Paracellin-1, a renal tight junction protein required for paracellular Mg2+ resorption.
Epithelia permit selective and regulated flux from apical to basolateral surfaces by transcellular passage through cells or paracellular flux between cells. Tight junctions constitute the barrier to paracellular conductance; however, little is known about the specific molecules that mediate paracellular permeabilities. Renal magnesium ion (Mg2+) resorption occurs predominantly through a paracellular conductance in the thick ascending limb of Henle (TAL). Here, positional cloning has identified a human gene, paracellin-1 (PCLN-1), mutations in which cause renal Mg2+ wasting. PCLN-1 is located in tight junctions of the TAL and is related to the claudin family of tight junction proteins. These findings provide insight into Mg2+ homeostasis, demonstrate the role of a tight junction protein in human disease, and identify an essential component of a selective paracellular conductance.


The dynamics of variation in individuals.
This paper examines the factors conditioning the production of linguistic variables in real time by individual speakers: the study of what we term the dynamics of variation in individuals. We propose a framework that recognizes three types of factors conditioning variation: sociostylistic (s-), internal linguistic (i-), and psychophysiological (p-). We develop two main points against this background. The first is that sequences of variants produced by individuals display systematic patterns that can be understood in terms of s-conditioning and p-conditioning (with a focus on the latter). The second main point is that p-conditioning and i-conditioning are distinct in their mental implementations; this claim has implications for understanding the locality of the factors conditioning alternations, for the universality and language-specificity of variation, and for the general question of whether grammar and language use are distinct. Throughout the paper, questions about the dynamics of variation in individuals are set against the typical community-centered variationist perspective, with an eye towards showing how findings in the two domains, though differing in explanatory focus, can ultimately be mutually informative.


Overexpression of anti-fibrotic factors ameliorates anti-fibrotic properties of Wharton's jelly derived mesenchymal stem cells under oxidative damage.
Transplantation with Wharton's jelly derived mesenchymal stem cells (WJ-MSCs) showed great benefits for restoring myocardial function. However, the outcome of WJ-MSCs transplantation was unsuccessful due to multiple factors including oxidative damage. The presence of oxidative stress due to myocardium injury influences fibrous tissue formation, which causes disability of cardiac muscle. Hepatocyte growth factor (HGF), insulin-like growth factor (IGF1), and sonic hedgehog (SHH) are well-known master regulators in anti-fibrosis when secreted by WJ-MSCs. They showed a beneficial role in the recovery of cardiac fibrosis after WJ-MSCs transplantation. This study hypothesizes whether the reduction of the anti-fibrosis property in WJ-MSCs from oxidative damage can be recovered by overexpression of the HGF, IGF1, or SHH gene. Overexpression was attained by transfection of WJ-MSCs with pCMV3-HGF, pCMV3-IGF1, or pCMV3-SHH followed by H2O2 exposure and co-culturing with cardiac fibroblasts. Myofibroblast specific markers comprised of alpha-smooth muscle actin (α-SMA) and collagen type 1 (COL1) were evaluated. The WJ-MSCs treated with H2O2 influenced the expression of myofibroblastic markers, whereas the overexpression of HGF, IGF1 or SHH reduced myofibroblastic formation. These results indicate that the oxidative stress impaired anti-fibrotic property of WJ-MSCs, leads to an increase of myofibroblasts. Overexpression of anti-fibrotic genes restored the endogenous HGF, IGF1, and SHH alleviating improvement of cardiac function.


Comparative effects of dimethoate and deltamethrin on reproductive system in male mice.
The effects of dimethoate (5, 15 and 28 mg kg(-1) day(-1)), deltamethrin (5 mg kg(-1) day(-1)) and their mixture (5 mg kg(-1) day(-1)) on male reproduction in mice were studied. The insecticides were given orally by gavage to male mice for 21 days. At the end of the treatment period, body, testes and epididymides weights and sperm parameters were determined. Alone mixture treatment has significantly decreased body weights. Dimethoate at 28 mg kg(-1) day(-1), deltamethrin at 5 mg kg(-1) day(-1) and their mixture at 5 mg kg(-1) day(-1) were associated with a significantly decreased sperm count, motility and viability and significantly increased percent morphologically abnormal spermatozoa compared with the controls. This study demonstrated the adverse effects of dimethoate at high dose, deltamethrin and their combining at 5 mg kg(-1) day(-1) on reproductive system and sperm parameters in male mice.


