H2-E transgenic class II-negative mice can distinguish self from nonself in susceptibility to heterologous thyroglobulins in autoimmune thyroiditis.
Susceptibility to experimental autoimmune thyroiditis (EAT) is linked to H2-A class II genes; k and s haplotypes are susceptible, while b and f are resistant. EAT is inducible with thyroglobulins (Tgs) from several mammalian species which share portions of identical sequences. But cross-activation and cross-tolerance studies with mouse (m), human (h), and porcine (p) Tg have indicated mTg-unique T-cell epitope(s), in addition to conserved, in EAT induction. The recent introduction of the HLA-DRB1*0301 (DR3) transgene rendered major histocompatibility complex (MHC) class II-negative (Ab(0)) mice susceptible to EAT induction by both hTg and mTg, suggesting usage of conserved epitopes. Here, we introduced the H2-Ea(k) transgene into resistant B10 (H2(b)) or Ab(0) mice with a defective Ea gene to provide functional surface H2E (b haplotype) expression. Surprisingly, both transgenic strains showed severe inflammation only after hTg, but not mTg, immunization, although the moderating influence of the A(b) gene in B10 was evident. In proliferative assays, hTg-primed cells did not respond to mTg, nor to conserved 12mer peptides from three primary hormonogenic sites, two of which can activate T cells for thyroiditis transfer and cytotoxicity. The vigorous response to hTg stimulation was reduced only by Ebeta(b)-specific monoclonal antibody. EAT induction with bovine and pTg showed responses similar to hTg, suggesting thyroiditogenic epitopes shared with hTg, but not mTg. This is the first demonstration of: (1) nonpermissiveness for EAT induction with mTg, normally the most thyroiditogenic Tg and the one with unique epitopes for susceptible mice, and (2) the separation of hTg from mTg in EAT induction in H2-E-transgenic mice.


USP33 is a Biomarker of Disease Recurrence in Papillary Thyroid Carcinoma.
To investigate the clinical significance and functional mechanisms of deubiquitinase USP33 in papillary thyroid carcinoma (PTC). Immunohistochemistry staining was conducted to explore the expression of USP33 in PTC tissues and adjacent normal thyroid tissues. Patients' prognosis was evaluated by disease-free survival. The prognostic role of USP33 was tested by univariate and multivariate analyses. To confirm the effect of USP33 in cell proliferation and invasion, overexpression and knockdown of USP33 were performed in two PTC cell lines. Besides, cell cycle, immunoprecipitation, and apoptosis experiments were conducted to further explore the signaling pathways. By analyzing series of 158 PTC tissues, we found that USP33 was down-regulated in tumor tissue compared with normal thyroid tissues, which was closely associated with lymph node metastasis (P<0.001). In particular, univariate and multivariate analyses indicated that USP33 was an independent prognostic biomarker for PTC, low USP33 expression indicated high recurrence risk. Cellular studies with TPC-1 and BCPAP cells demonstrated that USP33 can attenuate the cell capacities of proliferation, migration and invasion. Fluorescence-activated cell sorting experiment found no significant effect of USP33 on cell cycle, whereas the apoptotic caspase proteins were activated by USP33-overexpression. Moreover, the interaction between USP33 and Robo1 protein was identified, and knockdown of Robo1 enhancing the oncogenic effect upon USP33-knockdown, suggesting that USP33 may inhibit tumor progression through Robo1 signaling. Our data demonstrated that USP33 downregulation in PTC tissues was correlated with poor clinical outcome, which may serve as a novel biomarker and potential therapeutic target.


Environmental exposure does not explain putative maladaptation in road-adjacent populations.
While the ecological consequences of roads are well described, little is known of their role as agents of natural selection, which can shape adaptive and maladaptive responses in populations influenced by roads. This knowledge gap persists despite a growing appreciation for the influence of evolution in human-altered environments. There, insights indicate that natural selection typically results in local adaptation. Thus, populations influenced by road-induced selection should evolve fitness advantages in their local environment. Contrary to this expectation, wood frog tadpoles from roadside populations show evidence of a fitness disadvantage, consistent with local maladaptation. Specifically, in reciprocal transplants, roadside populations survive at lower rates compared to populations away from roads. A key question remaining is whether roadside environmental conditions experienced by early stage embryos induce this outcome. This represents an important missing piece in evaluating the evolutionary nature of this maladaptation pattern. Here, I address this gap using a reciprocal transplant experiment designed to test the hypothesis that embryonic exposure to roadside pond water induces a survival disadvantage. Contrary to this hypothesis, my results show that reduced survival persists when embryonic exposure is controlled. This outcome indicates that the survival disadvantage is parentally mediated, either genetically and/or through inherited environmental effects. This result suggests that roadside populations are either truly maladapted or potentially locally adapted at later life stages. I discuss these interpretations, noting that regardless of mechanism, patterns consistent with maladaptation have important implications for conservation. In light of the pervasiveness of roads, further resolution explaining maladaptive responses remains a critical challenge in conservation.


1-Iodobuta-1,3-diynes in Copper-Catalyzed Azide-Alkyne Cycloaddition: A One-Step Route to 4-Ethynyl-5-iodo-1,2,3-triazoles.
Cu-catalyzed 1,3-dipolar cycloaddition of iododiacetylenes with organic azides using iodotris(triphenylphosphine)copper(I) as a catalyst was found to be an efficient one-step synthetic route to 5-iodo-4-ethynyltriazoles. The reaction is tolerant to various functional groups in both butadiyne and azide moieties. The synthetic application of 5-iodo-4-ethynyl triazoles obtained was also evaluated: the Sonogashira coupling with alkynes resulted in unsymmetrically substituted triazole-fused enediyne systems, while the Suzuki reaction yielded the corresponding 5-aryl-4-ethynyl triazoles.


Ligelizumab for Chronic Spontaneous Urticaria.
In the majority of patients with chronic spontaneous urticaria, most currently available therapies do not result in complete symptom control. Ligelizumab is a next-generation high-affinity humanized monoclonal anti-IgE antibody. Data are limited regarding the dose-response relationship of ligelizumab and the efficacy and safety of ligelizumab as compared with omalizumab and placebo in patients who have moderate-to-severe chronic spontaneous urticaria that is inadequately controlled with H1-antihistamines at approved or increased doses, alone or in combination with H2-antihistamines or leukotriene-receptor antagonists. In a phase 2b dose-finding trial, we randomly assigned patients to receive ligelizumab at a dose of 24 mg, 72 mg, or 240 mg, omalizumab at a dose of 300 mg, or placebo, administered subcutaneously every 4 weeks for a period of 20 weeks, or a single 120-mg dose of ligelizumab. Disease symptoms of hives, itch, and angioedema were monitored by means of weekly activity scores. The main objective was to determine a dose-response relationship for the complete control of hives (indicated by a weekly hives-severity score of 0, on a scale from 0 to 21, with higher scores indicating greater severity); the primary end point of this response was assessed at week 12. Complete symptom control was indicated by a weekly urticaria activity score of 0 (on a scale from 0 to 42, with higher scores indicating greater severity). Safety was analyzed throughout the trial. A total of 382 patients underwent randomization. At week 12, a total of 30%, 51%, and 42% of the patients treated with 24 mg, 72 mg, and 240 mg, respectively, of ligelizumab had complete control of hives, as compared with 26% of the patients in the omalizumab group and no patients in the placebo group. A dose-response relationship was established. At week 12, a total of 30%, 44%, and 40% of the patients treated with 24 mg, 72 mg, and 240 mg, respectively, of ligelizumab had complete control of symptoms, as compared with 26% of the patients in the omalizumab group and no patients in the placebo group. In this small and short trial, no safety concerns regarding ligelizumab or omalizumab emerged. A higher percentage of patients had complete control of symptoms of chronic spontaneous urticaria with ligelizumab therapy of 72 mg or 240 mg than with omalizumab or placebo. (Funded by Novartis Pharma; ClinicalTrials.gov number, NCT02477332.).


Purification and characterization of 6-aminohexanoic-acid-oligomer hydrolase of Flavobacterium sp. Ki72.
6-Aminohexanoic-oligomer hydrolase of Flavobacterium sp. KI72 was purified to homogeneity by column chromatography three times, and by preparation polyacrylamide gel electrophoresis twice. The purified enzyme had the following characteristics. 1. The molecular weight was estimated to be 84000 by Sephadex G-200 molecular-sieve chromatography. The enzyme consisted of two homologous subunits of 42000, judged from sodium dodecylsulfate/polyacrylamide gel electrophoresis. 2. The optimum pH for activity was between 8 and 9, the optimum temperature was 40 degrees C for a 1-h reaction. The Michaelis-Menten constants and turnover numbers for the 6-aminohexanoic acid dimer and trimer were 5.9 mM and 2.4 s-1, and 6.2 mM and 2.0 s-1 respectively. 3. The enzyme was inhibited by 0.37 mM diisopropylfluorophosphate and by 0.013 mM p-chloromercuribenzoate. 4. The enzyme was active on 6-aminohexanoic acid oligomers from dimer to hexamer and icosamer but not on hectamer, and the activity decreased with the increase of the polymerization number of the oligomer. The oligomers were hydrolyzed so as to remove the 6-aminohexanoic acid residue successively from the amino terminus. The enzyme could not hydrolyze other linear amides, cyclic amides, dipeptides, tripeptides or casein. 5. 6-aminohexanoic-acid-oligomer hydrolase was classified as a new member of the linear amidases (EC 3.5.1.-).


Truly late onset of eating disorders: a study of 11 cases averaging 60 years of age at presentation.
To study late-onset cases of eating disorders in order to (1) document the occurrence of these cases as truly new onset, even if postmenopausal; (2) to alert clinicians to the category of late-onset eating disorders, especially clinical features and treatment response; (3) to challenge some prevailing assumptions of etiology. Selection of cases of eating disorders with first onset after age 40 that met DSM-IV criteria, by review of eating disorders admissions to three university hospital programs. Eleven patients, approximately 1% of all cases of eating disorders, had first onset of an eating disorder after age 40 and as late as 77, with an average onset of 56 and clinical presentation at 60 years. They met DSM-IV criteria for all subtypes of eating disorders. In general, concurrent medical and comorbid psychiatric symptoms made recognition and treatment more complex. Truly late-onset cases do occur, challenging etiological theories requiring adolescent age of onset, premenopausal endocrine functioning, or adolescent psychodynamic conflicts. Late-occurring cases, after accurate diagnosis, require an appreciation of psychological themes pertinent to this age group, such as bereavement or unresolved body image issues. Age by itself is no barrier to onset of eating disorders, which may occur whenever self-starvation and/or binge-purge behaviors become entrenched as sustaining behaviors for amelioration of psychodynamic conflicts, mood disorders, or interpersonal distress.


