D(s) meson as a quantitative probe of diffusion and hadronization in nuclear collisions.
The modifications of D(s)-meson spectra in ultrarelativistic heavy-ion collisions are identified as a quantitative probe of key properties of the hot nuclear medium. The unique valence-quark content of the D(s)=cs̄ couples the well-known strangeness enhancement with the collective-flow pattern of primordially produced charm quarks. This idea is illustrated utilizing a consistent strong-coupling treatment with hydrodynamic bulk evolution and nonperturbative T-matrix interactions for both heavy-quark diffusion and hadronization in the quark-gluon plasma (QGP). A large enhancement of the D(s) nuclear modification factor at Relativistic Heavy Ion Collider is predicted, with a maximum of ∼1.5-1.8 at transverse momenta around 2 GeV/c. This is a direct consequence of the strong coupling of the heavy quarks to the QGP and their hadronization via coalescence with strange quarks. We furthermore introduce the effects of diffusion in the hadronic phase and suggest that an increase of the D-meson elliptic flow compared to the D(s) can disentangle the transport properties of hadronic and QGP liquids.


Loneliness in women with multiple sclerosis.
Loneliness is an underrecognized component of the experience of chronic illness and disability. Women with multiple sclerosis (MS) may be at risk for loneliness due to social network changes that occur in the course of chronic illness. The purpose of this secondary analysis was to assess the extent of loneliness in a sample of 659 women with MS and examine the relationship of loneliness, social support, functional limitation, self-rated health status, social demands of illness, and marital status. More than half of the women in this sample reported feeling lonely. A significant association was found between loneliness and lower levels of social support, greater social demands of illness, greater functional limitation, lower self-rated health status, and marital status. These findings suggest that rehabilitation nurses and other health professionals should be aware that loneliness may be a common occurrence for women with MS and point to the need for screening procedures to identify loneliness in this population.


Social and economic aspects of malaria in Singapore.
Singapore is both receptive and vulnerable to the introduction of malaria because of its location in a malarious zone. The social and economic factors that determined the incidence of malaria included rapid urban and industrial developments which created suitable environmental conditions for the breeding of Anopheles vectors, influx of foreign workers from the endemic countries and increased travel to the malarious areas. The public health significance of the interactions of these factors was exemplified by the occurrence of periodic localized outbreaks. The problem of introduction of drug-resistant falciparum malaria by travellers into the country was another matter of concern. However, the health authorities were fully committed in bringing these problems under control, with political support as well as regional and international collaboration. The result of these efforts was realised in 1982 when Singapore was certified malaria-free by the WHO. The strategies implemented included epidemiological and vector surveillance, health education and environmental health supervision of all major development projects in receptive areas.


HIV-HBV coinfection in Southern Africa and the effect of lamivudine- versus tenofovir-containing cART on HBV outcomes.
This study assessed HIV-hepatitis B virus (HBV) coinfection in southern Africa in terms of prevalence, viral characteristics, occult HBV, and the effect of lamivudine- versus tenofovir-containing first-line combination antiretroviral treatment (cART) on HBV-related outcomes. A multicenter prospective cohort of HIV-infected adults in Zambia and South Africa who initiated cART. Outcomes by month 12 on cART were immunological recovery, hepatitis B surface antigen (HBsAg) loss, viral suppression, and drug resistance. We used descriptive statistics, logistic regression, and linear mixed models. Of the 1087 participants, 92 were HBsAg seropositive, yielding a sample-weighted prevalence of 7.4% (95% confidence interval: 5.6 to 9.2), with 76% genotype HBV-A1. The estimated CD4 recovery on cART was similar between HIV monoinfection and HIV-HBV coinfection groups and between lamivudine- and tenofovir-treated participants. HBsAg loss was documented in 20% (4/20) of lamivudine-treated and 18% (3/17) of tenofovir-treated participants (P = 0.305). Viral suppression (HBV-DNA < 20 IU/mL) was achieved in 61.5% (16/26) of lamivudine-treated and 71.4% (15/21) of tenofovir-treated participants (P = 0.477). HBV pol sequencing demonstrated M204I (n = 3) and N236T (n = 1) resistance-associated mutations in 4 of 8 (50%) lamivudine-treated participants and none in tenofovir-treated participants. Occult HBV infection was present in 13.3% before cART, but by month 12, HBV-DNA was below the limit of detection (<15 IU/mL) in 90.5% (19/21) of lamivudine-treated and 100% (18/18) of tenofovir-treated participants (P = 0.179). Tenofovir-containing first-line cART is preferred for HIV-HBV coinfection in Africa because of a superior resistance profile relative to lamivudine monotherapy. Extended follow-up will be needed to determine long-term complications of occult HBV coinfection. Improved access to HBsAg screening and tenofovir is needed.


Evaluation of bone scan by scintigraphy to detect subclinical invasion of the mandible by squamous cell carcinoma of the oral cavity.
A prospective study using scintigraphy was performed to compare the sensitivity of the Panorex roentgenogram and the bone scan in detecting subclinical invasion of the mandible by squamous cell carcinoma of the oral cavity and floor of the mouth. Twenty-five patients with squamous cell carcinoma of the floor of the mouth were evaluated preoperatively by both the Panorex and scintigraphic techniques and the results compared with postoperative pathologic findings. In 13 (52%) of the cases, Panorex and scintigraphic techniques were comparable in detecting tumor involvement in bone. In eight cases (32%) all three modalities had normal pathologic indications of the mandible. However, in four cases (16%) results of the scintigraphic techniques were abnormal and the Panorex, normal. In four separate cases, the extent of lesion demonstrable by scintiscanning was greater than delineated by Panorex; surgical specimen confirmed this finding. Pathologic examination of operative specimens confirmed tumor involvement. These data lead us to believe that the scintigraphic techniques may be more sensitive in detecting early mandibular involvement with squamous cell carcinoma than the Panorex technique and may help alter the therapeutic approach.


Generating and identifying axolotls with targeted mutations using Cas9 RNA-guided nuclease.
The CRISPR/Cas9 RNA-guided nuclease now enables a reverse genetics approach to investigate the function of genes of interest during regeneration in the axolotl. The process of generating the constructs necessary for targeting a gene of interest is considerably less labor intensive than for other methods of targeted mutagenesis such as Zinc finger nucleases or Transcription activator-like effector nucleases. Here, we describe the identification of targetable sequences in the gene of interest, the construction of unique guide RNAs, the microinjection of these RNAs with Cas9-encoding mRNA, the selection of well-injected animals, and an inexpensive, PCR-based method for identifying highly mutagenized animals.


Assessment of microbial quality of household water output from desalination systems by the heterotrophic plate count method.
Point-of-use household water desalination systems (HWDSs) are becoming popular in Iran because of the deterioration of drinking water. This study aimed to determine the microbial quality of output water from HWDSs in Qom, Iran by using the heterotrophic plate count (HPC) method. Samples of input and output water from 30 HWDSs were collected over a six-month period. Heterotrophic bacteria were tested using the pour plate technique. At the first sampling stage, the HPC level in 23% of samples exceeded the 500 CFU/ml threshold level. On average, for 50% of samples, the HPC level of input samples was 0-10 CFU/ml, for 42% it was 10-100 CFU/ml and for 8% it was 100-500 CFU/ml. For output samples, for 25%, the level of HPC was 0-10 CFU/ml, for 43% it was 10-100 CFU/ml, for 24% it was 100-500 CFU/ml and for 8% it exceeded 500 CFU/ml. For total coliforms the most probable number test was positive for the first and third stages of sampling (3% input samples). The comparison of the averages with national standard values shows that in some cases, the contamination of output water from HWDSs in the city of Qom has been above the standard values.


Early infantile presentation of 3-methylglutaconic aciduria type 1 with a novel mutation in AUH gene: A case report and literature review.
3-Methylglutaconic aciduria is a member of inborn errors of leucine metabolism pathway. 3-Methylglutaconic aciduria type I (MGA1) causes neurological problems which are present during infancy or childhood but the diagnosis may be delayed until adulthood. Here we report a 3years old patient with developmental delay from a relative parent's that his medical evaluations include analyses of urinary organic acid and blood acylcarnitine showed high level of 3-methylglutacoic acid, 3-hydroxyisovaleric acid and increased level of 3-hydroxyisovalerylcarnitine respectively. Further evaluation and genetic tests revealed a novel homozygous mutation of variant c.179del G (p.Gly60Valfs*12) in exon 1 of the AUH gene that was compatible with the diagnosis of MGA1. In segregation analysis of his family, both parents were heterozygous for the respective mutation, confirming obligate parental carrier status and segregation of the mutation.


Regulation of KATP channel expression and activity by the SUR1 nucleotide binding fold 1.
ATP-sensitive K(+) (K(ATP)) channels are oligomeric complexes of pore-forming Kir6 subunits and regulatory Sulfonylurea Receptor (SUR) subunits. SUR, an ATP-Binding Cassette (ABC) transporter, confers Mg-nucleotide stimulation to the channel via nucleotide interactions with its two cytoplasmic domains (Nucleotide Binding Folds 1 and 2; NBF1 and NBF2). Regulation of K(ATP) channel expression is a complex process involving subunit assembly in the ER, SUR glycosylation in the Golgi, and trafficking to the plasma membrane. Dysregulation can occur at different steps of the pathway, as revealed by disease-causing mutations. Here, we have addressed the role of SUR1 NBF1 in gating and expression of reconstituted channels. Deletion of NBF1 severely impairs channel expression and abolishes MgADP stimulation. Total SUR1 protein levels are decreased, suggestive of increased protein degradation, but they are not rescued by treatment with sulfonylureas or the proteasomal inhibitor MG-132. Similar effects of NBF1 deletion are observed in recombinant K(ATP) channels obtained by "splitting" SUR1 into two separate polypeptides (a N-terminal "half" and a C-terminal "half"). Interestingly, the location of the "splitting point" in the vicinity of NBF1 has marked effects on the MgADP stimulation of resulting channels. Finally, ablation of the ER retention motif upstream of NBF1 (in either "split" or full-length SUR1) does not rescue expression of channels lacking NBF1. These results indicate that, in addition to NBF1 being required for MgADP stimulation of the channel, it plays an important role in the regulation of channel expression that is independent of the ER retention checkpoint and the proteasomal degradation pathway.