Causation and melanoma classification.
In this article, I begin by giving a brief history of melanoma causation. I then discuss the current manner in which malignant melanoma is classified. In general, these systems of classification do not take account of the manner of tumour causation. Instead, they are based on phenomenological features of the tumour, such as size, spread, and morphology. I go on to suggest that misclassification of melanoma is a major problem in clinical practice. I therefore outline an alternative means of classifying these tumours based on causal factors. By analogy with similar systems that have recently emerged for other cancers, I suggest that this causal classification is likely to be both workable and helpful, even in the absence of a full causal-mechanistic understanding of the aetiology of the tumour.


Close association of water channel AQP1 with amyloid-beta deposition in Alzheimer disease brains.
Aquaporin-1 (AQP1), a membrane water channel protein, is expressed exclusively in the choroid plexus epithelium in the central nervous system under physiological conditions. However, AQP1 expression is enhanced in reactive astrocytes, accumulating in brain lesions of Creutzfeldt-Jakob disease and multiple sclerosis, suggesting a role of AQP1-expressing astrocytes in brain water homeostasis under pathological conditions. To clarify a pathological implication of AQP1 in Alzheimer disease (AD), we investigated the possible relationship between amyloid-beta (Abeta) deposition and astrocytic AQP1 expression in the motor cortex and hippocampus of 11 AD patients and 16 age-matched other neurological disease cases. In all cases, AQP1 was expressed exclusively in a subpopulation of multipolar fibrillary astrocytes. The great majority of AQP1-expressing astrocytes were located either on the top of or in close proximity to Abeta plaques in AD brains but not in non-AD cases, whereas those independent of Abeta deposition were found predominantly in non-AD brains. By Western blot, cultured human astrocytes constitutively expressed AQP1, and the levels of AQP1 protein expression were not affected by exposure to Abeta(1-42) peptide, but were elevated by hypertonic sodium chloride. By immunoprecipitation, the C-terminal fragment-beta (CTFbeta) of amyloid precursor protein interacted with the N-terminal half of AQP1 spanning the transmembrane helices H1, H2 and H3. These observations suggest the possible association of astrocytic AQP1 with Abeta deposition in AD brains.


Kidney, pancreas and liver allocation and distribution in the United States.
Kidney transplant and liver transplant are the treatments of choice for patients with end-stage renal disease and end-stage liver disease, respectively. Pancreas transplant is most commonly performed along with kidney transplant in diabetic end-stage renal disease patients. Despite a steady increase in the numbers of kidney and liver transplants performed each year in the United States, a significant shortage of kidneys and livers available for transplant remains. Organ allocation is the process the Organ Procurement and Transplantation Network (OPTN) uses to determine which candidates are offered which deceased donor organs. OPTN is charged with ensuring the effectiveness, efficiency and equity of organ sharing in the national system of organ allocation. The policy has changed incrementally over time in efforts to optimize allocation to meet these often competing goals. This review describes the history, current status and future direction of policies regarding the allocation of abdominal organs for transplant, namely the kidney, liver and pancreas, in the United States.


