a 0-3 scale at this time and at 2, 4, 6, and 24 hours. Patients were treated with droperidol as deemed necessary by the primary care nurse. Within 2-4 hours, transdermal scopolamine patients evidenced less nausea and vomiting and required less droperidol than their counterparts who did not receive transdermal scopolamine. A significant decline in the severity of nausea was noted in the transdermal scopolamine group between 2-24 hours; significant inter-group differences were noted for changes in nausea severity during the 0-6-hour and 0-24-hour intervals. Transdermal scopolamine patients evidenced a significant (P less than .05) decrease in the severity of vomiting during the first 2 hours, significantly different from the increase in the non-transdermal scopolamine patients. After the 4-hour assessment, no transdermal scopolamine patients required droperidol; nine doses were administered to the patients who were not given transdermal scopolamine (P less than .05). Thus, transdermal scopolamine therapy appears to be an effective means of treating the nausea and vomiting that are encountered after gynecologic surgery.


Toxicity of poly(rI).poly(rC) for interferon-treated cells. An ultrastructural evaluation.
The cytotoxic effect of poly(rI).poly(rC) for interferon-treated cells is preceded by discrete ultrastructural alterations in (1) the mitochondria: swelling of the cristae and increased density of their matrix, (2) the nucleus: disorganization of the structure, and (3) the endoplasmic reticulum: slight dilatation with disorganization and depletion of the filament bundles. Our results do not allow us to conclude whether any of these alterations is a primary effect of the treatment. Possibly they are secondary to some, as yet unknown, functional defect, and only reflect incipient cell death.


Geographic variation in cardiometabolic risk distribution: A cross-sectional study of 256,525 adult residents in the Illawarra-Shoalhaven region of the NSW, Australia.
Metabolic risk factors for cardiovascular disease (CVD) warrant significant public health concern globally. This study aims to utilise the regional database of a major laboratory network to describe the geographic distribution pattern of eight different cardiometabolic risk factors (CMRFs), which in turn can potentially generate hypotheses for future research into locality specific preventive approaches. A cross-sectional design utilising de-identified laboratory data on eight CMRFs including fasting blood sugar level (FBSL); glycated haemoglobin (HbA1c); total cholesterol (TC); high density lipoprotein (HDL); albumin creatinine ratio (ACR); estimated glomerular filtration rate (eGFR); body mass index (BMI); and diabetes mellitus (DM) status was used to undertake descriptive and spatial analyses. CMRF test results were dichotomised into 'higher risk' and 'lower risk' values based on existing risk definitions. Australian Census Statistical Area Level 1 (SA1) were used as the geographic units of analysis, and an Empirical Bayes (EB) approach was used to smooth rates at SA1 level. Choropleth maps demonstrating the distribution of CMRFs rates at SA1 level were produced. Spatial clustering of CMRFs was assessed using Global Moran's I test and Local Indicators of Spatial Autocorrelation (LISA). A total of 1,132,016 test data derived from 256,525 individuals revealed significant geographic variation in the distribution of 'higher risk' CMRF findings. The populated eastern seaboard of the study region demonstrated the highest rates of CMRFs. Global Moran's I values were significant and positive at SA1 level for all CMRFs. The highest spatial autocorrelation strength was found among obesity rates (0.328), and the lowest for albuminuria (0.028). LISA tests identified significant High-High (HH) and Low-Low (LL) spatial clusters of CMRFs, with LL predominantly in the less populated northern, central and southern regions of the study area. The study describes a range of CMRFs with different distributions in the study region. The results allow generation of hypotheses to test in future research concerning location specific population health approaches.


Syndrome of inappropriate secretion of antidiuretic hormone in Hodgkin's disease.
A patient with clinical Stage IV Hodgkin's disease of the nodular sclerosis type demonstrated all features of the syndrome of inappropriate secretion of antidiuretic hormone. Electroencephalogram, radiography of the skull, and computerized tomography of the brain and cardiac, renal, thyroid, and adrenal function tests were all normal. It is suggested that the patient presents a rare case of Hodgkin's disease with inappropriate secretion of antidiuretic hormone.


Female mate preference to maximize paternal care. II. Female competition leads to monogamy.
A two-step game model of female mate preference and paternal care is examined, with a particular focus on the case of two females and two males. In a mating season, females choose their mates, and in the following breeding season males invest in paternal care, knowing the likelihood of their paternity in chicks. If parental ability is the same between individuals of each sex, the evolutionarily stable mating pattern is always monogamy. If females differ in fecundity and males differ in paternal care capacity, monogamy with assortative mating is likely to be evolutionarily stable. If the male cost function increases at a strongly accelerating rate, however, polyandry is evolutionarily stable when the difference of female fecundity is very large, but the game may have no evolutionarily stable state when the difference of female fecundity is small. The care graph (in which females are connected to males giving paternal care to their chicks) is often much simpler than the mating graph (in which females are connected to males they accepted). To be exact, no "loop" should be included in the evolutionarily stable care graph for the general case of n females and m males. This prediction is in accord with the observed prevalence of social monogamy in spite of genetic promiscuity among altricial birds.