The novel neuropeptide phoenixin is highly co-expressed with nesfatin-1 in the rat hypothalamus, an immunohistochemical study.
The hypothalamus regulates a number of autonomic functions essential for homeostasis; therefore, investigations concerning hypothalamic neuropeptides and their functions and distribution are of great importance in contemporary neuroscience. Recently, novel regulatory factors expressed in the hypothalamus have been discovered, of which nesfatin-1 and phoenixin (PNX), show intriguing similarities in their brain distributions. There are currently few studies characterizing PNX expression, so it is imperative to accurately trace its localization, with particular attention to the hypothalamic nuclei and nesfatin-1 co-expression. Using fluorescence and classical immunohistochemical stainings on adult rat brain, we visualized the potential co-expression of nesfatin-1 and PNX immunoreactive cells. We have demonstrated a distinct PNX-immunoreactivity in 21-32% of cells in the arcuate nucleus, paraventricular nucleus, ventromedial and lateral hypothalamus. Nesfatin-1 expression reached 45-68% of all neurons in the same sites, while co-expression was strikingly seen in the vast majority (70-86%) of PNX-immunoreactive neurons in the rat hypothalamic nuclei. Our results demonstrate for the first time, a wide distribution of PNX in the hypothalamus which could implicate a potential functional relationship with nesfatin-1, possibly in the regulation of the hypothalamic-pituitary-gonadal axis or other autonomic functions, which require further study.


Perceived and mentally rotated contents are differentially represented in cortical depth of V1.
Primary visual cortex (V1) in humans is known to represent both veridically perceived external input and internally-generated contents underlying imagery and mental rotation. However, it is unknown how the brain keeps these contents separate thus avoiding a mixture of the perceived and the imagined which could lead to potentially detrimental consequences. Inspired by neuroanatomical studies showing that feedforward and feedback connections in V1 terminate in different cortical layers, we hypothesized that this anatomical compartmentalization underlies functional segregation of external and internally-generated visual contents, respectively. We used high-resolution layer-specific fMRI to test this hypothesis in a mental rotation task. We found that rotated contents were predominant at outer cortical depth bins (i.e. superficial and deep). At the same time perceived contents were represented stronger at the middle cortical bin. These results identify how through cortical depth compartmentalization V1 functionally segregates rather than confuses external from internally-generated visual contents. These results indicate that feedforward and feedback manifest in distinct subdivisions of the early visual cortex, thereby reflecting a general strategy for implementing multiple cognitive functions within a single brain region.


Finance issue brief: health care claims payment: prompt payment: year end report-2003.
Since the mid 1990's state legislators and regulators have worked to resolve the complex issue of timely payment of health care claims. They have been challenged with bridging the communication gap between provider and payor and forced to address such base problems as what determines a correctly billed service. As time has progressed it is ever apparent that the completion of payment for services is dependent on many variables, not just simply timely processing of a claim.


Size dependent charge separation and recombination in CsPbI3 perovskite quantum dots.
CsPbI3 perovskite quantum dots (QDs) have shown great potential in light-harvesting and light-emitting applications, which often involve the transfer of charge carriers in and out of these materials. Here, we studied size-dependent charge separation (CS) and charge recombination (CR) between CsPbI3 QDs and rhodamine B (RhB) molecules, using transient absorption spectroscopy. When the average size decreases from 11.8 nm to 6.5 nm, the average intrinsic CS time constant decreases from 872 ± 52 ps to 40.6 ± 4.3 ps and the corresponding charge recombination time constant decreases from 3829 ± 51 ns to 1384 ± 54 ns. The observed trend of size-dependent CS and CR rates can be well explained by Marcus theory using the theoretically calculated CS and CR driving forces (ΔGCS and ΔGCR), molecular reorganization energy (λRhB), and electronic coupling strength between QD and RhB (HCS and HCR). Unlike the extensively studied more strongly quantum confined Cd chalcogenide QDs, the CsPbI3 QDs are in a weak quantum confinement regime in which size-dependent coupling strength plays a dominant role in the size-dependent charge transfer properties.


Regulation of chymase production in human mast cell progenitors.
Although mature tryptase-positive mast cells (MCs) and tryptase and chymase double-positive MCs are recognized using in situ staining and are preferentially distributed in different tissues, recent findings suggest that tryptase-positive MCs can give rise to tryptase and chymase double-positive MCs. We investigated the regulation of chymase production in developing MCs. Human cord blood or peripheral blood cells were cultured in the presence of stem cell factor and IL-6 with or without IL-4 in methylcellulose or liquid medium. Intracellular chymase and tryptase were determined with immunocytochemistry, flow cytometry, and ELISA. Chymase messenger RNA expression was examined with 3 different methods, such as Northern blotting. Flow cytometric analysis always showed a unimodal histogram of chymase-positive, as well as tryptase-positive, cells in the presence of various cytokines, even when chymase was not detected in some MCs with immunocytochemistry. The chymase protein expression increased by culture duration and was enhanced by cytokines, such as a high concentration of stem cell factor or IL-4. Chymase messenger RNA was expressed higher in immature MCs than mature chymase protein-rich MCs. We generated macroscopic MC colonies in methylcellulose by culturing CD34(+) cells for 10 weeks and measured cellular chymase, tryptase, and histamine. The chymase/histamine ratio widely varied (0.07-1.01) depending on MC colony, even under the same culture conditions, including IL-4, whereas the tryptase/histamine ratio was relatively constant (1.02-1.89). All human MCs in culture are capable of producing chymase, and the production is clonally regulated at their progenitors by cytokine-independent mechanisms, as well as being totally controlled by cytokine-dependent mechanisms accompanied by maturation.


Plant ecology. Worldwide evidence of a unimodal relationship between productivity and plant species richness.
The search for predictions of species diversity across environmental gradients has challenged ecologists for decades. The humped-back model (HBM) suggests that plant diversity peaks at intermediate productivity; at low productivity few species can tolerate the environmental stresses, and at high productivity a few highly competitive species dominate. Over time the HBM has become increasingly controversial, and recent studies claim to have refuted it. Here, by using data from coordinated surveys conducted throughout grasslands worldwide and comprising a wide range of site productivities, we provide evidence in support of the HBM pattern at both global and regional extents. The relationships described here provide a foundation for further research into the local, landscape, and historical factors that maintain biodiversity.


Magnetocardiogram recordings in a nonshielded environment--reproducibility and ischemia detection.
Magnetocardiography (MCG) is a noninvasive technology that measures the magnetic field of the heart by superconducting quantum interference devices (SQUID) sensors. The novelty of the present system is that the sensors can be operated without electromagnetic shielding of the examination room, thus allowing the system to be easily installed in the emergency department or chest pain unit. Studies in shielded rooms, found that this imaging modality may have better sensitivity as compared to ECG in detecting ischemia. We aimed (1) to assess the reproducibility, intra-observer, and interobserver interpretation variability and (2) to assess the MCG maps in the presence of coronary narrowings. All measurements were performed in a nonshielded room. For the first part of the study, two MCG maps were recorded in 24 otherwise healthy volunteers (age 20-44 years, median 24, 16 male) in an interval ranging from 2 to 48 hours. The maps were interpreted using the CardioMag software for contour maps, averaged MCG time traces, and waveform morphology of repolarization by two observers blinded to each other. The parameters tested had low disagreement between repeated measurements. The correlations of the intra-observer and interobserver interpretation were excellent. Secondly, MCG maps were obtained in 29 patients referred for angiography due to suspected coronary artery disease. Nineteen of them had coronary narrowings defined as more than 50%. In this group, 16 (84.2%) had abnormal MCG maps as compared to only 5 (26.3%) who had abnormal ECGs (P < 0.01). MCG maps can be successfully obtained in a nonshielded room and allow feasible, accurate, and reproducible measurements with little intra-observer and interobserver variability. Ischemic changes in the heart's magnetic field may occur before electrical changes. Our pilot data suggests that this imaging modality may potentially offer better sensitivity as compared to rest ECG in detecting ischemia in a cohort of patients who had coronary narrowings identified by angiography.


Metastatic colorectal cancer: Therapeutic options.
For the past several decades, the therapy for metastatic colorectal cancer had modest benefits because of the limited therapeutic options. Bolus 5-fluorouracil (5FU) and leucovorin (LV) were the standard of care in the United States until 2002, with a response rate of 25% and a median survival of 10 to 12 months. However, with the advent of new agents, namely oxaliplatin and irinotecan, there has been a dramatic change in the way we treat metastatic colorectal cancer. Based on many well-conducted large randomized trials, we have evidence that combination chemotherapy incorporating oxaliplatin or irinotecan with infusional 5FU/LV is superior to 5FU/LV, with doubling of overall survival (OS) to approximately 20 months. There remains some uncertainty as to the best first-line regimen. This might be irrelevant because studies have shown that OS is dependent on exposure to all the active agents, regardless of the time period of exposure. Bevacizumab, which uses anti-angiogenic strategies, has improved disease-free survival (DFS) and OS when combined with standard chemotherapy and is a vital component of metastatic colorectal cancer therapy. However, there are no data supporting its use past progression. Cetuximab, an epithelial growth factor receptor inhibitor, is mainly used in irinotecan-refractory patients. In spite of all these advances, 5-year OS rates continue to be limited. Patients with curative resection of metastatic disease seem to have longer DFS and better 5-year OS rates. This should be a potential goal for responding patients with upfront unresectable, organ-limited disease.


The dynamic instability of microtubules is not modulated by alpha-tubulin tyrosinylation.
The tyrosinylation of chick brain alpha-tubulin and the effects of the tyrosinylation status on the assembly and dynamic instability of chick brain MAP2:tubulin microtubule protein have been examined. Each of the eight major alpha-isotypes can be tyrosinylated in vitro, irrespective of whether a C-terminal tyrosine is genetically encoded. The extent of tyrosinylation is however limited to congruent to 0.3 mol.mol-1. The tyrosinylation status (0 vs. 0.3 mol.mol-1) has no effect on either the assembly kinetics of chick brain microtubule protein or on the rate of length redistribution following assembly and shearing. It is therefore unlikely that the tyrosinylation status directly affects the intrinsic stability of assembled microtubules since the rate of length redistribution is both a sensitive assay and a function of the kinetic parameters governing dynamic instability.