The Influence of Sodium Fluoride on the Growth of Ameloblasts and Kidney Proximal Tubular Cells.
Fluoride has toxic potential particularly for teeth, bones, and kidney. This study was aimed to investigate the NaF exposure effects on the growth of ameloblasts and kidney proximal tubular cells. Adult male healthy rats were used as experiment models, divided into control and NaF-induced groups. The expression of amelogenin, Bcl-2, and caspase-3 were significantly different in the control and NaF-induced group (P < 0.05). There was no correlation among these proteins in the control group but significant correlation in the NaF-induced group (r = 0.694). There was a significant correlation in proximal tubular cells, as seen from the increase of caspase-3 in the NaF-induced group (r = 0.715).


Morphological changes to somatotroph cells and in vitro individual GH release, in male rats treated with recombinant human GH.
The effect of in vivo chronic administration of recombinant human growth hormone (rhGH) on morphology and individual GH release in somatotroph cells was evaluated in young male Wistar rats. Over an 18-day period, 30-day-old male rats were injected daily with 1.5 1U rhGH/kg (GPG group) or saline (VPG group) by subcutaneous injection. Electron-immunocytochemical, ultrastructural and morphometric studies of somatotroph cells were carried out. Additionally, rat pituitary cells were dispersed and overall and individual GH release was studied by radioimmunoassay and cell immunoblot assay (quantified by image analysis), respectively. The ultrastructure and size of somatotroph cells did not change, but volume density of secretion granules was reduced (p<0.01) by previous in vivo GH treatment. At four days, basal GH release of rat pituitary cell monolayer cultures was lower in the GPG group than in the VPG group (p<0.05); after 12 hours of culture, GHRH stimulation of GH release was lower in the GPG group than in the VPG group (p<0.05), and GHRH+SRIH inhibited GH release in the GPG group (p<0.05), but not in the VPG group. The percentage of somatotroph cells was not modified, but the ratio of strongly/weakly GH-immunostained cells had changed; weakly GH-immunostained cells increased from 34% to 55%. Moreover, in vitro treatment with GHRH, SRIH, and both, easily changed the strongly/weakly GH-immunostained cell ratio. Individual GH release, however, was not changed by previous in vivo GH treatment, although GHRH preferably stimulated a subpopulation of GH cells and SRIH did not inhibit individual GH release. These data suggest that exogenous chronic rhGH treatment down-regulates somatotroph function by modifying the proportion of GH cell subpopulation.


Plasma therapy for severe hemolytic-uremic syndrome in children in Atlantic Canada.
Clinical reports have suggested that therapy with fresh frozen plasma is a useful adjunct in the management of the hemolytic-uremic syndrome (HUS). We reviewed the charts of 36 children with severe HUS who were treated at the Izaac Walton Killam Hospital for Children, Halifax, over 10 years to assess the effectiveness of plasma therapy. All children who required specific supportive therapy for renal dysfunction, hemolysis or serious extrarenal complications were included. We compared the outcome of 18 children who received plasma therapy from 1982 to 1987 with that of 18 children who did not. The two groups were similar with regard to the severity of HUS, the length of hospital stay, the duration of renal dysfunction and the incidence of disease-related complications, such as seizures, enterocolitis and cardiomyopathy. At discharge the prevalence of hypertension was higher in the plasma therapy group than in the control group. Plasma therapy did not demonstrate any benefit that would outweigh the risk of fluid overload, hyperproteinemia and transmission of viral infection.


Hernias as a cause of chronic pelvic pain in women.
Chronic pelvic pain in women due to hernias may be misdiagnosed by practicing clinicians. These fascial defects, their symptoms, physical findings, and proper treatment must be known in order to help women experiencing this form of chronic pelvic pain. All procedures were performed by the primary author using standard laparoscopic tension-free mesh techniques. The study included 264 patients referred to a chronic pelvic pain clinic, who underwent 386 laparoscopic surgical repairs of hernial defects. Ninety percent of the patients underwent concomitant procedures appropriate for their multiple pain generators. Length of follow-up is 1.53 years (range, 2 months to 5.5 years). Evaluation of patients' pain component from the repaired hernia was recorded. There have been no recurrences. One persistent ilioinguinal neuropathy from an inguinal hernia repair (.4%) has occurred. All other patients received relief of their hernia pain (99.6%). Four complications from concomitant surgeries (1.5%) are reported. Laparoscopic treatment of hernia pain in women is effective in relieving chronic pain and has a low recurrence and complication rate in the hands of experienced laparoscopists.


A pilot study of macrophage responses to silk fibroin particles.
Silk fibroin (SF) shows promise for tissue engineering and other biomedical applications due to its excellent biocompatibility, unique biomechanical properties, and controllable biodegradability. The particulate form of SF materials may have many potential uses, including the use as a filler for tissue defects or as a controlled-release agent for drug delivery. However, many past in vivo and in vitro studies evaluating the biocompatibility and biodegradability of SF have involved bulk implants. It is essential to evaluate the inflammatory effects of SF particles before further use. In this study, two different sizes of SF particles were evaluated to assess their impact on the release of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6, in comparison with lipopolysaccharide positive control stimulation. The inflammatory processes were characterized using real-time reverse transcription polymerase chain reaction, enzyme-linked immunosorbent assay, and light microscopy evaluations. The results indicated that small silk fibroin particles and large silk fibroin particles, in culture with RAW 264.7 murine macrophage cells for 24 h, caused up-regulation of mRNA coding for TNF-α, which indicated that both size of particles have potential inflammatory effects. There was a statistically significant increase in this up-regulation under small silk fibroin stimulation. However, the immunosorbent assay suggested that there was virtually no observed release of IL-1β, IL-6, or TNF-α, relative to the control group. The results suggest that SF particles of the chosen dimensions may have good biocompatibility in culture with RAW 264.7 murine macrophages.


Effects of comsumption of high and low nicotine tobacco byManduca sexta (Lepidoptera: Sphingidae) on survival of gregarious endoparasitoidCotesia congregata (Hymenoptera: Braconidae).
The significance of nicotine in the three trophic level interaction involving tobacco (Nicotiana tabacum), the tobacco hornworm (Manduca sexta), and the parasitoidCotesia congregata was investigated in field plots of two varieties of tobacco which had about a 10-fold difference in their nicotine content. WhileM. sexta mortality, rates of parasitism byC. congregata, and the total number ofC. congregata larvae produced per host were similar on each of the two varieties, the number of parasitoids reaching adult-hood on the low nicotine treatment was nearly twice that on the high nicotine treatment. This difference was due to the significantly greater proportion of parasitoid larvae which failed to emerge from the host or that died prior to pupation after emerging from hosts which fed on the high nicotine variety. A greater proportion of larvae from hosts which fed on the low nicotine tobacco died as pupae. No treatment differences occurred for either sex of the parasitoid in individual dry weight, longevity, or pupal development time, except that female pupal duration was prolonged in the high nicotine treatment. These results support the suggestion that plant allelochemicals, which may function to provide plant resistance against pest herbivores, can be detrimental to natural enemies of the pest.


Cephamycins, a new family of beta-lactam antibiotics. 3. In vitro studies.
Cephamycins A, B, and C are naturally produced cephalosporin-type antibiotics. Although A and B were found to be more active than C against gram-positive organisms, they were not so active against such strains as are cephalosporin C or the semisynthetic antibiotics cephaloridine and cephalothin. Against gram-negative organisms, cephamycin C was more active than A or B and, in general, was as active as the cephalosporins. In addition, cephamycin C was active in vitro against clinically isolated strains resistant to the cephalosporins, such as Proteus, Providencia, and Escherichia coli. The in vitro antibacterial activity of cephamycin C, cephalothin, and cephaloridine is primarily bactericidal. A 10,000-fold increase in inoculum of a strain of Proteus mirabilis resulted in 200-fold or greater increases in minimal inhibitory and minimal bactericidal end points of cephalothin and cephaloridine, but only 10- and 16-fold increases, respectively, for cephamycin C. After 15 passages through antibiotic-containing broths, during which time a culture of E. coli showed an increase in minimal inhibitory concentrations of streptomycin of >1,000-fold, end points for cephamycin C increased 4-fold, for cephalothin, 1.5-to 6-fold, and for cephaloridine, 128-fold.


Experimental validation of single-iteration multigrid wavefront reconstruction at the Palomar Observatory.
Single-iteration multigrid (SIMG) wavefront reconstruction schemes were implemented and validated on the adaptive optics system at the Hale 5.1 m telescope at the Palomar Observatory. Results indicate that even the simplest such method produces a performance indistinguishable from that of the standard least-squares reconstructor for both bright and dim guide stars. SIMG provides a dramatic reduction in computational cost when compared to vector-matrix multiplication and can be implemented in parallel, making it the obvious choice for reconstruction in future large-scale adaptive optics systems.


Association analysis of the lymphocyte-specific protein tyrosine kinase (LCK) gene in type 1 diabetes.
Prior data associating the expression of lymphocyte-specific protein tyrosine kinase (LCK) with type 1 diabetes, its critical function in lymphocytes, and the linkage of the region to diabetes in the nonobese diabetic (NOD) mouse model make LCK a premier candidate for a susceptibility gene. Resequencing of LCK in 32 individuals detected seven single nucleotide polymorphisms (SNPs) with allele frequencies >3%, including four common SNPs previously reported. These and six other SNPs from dbSNP were genotyped in a two-stage strategy using 2,430 families and were all shown not to be significantly associated with type 1 diabetes. We conclude that a major role for the common LCK polymorphisms in type 1 diabetes is unlikely. However, we cannot rule out the possibility of there being a causal variant outside the exonic, intronic, and untranslated regions studied.


Hormone therapy and young-onset breast cancer.
Estrogen plus progestin hormone therapy (HT) is associated with an increased risk of postmenopausal breast cancer, but few studies have examined the impact of HT use on the risk of breast cancer in younger women. We assessed the association between estrogen plus progestin HT or unopposed estrogen HT and young-onset breast cancer using data from the Two Sister Study (2008-2010), a sister-matched study of 1,419 cases diagnosed with breast cancer before the age of 50 years and 1,665 controls. We assessed exposures up to a family-specific index age to ensure comparable opportunities for exposures and used propensity scores to control for birth cohort effects on HT use. Ever HT use was uncommon (7% and 11% in cases and controls, respectively). Use of estrogen plus progestin was not associated with an increased risk of young-onset breast cancer (odds ratio = 0.80, 95% confidence interval: 0.41, 1.59). Unopposed estrogen use was inversely associated with the risk of young-onset breast cancer (odds ratio = 0.58, 95% confidence interval: 0.34, 0.99). Duration of use, age at first use, and recency of use did not modify these associations.