Climate Effects on High Latitude Daphnia via Food Quality and Thresholds.
Climate change is proceeding rapidly at high northern latitudes and may have a variety of direct and indirect effects on aquatic food webs. One predicted effect is the potential shift in phytoplankton community structure towards increased cyanobacterial abundance. Given that cyanobacteria are known to be a nutritionally poor food source, we hypothesized that such a shift would reduce the efficiency of feeding and growth of northern zooplankton. To test this hypothesis, we first isolated a clone of Daphnia pulex from a permafrost thaw pond in subarctic Québec, and confirmed that it was triploid but otherwise genetically similar to a diploid, reference clone of the same species isolated from a freshwater pond in southern Québec. We used a controlled flow-through system to investigate the direct effect of temperature and indirect effect of subarctic picocyanobacteria (Synechococcus) on threshold food concentrations and growth rate of the high latitude clone. We also compared the direct effect of temperature on both Daphnia clones feeding on eukaryotic picoplankton (Nannochloropsis). The high latitude clone had a significantly lower food threshold for growth than the temperate clone at both 18 and 26°C, implying adaptation to lower food availability even under warmer conditions. Polyunsaturated fatty acids were present in the picoeukaryote but not the cyanobacterium, confirming the large difference in food quality. The food threshold for growth of the high latitude Daphnia was 3.7 (18°C) to 4.2 (26°C) times higher when fed Synechococcus versus Nannochloropsis, and there was also a significant negative effect of increased temperature and cyanobacterial food on zooplankton fatty acid content and composition. The combined effect of temperature and food quality on the performance of the high latitude Daphnia was greater than their effects added separately, further indicating the potentially strong indirect effects of climate warming on aquatic food web processes.


Calcium dynamics during fertilization in C. elegans.
Of the animals typically used to study fertilization-induced calcium dynamics, none is as accessible to genetics and molecular biology as the model organism Caenorhabditis elegans. Motivated by the experimental possibilities inherent in using such a well-established model organism, we have characterized fertilization-induced calcium dynamics in C. elegans. Owing to the transparency of the nematode, we have been able to study the calcium signal in C. elegans fertilization in vivo by monitoring the fluorescence of calcium indicator dyes that we introduce into the cytosol of oocytes. In C. elegans, fertilization induces a single calcium transient that is initiated soon after oocyte entry into the spermatheca, the compartment that contains sperm. Therefore, it is likely that the calcium transient is initiated by contact with sperm. This calcium elevation spreads throughout the oocyte, and decays monotonically after which the cytosolic calcium concentration returns to that preceding fertilization. Only this single calcium transient is observed. Development of a technique to study fertilization induced calcium transients opens several experimental possibilities, e.g., identification of the signaling events intervening sperm binding and calcium elevation, identifying the possible roles of the calcium elevation such as the completion of meiosis, the formation of the eggshell, and the establishing of the embryo's axis of symmetry.


Targeted deletion of the zebrafish actin-bundling protein L-plastin (lcp1).
Regulation of the cytoskeleton is essential for cell migration in health and disease. Lymphocyte cytosolic protein 1 (lcp1, also called L-plastin) is a hematopoietic-specific actin-bundling protein that is highly conserved in zebrafish, mice and humans. In addition, L-plastin expression is documented as both a genetic marker and a cellular mechanism contributing to the invasiveness of tumors and transformed cell lines. Despite L-plastin's role in both immunity and cancer, in zebrafish there are no direct studies of its function, and no mutant, knockout or reporter lines available. Using CRISPR-Cas9 genome editing, we generated null alleles of zebrafish lcp1 and examined the phenotypes of these fish throughout the life cycle. Our editing strategy used gRNA to target the second exon of lcp1, producing F0 mosaic fish that were outcrossed to wild types to confirm germline transmission. F1 heterozygotes were then sequenced to identify three unique null alleles, here called 'Charlie', 'Foxtrot' and 'Lima'. In silico, each allele truncates the endogenous protein to less than 5% normal size and removes both essential actin-binding domains (ABD1 and ABD2). Although none of the null lines express detectable LCP1 protein, homozygous mutant zebrafish (-/-) can develop and reproduce normally, a finding consistent with that of the L-plastin null mouse (LPL -/-). However, such mice do have a profound immune defect when challenged by lung bacteria. Interestingly, we observed reduced long-term survival of zebrafish lcp1 -/- homozygotes (~30% below the expected numbers) in all three of our knockout lines, with greatest mortality corresponding to the period (4-6 weeks post-fertilization) when the innate immune system is functional, but the adaptive immune system is not yet mature. This suggests that null zebrafish may have reduced capacity to combat opportunistic infections, which are more easily transmissible in the aquatic environment. Overall, our novel mutant lines establish a sound genetic model and an enhanced platform for further studies of L-plastin gene function in hematopoiesis and cancer.