Overall health impacts of a potential increase in cycle commuting in Stockholm, Sweden.
To estimate the overall health impact of transferring commuting trips from car to bicycle. In this study registry information on the location of home and work for residents in Stockholm County was used to obtain the shortest travel route on a network of bicycle paths and roads. Current modes of travel to work were based on travel survey data. The relation between duration of cycling and distance cycled was established as a basis for selecting the number of individuals that normally would drive a car to work, but have a distance to work that they could bicycle within 30 minutes. The change in traffic flows was estimated by a transport model (LuTrans) and effects on road traffic injuries and fatalities were estimated by using national hospital injury data. Effects on air pollution concentrations were modelled using dispersion models. Within the scenario, 111,000 commuters would shift from car to bicycle. On average the increased physical activity reduced the one-year mortality risk by 12% among the additional bicyclists. Including the number of years lost due to morbidity, the total number of disability adjusted life-years gained was 696. The amount of disability adjusted life-years gained in the general population due to reduced air pollution exposure was 471. The number of disability adjusted life-years lost by traffic injuries was 176. Also including air pollution effects among bicyclists, the net benefit was 939 disability adjusted life-years per year. Large health benefits were estimated by transferring commuting by car to bicycle.


Variability of carotenoids in a Musa germplasm collection and implications for provitamin A biofortification.
Bananas are important staples in tropical and sub-tropical regions and their potential as a source of provitamin A has recently attracted attention for biofortification. A collection of 189 banana genotypes (AAB-plantains, M. acuminata cultivars and bred hybrids) was screened to determine variability in fruit pulp provitamin A carotenoid (pVAC) content using high performance liquid chromatography. Total carotenoid content in tested genotypes varied from 1.45 µg/g for hybrid 25447-S7 R2P8 to 36.21 µg/g for M. acuminata cultivar ITC.0601 Hung Tu with a mean of 8.00 µg/g fresh weight. Predominant carotenoids identified were α-carotene (38.67%), trans-β-carotene (22.08%), lutein (22.08%), 13-cis-β-carotene (14.45%) and 9-cis-β-carotene (2.92%), indicating that about 78% of the carotenoids in bananas are pVAC. High pVAC genotypes were identified for integration into biofortification strategies to combat vitamin A deficiency in developing countries.


A lower threshold for revision of aseptic unicompartmental vs total knee arthroplasty.
It has been hypothesized that a unicompartmental knee arthroplasty (UKA) is more likely to be revised than a total knee arthroplasty (TKA) because conversion surgery to a primary TKA is a less complicated procedure. The purpose of this study was to determine if there is a lower threshold for revising a UKA compared with TKA based on Oxford Knee Scores (OKSs) and range of movement (ROM) at the time of revision. We retrospectively reviewed 619 aseptic revision cases performed between December 1998 and October 2018. This included 138 UKAs that underwent conversion to TKA and 481 initial TKA revisions. Age, body mass index (BMI), time in situ, OKS, and ROM were available for all patients. There were no differences between the two groups based on demographics or time to revision. The top reasons for aseptic TKA revision were loosening in 212 (44%), instability in 88 (18%), and wear in 69 (14%). UKA revision diagnoses were primarily for loosening in 50 (36%), progression of osteoarthritis (OA) in 50 (36%), and wear in 17 (12%). Out of a maximum 48 points, the mean OKS of the UKAs before revision was 23 (SD 9.3), which was significantly higher than the TKAs at 19.2 (SD 9.8; p < 0.001). UKA patients scored statistically better on nine of the 12 individual OKS questions. The UKA cases also had a larger pre-revision mean ROM (114°, SD 14.3°) than TKAs (98°, SD 25°) ; p < 0.001). At revision, the mean UKA OKSs and ROM were significantly better than those of TKA cases. This study suggests that at our institution there is a difference in preoperative OKS between UKA and TKA at the time of revision, demonstrating a revision bias. Cite this article: Bone Joint J 2020;102-B(6 Supple A):91-95.


Fetal and early-postnatal developmental patterns of obese-genotype piglets exposed to prenatal programming by maternal over- and undernutrition.
The present study evaluated the effect of nutritional imbalances during pregnancy, either by excess or deficiency, on fertility and conceptus development in obese-genotype swine (Iberian pig). Twenty-five multiparous sows were fed, from mating to farrowing, with a standard diet fulfilling either 1.6 folds their daily maintenance requirements for pregnancy (overfed group, n = 12) or only the 50% of such requirements (underfed group, n = 13). Ten out of 12 overfed but only two out of 13 underfed sows became pregnant (P<0.005). Fetal development was determined in the pregnant females at Days 35, 50, 75 and 90 of pregnancy. The embryos from undernourished sows were smaller than the embryos from overfed females as early as at 35 days of pregnancy (P<0.05) and remained smaller until Day 90 of gestation. However, at the end of pregnancy, there were significant changes in the developmental patterns of fetuses. Thus, weight and size of the offspring from both nutritional treatments were finally similar at delivery; the same was found at weaning. There was thereafter a sex-related effect on the growth during the early-postnatal period, with male piglets of both nutritional origins being significantly heavier and more corpulent at weaning that their sisters (P<0.05). In conclusion, fetal growth conditioned by malnutrition from periconceptional stages is mainly regulated at the end of the pregnancy, so that ensure an adequate body-weight and size and, therefore, the survival of the offspring. Afterwards, the early-postnatal development of the offspring is affected by sex, independently from nutritional origin, with male piglets growing faster than females.


Medical students and radiology residents: can they learn as effectively with the same educational materials?
The purpose of this study was to evaluate the effectiveness of resident-prepared, independent-learning materials for teaching chest radiology to medical students. Students from three U.S. medical schools enrolled in radiology clerkships between March 1998 and June 1998 were randomly divided into control (n = 27) and experimental (n = 31) groups. The experimental group studied 12 chest radiology independent-learning cases (intervention) used to teach radiology residents in a previous study. Both groups took a 36-item, multiple-choice test (previously used to test radiology residents) on three occasions (before intervention [pretest], 1 day after intervention [posttest], and 2-4 weeks after intervention [final examination]). Student scores were then compared with resident scores. Mean scores were similar across institutions at pretest, but increases at posttest and final examination scores differed across time, school, and group (P < .005). Mean differences in scores between experimental and control groups at pretest, posttest, and 2-4-week final examination were -0.22, 9.79, and 9.14, respectively, demonstrating increased performance at posttesting that remained present (though slightly attenuated) 2-4 weeks later. Comparing performance, residents had mean pretest scores of 19.2 and students of 14.1, a five-point difference attributable to the residents' greater experience. Both residents and students gained approximately nine points at posttest. At final examination, the difference between residents and students was only 1.4 points, suggesting the experimental program (teaching materials) brought students close to the long-term retention shown by residents. Independent study of resident-prepared chest radiology teaching cases increased medical student knowledge for at least 2 or 4 weeks after instruction. Although starting at lower knowledge levels, students experienced gains in knowledge comparable to those of residents, suggesting the same materials can be used to teach both students and residents.


Zika Virus Associated Pathology and Antigen Presence in the Testicle in the Absence of Sexual Transmission During Subacute to Chronic Infection in a Mouse Model.
Zika virus (ZIKV) is an arboviral infection that has been shown to be sexually transmitted. The study outlined herein aims to determine if accessory sex glands and epididymal epithelial cells are sources of viral persistence in subacute and chronic ZIKV infection, and if infection of these organs is important in sexual transmission during long-term (chronic) infection. Male interferon type I receptor knockout (Ifnar-/-) mice were challenged with ZIKV and reproductive tissues were harvested 14 and 35 days post infection (DPI) for inoculation studies and 14, 35 and 70 DPI for histopathology. Artificial insemination fluid derived from epididymal flush and seminal plasma from the prostate and seminal vesicle was obtained from ZIKV inoculated and sham-infected males. Naïve interferon type I and II receptor knockout (AG129) female mice were pre-treated with progesterone and inoculated intravaginally with artificial insemination fluid from ZIKV-infected males. ZIKV RNA was detected in the artificial insemination fluid generated from epididymal flush or seminal plasma from ZIKV infected males at 14 and 35 DPI. ZIKV antigens were only detected in seminiferous tubules at 14 DPI. Epididymal epithelial cells did not show ZIKV antigen immunoreactivity at 14, 35 or 70 DPI. Severe fibrosing orchitis (end stage orchitis) was observed at 35 and 70 DPI. Mild inflammation and peri-tubular fibrosis were observed in the epididymis following clearance of virus. Viral RNA was not detected by PCR in whole blood samples of females from any intravaginal experimental group and only detected in 20% of subcutaneously inoculated animals (derived from 1 experimentally infected male) at 35 DPI. While ZIKV RNA and antigens can be detected in the male reproductive tract at 14 DPI and RNA can also be detected at 35 DPI, intravaginal inoculation of artificial insemination fluid from these time-points failed to result in viremia in naïve females inoculated intravaginally. These studies support the hypothesis that epididymal epithelial cells are critical to sexual transmission in immunocompromised mice. Additionally, acute but not chronic male reproductive tract infection with ZIKV results in infectious virus capable of being sexually transmitted in mice.


Vitamin A levels and immunity in humans.
In animal studies, vitamin A deficiency induces a shift from type 2 (humoral) to type 1 (cellular) cytokines; there are no similar data for humans. Control of human immunodeficiency virus (HIV) and Mycobacterium tuberculosis infections requires type 1 cytokine (cellular) immunity. These infections and vitamin A deficiency are highly prevalent in Africa. We therefore examined the interactions among serum vitamin A levels, immune parameters, HIV infection status, Mycobacterium bovis BCG vaccine scarring (as an indicator of a type 1 cytokine profile), and clinical findings for 70 hospitalized children in Malawi, Africa. Directly conjugated monoclonal antibodies and flow cytometry were used to assess cell-specific cytokine production by peripheral blood monocytes and lymphocyte subpopulations. The statistical techniques employed included nonparametric statistics and logistic regression analyses. Thirty percent of the participants had severe vitamin A deficiency (<10 microg/dl), 34% had moderate deficiency (10 to <20 microg/dl), and 36% had normal levels (> or = 20 microg/dl). Vitamin A levels were lower for HIV-positive than for HIV-negative children (median, 10 and 17 microg/dl, respectively). Vitamin A-deficient children (<20 microg/dl) were more likely than non-vitamin A-deficient children to have higher proportions of natural killer (NK) cells (median, 8.3 and 5.2%, respectively) and lower ratios of interleukin-10-producing monocytes to tumor necrosis factor alpha-producing monocytes after induction (median, 1.0 and 2.3, respectively). Vitamin A-deficient children were also more likely than non-vitamin A-deficient children to exhibit respiratory symptoms (47% versus 12%) and visible BCG vaccine scars (83% versus 48%), which are indicative of a type 1 response to vaccination. Vitamin A status did not vary with gender, age, incidence of malaria parasitemia, blood culture positivity, or rates of mortality (6% of vitamin A-deficient children died versus 20% of non-vitamin A-deficient children). Lower vitamin A levels were associated with a relative type 1 cytokine dominance and proportionately more NK cells, both of which may be somewhat beneficial to persons who are exposed to HIV, M. tuberculosis, or other type 1 pathogens.