Dental caries diagnosis.
As with any disease process, the early initiation of therapy for dental caries is often the most effective means to ensure resolution. However, for any therapy to be effective, early diagnosis is paramount to success. Unfortunately, current tools used in dental caries detection are not sensitive enough to diagnose the disease process in its early stages and, frequently, once a diagnosis is made restoration is the only effective means of treatment. Quantitative light-induced fluorescence, electrical conductance measurements, direct digital radiography, and direct fiber optic transillumination are methods that may offer promise as effective tools in diagnosing early dental caries. This article describes some of the research that has been performed to make these methods a reality.


Active transepithelial potassium transport in frog skin via specific potassium channels in the apical membrane.
In frog skin bathed in Cl--Ringer's solution the short circuit current (SCC) is equal to the net Na+ flux. In the present study Na+ and K+ transport across frog skin have been investigated in skins bathed in a solution where all Cl- has been substituted by the impermeable anion gluconate. In this solution the net Na+ flux (9.22 +/- 0.72 nmole/cm2/min) was significantly higher than the SCC (7.61 +/- 0.63) nmole/cm2/min). Measurement of the transepithelial K+ influx and K+ efflux showed that the discrepancy between the net Na+ flux and the SCC was caused by an active outwards going transepithelial K+ transport. The K+ but not the Na+ transport could be blocked by adding the K+ channel blocking agent Ba++ to the apical solution. Thus, the K+ transport occurs via a K+ specific pathway in the apical membrane. Ouabain blocked both the Na+ and the K+ transport, whereas the presence of the Na+ channel blocking agent amiloride in the apical solution blocked the Na+ transport and reduced the K+ transport. In the presence of amiloride in the apical solution the SCC and the transepithelial potential difference (PD) reversed so that the outside (the apical side) of the frog skin became positive with respect to the basolateral side. The inverted SCC was carried by an active transepithelial K+ transport, this K+ transport required the presence of Na+ in the basolateral solution. The experiments show that frog skin can insert or activate K+ channels in the apical membrane, indicating that the frog may regulate its K+ content by varying the K+ permeability of the apical membrane.


Asbestos-induced autoimmunity in C57BL/6 mice.
Environmental impacts on autoimmunity have significant public health implications. Epidemiological studies have shown associations between exposure to airborne silicates, such as crystalline silica or asbestos, and autoimmunity, but the etiology remains unclear. The purpose of this study was to test the hypothesis that asbestos could lead to a specific pattern of autoantibodies and pathology indicative of systemic autoimmune disease (SAID). Female C57Bl/6 mice were instilled intratracheally with 2 doses x 60 microg/mouse of amphibole asbestos (tremolite), wollastonite (a non-fibrogenic control fiber), or saline alone. Serum samples were collected and urine was checked for protein bi-weekly for 7 months. By 26 weeks, the asbestos-instilled animals had a significantly higher frequency of positive anti-nuclear antibody (ANA) tests compared to wollastonite and saline groups. The majority of positive ANAs showed homogeneous or combined homogeneous/speckled patterns, and tested positive for antibodies to dsDNA and SSA/Ro 52. Serum isotyping showed no significant changes in IgM, IgA, or IgG subclasses. However, there was an overall decrease in the mean IgG serum concentration in asbestos-instilled mice. IgG immune complex deposition was demonstrated in the kidneys of asbestos-instilled mice, with evidence of glomerular and tubule abnormalities suggestive of glomerulonephritis. Flow cytometry demonstrated moderate changes in the percentages of CD25+ T-suppressor cells and B1a B-cells in the superficial cervical lymph nodes of the asbestos-instilled mice. These data demonstrate that asbestos leads to immunologic changes consistent with the development of autoimmunity. This study provides a non-autoimmune prone murine model for use in future elucidation of mechanisms involved in asbestos-induced autoimmune disease.


Impact of the number of treatment courses on the clinical response of patients who receive high-dose bolus interleukin-2.
To determine the impact of treatment with successive courses of high-dose bolus interleukin-2 (IL-2) on the incidence of clinical responses in patients with metastatic melanoma or renal cell cancer. A consecutive series of 350 patients with either metastatic melanoma or renal cell cancer who were treated with high-dose bolus IL-2 in the Surgery Branch, National Cancer Institute, between September 1985 and November 1996 was analyzed, with a median potential follow-up of 7.1 years. All patients were treated with 720,000 IU/kg of IL-2 administered by a 15-minute intravenous infusion every 8 hours for up to 5 days, as clinically tolerated per cycle. Patients were retreated according to clinical response and tolerance to the IL-2 therapy. Of the 149 patients with melanoma, 10 achieved complete responses (CRs) and 13 partial responses (PRs), for an overall response rate of 15.4%. Of the 201 patients with renal cell cancer, 18 achieved CRs and 20 PRs, for an overall response rate of 19.0%. Among responding patients, 21 of 23 with melanoma and 34 of 38 with renal cell cancer developed at least PRs after the first course of IL-2. Most patients with metastatic melanoma and renal cell cancer who achieved PRs or CRs to intravenous high-dose bolus IL-2 were identified after the first course of therapy. Those who demonstrated no response after two treatment courses failed to respond to additional IL-2 therapy. Based on this retrospective analysis, we recommend that patients who exhibit objective responses to treatment with high-dose bolus IL-2 receive additional treatment courses until either CR or IL-2 intolerance develops. Patients who do not achieve objective responses after two courses of IL-2 should receive no further treatment with this regimen.


Tuning Structure and Excitonic Properties of 2D Ruddlesden-Popper Germanium, Tin, and Lead Iodide Perovskites via Interplay between Cations.
The compositional tunability of 2D metal halide perovskites enables exploration of diverse semiconducting materials with different structural features. However, rationally tuning the 2D perovskite structures to target physical properties for specific applications remains challenging, especially for lead-free perovskites. Here, we study the effect of the interplay of the B-site (Ge, Sn, and Pb), A-site (cesium, methylammonium, and formamidinium), and spacer cations on the structure and optical properties of a new series of 2D Ruddlesden-Popper perovskites using the previously unreported spacer cation 4-bromo-2-fluorobenzylammonium (4Br2FBZ). We report eight new crystal structures and study the consequence of varying the B-site (Pb, Sn, Ge) and dimension (n = 1, 2, vs 3D). Dimension strongly influences local distortion and structural symmetry, and the increased octahedral tilting and lone pair effects in Ge perovskites lead to a polar n = 2 perovskite that exhibits second harmonic generation, (4Br2FBZ)2(Cs)Ge2I7. In contrast, the analogous Sn and Pb perovskites remain centrosymmetric, but the B-site metal influences the photoluminescence properties. The Pb perovskites exhibit broad, defect-mediated emission at low temperature, whereas the Sn perovskites show purely excitonic emission over the entire temperature range, but the carrier recombination dynamics depend on dimensionality and dark excitonic states. Wholistic understanding of these differences that arise based on cations and dimensionality can guide the rational materials design of 2D perovskites for targeting physical properties for optoelectronic applications based on the interplay of cations and the connectivity of the inorganic framework.


Apoptotic markers in a prostate cancer cell line: effect of ellagic acid.
Ellagic acid (EA) inhibits cell growth and induces apoptosis in cultured cells; however, the precise molecular mechanism involved in EA-induced apoptosis in prostate cancer cells is unknown. The aim of the present study was to delineate possible apoptotic pathway(s) involved in the EA-mediated chemotherapeutic effects in the LNCaP human prostatic cancer cell line. EA produced anti-proliferative effects through inhibition of rapamycin (mTOR) activation and a reduction in intracellular levels of β-catenin. Moreover, we demonstrated that EA induced apoptosis via downregulation of the anti-apoptotic proteins, silent information regulator 1 (SIRT1), human antigen R (HuR) and heme oxygenase-1 (HO-1). EA modulated the expression of apoptosis-inducing factor (AIF) resulting in a significant increase in reactive oxygen species (ROS) levels and the activation of caspase-3. Finally, we demonstrated that EA reduced both transforming growth factor-β (TGF-β) and interleukin-6 (IL-6) levels. EA treatment resulted in the increased expression of the tumor suppressor protein p21 and increased the percentage of apoptotic cells. In conclusion, the results suggest that EA treatment represents a new and highly effective strategy in reducing prostate cancer carcinogenesis.


Dietary fat and growth promotion of rat mammary tumors.
A hypothesis is presented that explains the mammary tumor-promoting effects of high fat diets on the basis of alteration in the hormonal milieu, namely the relative concentrations of circulating prolactin to estrogen. Evidence from in vivo and in vitro studies drawn from work in our laboratory and others is reviewed in light of this hypothesis. It is postulated that mammary tumor cell proliferation is stimulated when the prolactin:estrogen ratio is high and is inhibited when the ratio is low. Chronic high fat intake elevates serum prolactin levels, thus raises the prolactin:estrogen ratio, and thereby promotes mammary tumor cell growth.


Excitons in Bilayer MoS_{2} Displaying a Colossal Electric Field Splitting and Tunable Magnetic Response.
van der Waals heterostructures composed of transition metal dichalcogenide monolayers (TMDCs) are characterized by their truly rich excitonic properties which are determined by their structural, geometric, and electronic properties: In contrast to pure monolayers, electrons and holes can be hosted in different materials, resulting in highly tunable dipolar many-particle complexes. However, for genuine spatially indirect excitons, the dipolar nature is usually accompanied by a notable quenching of the exciton oscillator strength. Via electric and magnetic field dependent measurements, we demonstrate that a slightly biased pristine bilayer MoS_{2} hosts strongly dipolar excitons, which preserve a strong oscillator strength. We scrutinize their giant dipole moment, and shed further light on their orbital and valley physics via bias-dependent magnetic field measurements.