Unnatural deaths in reindeer-herding Sami families in Sweden, 1961-2001.
Unnatural deaths among Indigenous populations, including the Swedish Sami, occur more often than among the general population. To find prevention strategies, we explored the circumstances of the unnatural deaths of members of reindeer-herding Sami families. The number of deaths from among a cohort of 7,482 members of reindeer-herding Sami families were retrieved from the National Board of Health and Welfare for the years 1961- 2001. An evaluation of the information from autopsy records at the National Board of Forensic Medicine, police reports, and available medical records identified 158 unnatural deaths. These were then analysed in detail. Transport-related deaths and suicides were the most common unnatural deaths among Swedish reindeer-herding Sami family members. Suicides contributed to 23% of all deaths, road traffic accidents to 16%, and snowmobile fatalities to 11%. The accidents generally reflected an "outdoor lifestyle" and the working conditions were characterized by the use of off-road vehicles such as snowmobiles. Half of the number of victims tested positive for alcohol and alcohol abuse was documented in 15% of all victims. The results indicate that alcohol is an important factor in preventing unnatural deaths among reindeer-herding Sami, together with increased safety of both on-road and off-road transportation.


First report of Lyme borreliosis leading to cardiac bradydysrhythmia in two cats.
Two cats were presented for investigation of bradyarrhythmia detected by their referring veterinarians during routine examination. Both cats had extensive investigations, including haematology, serum biochemistry with electrolytes and thyroxine concentrations, systolic blood pressure measurement, echocardiography, electrocardiography and infectious disease testing. Infectious disease testing included serology for Toxoplasma gondii, Ehrlichia canis, Anaplasma phagocytophilum and Borrelia burgdorferi, and PCR for B burgdorferi antigen in both cats. Case 1 was also assessed by PCR for Bartonella henselae antigen and case 2 was assessed for Dirofilaria immitis by serology. All infectious disease tests, other than for B burgdorferi, were negative. Case 1 was diagnosed with Lyme carditis based on marked bradydysrhythmia, positive B burgdorferi serology, a structurally normal heart and clinical resolution with appropriate treatment with a 4-year follow-up. Case 2 was diagnosed with Lyme carditis based on marked bradydysrhythmia and positive B burgdorferi PCR; however, this cat had structural heart disease that did not resolve with treatment. This small case series describes two B burgdorferi positive cats presenting with newly diagnosed cardiac abnormalities consistent with those found in humans and dogs with Lyme carditis. Both cats were asymptomatic as perceived by their owners; the arrhythmia was detected by their veterinarians.


The inorganic perspective of nerve growth factor: interactions of Cu2+ and Zn2+ with the N-terminus fragment of nerve growth factor encompassing the recognition domain of the TrkA receptor.
There is a significant overlap between brain areas with Zn(2+) and Cu(2+) pathological dys-homeostasis and those in which the nerve growth factor (NGF) performs its biological role. The protein NGF is necessary for the development and maintenance of the sympathetic and sensory nervous systems. Its flexible N-terminal region has been shown to be a critical domain for TrkA receptor binding and activation. Computational analyses show that Zn(2+) and Cu(2+) form pentacoordinate complexes involving both the His4 and His8 residues of the N-terminal domain of one monomeric unit and the His84 and Asp105 residues of the other monomeric unit of the NGF active dimer. To date, neither experimental data on the coordination features have been reported, nor has one of the hypotheses according to which Zn(2+) and Cu(2+) may have different binding environments or the Ser1 α-amino group could be involved in coordination been supported. The peptide fragment, encompassing the 1-14 sequence of the human NGF amino-terminal domain (NGF(1-14)), blocked at the C terminus, was synthesised and its Cu(2+) and Zn(2+) complexes characterized by means of potentiometric and spectroscopic (UV/Vis, CD, NMR, and EPR) techniques. The N-terminus-acetylated form of NGF(1-14) was also investigated to evaluate the involvement of the Ser1 α-amino group in metal-ion coordination. Our results demonstrate that the amino group is the first anchoring site for Cu(2+) and is involved in Zn(2+) coordination at physiological pH. Finally, a synergic proliferative activity of both NGF(1-14) and the whole protein on SHSY5Y neuroblastoma cell line was found after treatment in the presence of Cu(2+). This effect was not observed after treatment with the N-acetylated peptide fragment, demonstrating a functional involvement of the N-terminal amino group in metal binding and peptide activity.