Ginkgo biloba induces different gene expression signatures and oncogenic pathways in malignant and non-malignant cells of the liver.
Ginkgo biloba (EGb761) is a widely used botanical drug. Several reports indicate that EGb761 confers preventive as well as anti-tumorigenic properties in a variety of tumors, including hepatocellular carcinoma (HCC). We here evaluate functional effects and molecular alterations induced by EGb761 in hepatoma cells and non-malignant hepatocytes. Hepatoma cell lines, primary human HCC cells and immortalized human hepatocytes (IH) were exposed to various concentrations (0-1000 μg/ml) of EGb761. Apoptosis and proliferation were evaluated after 72h of EGb761 exposure. Response to oxidative stress, tumorigenic properties and molecular changes were further investigated. While anti-oxidant effects were detected in all cell lines, EGb761 promoted anti-proliferative and pro-apoptotic effects mainly in hepatoma cells. Consistently, EGb761 treatment caused a significant reduction in colony and sphere forming ability in hepatoma cells and no mentionable changes in IH. Transcriptomic changes involved oxidative stress response as well as key oncogenic pathways resembling Nrf2- and mTOR signaling pathway. Taken together, EGb761 induces differential effects in non-transformed and cancer cells. While treatment confers protective effects in non-malignant cells, EGb761 significantly impairs tumorigenic properties in cancer cells by affecting key oncogenic pathways. Results provide the rational for clinical testing of EGb761 in preventive and therapeutic strategies in human liver diseases.


DNA replication through hard-to-replicate sites, including both highly transcribed RNA Pol II and Pol III genes, requires the S. pombe Pfh1 helicase.
Replication forks encounter impediments as they move through the genome, including natural barriers due to stable protein complexes and highly transcribed genes. Unlike lesions generated by exogenous damage, natural barriers are encountered in every S phase. Like humans, Schizosaccharomyces pombe encodes a single Pif1 family DNA helicase, Pfh1. Here, we show that Pfh1 is required for efficient fork movement in the ribosomal DNA, the mating type locus, tRNA, 5S ribosomal RNA genes, and genes that are highly transcribed by RNA polymerase II. In addition, converged replication forks accumulated at all of these sites in the absence of Pfh1. The effects of Pfh1 on DNA replication are likely direct, as it had high binding to sites whose replication was impaired in its absence. Replication in the absence of Pfh1 resulted in DNA damage specifically at those sites that bound high levels of Pfh1 in wild-type cells and whose replication was slowed in its absence. Cells depleted of Pfh1 were inviable if they also lacked the human TIMELESS homolog Swi1, a replisome component that stabilizes stalled forks. Thus, Pfh1 promotes DNA replication and separation of converged replication forks and suppresses DNA damage at hard-to-replicate sites.


Removal of cyanobacterial metabolites through wastewater treatment plant filters.
Wastewaters have the potential to proliferate excessive numbers of cyanobacteria due to high nutrient levels. This could translate to the production of metabolites, such as the saxitoxins, geosmin and 2-methylisoborneol (MIB), which can impair the quality of wastewater destined for re-use. Biological sand filtration was assessed for its ability to remove these metabolites from a wastewater. Results indicated that the sand filter was incapable of effectively removing the saxitoxins and in some instances, the effluent of the sand filter displayed greater toxicity than the influent. Conversely, the sand filter was able to effectively remove geosmin and MIB, with removal attributed to biodegradation. Granular activated carbon was employed as an alternative filter medium to remove the saxitoxins. Results showed similar removals to previous drinking water studies, where efficient removals were initially observed, followed by a decrease in the removal; a consequence of the presence of competing organics which reduced adsorption of the saxitoxins.


Hypoxia inducible factor-1 (HIF-1)-mediated repression of cystic fibrosis transmembrane conductance regulator (CFTR) in the intestinal epithelium.
Diarrhea is widespread in intestinal diseases involving ischemia and/or hypoxia. Since hypoxia alters stimulated Cl(-) and water flux, we investigated the influence of such a physiologically and pathophysiologically important signal on expression of the cystic fibrosis transmembrane conductance regulator (CFTR). Located on the apical membrane, this cAMP-activated Cl(-) channel determines salt and fluid transport across mucosal surfaces. Our studies revealed depression of CFTR mRNA, protein, and function in hypoxic epithelia. Chromatin immunoprecipitation identified a previously unappreciated binding site for the hypoxia inducible factor-1 (HIF-1), and promoter studies established its relevance by loss of repression following point mutation. Consequently, HIF-1 overexpressing cells exhibited significantly reduced transport capacity in colorimetric Cl(-) efflux studies, altered short circuit measurements, and changes in transepithelial fluid movement. Whole-body hypoxia in wild-type mice resulted in significantly reduced small intestinal fluid and HCO(3)(-) secretory responses to forskolin. Experiments performed in Cftr(-/-) and Nkcc1(-/-) mice underlined the role of altered CFTR expression for these functional changes, and work in conditional Hif1a mutant mice verified HIF-1-dependent CFTR regulation in vivo. In summary, our study clarifies CFTR regulation and introduces the concept of a HIF-1-orchestrated response designed to regulate ion and fluid movement across hypoxic intestinal epithelia.


The scoring of poses in protein-protein docking: current capabilities and future directions.
Protein-protein docking, which aims to predict the structure of a protein-protein complex from its unbound components, remains an unresolved challenge in structural bioinformatics. An important step is the ranking of docked poses using a scoring function, for which many methods have been developed. There is a need to explore the differences and commonalities of these methods with each other, as well as with functions developed in the fields of molecular dynamics and homology modelling. We present an evaluation of 115 scoring functions on an unbound docking decoy benchmark covering 118 complexes for which a near-native solution can be found, yielding top 10 success rates of up to 58%. Hierarchical clustering is performed, so as to group together functions which identify near-natives in similar subsets of complexes. Three set theoretic approaches are used to identify pairs of scoring functions capable of correctly scoring different complexes. This shows that functions in different clusters capture different aspects of binding and are likely to work together synergistically. All functions designed specifically for docking perform well, indicating that functions are transferable between sampling methods. We also identify promising methods from the field of homology modelling. Further, differential success rates by docking difficulty and solution quality suggest a need for flexibility-dependent scoring. Investigating pairs of scoring functions, the set theoretic measures identify known scoring strategies as well as a number of novel approaches, indicating promising augmentations of traditional scoring methods. Such augmentation and parameter combination strategies are discussed in the context of the learning-to-rank paradigm.


A Copolymer Scaffold Functionalized with Nanodiamond Particles Enhances Osteogenic Metabolic Activity and Bone Regeneration.
Functionalizing polymer scaffolds with nanodiamond particles (nDPs) has pronounced effect on the surface properties, such as improved wettability, an increased active area and binding sites for cellular attachment and adhesion, and increased ability to immobilize biomolecules by physical adsorption. This study aims to evaluate the effect of poly(l-lactide-co-ε-caprolactone) (poly(LLA-co-CL)) scaffolds, functionalized with nDPs, on bone regeneration in a rat calvarial critical size defect. Poly(LLA-co-CL) scaffolds functionalized with nDPs are also compared with pristine scaffolds with reference to albumin adsorption and seeding efficiency of bone marrow stromal cells (BMSCs). Compared with pristine scaffolds, the experimental scaffolds exhibit a reduction in albumin adsorption and a significant increase in the seeding efficiency of BMSCs (p = 0.027). In the calvarial defects implanted with BMSC-seeded poly(LLA-co-CL)/nDPs scaffolds, live imaging at 12 weeks discloses a significant increase in osteogenic metabolic activity (p = 0.016). Microcomputed tomography, confirmed by histological data, reveals a substantial increase in bone volume (p = 0.021). The results show that compared with conventional poly(LLA-co-CL) scaffolds those functionalized with nDPs promote osteogenic metabolic activity and mineralization capacity. It is concluded that poly(LLA-co-CL) composite matrices functionalized with nDPs enhance osteoconductivity and therefore warrant further study as potential scaffolding material for bone tissue engineering.


The epidemiology and socioeconomic associations of retinal detachment in Scotland: a two-year prospective population-based study.
Rhegmatogenous retinal detachment (RRD) is a common ophthalmic emergency. Population-based data on primary RRD incidence has been variable, with large differences reported. This study is the first large-scale prospective examination of the incidence of primary RRD in the United Kingdom. The authors established a two-year prospective, population-based observational study recruiting all cases of primary RRD in Scotland. The annual incidence was calculated and analyzed in relation to age, sex, refractive error, and lens status. A national, population-based tool, the Scottish Index of Multiple Deprivation (SIMD), was used to examine the socioeconomic distribution of all incident cases. A total of 1244 cases were identified during the study period from a population of 5,168,500 yielding an annual incidence of 12.05 per 100,000 population (95% confidence interval, 11.35-12.70). The age-specific incidence increased to a peak in both sexes in the 60- to 69-year age group. RRD was significantly more frequent in males than in females (14.70 vs. 8.75 per 100,000; P < 0.001). Of the cases without previous intraocular surgery, 53.2% were myopic, with a spherical equivalent refractive error > -1 D, 23.4% had undergone cataract surgery, and 10.4% had sustained traumatic injury. A strong association was found between RRD incidence and affluence, with a significant rising trend across quintiles of deprivation. The estimated annual incidence of primary RRD in Scotland is 12.05 per 100,000. Based on this estimate, there are approximately 7300 new cases annually in the United Kingdom. RRD incidence increases with age, is more common in men and right eyes, and is strongly associated with affluence.


Analysis of radiographically confirmed blunt-mechanism facial fractures.
Facial fractures resulting from blunt-mechanism trauma, although common, have been infrequently evaluated in large studies that do not include confirmation of fractures based on author review of available patient radiographic studies. An 8-year review (1998-2006) of the R Adams Cowley Shock Trauma Registry was performed with institutional review board approval. Patients diagnosed with blunt-mechanism facial fractures were identified by the International Classification of Diseases, Ninth Revision (ICD-9) codes and their facial fractures confirmed by author review of computed tomographic scans. Individual fractures were classified and grouped according to the facial thirds. Intra- and interreader variability was calculated, and confirmed fracture patterns were compared to fracture patterns listed by ICD-9 codes. Concomitant injuries and demographic data were additionally evaluated. Four thousand three hundred ninety-eight patients with 8127 fractures were identified. Intra- and interreader variability was 2% and 7%, respectively. ICD-9 coding misdiagnosed 12.5% of all fractures. Eighty-two percent of patients sustained associated injuries, including basilar skull fractures (7.6%) and cervical spine fractures (6.6%). 1.1% had at least one fracture in each facial third (panfacial fracture pattern). Significant relationships were found between demographic parameters, concomitant injuries, specific fractures, and fracture patterns. Studies investigating facial fractures should report fracture patterns confirmed by author review of available radiographic imaging. Large retrospective data sets containing confirmed fractures and capable of addressing rare fracture patterns can be compiled with low inter- and intrauser variability, and are useful for generating mechanistic hypotheses suitable for evaluation in prospective series or by directed biomechanical studies.