Diet and aging: bearing on thrombosis and hemostasis.
Diet is one of the environmental factors that influences thrombosis and hemostasis. Macronutrients, micronutrients, and other bioactive food components alter the predisposition to thrombosis. The type and amount of dietary fat has been shown to alter thromboxane A2 production and platelet aggregation, bleeding time, factor VII, fibrinogen, tissue plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1). Both epidemiological studies and clinical trials indicate that the very long chain n-3 fatty acids lower thrombotic tendency and risk of heart disease. Other polyunsaturated fats and monounsaturated fat appear to have antithrombotic properties, but further studies are indicated. Hypercaloric diets and those with high glycemic loads are associated with elevations of PAI-1. Moderate consumption of alcohol is associated with decreased platelet aggregation. Low intakes of folate, vitamin B12, and vitamin B6 predispose to hyperhomocysteinemia, and the benefits of supplementation in decreasing vascular disease are under investigation. In a limited number of clinical and laboratory studies, vitamin E has been shown to decrease platelet aggregation and the concentration of PAI-1. Flavonoids and isoflavones appear to inhibit platelet aggregation at pharmacologic concentrations only. Nutritional status frequently declines with aging and may exacerbate the already increased risk for thrombosis. Diet presents an interesting area for research into thrombophilia, but additional work is indicated before specific recommendations are made.


Silicon-Induced Systemic Defense Responses in Perennial Ryegrass Against Infection by Magnaporthe oryzae.
Sustainable integrated disease management for gray leaf spot of perennial ryegrass may involve use of plant defense elicitors with compatible traditional fungicides to reduce disease incidence and severity. Silicon (Si) has previously been identified as a potential inducer or modulator of plant defenses against different fungal pathogens. To this end, perennial ryegrass was inoculated with the causal agent of gray leaf spot, Magnaporthe oryzae, when grown in soil that was nonamended or amended with three different levels of calcium silicate (1, 5, or 10 metric tons [t]/ha). When applied at a rate of 5 t/ha, calcium silicate was found to significantly suppress gray leaf spot in perennial ryegrass, including a significant reduction of disease incidence (39.5%) and disease severity (47.3%). Additional studies observed nonpenetrated papillae or cell-wall appositions harboring callose, phenolic autofluorogens, and lignin-associated polyphenolic compounds in grass grown in the Si-amended soil. Regarding defense-associated enzyme levels, only following infection did grass grown in Si-amended soil exhibit greater activities of peroxidase and polyphenol oxidase than equivalent inoculated control plants. Also following infection with M. oryzae, grass levels of several phenolic acids, including chlorogenic acid and flavonoids, and relative expression levels of genes encoding phenylalanine ammonia lyase (PALa and PALb) and lipoxygenase (LOXa) significantly increased in Si-amended plants compared with that of nonamended control plants. These results suggest that Si-mediated increase of host defense responses to fungal pathogens in perennial ryegrass has a great potential to be part of an effective integrated disease management strategy against gray leaf spot development.


Ceruloplasmin as an indicator of copper reserves in wild ruminants at high latitudes.
Northern ungulates must establish trace mineral reserves when forage is available in spring and summer to sustain biochemical activities through the long winter. Copper (Cu), zinc (Zn) and iron (Fe) reserves were measured in the serum, digestive tract, liver, and kidney of six male caribou and reindeer (Rangifer tarandus) fed a complete pelleted ration. Dry matter content and absolute amounts of Cu, Zn and Fe were highest in the liver. Digesta contents of Cu and Zn were greatest in the rumen but dry matter concentrations were greatest in the cecum reflecting the high levels of Cu and Zn in the diet. Serum ceruloplasmin (an oxidase containing Cu) activity was related to liver copper in captive reindeer and caribou (P < 0.05, r2 = 0.82) during spring and winter but not during the rut. Michaelis-Menten kinetics of ceruloplasmin were measured in sera from captive reindeer, muskox (Ovibos moschatus) and moose (Alces alces) (n = 3/species). Maximum velocities (VMAX) were 42, 20 and 9 (IU x L(-1)); kM were 0.38, 0.55 and 0.62 (mM) for muskox, reindeer and moose respectively. Wild caribou (n = 3) from the Teshekpuk herd and moose (n = 3) from the Colville River had lower VMAX (7 IU x L(-1)) and higher km (1.9 mM) than their captive conspecifics. These kinetic parameters probably reflect differences in ceruloplasmin structure between species as well as differences in tissue reserves between populations within each species. Serum ceruloplasmin activity and kinetics can provide a non-lethal alternative to direct measures of hepatic Cu reserves in wild and captive populations. However, the method requires validation for the effects of sex, season, development and disease in each species.


Marangoni Flow Induced Evaporation Enhancement on Binary Sessile Drops.
The evaporation processes of pure water, pure 1-butanol, and 5% 1-butanol aqueous solution drops on heated hydrophobic substrates are investigated to determine the effect of temperature on the drop evaporation behavior. The evolution of the parameters (contact angle, diameter, and volume) during evaporation measured using a drop shape analyzer and the infrared thermal mapping of the drop surface recorded by an infrared camera were used in investigating the evaporation process. The pure 1-butanol drop does not show any thermal instability at different substrate temperatures, while the convection cells created by the thermal Marangoni effect appear on the surface of the pure water drop from 50 °C. Because 1-butanol and water have different surface tensions, the infrared video of the 5% 1-butanol aqueous solution drop shows that the convection cells are generated by the solutal Marangoni effect at any substrate temperature. Furthermore, when the substrate temperature exceeds 50 °C, coexistence of the thermal and solutal Marangoni flows is observed. By analyzing the relation between the ratio of the evaporation rate of pure water and 1-butanol aqueous solution drops and the Marangoni number, a series of empirical equations for predicting the evaporation rates of pure water and 1-butanol aqueous solution drops at the initial time as well as the equations for the evaporation rate of 1-butanol aqueous solution drop before the depletion of alcohol are derived. The results of these equations correspond fairly well to the experimental data.


A rapid bioassay for recombinant interleukin-22.
Interleukin-22 is a novel cytokine produced mainly in activated T cells. The elaborate biological functions of IL-22 in vivo are still widely unknown. In this report, we describe a rapid, simple, and reproducible in vitro cell-based bioassay for measuring the bioactivity of recombinant interleukin-22 (IL-22) to study the primary function of IL-22 in vivo. Human hepatocyte cell line (HepG2) was transfected with pSTAT3-Luc, a plasmid carrying the luciferase gene under the control of STAT3. After screening and selection, one stable clone was established which generates a strong response to recombinant human IL-22 (rhIL-22) stimulation in a dose-dependent manner. The cell showed ED50 of 17.0 +/- 1.4 ng/mL (n = 15) to recombinant human IL-22. Pre-incubation of anti-IL-22 mAb with IL-22 recombinant proteins completely blocked the bioactivities. The assay can be completed within one day. The current assay provides a rapid analytical method to measure the biological activity of IL-22 in vitro.


A single exposure to amphetamine is sufficient to induce long-term behavioral, neuroendocrine, and neurochemical sensitization in rats.
Repeated treatment with psychostimulant drugs causes long-lasting behavioral sensitization and associated neuroadaptations. Although sensitization induced by a single psychostimulant exposure has also been reported, information on the behavioral and neurochemical consequences of a single psychostimulant exposure is sparse. Therefore, to evaluate whether behavioral sensitization evoked by single and repeated psychostimulant pretreatment regimens represent the same neurobiological phenomenon, the time-dependent expression of behavioral, neurochemical, and neuroendocrine sensitization after a single exposure to amphetamine was investigated in rats. A single exposure to amphetamine (5 mg/kg, i.p.) caused context-independent sensitization of the locomotor effects of amphetamine, which intensified over time. Thus, sensitization to amphetamine was marginal at 3 d after treatment and more evident after 1 week, whereas 3 weeks after treatment, profound sensitization, as well as cross-sensitization, to cocaine was observed. Amphetamine pretreatment caused an increase in the electrically evoked release of [(3)H]dopamine from nucleus accumbens, caudate putamen, and medial prefrontal cortex slices and of [(14)C]acetylcholine from accumbens and caudate slices. The hyperreactivity of dopaminergic nerve terminals appeared to parallel the development of locomotor sensitization, i.e., whereas hyperreactivity of accumbens dopaminergic terminals increased between 3 d and 3 weeks after treatment, the hyperreactivity of medial prefrontal dopaminergic terminals decreased. Pre-exposure to amphetamine also sensitized the hypothalamus-pituitary-adrenal axis response to amphetamine at 1 and 3 weeks, but not at 3 d after treatment. Because these data closely resemble those reported previously for repeated amphetamine pretreatment, it is concluded that a single exposure to amphetamine is sufficient to induce long-term behavioral, neurochemical, and neuroendocrine sensitization in rats.


Type, content, and source of social support perceived by women during pregnancy: evidence from Matlab, Bangladesh.
Specific and contextualized data on social support during distinct health events are needed to improve social support interventions. This study identified the type, content, and source of social support perceived by women during pregnancy. In-depth interviews with 25 women, aged 18-49 years, living in Matlab, Bangladesh, were conducted. The findings demonstrated that women perceived, the receipt of eight distinct types of support. The four most frequently-mentioned types included: practical help with routine activities, information/advice, emotional support and assurance, as well as the provision of resources and material goods. Sources varied by type of support and most frequently included-mothers, mothers-in-law, sisters-in-law, and husbands. Examples depicting the content of each type of support revealed culturally-specific issues that can inform community-based social support interventions.


Impact of two recent extreme heat episodes on morbidity and mortality in Adelaide, South Australia: a case-series analysis.
Extreme heatwaves occurred in Adelaide, South Australia, in the summers of 2008 and 2009. Both heatwaves were unique in terms of their duration (15 days and 13 days respectively), and the 2009 heatwave was also remarkable in its intensity with a maximum temperature reaching 45.7 °C. It is of interest to compare the health impacts of these two unprecedented heatwaves with those of previous heatwaves in Adelaide. Using case-series analysis, daily morbidity and mortality rates during heatwaves (≥ 35 °C for three or more days) occurring in 2008 and 2009 and previous heatwaves occurring between 1993 and 2008 were compared with rates during all non-heatwave days (1 October to 31 March). Incidence rate ratios (IRRs) were established for ambulance call-outs, hospital admissions, emergency department presentations and mortality. Dose response effects of heatwave duration and intensity were examined. Ambulance call-outs during the extreme 2008 and 2009 events were increased by 10% and 16% respectively compared to 4.4% during previous heatwaves. Overall increases in hospital and emergency settings were marginal, except for emergency department presentations in 2008, but increases in specific health categories were observed. Renal morbidity in the elderly was increased during both heatwaves. During the 2009 heatwave, direct heat-related admissions increased up to 14-fold compared to a three-fold increase seen during the 2008 event and during previous heatwaves. In 2009, marked increases in ischaemic heart disease were seen in the 15-64 year age group. Only the 2009 heatwave was associated with considerable increases in total mortality that particularly affected the 15-64 year age group (1.37; 95% CI, 1.09, 1.71), while older age groups were unaffected. Significant dose-response relationships were observed for heatwave duration (ambulance, hospital and emergency setting) and intensity (ambulance and mortality). While only incremental increases in morbidity and mortality above previous findings occurred in 2008, health impacts of the 2009 heatwave stand out. These findings send a signal that the intense and long 2009 heatwave may have exceeded the capacity of the population to cope. It is important that risk factors contributing to the adverse health outcomes are investigated to further improve preventive strategies.