First Report of Fusarium verticillioides Causing Stalk and Root Rot of Sorghum in Spain.
Sweet sorghum (Sorghum bicolor L.) is considered one of the most promising crops for bioethanol production in many countries and is a focus of bioenergy research worldwide. In July 2011, plants of the sweet sorghum cv. Suchro 506 in Oropesa (Toledo, Spain, 40.048577°N, 5.360298°W) (European Datum 1950 UTM zone 30 N) were observed with severe wilting. Upon examination, the lower internodes were found to be straw colored. When the plant was split, the internal pith was reddish, soft, and disintegrating. Small pieces of symptomatic stems and roots were surface disinfected in sodium hypochlorite (0.5% wt/vol) for 2 min and air dried. The sections were then placed on either PDA (potato dextrose agar) medium or Komada agar and incubated for 5 days at 25°C. Isolations from diseased stem and root tissue consistently yielded Fusarium verticillioides (Sacc.) Nirenberg (3). The small, hyaline, mostly single-celled, oval to club-shaped microconidia of F. verticillioides were produced in long catenate chains arising from monophialides. PCR amplification of the ITS1-5.8S-ITS2 was performed using the primers and protocols described elsewhere (4) and the fragments obtained were subsequently sequenced in both directions. Sequences were deposited in the EMBL Sequence Database (Accession Nos. HE652878, HE652879, HE652880, and HE652881). Four of the recovered F. verticilliodes isolates were tested in pathogenicity assays. One-week-old cultures of each isolate were homogenized in 400 ml of sterile water and 200 ml were used to inoculate water-growth-chamber-grown plants in 500-ml pots. Two pots each with three plants of cv. Suchro 506 were inoculated for each isolate. Water with sterile PDA was used as a control. All plants were kept at 20 to 25°C under a photoperiod of 14 h at 12,000 lux. After 21 days, above- and belowground parts were dried for 24 h at 60°C. Total length and dry weight of both sections were obtained. Inoculated plants produced root rot symptoms characteristic of F. verticillioides with dark red discolorations of the cortex of seedling roots (1), whereas the plants watered with water containing only PDA did not produce symptoms. Inoculated plants also had a decrease in dry weight for above- and belowground sections (P = 0.05) compared with the control with 43 and 47% reductions, respectively. The length of aerial parts was approximately 5% less in inoculated plants compared with control plants. F. verticillioides was reisolated from all inoculated plants. Sorghum stalk and root rot caused by F. verticillioides has been reported in different countries including India (2) and the United States (3). To our knowledge, this is the first report of F. verticillioides causing stalk and root rot of sorghum in Spain. An increase of production of this crop is expected to meet targets of the renewable energy share in Spain and any disease compromising yield may be a threat to this endeavour. References: (1) R. A. Frederiksen and G. N. Odvody. Compendium of Sorghum Diseases. The American Phytopathological Society. St. Paul, MN, 2000. (2) N. N. Khune et al. Indian Phytopathol. 37:316, 1984. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, New York, 1990.