[Bilingual guides: a strategy to decrease cultural barriers to health care access in the Wayuu communities of Maicao, Colombia].
The article examines the use of bilingual guides to decrease cultural barriers to health care access in the Wayuu indigenous communities of Colombia. Within a larger project on HIV carried out between 2012 and 2014, 24 interviews were conducted with key actors in the administrative and health areas, including Wayuu bilingual guides. As a result of the qualitative analysis, the study identified three cultural barriers to health care access: a) language; b) the Wayuu worldview regarding the body, health, and illness; and c) information about sexual and reproductive health and HIV not adapted to the Wayuu culture. The study identifies the bilingual guides as key actors in reducing these barriers and concludes with a discussion of the role of the guides, the tensions inherent to their work, and the complexity of their contributions as cultural mediators.


Hypertriglyceridemia-Associated Drug-Induced Acute Pancreatitis.
The aim of our study was to investigate the cases of drug-induced acute pancreatitis (DIAP) with hypertriglyceridemia as the mechanism of injury. A MEDLINE search (1963-2018) of the English language literature was performed looking for all human case reports of adults (>18 years old) with hypertriglyceridemia as the mechanism of DIAP. The latest search date was February 28, 2018. Drugs were classified into probability groups based on a classification used by Badalov et al (Clin Gastroenterol Hepatol. 2007;5:648-661). The search revealed a total of 76 cases in 59 published reports. A total of 25 medications were found to be implicated in DIAP secondary to hypertriglyceridemia mechanism. Most of acute pancreatitis cases were mild or moderately severe with favorable outcomes. In 3 cases involving tamoxifen, pagaspargase, and quetiapine, patient death was the outcome. Plasmapheresis was only used in 9 cases in an effort to decrease triglyceride levels. Hypertriglyceridemia-associated DIAP is a rare phenomenon, and the current systematic review provides a summary of drugs that have been implicated in this phenomenon, which allow physicians to be oriented about this adverse effect when these drugs are used.


Re-evaluation of plant sulpholipid labelling from UDP-[14C]glucose in pea chloroplasts.
The sulphoquinovosyldiacylglycerol (sulpholipid) zone obtained by TLC of pea chloroplast lipids yields, on hydrolysis, not only sulphoquinovose but also galactose and glucose. Following incorporation from UDP-[(14)C]glucose, the percentages of the total radioactivity in these three sugars were typically 1, 5 and 85%, respectively. The occurrence of the glucolipid causes difficulties in elucidating further the biosynthetic pathway for sulphoquinovose.


Biological activities and receptor binding of two human recombinant interferons and their hybrids.
Two human recombinant lymphoblastoid interferon-alpha subtypes, LyIFN-B (alpha 8) and LyIFN-D (alpha 1), and 10 hybrids generated therefrom were produced in Escherichia coli and purified. The antiviral and antiproliferative activities and the induction of (2'-5')oligoadenylate synthetase were compared to their receptor binding affinities. The IFN subtypes and their hybrids had similar specific antiviral activities on bovine cells. On human cells both the specific antiviral and antiproliferative activities of LyIFN-B were about 30-fold higher than those of LyIFN-D. This difference in activity could be attributed partly to the N-terminal amino acids 1 to 60 and partly to amino acids 61 to 92. A third domain affecting the biological activities was found within the carboxy-proximal segment from amino acids 93 to 150. The differences in these activities were found to correlate with their ability to bind the receptor, suggesting that the differences in activity might be due to altered binding of the IFNs to the cellular receptors. In contrast, the induction of (2'-5')oligoadenylate synthetase did not follow the same activity profile. On mouse cells, the efficiency of the hybrids was affected by at least four sites on the IFN protein. A hybrid with the N-terminal segment 1 to 60 from IFN-B and amino acids 61 to 166 from IFN-D had a specific antiviral activity on mouse cells as high as on human cells corresponding to a 500- and 5000-fold increase in specific activity compared to IFN-D and IFN-B, respectively. We suggest that on mouse cells the IFN activity may be more dependent on conformational differences than on human cells, which in turn might reflect a less precise fit to the mouse receptor than to the human receptor.


PARP2 deficiency affects invariant-NKT-cell maturation and protects mice from concanavalin A-induced liver injury.
Excessive or persistent inflammation and hepatocyte death are the key triggers of liver diseases. The poly(ADP-ribose) polymerase (PARP) proteins induce cell death and inflammation. Chemical inhibition of PARP activity protects against liver injury during concanavalin A (ConA)-induced hepatitis. In this mice model, ConA activates immune cells, which promote inflammation and induce hepatocyte death, mediated by the activated invariant natural killer T (iNKT) lymphocyte population. We analyzed immune cell populations in the liver and several lymphoid organs, such as the spleen, thymus, and bone marrow in Parp2-deficient mice to better define the role of PARP proteins in liver immunity and inflammation at steady state and during ConA-induced hepatitis. We show that 1) the genetic inactivation of Parp2, but not Parp1, protected mice from ConA hepatitis without deregulating cytokine expression and leucocyte recruitment; 2) cellularity was lower in the thymus, but not in spleen, liver, or bone marrow of Parp2-/- mice; 3) spleen and liver iNKT lymphocytes, as well as thymic T and NKT lymphocytes were reduced in Parp2 knockout mice. In conclusion, our results suggest that the defect of T-lymphocyte maturation in Parp2 knockout mice leads to a systemic reduction of iNKT cells, reducing hepatocyte death during ConA-mediated liver damage, thus protecting the mice from hepatitis.NEW & NOTEWORTHY The genetic inactivation of Parp2, but not Parp1, protects mice from concanavalin A hepatitis. Immune cell populations are lower in the thymus, but not in the spleen, liver, or bone marrow of Parp2-deficient mice compared with wild-type mice. Spleen and liver invariant natural killer T (NKT) lymphocytes, as well as thymic T and NKT lymphocytes, are reduced in Parp2-deficient mice.


Treatment of stage I and II oral tongue cancer.
We reviewed 156 previously untreated patients with squamous cell carcinoma of the oral tongue staged T1 and T2 to determine the incidence of nodal metastasis, and if elective neck dissection affected local/regional control or survival. Patients were divided into two nonrandomized groups: group 1, intraoral glossectomy only (102 patients); and group 2, intraoral glossectomy plus neck dissection (54 patients). Analysis revealed no significant differences for tumor location, histologic differentiation, status of margins, or clinical appearance; however, perineural invasion significantly adversely affected survival and local/regional control. In group 1 patients, 16.5% subsequently developed cervical metastasis, and 20.4% of patients in group 2 had occult nodal disease. The survival and local/regional control for group 1 patients subsequently developing nodes was 33% and 50%, respectively. The survival and local/regional control for group 2 patients with occult metastasis was 55% and 91%, respectively. We believe elective neck dissection is indicated for early staged oral tongue cancer.


Synthesis of diaryl ether components of ellagitannins using ortho-quinone with consonant mesomeric effects.
Methods for synthesizing C-O digallate structures, the basic unit of diaryl ether components of natural ellagitannins, are described. In the designed building block derived from gallic acid, consonantly overlapped mesomeric effects enhanced its electrophilicity. This building block demonstrated substantial reactivity to improve the synthesis of dehydrodigalloyl, tergalloyl, and valoneoyl groups.


Heat shock protein 70 confers cardiovascular protection during endotoxemia via inhibition of nuclear factor-kappaB activation and inducible nitric oxide synthase expression in the rostral ventrolateral medulla.
Overproduction of nitric oxide (NO) by inducible NO synthase (iNOS) in the rostral ventrolateral medulla (RVLM), where sympathetic premotor neurons are located, plays a pivotal role in the manifestation of fatal cardiovascular depression during endotoxemia. The iNOS gene is regulated transcriptionally by nuclear factor-kappaB (NF-kappaB) activation. The present study tested the hypothesis that heat shock protein 70 (HSP70) may confer protection against sepsis-induced circulatory fatality via inhibition of iNOS gene expression in the RVLM through prevention of NF-kappaB activation. Adult male Sprague-Dawley rats subjected to a brief hyperthermic heat shock (42 degrees C for 15 minutes) exhibited significant upregulation of HSP70 in the RVLM. Brief heat shock preconditioning also significantly suppressed iNOS mRNA or protein surge and alleviated hypotension, bradycardia, and reduction in neurogenic sympathetic vasomotor activity manifested during experimental endotoxemia induced by intravenous administration of Escherichia coli lipopolysaccharide. An increase in DNA binding activity and nuclear translocation of transcription factor NF-kappaB were detected during endotoxemia. Heat shock preconditioning significantly decreased DNA binding activity of NF-kappaB, which was reversed by microinjection of an hsp70 antisense oligonucleotide bilaterally into the RVLM. Heat shock preconditioning also blocked inhibitory kappaB (IkappaB) kinase activity or degradation of IkappaB in the RVLM during endotoxemia. We conclude that HSP70 confers protection against sepsis-related circulatory fatality via inhibition of iNOS gene expression in the RVLM through prevention of NF-kappaB activation in cellular processes that include prevention of IkappaB kinase activation and inhibition of IkappaBalpha degradation.