Profiling of Aroma-Active Compounds in Ylang-Ylang Essential Oils by Aroma Extract Dilution Analysis (AEDA) and Chemometric Methods.
The aroma-active compounds in the extra, first, and third grades of ylang-ylang essential oils (YYEO) from Comoros and Madagascar were identified by gas chromatography-mass spectrometry with olfactometry (GC-MS/O) using an aroma extract dilution analysis (AEDA) technique. In the previous study, the authors investigated differences in volatile compound profiles between YYEO of different grades and regions using GC coupled with a flame ionization detector (FID) and GC-MS. This study follows up with identification of the aroma-active compounds present in YYEO of various grades from both origins and to profile the aroma of those oils. For the first time, principal component analysis (PCA) on AEDA logarithmic flavor dilution (LFD) data was performed, in comparison with the corresponding PCA on GC-FID-MS data. Based on AEDA data, 21 aroma-active compounds were found across all samples and grades of YYEO, with 8 common ones previously identified by GC-FID. Linalool had the highest odor activity and is the major component of YYEO, followed by geraniol, although the latter only appeared as a much smaller peak in the chromatogram. Other trace compounds such as eugenol and vanillin were also found to be significant to the aroma of YYEO. Using PCA on resulting LFD data, YYEO from Comoros were found to have spicier odor qualities as compared to those from Madagascar. The main contributors that determine the difference in a spicy aroma profile of Comoros and Madagascar oils are vanillin, methyl eugenol, and trans-cinnamyl acetate.


A pilot study of a subsurface-flow constructed wetland treating membrane concentrate produced from reclaimed water.
A pilot study was conducted for 7 months for the City of Oxnard, California, on the use of constructed wetlands to treat concentrate produced by microfiltration and reverse osmosis (RO) of reclaimed wastewater. The treatment performance of a transportable subsurface-flow wetland was investigated by monitoring various forms of nitrogen, orthophosphate, oxygen demand, organic carbon, and selenium. Significant mass removal of constituents was measured under two hydraulic residence times (HRTs) (2.5 and 5 days). Inflow and outflow concentrations of nitrate-N and ammonia-N were significantly different for both HRTs, whereas nitrite-N and total organic carbon (TOC) were significantly different during HRT2. Mass removal by the constructed wetland averaged 61% of nitrate-N, 32% of nitrite-N, 42% of ammonia-N, 43% of biochemical oxygen demand, 19% of orthophosphate as P, 18% of TOC and 61% of selenium. Mass removal exceeded concentration reductions through water volume loss through evapotranspiration. Calibrated first-order area-based removal rates were consistent with literature ranges, and were greater during HRT1 consistent with greater mass loads, higher hydraulic loading and shorter HRTs. The rate constants may provide a basis for sizing a full-scale wetland receiving a similar quality of water. The results indicated that engineered wetlands can be useful in the management of RO membrane concentrate for reclaimed water reuse.


Isolation and characterization of a conserved sequence highly expressed in tumors and growing cells.
We isolated a cDNA clone, pAH1005, originating from the 6.8-kilobases mRNA of the rat ascites hepatoma cell line, AH60C. The sequence was highly expressed in various tumor tissues and tumor lines in rats, mice and humans. In rats, the expression of the sequence was also observed in fetal liver of 7 and 10 days of gestation, to some extent in the regenerating liver, yet was sparse in normal adult tissues. Additionally, this expression was induced when adult rat hepatocytes in primary monolayer cultures proliferated by the addition of epidermal growth factor and insulin. Southern blot analysis showed no major gene amplification and rearrangement between DNAs from hepatoma and normal liver of the same species. In addition, in vitro nuclear transcription assay revealed that a genomic clone corresponding to the pAH1005 hybridized to the nascent RNA from isolated nuclei of AH60C but not from those of normal rat liver. All these observations suggest that the expression of the conserved sequence we isolated is closely associated with cell growth and is regulated at the transcriptional level.


[Introduction of Support for Thoracoscopic Surgery by Clinical Engineers:Task Shift as Scopist by Clinical Engineers].
On October 1, 2021, the Clinical Engineer Law was revised and expanded the task of clinical engineers (CEs). After that, they can hold and operate a endoscope in endoscopic surgery. On June 9, 2022, our hospital asked CEs to directly participate in thoracoscopic pleural biopsy as scopist( scope operator) for the first time, and since then, a total of 54 thoracoscopic surgery cases were performed by CEs as scopist over the course of one year. In the CE-supported lung surgery of lobectomy and segmentectomy cases, there was a trend toward an increase in operating time of about 15 minutes, although there was no significant difference in operating time compared with conventional surgery. Other than that, however, there were no particular problems, and we expect that further CE education will provide a favorable surgical environment.


Replacement lipomatosis: CT and MRI findings of a rare renal mass.
Replacement lipomatosis of the kidney (RLK) is an advanced form of renal sinus lipomatosis, in which infection, renal calculi and long-standing hydronephrosis are accompanied by renal parenchymal atrophy. The kidneys are usually poor or non-functioning. We present CT and MRI findings of an unusual focal RLK of a 52-year-old male, who was examined with the suspicion of renal malignancy.


Von Willebrand disease type 2N: An update.
Quantitative or qualitative defects of von Willebrand factor (VWF) are responsible for the most common inherited bleeding disorder, von Willebrand disease (VWD). Type 2N VWD is an uncommon recessive disorder that results from gene mutations located in the region coding for the binding site of VWF for factor VIII (FVIII). This narrative review describes the pathophysiology, diagnostic procedures and treatment as well as the molecular biology of type 2N VWD. Although other VWF-dependent functions like binding to platelets and collagen are preserved, FVIII plasma levels are low due to the rapid clearance of this moiety in the absence or reduction of its binding to VWF. The diagnosis of type 2N should be considered in patients with low FVIII coagulant activity (FVIII:C) and disproportionally higher VWF antigen, especially when they present with an autosomal recessive pattern of inheritance. Because an accurate diagnosis is essential for genetic counseling and optimal treatment, type 2N must be distinguished from mild/moderate hemophilia A and its carrier state. This differential diagnosis can be obtained by using the laboratory assay of the FVIII binding capacity of VWF (VWF:FVIIIB) or analysis of the FVIII binding site on the VWF gene.


Laser-induced fluorescence in high pressure solid propellant flames.
The application of laser-induced fluorescence (LIF) to the study of high pressure solid propellant flames is described. The distribution of the OH and CN radicals was determined in several solid propellant flames at pressures up to 3.5 MPa. The greatest difficulty in these measurements was the separation of the desired LIF signals from the large scattering at the laser wavelength from the very optically thick propellant flames. Raman experiments using 308-nm excitation were also attempted in the propellant flames but were unsuccessful due to LIF interferences from OH and NH.


Stigmatization of illicit drug use among Puerto Rican health professionals in training.
Social stigma continues to be a barrier for health promotion in our society. One of the most stigmatized health conditions in our time continues to be addiction to illicit drug use. Although it has been widely recognized as a health concern, criminalizing approaches continue to be common in Puerto Rico. Health professionals need to engage in challenging the stigma of illicit drug use in order to foster policies and government efforts with health-oriented approaches. Still, personal stigmatizing attitudes among them continue to be a barrier for the implementation of this agenda. Therefore, the main objectives of this study were to document stigma towards illicit drug use among a sample of health professionals in training, and explore differences in such attitudes among participants from different areas of training. In order to achieve this objective we carried out a sequential mixed method approach with a sample of 501 health professionals in training or practice from the disciplines of medicine, nursing, psychology and social work. Results evidence the continued existence of stigmatizing attitudes among this population. We discuss some of the implications for public health and potential strategies for action.


Neonatal Exposure to BPA, BDE-99, and PCB Produces Persistent Changes in Hepatic Transcriptome Associated With Gut Dysbiosis in Adult Mouse Livers.
Recent evidence suggests that complex diseases can result from early life exposure to environmental toxicants. Polybrominated diphenyl ethers (PBDEs), and polychlorinated biphenyls (PCBs) are persistent organic pollutants (POPs) and remain a continuing risk to human health despite being banned from production. Developmental BPA exposure mediated-adult onset of liver cancer via epigenetic reprogramming mechanisms has been identified. Here, we investigated whether the gut microbiome and liver can be persistently reprogrammed following neonatal exposure to POPs, and the associations between microbial biomarkers and disease-prone changes in the hepatic transcriptome in adulthood, compared with BPA. C57BL/6 male and female mouse pups were orally administered vehicle, BPA, BDE-99 (a breast milk-enriched PBDE congener), or the Fox River PCB mixture (PCBs), once daily for three consecutive days (postnatal days [PND] 2-4). Tissues were collected at PND5 and PND60. Among the three chemicals investigated, early life exposure to BDE-99 produced the most prominent developmental reprogramming of the gut-liver axis, including hepatic inflammatory and cancer-prone signatures. In adulthood, neonatal BDE-99 exposure resulted in a persistent increase in Akkermansia muciniphila throughout the intestine, accompanied by increased hepatic levels of acetate and succinate, the known products of A. muciniphila. In males, this was positively associated with permissive epigenetic marks H3K4me1 and H3K27, which were enriched in loci near liver cancer-related genes that were dysregulated following neonatal exposure to BDE-99. Our findings provide novel insights that early life exposure to POPs can have a life-long impact on disease risk, which may partly be regulated by the gut microbiome.


Communication About Dying, Death, and Bereavement: A Systematic Review of Children's Literature.
Children's books have the potential to facilitate communication about death for children living with a serious illness and for children coping with the death of a loved one. This study examines the content of children's literature relevant to the topic of dying and death and identifies books providers can share with children and their families. A search of children's literature was conducted using four electronic databases and one additional search engine using the word "Death" or "Dying." Storybooks about dying, death, and bereavement published in English, French, or Spanish between 1995 and 2015 were included. Each book underwent content analysis by at least two independent reviewers. Strict PRISMA standard was followed. Full protocol is available as PROSPERO #CRD42016042129. Two hundred ten books met inclusion criteria. The dying subject was primarily a grandparent (n = 78) or pet (n = 44). Books on the experience of a child dying were scarce (n = 5). The word death or dying was used in 75% of the books (n = 158), while others utilized euphemisms. The majority of books featured animals (n = 40) or Caucasian subjects (n = 122) and included spiritual elements such as heaven (n = 122). Less than one-quarter of the books included tools for readers to address the topic of death. Storybooks can be a helpful tool to introduce communication about dying and death with children. Gaps exist in current children's literature to effectively enable children to reflect on their own dying process. A general summary of available books is provided to assist those caring for children and families facing end-of-life issues.