Deformation-based Statistical Shape Analysis of the Corpus Callosum in Mild Cognitive Impairment and Alzheimer's Disease.
In this study, we investigated the influence that the pathology of Alzheimer's disease (AD) exerts upon the corpus callosum (CC) using a total of 325 mild cognitive impairment (MCI) subjects, 155 AD subjects, and 185 healthy control (HC) subjects. Regionally-specific morphological CC abnormalities, as induced by AD, were quantified using a large deformation diffeomorphic metric curve mapping based statistical shape analysis pipeline. We also quantified the association between the CC shape phenotype and two cognitive measures; the Mini Mental State Examination (MMSE) and the Alzheimer's Disease Assessment Scale-Cognitive Behavior Section (ADAS-cog). To identify AD-relevant areas, CC was sub-divided into three subregions; the genu, body, and splenium (gCC, bCC, and sCC). We observed significant shape compressions in AD relative to that in HC, mainly concentrated on the superior part of CC, across all three sub-regions. The HC-vs-MCI shape abnormalities were also concentrated on the superior part, but mainly occurred on bCC and sCC. The significant MCI-vs-AD shape differences, however, were only detected in part of sCC. In the shape-cognition association, significant negative correlations to ADAS-cog were detected for shape deformations at regions belonging to gCC and sCC and significant positive correlations to MMSE at regions mainly belonging to sCC. Our results suggest that the callosal shape deformation patterns, especially those of sCC, linked tightly to the cognitive decline in AD, and are potentially a powerful biomarker for monitoring the progression of AD.


Genetic and functional analysis of KIF5A variants in Japanese patients with sporadic amyotrophic lateral sclerosis.
Two recent genetic studies reported that loss-of-function mutation of the C-terminal cargo-binding tail domain of the KIF5A gene cause amyotrophic lateral sclerosis (ALS). The aim of this study is to investigate the frequency of KIF5A variants in Japanese patients with sporadic ALS. In total, 807 sporadic ALS patients and 191 normal controls from a multicenter ALS cohort in Japan were included. Whole exome sequencing on an Illumina HiSeq 2000/2500 sequencer was used to identify and select variants within the KIF5A gene. Thirteen patients harbored a nonsynonymous variant in the KIF5A gene; These were considered variants of uncertain significance. One patient harbored a novel splice-site variant (c.2993-3C>A) in the C-terminal cargo-binding tail domain of the KIF5A gene. Functional analysis of this variant revealed that it caused skipping of exon 27. The frequency of KIF5A mutations in Japanese patients with sporadic ALS was 0.12% (1/807). This study reports a novel loss-of-function variant in KIF5A, and indicates that loss-of-function variant in KIF5A is a rare cause of sporadic ALS in Japanese patients.


A systems genetics approach to dyslipidemia in children and adolescents.
Elevated triglycerides (TG) or low high density lipoprotein cholesterol (HDL-C) levels are common cardiometabolic risk factors in children. From a systems genetics standpoint, Visualization of Statistical Epistasis Networks (ViSEN) is a nonparametric entropy-based method that can characterize the global structure of interacting genetic factors. We identified a novel set of connected genetic and cardiometabolic risk factors with strong and significant interaction effects on two important dyslipidemia phenotypes (low HDL-C and high TG) in children and adolescents. Study participants were recruited from five schools in Istanbul, Turkey (n=360; 170 boys, 190 girls). Participants with TG levels≥75th and HDL-C levels≤25th percentile were defined as 'high TG' and 'low HDL-C', respectively. We genotyped participants for six single nucleotide polymorphisms (SNPs) in five genes with known associations to lipid levels (rs328 in LPL, rs708272 in CETP, rs1800588 in LIPC, rs1800977 in ABCA1, rs1799941 and rs6257 in SHBG gene). ViSEN was used to identify associations with dyslipidemia phenotypes. There were 71 (50 males, 21 females) and 93 (60 males and 33 females) subjects with low HDL-C and high TG, respectively. Biological variables including age, gender, and BMI were significantly associated with both phenotypes (p<0.001). Importantly, a single SNP, rs708272, was associated with low HDL-C (IG=2.24%, p=0.026). Pairwise and higher order interaction analyses in the full dataset for low HDL-C and high TG revealed the largest effects in the models containing rs1800977, rs708272, age (IG=6.20%, p=0.046) and rs1800588, age, BMI (IG, 3.06%, p=0.022), respectively. In conclusion, the present study brings us a step closer to a systems genetic approach in understanding lipid phenotypes in children. Further efforts can integrate population and laboratory-based studies, hence accelerate the preventive medicine efforts.