Relationship between work rate and oxygen uptake in mitochondrial myopathy during ramp-incremental exercise.
We determined the response characteristics and functional correlates of the dynamic relationship between the rate (Δ) of oxygen consumption (VO(2)) and the applied power output (work rate = WR) during ramp-incremental exercise in patients with mitochondrial myopathy (MM). Fourteen patients (7 males, age 35.4 ± 10.8 years) with biopsy-proven MM and 10 sedentary controls (6 males, age 29.0 ± 7.8 years) took a ramp-incremental cycle ergometer test for the determination of the VO(2) on-exercise mean response time (MRT) and the gas exchange threshold (GET). The ΔVO(2)/ΔWR slope was calculated up to GET (S(1)), above GET (S(2)) and over the entire linear portion of the response (S(T)). Knee muscle endurance was measured by isokinetic dynamometry. As expected, peak VO(2) and muscle performance were lower in patients than controls (P < 0.05). Patients had significantly lower ΔVO(2)/ΔWR than controls, especially the S(2) component (6.8 ± 1.5 vs 10.3 ± 0.6 mL·min(-1)·W(-1), respectively; P < 0.001). There were significant relationships between ΔVO(2)/ΔWR (S(T)) and muscle endurance, MRT-VO(2), GET and peak VO(2) in MM patients (P < 0.05). In fact, all patients with ΔVO(2)/ΔWR below 8 mL·min(-1)·W(-1) had severely reduced peak VO(2) values (<60% predicted). Moreover, patients with higher cardiopulmonary stresses during exercise (e.g., higher Δ ventilation/carbon dioxide output and Δ heart rate/ΔVO(2)) had lower ΔVO(2)/ΔWR (P < 0.05). In conclusion, a readily available, effort-independent index of aerobic dysfunction during dynamic exercise (ΔVO(2)/ΔWR) is typically reduced in patients with MM, being related to increased functional impairment and higher cardiopulmonary stress.


Novel phosphorylation of c-ras p21 by protein kinases.
Novel p21 phosphorylation was found in cells expressing high levels of this product of c-H- and c-K-ras genes. Phorbol 12,13-dibutyrate, a protein kinase C (PKC) activator, and permeable c-AMP derivatives, which activate protein kinase A (PKA), stimulated phosphorylation of K-ras(4B) p21 in 416B cells 3 to 5 fold. By tryptic peptide mapping, it was found that both PKC and PKA phosphorylated in vitro the K-ras p21 at the same site as was found in p21 from cells labeled with [32P]orthophosphate in vivo. A common site of H-ras p21 was also phosphorylated by both PKC and PKA, although phosphopeptides of H-ras p21 were distinct from those of K-ras p21. The construction of a mutant by site-directed mutagenesis allowed the identification of serine-177 as the phosphorylation site of H-ras p21. This novel phosphorylation site lies in the hypervariable region, which links the globular catalytic domain of p21 to the membrane-anchoring site at the C-terminus, a location suggesting that this phosphorylation plays a role in modulating transmembrane signaling.


Selective binding of N-acetylglucosamine to the chicken hepatic lectin.
Among Ca2+-dependent (C-type) animal lectins, the chicken hepatic lectin (CHL) is unique in displaying almost complete selectivity for N-acetylglucosamine over other monosaccharide ligands. The crystal structures of the carbohydrate-recognition domain (CRD) from serum mannose-binding protein (MBP) and of a complex between the CRD from liver MBP and the methyl glycoside of N-acetylglucosamine were used to model the binding site in CHL. Substitution of portions of CHL into the MBP framework did not substantially increase selectivity. A bacterial expression system for the CRD of CHL was developed so that specific residues predicted to be near the 2-acetamido substituent of N-acetylglucosamine could be altered by site-directed mutagenesis. The results indicate that the ligand is bound to CHL in the same orientation as it binds to liver MBP. A tyrosine and a valine residue that probably contact the the N-acetyl group have been identified. These results, together with studies of ligand-binding selectivity, suggest that these residues form part of a binding pocket for the N-acetyl group, which confers selective binding of N-acetylglucosamine.


Incidence and risk of treatment-related mortality with mTOR inhibitors everolimus and temsirolimus in cancer patients: a meta-analysis.
Two novel mammalian targets of rapamycin (mTOR) inhibitors everolimus and temsirolimus are now approved by regulatory agencies and have been widely investigated among various types of solid tumors, but the risk of fatal adverse events (FAEs) with these drugs is not well defined. We searched PubMed, EMBASE, and Cochrane library databases for relevant trials. Eligible studies included prospective phase II and III trials evaluating everolimus and temsirolimus in patients with all malignancies and data on FAEs were available. Statistical analyses were conducted to calculate the summary incidence, RRs and 95% confidence intervals (CIs) by using either random effects or fixed effect models according to the heterogeneity of the included studies. A total of 3322 patients with various advanced solid tumors from 12 trials were included. The overall incidence of mTOR inhibitors associated FAEs was 1.8% (95%CI: 1.3-2.5%), and the incidences of everolimus related FAEs were comparable to that of temsirolimus (1.7% versus 1.8%). Compared with the controls, the use of mTOR inhibitors was associated with an increased risk of FAEs, with a RR of 3.24 (95%CI: 1.21-8.67, p = 0.019). On subgroup analysis, a non-statistically significant increase in the risk of FAEs was found according to different mTOR inhibitors, tumor types or controlled therapy. No evidence of publication bias was observed. With the present evidence, the use of mTOR inhibitors seems to increase the risk of FAEs in patients with advanced solid tumors. More high quality trials are still needed to investigate this association.


Prominent mental (chin) crease: a new sign of pseudoxanthoma elasticum.
Pseudoxanthoma elasticum (PXE) is an inherited disease with many systemic complications. Its diagnosis can be easily overlooked. The purpose of this study was to assess a new clinical sign of PXE. A total of 47 consecutive patients with PXE were examined for mental creases and compared with 47 age-matched control patients. Photographs were evaluated by a blinded investigator. Mental creases were present in 43 of 47 patients with PXE, affected the majority of individuals in every age group, and were significantly more common than in control patients (P <.0001). Under the age of 30 years, two thirds of patients with PXE had mental creases whereas mental creases were not present in any age-matched control patients. Mental creases commonly occur in patients with PXE. In young patients they are a sensitive and highly specific physical finding of this disorder. The presence of exaggerated mental creases is, thus, a useful physical marker for PXE.


Does Selection and Management of Patients with Chronic Kidney Disease In Government Run and Private Hospitals Differ?
Globally, incidence of Chronic Kidney Disease (CKD) is rapidly rising with huge burden on the life expectancy of the patients. Regular haemodialysis improves the quality of life in these patients. They get treatment at either government run or private sector hospitals. A difference in disease pattern, comorbidity, patient management and number of access failures can be observed in these set ups. The present study was carried out to find out selection, management and disease pattern of CKD patients admitted for dialysis in government run and private hospital. A cross-sectional study on patients (18-90 years) admitted and undergoing dialysis at government run (N=129) and private hospital (N=182) was undertaken in Karnataka, India. Parameters like comorbidity (diabetes), number of dialysis per week, number of access failures, and follow up visits were compared between these patients. Chi- squared test was used to compare the data. All tests were two-tailed and p< 0.05 was considered as significant. More number of younger patients and associated comorbidity, were seen in patients admitted in government run hospital (p<0.001), with no gender bias in selection of patients for dialysis between the two hospitals. Similarly, follow-ups with nephrologist, number of dialysis done per week and erythropoietin supplements administered were significantly more among private hospital patients (p<0.001). Number of dialysis sessions and mean haemoglobin level was less in government run hospital patients, as compared to those in private hospital. No statistical difference was seen with access failure in both these setups. No bias in management of CKD patient was seen among the two sets of hospitals though available facilities seemed to vary.


[Bleaching of teeth: a review].
Bleaching of vital and non-vital teeth is becoming a routine procedure in esthetic dentistry. In a relative safe way many types of discoloration can be treated. Four treatment systems are available at present. For an appropriate choice it is essential to know the cause of the discoloration. Long term results are still scarce. Most treatment systems show slight to moderate side-effects, varying from demineralisation of enamel to soft tissue problems.


Predicting Deep Body Temperature (Tb) from Forehead Skin Temperature: Tb or Not Tb?
There is a need to rapidly screen individuals for heat strain and fever using skin temperature (Tsk) as an index of deep body temperature (Tb). This study's aim was to assess whether Tsk could serve as an accurate and valid index of Tb during a simulated heatwave. Seven participants maintained a continuous schedule over 9-days, in 3-day parts; pre-/post-HW (25.4 °C), simulated-HW (35.4 °C). Contact thermistors measured Tsk (Tforehead, Tfinger); radio pills measured gastrointestinal temperature (Tgi). Proximal-distal temperature gradients (ΔTforehead-finger) were also measured. Measurements were grouped into ambient conditions: 22, 25, and 35 °C. Tgi and Tforehead only displayed a significant relationship in 22 °C (r: 0.591; p < 0.001) and 25 °C (r: 0.408; p < 0.001) conditions. A linear regression of all conditions identified Tforehead and ΔTforehead-finger as significant predictors of Tgi (r2: 0.588; F: 125.771; p < 0.001), producing a root mean square error of 0.26 °C. Additional residual analysis identified Tforehead to be responsible for a plateau in Tgi prediction above 37 °C. Contact Tforehead was shown to be a statistically suitable indicator of Tgi in non-HW conditions; however, an error of ~1 °C makes this physiologically redundant. The measurement of multiple sites may improve Tb prediction, though it is still physiologically unsuitable, especially at higher ambient temperatures.


Anaemia among patients with heart failure and preserved or reduced ejection fraction: results from the SENIORS study.
Anaemia is a co-morbidity frequently seen in patients with heart failure (HF) and reduced left ventricular ejection fraction (LVEF). Its presence carries adverse prognostic effects. The effects of anaemia have not been extensively investigated in patients with preserved or only mildly reduced LVEF. We sought to investigate prevalence and incidence of anaemia in patients with HF irrespective of whether reduced or preserved ejection fraction are present. In addition, we sought to study the effects of nebivolol on the development of anaemia. We analysed data from 2069 patients randomized to nebivolol or placebo in the SENIORS study, 391 (10.0%) of whom were anaemic. Anaemia was similarly common in patients with LVEF≤35% and those with LVEF>35% (19.0 vs. 18.7%, P= 0.89). Anaemic patients were older, had lower diastolic blood pressure, and worse kidney function (all P< 0.05). No difference was found between patients on nebivolol or placebo with regard to the presence of anaemia. A total of 348 (16.8%) patients died during follow-up. The combined primary endpoint of all-cause mortality and cardiovascular hospital admission was reached by 687 (33.2%) patients during follow-up, 164 (23.4%) of whom were anaemic. Anaemic patients had a higher risk of reaching a primary endpoint than non-anaemics [LVEF≤35%: hazard ratio (HR) 1.47, 95% confidence interval (CI) 1.18-1.82, P< 0.001; LVEF>35%: HR 1.47, 95% CI 1.09-2.00, P= 0.012]. After multivariable adjustment, haemoglobin remained an independent predictor of the primary outcome in this cohort of patients (HR 0.94 per 1 g/dL increase, 95% CI 0.89-0.99, P= 0.017). Anaemia is an independent predictor of death or hospitalization for cardiovascular reasons among elderly patients with chronic HF and reduced or preserved/mildly reduced LVEF. Nebivolol does not affect haemoglobin values during follow-up.