Correlation between Myocardial Velocity Measured using Tissue Doppler Imaging in the Left Ventricular Lead-Implanted Segment and Response to Cardiac Resynchronization Therapy.
This study investigated whether tissue Doppler imaging parameters, especially the peak systolic velocity of the left ventricular lead-implanted segment (Ss), affect cardiac resynchronization therapy response. In this case-control study, 110 enrolled patients were divided into cases (responder group, n=65) and controls (nonresponder group, n=45) based on whether their left ventricular end-systolic volume was reduced by ≥15% at 6 months after surgery. Preoperative clinical and echocardiographic data were collected. Multivariate logistic regression models were used to analyze the factors affecting the response to cardiac resynchronization therapy, and receiver operating characteristic curves were plotted to evaluate their diagnostic values. The proportion of patients with left bundle branch block in the case group was higher than that in the control group. The control group showed a higher left atrial volume index, E/A ratio and E/Em ratio but lower Ss than that of the case group. A multivariate regression analysis showed that left bundle branch block, Ss, and an E/Em ratio>14 were independent risk factors affecting the response to cardiac resynchronization therapy. Ss=4.1 cm/s was the best diagnostic threshold according to the receiver operating characteristic curve. Ss is an important factor affecting the response to cardiac resynchronization therapy. Patients with heart failure associated with Ss<4.1 cm/s have a higher risk of nonresponse.


Hydroxyurea-induced augmentation of fetal hemoglobin production in patients with sickle cell anemia.
Five patients with sickle cell anemia were treated with hydroxyurea (HU), in hopes of augmenting their production of fetal hemoglobin. Laboratory responses in two patients treated for more than 2 years were encouraging and there were suggestions of clinical improvement. Long-term HU therapy should be considered for severely affected adults with sickle cell anemia who are willing to accept what is probably a small risk of carcinogenesis. Preliminary chromosomal analysis and knowledge of the clastogenic properties of HU suggest that conception and pregnancy should be avoided. Pharmacokinetic studies will probably be necessary to adjust individual dosage schedules so that cytotoxicity is avoided. F cell responses can be seen in 2 to 3 weeks if the HU dose is optimal, but establishment of a large number of F cells in the circulation may take a month or longer.


Galectin-3 acts as an angiogenic switch to induce tumor angiogenesis via Jagged-1/Notch activation.
Angiogenesis is a coordinated process tightly regulated by the balance between Delta-like-4 (DLL4) and Jagged-1 (JAG1) in endothelial cells. Here we show that galectin-3 (gal-3), a glycan-binding protein secreted by cancer cells under hypoxic conditions, triggers sprouting angiogenesis, assisted by hypoxic changes in the glycosylation status of endothelial cells that enhance binding to gal-3. Galectin-3's proangiogenic functions were found to be predominantly dependent on the Notch ligand JAG1. Differential direct binding to JAG1 was shown by surface plasmon resonance assay. Upon binding to Notch ligands, gal-3 preferentially increased JAG1 protein half-life over DLL4 and preferentially activated JAG1/Notch-1 signaling in endothelial cells. JAG1 overexpression in Lewis lung carcinoma cells accelerated tumor growth in vivo, but this effect was prevented in Lgals3-/- mice. Our findings establish gal-3 as a molecular regulator of the JAG1/Notch-1 signaling pathway and have direct implications for the development of strategies aimed at controlling tumor angiogenesis.


Dynamic chromosomal interactions and control of heterochromatin positioning by Ki-67.
Ki-67 is a chromatin-associated protein with a dynamic distribution pattern throughout the cell cycle and is thought to be involved in chromatin organization. The lack of genomic interaction maps has hampered a detailed understanding of its roles, particularly during interphase. By pA-DamID mapping in human cell lines, we find that Ki-67 associates with large genomic domains that overlap mostly with late-replicating regions. Early in interphase, when Ki-67 is present in pre-nucleolar bodies, it interacts with these domains on all chromosomes. However, later in interphase, when Ki-67 is confined to nucleoli, it shows a striking shift toward small chromosomes. Nucleolar perturbations indicate that these cell cycle dynamics correspond to nucleolar maturation during interphase, and suggest that nucleolar sequestration of Ki-67 limits its interactions with larger chromosomes. Furthermore, we demonstrate that Ki-67 does not detectably control chromatin-chromatin interactions during interphase, but it competes with the nuclear lamina for interaction with late-replicating DNA, and it controls replication timing of (peri)centromeric regions. Together, these results reveal a highly dynamic choreography of genome interactions and roles for Ki-67 in heterochromatin organization.


Modified versus producer milk calibration: mid-infrared analyzer performance validation.
Our objective was to determine the validation performance of mid-infrared (MIR) milk analyzers, using the traditional fixed-filter approach, when the instruments were calibrated with producer milk calibration samples vs. modified milk calibration samples. Ten MIR analyzers were calibrated using producer milk calibration sample sets, and 9 MIR milk analyzers were calibrated using modified milk sample sets. Three sets of 12 validation milk samples with all-laboratory mean chemistry reference values were tested during a 3-mo period. Calibration of MIR milk analyzers using modified milk increased the accuracy (i.e., better agreement with chemistry) and improved agreement between laboratories on validation milk samples compared with MIR analyzers calibrated with producer milk samples. Calibration of MIR analyzers using modified milk samples reduced overall mean Euclidian distance for all components for all 3 validation sets by at least 24% compared with MIR analyzers calibrated with producer milk sets. Calibration with modified milk sets reduced the average Euclidian distance from all-laboratory mean reference chemistry on validation samples by 40, 25, 36, and 27%, respectively for fat, anhydrous lactose, true protein, and total solids. Between-laboratory agreement was evaluated using reproducibility standard deviation (s(R)). The number of single Grubbs statistical outliers in the validation data was much higher (53 vs. 7) for the instruments calibrated with producer milk than for instruments calibrated with modified milk sets. The s(R) for instruments calibrated with producer milks (with statistical outliers removed) was similar to data collected in recent proficiency studies, whereas the s(R) for instruments calibrated with modified milks was lower than those calibrated with producer milks by 46, 52, 61, and 55%, respectively for fat, anhydrous lactose, true protein, and total solids.


ITS1 metabarcoding highlights low specificity of lichen mycobiomes at a local scale.
As self-supporting and long-living symbiotic structures, lichens provide a habitat for many other organisms beside the traditionally considered lichen symbionts-the myco- and the photobionts. The lichen-inhabiting fungi either develop diagnostic phenotypes or occur asymptomatically. Because the degree of specificity towards the lichen host is poorly known, we studied the diversity of these fungi among neighbouring lichens on rocks in an alpine habitat. Using a sequencing metabarcoding approach, we show that lichen mycobiomes clearly reflect the overlap of multiple ecological sets of taxa, which differ in their trophic association with lichen thalli. The lack of specificity to the lichen mycobiome is further supported by the lack of community structure observed using clustering and ordination methods. The communities encountered across samples largely result from the subsampling of a shared species pool, in which we identify three major ecological components: (i) a generalist environmental pool, (ii) a lichenicolous/endolichenic pool and (iii) a pool of transient species. These taxa majorly belong to the fungal classes Dothideomycetes, Eurotiomycetes and Tremellomycetes with close relatives in adjacent ecological niches. We found no significant evidence that the phenotypically recognized lichenicolous fungi influence the occurrence of the other asymptomatic fungi in the host thalli. We claim that lichens work as suboptimal habitats or as a complex spore and mycelium bank, which modulate and allow the regeneration of local fungal communities. By performing an approach that minimizes ambiguities in the taxonomic assignments of fungi, we present how lichen mycobiomes are also suitable targets for improving bioinformatic analyses of fungal metabarcoding.


Is there a need for audiologic evaluation in patients with Behçet disease?
Behçet disease is known to be a multisystem condition. We conducted a study to determine the prevalence of hearing loss in patients with Behçet disease and to identify any associations between audiologic findings and other clinical manifestations and treatment. Our study group was made up of 41 adults with Behçet disease and 41 healthy sex- and age-matched controls. All patients and controls underwent a complete clinical otolaryngologic examination, which included pure-tone audiometry, acoustic impedance testing, and otoacoustic emissions testing. Audiology revealed that the prevalence of sensorineural hearing loss (SNHL) was significantly higher in the Behçet patients than in the controls-68.3 vs. 22.0% (p < 0.002). The duration of Behçet disease had no significant impact on whether patients did or did not experience hearing loss. Hearing loss was the fourth most common clinical finding in the Behçet group, after oral ulcers, genital ulcers, and skin lesions. We conclude that SNHL is present in a significant number of Behçet patients, and we suggest the need for an adequate investigation of hearing in the routine follow-up of these patients.


Setting and prioritizing evidence-informed policies to control childhood obesity in Iran: a mixed Delphi and policy dialogue approach.
The prevalence of childhood obesity (CO) and related complications is high and alarmingly increasing in Iran. This study applied a mixed Delphi & Policy Dialogue approach to exploring and prioritizing policy options to control childhood obesity in Iran. This study is organized in three Delphi phases followed by a policy dialogue session. This study applied the advocacy collation framework and evidence-informed policy-making approach to enhance the chance of a feasible and acceptable policy package. The first step consisted of interviews with 30 experts and primary stakeholders. Based on their answers and a comprehensive literature review, a list of presumed effective policy options to combat CO in Iran was made. Then, panelists were asked to score each policy option using a five-point Likert scale in seven constructs. To maximize the spread of opinions, panelists were chosen to represent three perspectives: policy-makers at different levels, presidents of various organizations who would implement potential policy options, and academics. Twenty-one stakeholders were invited to discuss the policy options in a policy dialogue section. We introduced 27 policy options and asked stakeholders to rank them using seven criteria on a five-level Likert scale. Totally, 41 experts participated in round 2 (66.2% response rate), and 33 experts took part in round 3 (72% response rate). Participants believed that healthy schools, creating healthy environments in kindergartens and other child care centers, subsidizing healthy foods, educating healthy lifestyles in mass media, and increasing access to physical activity facilities are the most effective and feasible policies in controlling CO. After the policy dialogue, the healthy school remained the most prioritized policy. a policy package to combat CO in Iran was designed with the participation of all stakeholders. The advocacy collation framework and the evidence-informed policy-making approach were used to draft a policy package to combat CO, increasing the acceptability and feasibility of the developed policy package.