Novel carbon dot coated alginate beads with superior stability, swelling and pH responsive drug delivery.
Novel carbon dot coated alginate beads (CA-CD) exhibiting superior stability and swelling properties have been successfully prepared. CA-CD show exceptional stability in ambient condition and are stable at room atmosphere and temperature even after 60 days. Moreover, CA-CD show excessive swelling in comparison to calcium alginate (CA) beads. The beads were characterized by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), scanning electron microscopy (SEM) and optical microscopy. The CA and CA-CD beads were investigated for their use as pH dependent sustained drug delivery vehicles taking tetracycline (TC) and tetracycline associated with β-cyclodextrin (β-TC) as model drug systems. It was observed that TC loading was 35% and 77% with CA and CA-CD, respectively. Tetracycline associated with β-cyclodextrin (β-TC) shows 48% loading for CA and much greater loading (as high as 90%) for CA-CD. At pH 1, CA-CD and CA beads show maximum drug release with TC cumulative release of 70% and 37% at 96 h, respectively. However, the delivery rates at pH 1 were slower in case of tetracycline associated with β-cyclodextrin (β-TC) loading showing 61% release for CA-CD and 22% for CA after 96 h. Thus, CA-CD can be suitably used as an effective drug delivery vehicle with maximum release obtained at pH 1 emphasizing its use in the gastrointestinal tract where pH is low. Also, the use of β-cyclodextrin with the drug as an inclusion complex renders the CA and CA-CD beads useful for slow and long-term drug administration.


Correlation of amine number and pDNA binding mechanism for trehalose-based polycations.
Glycopolymers with repeat units comprised of the disaccharide trehalose and an oligoamine of increasing amine have been previously synthesized by our group and shown to efficiently deliver pDNA (plasmid DNA) to HeLa cells while remaining relatively nontoxic. Complexes formed between the most amine-dense of these polycations and pDNA were also found to be relatively stable in serum and have low aggregation, which is desirable for in vivo gene delivery. To lend insight into these interesting results, this study was aimed at investigating the binding strength and mechanism of interaction between these macromolecules, via isothermal titration calorimetry (ITC) and ethidium bromide exclusion assays. The size of these pDNA-polymer complexes, or polyplexes, at various states of formation was determined through light scattering and zeta-potential measurements. Varying degrees of pDNA secondary structure change occurred upon interaction with the polymers, as evidenced by circular dichroism spectra through increasing molar ratios of polymer amine to DNA phosphate, and Fourier transform infrared (FT-IR) results demonstrated stronger electrostatic binding with the phosphate backbone with the least amine-dense of the series. It was concluded that, depending on the number of secondary amines in the repeat unit, these polymers interact with pDNA via different mechanisms with varying extents of electrostatic interaction and hydrogen bonding. These differing mechanisms may affect the ability of trehalose to serve as a deterrent against aggregation in serum conditions and lend insight into the roles of polymer-pDNA binding during the complex transfection process.


Diagnostic accuracy of optical coherence tomography for surgical margin assessment of feline injection-site sarcoma.
The invasive, locally aggressive nature of feline injection-site sarcomas (FISSs) poses a unique challenge for surgeons to obtain complete margins with surgical excision. Optical coherence tomography (OCT), an imaging technology that uses light waves to generate real-time views of tissue architecture, provides an emerging solution to this dilemma by allowing fast, high-resolution scanning of surgical margins. The purpose of this study was to use OCT to assess surgical margins of FISS and to evaluate the diagnostic accuracy of OCT for detecting residual cancer using six evaluators of varying experience. Five FISSs were imaged with OCT to create a training set of OCT images that were compared with histopathology. Next, 25 FISSs were imaged with OCT prior to histopathology. Six evaluators of varying experience participated in a training session on OCT imaging after which each of the evaluators was given a dataset that included OCT images and videos to score on a scale from cancerous to non-cancerous. Diagnostic accuracy statistics were calculated. The overall sensitivity and specificity for classification of OCT images by evaluators were 78.9% and 77.6%, respectively. Correct classification rate of OCT images was associated with experience, while individual sensitivities and specificities had more variation between experience groups. This study demonstrates the ability of evaluators to correctly classify OCT images with overall low levels of experience and training and also illustrates areas where increased training can improve accuracy of evaluators in interpretation of OCT surgical margin images.


Calcium phosphate in catheter encrustation.
Encrusted catheters from nine female patients were the source of samples of deposits which were examined by X-ray diffraction, atomic absorption spectroscopy, infra-red spectroscopy and extended X-ray absorption fine structure (EXAFS) spectroscopy. In eight samples the only crystalline phase which could be clearly distinguished by X-ray diffraction was ammonium magnesium orthophosphate hexahydrate, NH4MgPO4 X 6H2O, which occurs naturally as the mineral struvite. However, atomic absorption spectroscopy revealed an appreciable concentration of calcium in all samples. Calcium phosphates have previously been detected in catheter deposits. Infra-red and EXAFS spectra were consistent with the calcium phosphate being present as a poorly crystalline hydroxyapatite. Thus the deposits appear to consist of a mixture of crystalline struvite and a form of hydroxyapatite which is not fully crystalline.


Patient attitudes toward resident involvement in cataract surgery.
To assess how patients in an academic ophthalmology practice feel regarding the involvement of residents in their cataract surgery. Using an anonymous survey, we asked patients with cataracts about issues related to resident involvement in cataract surgery, including informed consent, how likely they were to agree to resident involvement in their surgery, and the likelihood that they would seek care elsewhere if residents were to be involved in their surgery. Participants indicated they should be asked in advance if a resident may assist in (83%) or perform (96%) their surgery. The person asking permission should be the attending surgeon. Most participants would agree to resident assistance (83%), and nearly half would agree to resident performance (49%) of their cataract surgery. Participants indicated that they would be upset if the resident assisted in (45%) or performed (74%) their surgery without their expressed permission. Few would seek treatment in a setting without residents if a resident were to assist in (7%) or perform (26%) their cataract surgery. Most individuals would accept resident involvement in their cataract surgery provided full disclosure was provided by their attending surgeon.