The ecological risks of transgenic plants.
Biotechnologies have been utilized "ante litteram" for thousands of years to produce food and drink and genetic engineering techniques have been widely applied to produce many compounds for human use, from insulin to other medicines. The debate on genetically modified (GM) organisms broke out all over the world only when GM crops were released into the field. Plant ecologists, microbiologists and population geneticists carried out experiments aimed at evaluating the environmental impact of GM crops. The most significant findings concern: the spread of transgenes through GM pollen diffusion and its environmental impact after hybridisation with closely related wild species or subspecies; horizontal gene transfer from transgenic plants to soil microbes; the impact of insecticide proteins released into the soil by transformed plants on non-target microbial soil communities. Recent developments in genetic engineering produced a technology, dubbed "Terminator", which protects patented genes introduced in transgenic plants by killing the seeds in the second generation. This genetic construct, which interferes so heavily with fundamental life processes, is considered dangerous and should be ex-ante evaluated taking into account the data on "unexpected events", as here discussed, instead of relying on the "safe until proven otherwise" claim. Awareness that scientists, biotechnologists and genetic engineers cannot answer the fundamental question "how likely is that transgenes will be transferred from cultivated plants into the natural environment?" should foster long-term studies on the ecological risks and benefits of transgenic crops.


[Invagination vasoepididymoanastomosis with application of microsurgical tools].
A variant of microsurgical invagination vasoepididymostomy (patent of the Russian Federation N 2195205, 2002) was developed in 2001-2004 in experiment on 21 mongrel dogs. Fourteen dogs have undergone 22 vasoepididymostomies according to the novel technique and 5 control vasoepididymoanastomoses side-to-side. The proposed operation implies deep antegrade submersion and fixation of the stump of the epididymis duct in the lumen of the deferent duct through a small slit wound of the latter. Duration of the operation is 62 +/- 3 min. Passability of the anastomosis was assessed 6 months after the operation with ejaculate microscopy, retrograde staining of the deferent ducts on the preparations and with microscopy of stained histological sections. Spermatozoa were detected in ejaculate of all the operated on dogs. By microscopy, 20 anastomoses were capable. This operation technique was introduced into clinical practice: 3 patients with epididymic obstruction were operated on. Passability recovered 1.5-2 months after the operation in two of them. Thus, this variant of invagination vasoepididymostomy is perspective for wide clinical practice.


Daily oscillation of odorant detection in rat olfactory epithelium.
Most of biological variables follow a daily rhythm. It holds true as well for sensory capacities as two decades of research have demonstrated that the odorant induced activity in the olfactory bulbs oscillates during the day. Olfactory bulbs are the first central nervous system structures, which receive inputs from the olfactory neurons located in the nose olfactory epithelium in vertebrates. So far, data on variation in odorant detection in the olfactory epithelium throughout the day are missing. Using electroolfactogram recordings in rats housed under daily light and dark cycles, we found that the olfactory epithelium responsiveness varies during the day with a maximum in the beginning of the light phase. This fluctuation was consistent with cycling of transduction pathway gene expression in the olfactory epithelium examined by qPCR. It was also consistent with the levels of two transduction pathway proteins (olfactory-type G protein and adenylyl cyclase III) examined by western blot. Daily variations were also observed at the level of olfactory sensory neurons responses recorded by patch-clamp. To rule out a potential effect of the feeding status of the animal, we examined the variation in odorant response in starved animals during the day. We observed a similar pattern to ad libidum fed animals. Taken together, our results reveal that the olfactory epithelium sensitivity varies during the day in part due to modulation of the very first step of odorant detection.


Identification and pharmacological characterization of cholesterol-5,6-epoxide hydrolase as a target for tamoxifen and AEBS ligands.
The microsomal antiestrogen binding site (AEBS) is a high-affinity target for the antitumor drug tamoxifen and its cognate ligands that mediate breast cancer cell differentiation and apoptosis. The AEBS, a hetero-oligomeric complex composed of 3beta-hydroxysterol-Delta8-Delta7-isomerase (D8D7I) and 3beta-hydroxysterol-Delta7-reductase (DHCR7), binds different structural classes of ligands, including ring B oxysterols. These oxysterols are inhibitors of cholesterol-5,6-epoxide hydrolase (ChEH), a microsomal epoxide hydrolase that has yet to be molecularly identified. We hypothesized that the AEBS and ChEH might be related entities. We show that the substrates of ChEH, cholestan-5alpha,6alpha-epoxy-3beta-ol (alpha-CE) and cholestan-5beta,6beta-epoxy-3beta-ol (beta-CE), and its product, cholestane-3beta,5alpha,6beta-triol (CT), are competitive ligands of tamoxifen binding to the AEBS. Conversely, we show that each AEBS ligand is an inhibitor of ChEH activity, and that there is a positive correlation between these ligands' affinity for the AEBS and their potency to inhibit ChEH (r2=0.95; n=39; P<0.0001). The single expression of D8D7I or DHCR7 in COS-7 cells slightly increased ChEH activity (1.8- and 2.6-fold), whereas their coexpression fully reconstituted ChEH, suggesting that the formation of a dimer is required for ChEH activity. Similarly, the single knockdown of D8D7I or DHCR7 using siRNA partially inhibited ChEH in MCF-7 cells, whereas the knockdown of both D8D7I and DHCR7 abolished ChEH activity by 92%. Taken together, our findings strongly suggest that the AEBS carries out ChEH activity and establish that ChEH is a new target for drugs of clinical interest, polyunsaturated fatty acids and ring B oxysterols.


Transcriptional induction of cyclooxygenase-2 in osteoclast precursors is involved in RANKL-induced osteoclastogenesis.
Regulation of osteoclast differentiation is key to understanding the pathogenesis and to developing treatments for bone diseases such as osteoporosis. To gain insight into the mechanism of the receptor activator of nuclear factor (NF)-kappaB ligand (RANKL)-specific induction of the osteoclast differentiation program, we took a suppression-subtractive hybridization screening approach to identify genes specifically induced via the RANKL-Rac1 signaling pathway. Among identified targets, we show that RANKL selectively induces cyclooxygenase (COX) 2 expression via Rac1 that results in turn in production of prostaglandin E2 (PGE2) in RAW 264.7 cells. By using transient transfection assays, we found that the -233/-206 region of the COX-2 promoter gene was critical for RANKL-induced promoter activity. This RANKL-responsive region contained an NF-kappaB site that, when mutated, completely abolished the induction of NF-kappaB DNA-binding activity by RANKL. Blockade of COX-2 by celecoxib inhibits differentiation of bone marrow-derived monocyte/macrophage precursor cells (BMMs) into tartrate-resistant acid phosphatase-positive (TRAP+) osteoclastic cells. This inhibition can be rescued by the addition of exogenous PGE2, suggesting that COX-2-dependent PGE2 induction by RANKL in osteoclast precursors is required for osteoclast differentiation.


High-definition white-light (HDWL) colonoscopy and higher adenoma detection rate and the potential for paradoxical over surveillance.
Patients undergoing high-definition white-light colonoscopy by a high adenoma detector may have a lower risk of interval adenoma detection on follow-up colonoscopy and may require less frequent follow-up but may paradoxically be assigned to more frequent surveillance when more adenomas are detected. To evaluate whether high-definition white-light colonoscopy (vs. standard-definition white-light colonoscopy) and endoscopist adenoma detection rate (ADR) at index colonoscopy are associated with increased likelihood of adenomas at follow-up. Longitudinal follow-up of prior cross-section cohort study of patients who underwent colonoscopy at baseline with at least one detected adenoma was included. Associations of type of white-light at index colonoscopy and the ADR of the endoscopist at index colonoscopy (high vs. low adenoma detector) were evaluated with various adenoma and polyp detection endpoints. Eighteen endoscopists were classified as high and low adenoma detectors based on the median ADR of 0.255. There were no significant differences in subsequent interval adenoma or polyp detection endpoints with regard to whether baseline exam was performed with high-definition white-light or standard-definition white-light colonoscopy nor between high and low ADR after adjusting for multiple testing (P ≤ 0.0029 considered significant). Prior to multiple testing adjustment, there was a significantly lower detection rate of hyperplastic polyps in the left colon (24 vs. 35 %, OR: 0.56, P = 0.033) at follow-up colonoscopy when baseline exam was performed with high-definition white-light index colonoscopy. The results of this study do not support adjusting colonoscopy surveillance guidelines based on type of colonoscopy performed or the endoscopist's ADR.


[Effect of cholesterol reducing therapy with 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor on secondary prevention after myocardial infarction in Japanese patients].
Lowering the blood cholesterol level is a safe method to improve survival for primary and secondary prevention of coronary heart disease. However, there is no evidence for any effectiveness in Japanese. This study was designed to evaluate the effect of cholesterol lowering therapy with 3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA) reductase inhibitor on cardiac events(death and reinfarction) in Japanese patients after myocardial infarction. A total of 290 patients after myocardial infarction were studied retrospectively. The patients were divided into 2 groups with or without HMG-CoA reductase inhibitor therapy for lowering blood cholesterol levels. The cumulative cardiac events and percentage change of cholesterol levels[total cholesterol and low-density lipoprotein (LDL) cholesterol level] were compared between the 2 groups. HMG-CoA reductase inhibitor therapy lowered plasma cholesterol levels significantly (total cholesterol level--11 +/- 20%, LDL cholesterol level--23 +/- 26%) in patients with hypercholesterolemia, whereas there was no change(total cholesterol level 4.3 +/- 22%, LDL cholesterol level--7.2 +/- 24%) in patients without hypercholesterolemia. HMG-CoA reductase inhibitor therapy reduced cardiac events significantly compared in patients with hypercholesterolemia(p = 0.0008), but there was no benefit in patients without hypercholesterolemia. We suggest that treatment with HMG-CoA reductase inhibitor therapy for lowering cholesterol levels was effective for secondary prevention after myocardial infarction in Japanese patients with hypercholesterolemia.