The Nrf2/ARE pathway as a potential therapeutic target in neurodegenerative disease.
Nuclear factor E2-related factor 2 (Nrf2) is a transcription factor known to induce expression of a variety of cytoprotective and detoxification genes. Several of the genes commonly regulated by Nrf2 have been implicated in protection from neurodegenerative conditions. Work from several laboratories has uncovered the potential for Nrf2-mediated transcription to protect from neurodegeneration resulting from mechanisms involving oxidative stress. For this reason, Nrf2 may be considered a therapeutic target for conditions that are known to involve free radical damage. Because common mechanisms of neurodegeneration, such as mitochondrial dysfunction and build-up of reactive oxygen species, are currently being uncovered, targeting Nrf2 may be valuable in combating conditions with variable causes and etiologies. Most effectively to target this protein in neurodegenerative conditions, a description of the involvement of Nrf2 and potential for neuroprotection must come from laboratory models. Herein, we review the current literature that suggests that Nrf2 may be a valuable therapeutic target for neurodegenerative disease, as well as experiments that illustrate potential mechanisms of protection.


Systemic consequences of acute kidney injury.
Acute kidney injury (AKI) is a frequent and serious event associated with a high rate of complications, with an increased risk of progression to multiple organ dysfunction and excessive 'attributable' mortality. AKI affects all physiologic functions and organ systems with interrelated mechanisms, including the 'classical' consequences of the uremic state, the inflammatory nature of AKI per se and resulting systemic effects, the modulating effect of AKI in the presence of an (inflammatory) underlying disease process and the multiple untoward effects induced by renal replacement therapy (RRT) and anticoagulation. A rapidly increasing body of evidence is clarifying these systemic effects that are the reflection of a broad common pathology that ultimately results in an 'augmented' inflammation and impairment of immunocompetence. This includes the release of cytokines and inflammatory mediators, increase in oxidative stress, activation of various immune cells, neutrophil extravasation, generalized endothelial injury, increased vascular permeability and tissue oedema formation. These systemic phenomena associated with AKI induce distant organ injury affecting all organ systems with clinically the most relevant effects being exerted on the lungs, the intestines and liver and the heart and predispose the progression to multiple organ dysfunction syndrome and death. Currently available renal replacement therapy modalities are incapable of compensating for these systemic consequences of AKI.


Holding on to what you have got: keeping hospice palliative care volunteers volunteering.
In all, 119 hospice palliative care volunteers from 3 community-based hospice programs completed the Volunteer Retention Questionnaire (VRQ), a 33-item survey designed for this study. The VRQ asks volunteers to rate the importance of each item to their decision to continue volunteering. The items that received the highest mean importance ratings included enjoying the work they do, feeling adequately prepared/trained to perform their role, and learning from their patients' experiences/listening to their patients' life stories. Being recognized (eg, pins for years of service or being profiled in the hospice newsletter), receiving phone calls/cards from their volunteer coordinator on special occasions, and being reimbursed for out-of-pocket expenses were among the items that received the lowest mean importance ratings. Suggestions for improving volunteer retention are provided.


Silencing of tumor-suppressive NR_023387 in renal cell carcinoma via promoter hypermethylation and HNF4A deficiency.
Dysregulation of the epigenetic status of long noncoding RNAs (lncRNAs) has been linked to diverse human diseases including human cancers. However, the landscape of the whole-genome methylation profile of lncRNAs and the precise roles of these lncRNAs remain elusive in renal cell carcinoma (RCC). We first examined lncRNA expression profiles in RCC tissues and corresponding adjacent normal tissues (NTs) to identify the lncRNA signature of RCC, then lncRNA Promoter Microarray was performed to depict the whole-genome methylation profile of lncRNAs in RCC. Combined analysis of the lncRNAs expression profiles and lncRNAs Promoter Microarray identified a series of downregulated lncRNAs with hypermethylated promoter regions, including NR_023387. Quantitative real-time polymerase chain reaction (RT-PCR) implied that NR_023387 was significantly downregulated in RCC tissues and cell lines, and lower expression of NR_023387 was correlated with shorter overall survival. Methylation-specific PCR, MassARRAY, and demethylation drug treatment indicated that hypermethylation in the NR_023387 promoter contributed to its silencing in RCC. Besides, HNF4A regulated the expression of NR_023387 via transcriptional activation. Functional experiments demonstrated NR_023387 exerted tumor-suppressive roles in RCC via suppressing the proliferation, migration, invasion, tumor growth, and metastasis of RCC. Furthermore, we identified MGP as a putative downstream molecule of NR_023387, which promoted the epithelial-mesenchymal transition of RCC cells. Our study provides the first whole-genome lncRNA methylation profile in RCC. Our combined analysis identifies a tumor-suppressive and prognosis-related lncRNA NR_023387, which is silenced in RCC via promoter hypermethylation and HNF4A deficiency, and may exert its tumor-suppressive roles by downregulating the oncogenic MGP.


Developing an interactive STD-prevention program for youth: lessons from a north Indian slum.
An intervention developed to teach young men in the urban slums of Lucknow, India, about sexually transmitted diseases is described in detail. This community-based intervention was designed to impart core educational messages and to address preexisting ideas about sexual health and STD-prevention practices among participants. Indicators of knowledge recorded before and after the intervention are presented and evaluated against the scores of a control group. Levels of sexual activity and factors associated with risky sex are discussed. The intervention was successful in raising the young men's awareness of STDs significantly in all areas except for the length of time that symptoms take to manifest following risky sex and the ineffectiveness of washing one's genitals after sex to avoid acquiring STDs. Lessons learned during the intervention are described as a means of informing future STD-education programs, and issues requiring prompt attention are identified.


Magnetic resonance T2 image signal intensity ratio and clinical manifestation predict prognosis after surgical intervention for cervical spondylotic myelopathy.
A retrospective imaging study of 73 patients who underwent surgery for cervical spondylotic myelopathy (CSM) between April 2005 and July 2007. To investigate whether magnetic resonance (MR) T2 image signal intensity (SI) ratio and clinical manifestation can assess the prognosis in patients with CSM. The association between intramedullary high SI on T2-weighted MR images and surgical outcome in CSM remains controversial. The means of quantizing SI ratio for the disease has not been discussed. A total of 73 patients with cervical compressive myelopathy were retrospectively enrolled and were treated with anterior, posterior, and posterior-anterior united decompression. A total of 1.5-T magnetic resonance imaging was performed in all patients before surgery. T2-weighted images of sagittal increased SI on the cervical spinal cord were obtained, and the regions of interest (ROIs) are taken by 0.05 cm. T2-weighted MR images of sagittal normal cord SI on the cervical between C7-T1 disc levels were obtained, and the ROIs are taken by 0.3 cm. SI value is measured by computer, and the SI ratio between the regions 0.05 cm and 0.3 cm has been calculated. If no intramedullary high SI was noted on T2-weighted MR images, the ROIs were taken by 0.05 cm of the severe compression cord. All patients had been divided into 3 groups by hierarchical clustering analysis with SI ratio (Group 1: low SI ratio, Group 2: middle SI ratio, and Group 3: high SI ratio). Statistical analyses were performed with SPSS 11.0. There are significant differences between 3 groups by comparing the recovery rate (P < 0.001), age (P = 0.003), duration of disease (P = 0.001), Babinski sign (P < 0.001), preoperative JOA score (P = 0.006), and postoperative JOA score (P < 0.001). There are no significant differences on sex among 3 groups (P = 0.387). By using the multiple comparison analysis, the above results are further shown. Patients with low SI ratio who were not too old and had a shorter duration of disease experienced a good surgical outcome. However, with the increase of SI ratio and the occurrence of pyramidal sign, a poor prognosis after surgery will show. SI ratio and clinical manifestation can be a predictor of surgical outcome.


Trauma in the elderly.
As technology and medical care improve, older patients will be more active and productive at later ages. Falls and motor vehicle crashes will remain major mechanisms of injury in the elderly. Treatment of the elderly trauma patient must focus on their complex acute medical needs, their rehabilitation and social needs post trauma, and most of all prevention.


Approximation of continuous growth of Cephalotaxus harringtonia plant cell cultures using fed-batch operation.
In Cephalotaxus harringtonia plant cell cultures, periods of batch growth that are limited by hexose uptake are too short to make an accurate estimate of the Monod saturation constant. Continuous cultures are infeasible on a laboratory scale, and semicontinuous cultures require too frequent sampling. Fed-batch operation, consisting of intermittent removal from a culture that is fed continuously, was investigated as a possible solution to these problems. For a constant feed rate, computer simulations showed that a steady state can be achieved which is useful for studying growth at different specific growth rates. In terms of the dilution rate it was confirmed that the operation is essentially equivalent to continuous culture when the samples represent a small fraction of the total culture volume. Experiments with glucose or fructose as the carbon source were carried out in shake flasks fed by a multichannel syringe pump. Results indicate that Monod kinetics based on medium glucose levels cannot adequately describe growth under these conditions. Monod's expression for specific growth rate using internal glucose concentration gives an improved correlation.


Analysis of the weak interactions of ADP-Unc104 and ADP-kinesin with microtubules and their inhibition by MAP2c.
Microtubule based motors like conventional kinesin (Kinesin-1) and Unc104 (Kinesin-3), and classical microtubule associated proteins (MAPs), including MAP2, are intimately involved in neurite formation and organelle transport. The processive motility of both these kinesins involves weak microtubule interactions in the ADP-bound states. Using cosedimentation assays, we have investigated these weak interactions and characterized their inhibition by MAP2c. We show that Unc104 binds microtubules with five-fold weaker affinity and two-fold higher stoichiometry compared with conventional kinesin. Unc104 and conventional kinesin binding affinities are primarily dependent on positively charged residues in the Unc104 K-loop and conventional kinesin neck coiled-coil and removal of these residues affects Unc104 and conventional kinesin differently. We observed that MAP2c acts primarily as a competitive inhibitor of Unc104 but a mixed inhibitor of conventional kinesin. Our data suggest a specific model in which MAP2c differentially interferes with each kinesin motor by inhibiting its weak attachment to the tubulin C-termini. This is reminiscent of the defects we have observed in Unc104 and kinesin mutants in which the positively charged residues in K-loop and neck coiled-coil domains were removed.


Early use of omeprazole benefits patients with acute myocardial infarction.
Bleeding complications are not uncommon in patients with acute myocardial infarction (AMI) during treatments. How to prevent the occurrence of upper gastrointestinal bleeding in AMI patients has become one of the most intractable problems. And there are conflicting data on the efficacy and complication rate of omeprazole treatment. We conducted an intervention study to determine whether using omeprazole could benefit AMI patients. A total of 237 patients with AMI were divided into two groups at random: omeprazole group including 114 patients and control group including 123 patients. Omeprazole 40 mg by intravenous drip was given to the patients in omeprazole group when they were admitted to the hospitals. From the second day they were given omeprazole 20 mg per day by oral administration for 7 days. In contrast, no gastric acid inhibitor was given to the patients in control group. The incidence of upper gastrointestinal bleeding, the recanalization rate and overall mortality in both groups were observed. The incidence of upper gastrointestinal bleeding in omeprazole group was 5.3% (6/114) which was much lower than 14.6% (18/123) in control group (P = 0.017), but the recanalization rate had no significant difference between the two groups (P = 0.681). The overall mortality in omeprazole group was lower than that of control group (3.5% vs. 10.6%, P = 0.035). Our findings suggest that early use of omeprazole in AMI patients could decrease the incidence of upper gastrointestinal bleeding and the overall mortality, without influencing the recanalization rate. Early use of omeprazole might benefit AMI patients.


11 beta-hydroxysteroid dehydrogenase (11 beta-HSD-II) activity in human placenta: its relationship to placental weight and birth weight and its possible role in hypertension.
It has been assumed that low birth weight and high placenta weight were key factors for predicting hypertension in human adulthood. A deficiency in placental 11 beta-HSD-II enzyme activity was supposed to be the underlying cause. To possibly establish 11 beta-HSD-II as a leading factor, we determined 11 beta-HSD-II activities in 133 healthy pregnancies, 21 proteinuric pregnancies complicated by pregnancy-induced hypertension (PIH), 26 non proteinuric PIH pregnancies and 15 pregnancies complicated by fetal growth restriction (32nd-41st gestational week). We could not identify differences in 11 beta-HSD-II activity between pregnancies with the rare combination of small babies with big placentas and others (p = 0.59; Kruskal-Wallis test). And although there was no correlation between 11 beta-HSD-II activity and birth weight, in the control gestational age correlated with 11 beta-HSD-II activity (r = 0.22; p < 0.05; Spearman). 11 beta-HSD-II activity in the proteinuric PIH group was significantly higher than in the controls (11.7 pmol/min/mg prot.; range 10-13.2 vs. 7.9; range 7.0-9.1; p < 0.05). The lowest, but not significant, enzyme activities were in the IUGR group (5.8 pmol/min/mg prot.; range 4.0-9.2). In this group, analysis of variance detected a correlation between enzyme activity and placental weight. In conclusion, we could not confirm that placental 11 beta-HSD-II deficiencies act as an indicator for the risk of adult hypertension in small fetuses with large placentas. However, in growth restriction 11 beta-HSD-II activity might play a role. To clarify the influence in this group, further research is needed. Increased 11 beta-HSD-II activities with gestational age in the control may serve to sustain fetal adrenal steroid genesis and to prepare the fetus for autonomic life.


Characterization of mRAD18Sc, a mouse homolog of the yeast postreplication repair gene RAD18.
The RAD18 gene of the yeast Saccharomyces cerevisiae encodes a protein with ssDNA binding activity that interacts with the ubiquitin-conjugating enzyme RAD6 and plays an important role in postreplication repair. We identified and characterized the putative mouse homolog of RAD18, designated mRAD18Sc. The mRAD18Sc open reading frame encodes a 509-amino-acid polypeptide that is strongly conserved in size and sequence between yeast and mammals, with specific conservation of the RING-zinc-finger and the classic zinc-finger domain. The degree of sequence conservation between mRAD18Sc, RAD18, and homologous sequences identified in other species (NuvA from Aspergillus nidulans and Uvs-2 from Neurospora crassa) is entirely consistent with the evolutionary relationship of these organisms, strongly arguing that these genes are one another's homologs. Consistent with the presence of a nuclear translocation signal in the amino acid sequence, we observed the nuclear localization of GFP-tagged mRAD18Sc after stable transfection to HeLa cells. mRNA expression of mRAD18Sc in the mouse was observed in thymus, spleen, brain, and ovary, but was most pronounced in testis, with the highest level of expression in pachytene-stage primary spermatocytes, suggesting that mRAD18Sc plays a role in meiosis of spermatogenesis. Finally, we mapped the mRAD18Sc gene on mouse chromosome 6F.


Intestinal cell kinase, a protein associated with endocrine-cerebro-osteodysplasia syndrome, is a key regulator of cilia length and Hedgehog signaling.
Endocrine-cerebro-osteodysplasia (ECO) syndrome is a recessive genetic disorder associated with multiple congenital defects in endocrine, cerebral, and skeletal systems that is caused by a missense mutation in the mitogen-activated protein kinase-like intestinal cell kinase (ICK) gene. In algae and invertebrates, ICK homologs are involved in flagellar formation and ciliogenesis, respectively. However, it is not clear whether this role of ICK is conserved in mammals and how a lack of functional ICK results in the characteristic phenotypes of human ECO syndrome. Here, we generated Ick knockout mice to elucidate the precise role of ICK in mammalian development and to examine the pathological mechanisms of ECO syndrome. Ick null mouse embryos displayed cleft palate, hydrocephalus, polydactyly, and delayed skeletal development, closely resembling ECO syndrome phenotypes. In cultured cells, down-regulation of Ick or overexpression of kinase-dead or ECO syndrome mutant ICK resulted in an elongation of primary cilia and abnormal Sonic hedgehog (Shh) signaling. Wild-type ICK proteins were generally localized in the proximal region of cilia near the basal bodies, whereas kinase-dead ICK mutant proteins accumulated in the distal part of bulged ciliary tips. Consistent with these observations in cultured cells, Ick knockout mouse embryos displayed elongated cilia and reduced Shh signaling during limb digit patterning. Taken together, these results indicate that ICK plays a crucial role in controlling ciliary length and that ciliary defects caused by a lack of functional ICK leads to abnormal Shh signaling, resulting in congenital disorders such as ECO syndrome.


Revised norms and decision rules for the Luria-Nebraska Neuropsychological Battery, Form II.
All control subjects from a previous Luria-Nebraska Neuropsychological Battery, Form II (LNNB2) normative study were combined with a considerably larger normative sample to produce a composite normative sample of 392 subjects with a mean high school educational level. Revised norms for the LNNB2 were developed based on this sample and were scaled with uniform T-scores to make scale elevations comparable across LNNB2 measures. Differences between the scalewise LNNB2 normative values and published alternatives were highly significant. A critical level and actuarial decision rules for differentiation of normal from neurologically impaired performance level were developed for the LNNB2 using the current empirical norms (86% accuracy with current sample empirical critical level; 89% accuracy with LNNB1 critical level). The normative prediction values of Golden, Purisch, and Hammeke for the LNNB2 were not satisfactory (69% overall, with 45% accuracy among brain damaged sample). There was no significant effect of sex of subject on LNNB2 performance level among the control subjects.


Neuromodulation and Strategic Action Choice in Drosophila Aggression.
In this review, I discuss current knowledge and outstanding questions on the neuromodulators that influence aggressive behavior of the fruit fly Drosophila melanogaster. I first present evidence that Drosophila exchange information during an agonistic interaction and choose appropriate actions based on this information. I then discuss the influence of several biogenic amines and neuropeptides on aggressive behavior. One striking characteristic of neuromodulation is that it can configure a neural circuit dynamically, enabling one circuit to generate multiple outcomes. I suggest a consensus effect of each neuromodulatory molecule on Drosophila aggression, as well as effects of receptor proteins where relevant data are available. Lastly, I consider neuromodulation in the context of strategic action choices during agonistic interactions. Genetic components of neuromodulatory systems are highly conserved across animals, suggesting that molecular and cellular mechanisms controlling Drosophila aggression can shed light on neural principles governing action choice during social interactions.


Lysine-specific demethylase 1 as a therapeutic cancer target: observations from preclinical study.
Lysine-specific histone demethylase 1A (KDM1A/LSD1) has emerged as an important therapeutic target in various cancer types. LSD1 regulates a wide range of biological processes that influence cancer development, progression, metastasis, and therapy resistance. However, recent studies have revealed novel aspects of LSD1 biology, shedding light on its involvement in immunogenicity, antitumor immunity, and DNA damage response. These emerging findings have the potential to be leveraged in the design of effective LSD1-targeted therapies. This paper discusses the latest developments in the field of LSD1 biology, focusing on its role in regulating immunogenicity, antitumor immunity, and DNA damage response mechanisms. The newfound understanding of these mechanisms has opened possibilities for the development of novel LSD1-targeted therapies for cancer treatment. Additionally, the paper provides an overview of LSD1 inhibitor-based combination therapies for the treatment of cancer. Exploiting LSD1 role in antitumor immunity and DNA damage response provides cues to not only understand the LSD1-resistant mechanisms but also rationally design new combination therapies that are more efficient and less toxic than monotherapy. The exploration of LSD1 biology and the development of LSD1-targeted therapies hold great promise for the future of cancer treatment.


Antibiotic Misuse in Wound Care: Can Bacterial Localization through Fluorescence Imaging Help?
(1) Background: Systemic antibiotic use in chronic wounds is alarmingly high worldwide. Between 53% to 71% of patients are prescribed at least one course per chronic wound. Systemic antibiotic use should follow antibiotic stewardship guidelines and ought to be reserved for situations where their use is deemed supported by clinical indications. Unfortunately, in the field of wound care, indiscriminate and often inadequate use of systemic antibiotics is leading to both patient complications and worsening antibiotic resistance rates. Implementing novel tools that help clinicians prevent misuse or objectively determine the true need for systemic antibiotics is essential to reduce prescribing rates. (2) Methods: We present a compendium of available systemic antibiotic prescription rates in chronic wounds. The impact of various strategies used to improve these rates, as well as preliminary data on the impact of implementing fluorescence imaging technology to finesse wound status diagnosis, are presented. (3) Results: Interventions including feedback from wound care surveillance and treatment data registries as well as better diagnostic strategies can ameliorate antibiotic misuse. (4) Conclusions: Interventions that mitigate unnecessary antibiotic use are needed. Effective strategies include those that raise awareness of antibiotic overprescribing and those that enhance diagnosis of infection, such as fluorescence imaging.