Importance and Feasibility of Transitional Care for Children With Medical Complexity: Results of a Multistakeholder Delphi Process.
Children with medical complexity (CMC) account for disproportionate hospital utilization and adverse outcomes after discharge, and several gaps exist regarding the quality of hospital to home transitional care for this population. We conducted an expert elicitation process to identify important and feasible hospital to home transitional care interventions for CMC from the perspectives of parents and health care professionals. We conducted a 2-round electronic Delphi process to identify important and feasible transitional care interventions. Panelists included parents of CMC and multidisciplinary health care professionals. In the first round, panelists rated the importance and feasibility of 39 transitional care interventions on a 9-point Likert scale; agreement between panelists was defined according to RAND/UCLA Appropriateness Methods. The second round of data collection evaluated 16 interventions that panelists did not agree on in the first round and 8 new or revised interventions, accompanied by quantitative and qualitative data summaries. A total of 29 parents of CMC and 37 health care professionals participated in the Delphi process (response rate 75%). Both stakeholder panels endorsed most interventions as important; health care professionals were less likely to rate several interventions as feasible compared with the parent panel. Over 2 rounds of data collection, the 2 stakeholder panels endorsed 25 interventions as important as well as feasible. These interventions related to family engagement during the hospitalization, care coordination and social support assessment, predischarge education, and written materials. Parents and health care professionals considered several transitional care interventions important as well as feasible. This research might inform hospitals' transitional care programs and policies.


Plasticity of parental care under the risk of predation: how much should parents reduce care?
Predation can be an important agent of natural selection shaping parental care behaviours, and can also favour behavioural plasticity. Parent birds often decrease the rate that they visit the nest to provision offspring when perceived risk is high. Yet, the plasticity of such responses may differ among species as a function of either their relative risk of predation, or the mean rate of provisioning. Here, we report parental provisioning responses to experimental increases in the perceived risk of predation. We tested responses of 10 species of bird in north temperate Arizona and subtropical Argentina that differed in their ambient risk of predation. All species decreased provisioning rates in response to the nest predator but not to a control. However, provisioning rates decreased more in species that had greater ambient risk of predation on natural nests. These results support theoretical predictions that the extent of plasticity of a trait that is sensitive to nest predation risk should vary among species in accordance with predation risk.


The long-term effectiveness of different regimens of occlusion on recovery from early monocular deprivation in kittens.
Although the behavioural effects of an early period of monocular deprivation imposed on kittens can be very severe, resembling an extreme form of the human clinical condition deprivation amblyopia, they are not necessarily irreversible. Considerable behavioural as well as physiological recovery can occur if normal visual input is restored to the deprived eye sufficiently early, particularly if the other (initially non-deprived) eye is occluded at the same time (reverse occlusion). However, past work has shown that in many situations the improvement in the vision of the initially deprived eye that occurs during reverse occlusion is not retained following the subsequent introduction of binocular visual input. Furthermore, the vision of the other eye is often reduced as well, with the result that the eventual outcome is a condition of bilateral amblyopia. This study first examines the consequences of several periods of reverse occlusion whose onset and duration would be thought to maximize the opportunity for good and long-standing recovery of vision in the initially deprived eye. However, only in a very restricted set of exposure conditions did animals acquire good vision in one or both eyes; in most situations the final outcome was one of bilateral amblyopia. A second set of experiments examined the consequences of various regimens of part-time reverse occlusion, where the initially non-deprived eye was occluded for only part of each day to allow a period of binocular visual exposure, on kittens that had been monocularly deprived until 6, 8, 10 or 12 weeks of age. Whereas short or long daily periods of occlusion of the initially non-deprived eye resulted eventually in amblyopia in one, or usually both, eyes, certain intermediate occlusion times (3.5 or 5 h each day) resulted in recovery of normal acuities, contrast sensitivity and vernier acuity in both eyes, in animals that had been monocularly deprived until 6, 8 or 10 weeks of age, but not in animals deprived for longer periods. Experiments were done to establish some of the factors that contributed to the successful outcome associated with certain of the regimens of part-time reverse occlusion. It was established that recovery was just as good in animals in which the visual axes were vertically misaligned by means of prisms during the daily period of binocular visual exposure, thereby indicating that the visual input to the two eyes need not be concordant. However, animals that received equivalent visual exposure of the two eyes each day, but successively rather than simultaneously, all developed very severe bilateral amblyopia.(ABSTRACT TRUNCATED AT 400 WORDS)


High total histone deoxyribonucleic acid ratios for rat liver nuclei.
The ratios of total histone to DNA for rat liver nuclei isolated by four methods as well as for calf liver nuclei isolated by one method were determined by obtaining the ratios of the total areas of the electrophoretic histone peaks for the liver nuclei to the corresponding total area given by a known amount of standard calf thymus histone. Ratios of total histone to DNA of approx. 2 for rat liver nuclei isolated at pH3.8 or 5.8 and for calf liver nuclei isolated at pH3.8 were confirmed twice by the above procedure and also by direct measurement, by the method of Lowry et al. (1951), of histone extracted in 0.2m-H(2)SO(4). The histones of calf thymus, calf liver and rat liver were characterized by their amino acid compositions and by polyacrylamide-gel electrophoresis.


Age and micronutrient effects on the microbiome in a mouse model of zinc depletion and supplementation.
Older adult populations are at risk for zinc deficiency, which may predispose them to immune dysfunction and age-related chronic inflammation that drives myriad diseases and disorders. Recent work also implicates the gut microbiome in the onset and severity of age-related inflammation, indicating that dietary zinc status and the gut microbiome may interact to impact age-related host immunity. We hypothesize that age-related alterations in the gut microbiome contribute to the demonstrated zinc deficits in host zinc levels and increased inflammation. We tested this hypothesis with a multifactor two-part study design in a C57BL/6 mouse model. The two studies included young (2 month old) and aged (24 month old) mice fed either (1) a zinc adequate or zinc supplemented diet, or (2) a zinc adequate or marginal zinc deficient diet, respectively. Overall microbiome composition did not significantly change with zinc status; beta diversity was driven almost exclusively by age effects. Microbiome differences due to age are evident at all taxonomic levels, with more than half of all taxonomic units significantly different. Furthermore, we found 150 out of 186 genera were significantly different between the two age groups, with Bacteriodes and Parabacteroides being the primary taxa of young and old mice, respectively. These data suggest that modulating individual micronutrient concentrations does not lead to comprehensive microbiome shifts, but rather affects specific components of the gut microbiome. However, a phylogenetic agglomeration technique (ClaaTU) revealed phylogenetic clades that respond to modulation of dietary zinc status and inflammation state in an age-dependent manner. Collectively, these results suggest that a complex interplay exists between host age, gut microbiome composition, and dietary zinc status.


The effect of rest breaks on time to injury - a study on work-related ladder-fall injuries in the United States.
Rest breaks and other work-related temporal factors, such as time spent on task, influence the accumulation of fatigue, and thus impact occupational injury risk. The aim of this study was to investigate the effect of rest breaks on "time to injury" (the time between start of work and injury) for injured workers treated in a nationally representative sample of US emergency departments. Using the National Electronic Injury Surveillance System (NEISS), we identified 629 workers who had experienced a work-related ladder-fall. Of these, 306 were interviewed by telephone using a standardized questionnaire about the circumstances surrounding the injury. Survival analyses were used to estimate time to injury, and hazard ratios (HR) for time to injury were compared between workers who reported no rest break (reference) and workers who reported rest break(s) prior to the injury (accumulated break time categorized into 0, 1-15, 16-30, and >30 minutes). Age, gender, time of work start, injury time of day, and workload were included as covariates to control for demographic, circadian, and work-related factors, respectively. A clear dose-response relation indicated that longer accumulated break time was associated with a significantly longer time to injury when compared to workers without rest breaks [total break time 1-15 minutes: HR 0.60, 95% confidence interval (95% CI) 0.44-0.83; 16-30 minutes: HR 0.50, 95% CI 0.33-0.75; >30 minutes: HR 0.34, 95% CI 0.23-0.51], adjusted for all covariates. The results showed that longer total rest break time allowed for a significantly prolonged time spent on task without an injury. These findings suggest that rest break design could be used as a tool to enhance fatigue management and workplace safety.


Use of analogues of methionine and methionyl adenylate to sample conformational changes during catalysis in Escherichia coli methionyl-tRNA synthetase.
Binding of methionine to methionyl-tRNA synthetase (MetRS) is known to promote conformational changes within the active site. However, the contribution of these rearrangements to enzyme catalysis is not fully understood. In this study, several methionine and methionyl adenylate analogues were diffused into crystals of the monomeric form of Escherichia coli methionyl-tRNA synthetase. The structures of the corresponding complexes were solved at resolutions below 1.9A and compared to those of the enzyme free or complexed with methionine. Residues Y15 and W253 play key roles in the strength of the binding of the amino acid and of its analogues. Indeed, full motions of these residues are required to recover the maximum in free energy of binding. Residue Y15 also controls the size of the hydrophobic pocket where the amino acid side-chain interacts. H301 appears to participate to the specific recognition of the sulphur atom of methionine. Complexes with methionyl adenylate analogues illustrate the shielding by MetRS of the region joining the methionine and adenosine moieties. Finally, the structure of MetRS complexed to a methionine analogue mimicking the tetrahedral carbon of the transition state in the aminoacylation reaction was solved. On the basis of this model, we propose that, in response to the binding of the 3'-end of tRNA, Y15 moves again in order to deshield the anhydride bond in the natural adenylate.


Serum tumor necrosis factor-alpha in neonatal sepsis.
Tumor necrosis factor-alpha (TNF-alpha) is thought to be an important mediator in the pathogenesis of neonatal septicemia. To assess the role of TNF-alpha in neonatal sepsis, serum levels of TNF-alpha were measured in a group of neonates with septicemia and compared with the levels of gestational-postnatal, age-matched healthy controls. The relationships between severity of infection, the nature of causative microorganisms, and TNF-alpha levels were also investigated in this prospective study. A total of 49 infants (25 full-term, 24 preterm) with proven sepsis and 40 healthy infants (20 full-term, 20 preterm) were included. Serum TNF-alpha levels were measured using the TNF-alpha immunoradiometric assay. The median level of TNF-alpha was found to be significantly higher in infants suffering from sepsis (154 pg/mL) particularly in those with septic shock (242.5 pg/mL) as compared to healthy controls (61.5 pg/mL) (p < 0.001). No correlation was found between TNF-alpha and postnatal ages, gestational ages or birth weights of the infants. TNF-alpha levels were not different in surviving and terminal neonates. Although serum, TNF-alpha levels were found to be slightly higher in gram-negative septicemia, the difference was not significant. These findings suggest that TNF-alpha plays an important role in the pathophysiology of neonatal septicemia, but its importance as a prognostic factor is not yet clear.