Perceived Facilitators and Barriers in Response to a Walking Intervention in Rural Cancer Survivors: A Qualitative Exploration.
Physical activity has numerous associated benefits for cancer survivors. Compared to their urban counterparts, rural and remote Australians experience a health disadvantage, including poorer survival rate after the diagnosis of cancer. The purpose of this qualitative study was to (a) investigate factors that motivated or inhibited walking in rural participants during a 12-week intervention and (b) to investigate factors that motivated or inhibited physical activity behavior change three months post-intervention. Ten cancer survivors living in rural areas of South Australia participated in a 12-week computer-delivered walking-based intervention during which they reported daily steps, daily affect, and ratings of perceived exertion. Based on this information, individualized daily step goals were sent to them to increase walking. Following the intervention, participants engaged in face-to-face semi-structured interviews. Interviews were recorded, transcribed and coded using thematic analysis. Participants identified a range of physical, psychological, social, environmental, and organizational motivators and barriers. Participants appreciated the monitoring and support from the research team, but some voiced a need for better transition to post-program and many desired ongoing support to maintain their motivation. Future studies should incorporate strategies to help walking behavior to become more intrinsically motivated and therefore sustained.


Marital fertility and income: moderating effects of the Church of Jesus Christ of Latter-day Saints religion in Utah.
Utah has the highest total fertility of any state in the United States and also the highest proportion of population affiliated with the Church of Jesus Christ of Latter-day Saints (the LDS or Mormon Church). Data were used from the 1996 Utah Health Status Survey to investigate how annual household income, education and affiliation with the LDS Church affect fertility (children ever born) for married women in Utah. Younger age and higher education were negatively correlated with fertility in the sample as a whole and among non-LDS respondents. Income was negatively associated with fertility among non-LDS respondents. However, income was positively correlated with fertility among LDS respondents. This association persisted when instrumental variables were used to address the potential simultaneous equations bias arising from the potential endogeneity of income and fertility. The LDS religion's pronatalist stance probably encourages childbearing among those with higher income.


Intercellular transfer of carcinoembryonic antigen from tumor cells to NK cells.
The inhibition of NK cell killing is mainly mediated via the interaction of NK inhibitory receptors with MHC class I proteins. In addition, we have previously demonstrated that NK cells are inhibited in a class I MHC-independent manner via homophilic carcinoembryonic Ag (CEA) cell adhesion molecules (CEACAM1)-CEACAM1 and heterophilic CEACAM1-CEA interactions. However, the cross-talk between immune effector cells and their target cells is not limited to cell interactions per se, but also involves a specific exchange of proteins. The reasons for these molecular exchanges and the functional outcome of this phenomenon are still mostly unknown. In this study, we show that NK cells rapidly and specifically acquire CEA molecules from target cells. We evaluated the role of cytotoxicity in the acquisition of CEA and demonstrated it to be mostly killing independent. We further demonstrate that CEA transfer requires a specific interaction with an unknown putative NK cell receptor and that carbohydrates are probably involved in CEA recognition and acquisition by NK cells. Functionally, the killing of bulk NK cultures was inhibited by CEA-expressing cells, suggesting that this putative receptor is an inhibitory receptor.


Intravenous flumazenil following acute and repeated exposure to lorazepam in healthy volunteers: antagonism and precipitated withdrawal.
The effects of i.v. administered flumazenil (3.0 mg) were studied in healthy male subjects who received pretreatment with p.o. placebo or lorazepam. The duration of placebo or lorazepam (3.0 mg single p.o. daily dose) pretreatment before a flumazenil or placebo injection was 1, 3, 7 or 14 days in four sequential groups of subjects. Initial administration of lorazepam produced a classic sedative profile of effects on various psychomotor/behavioral performance, observer-rated and subject-rated measures. Tolerance to repeated daily administration of lorazepam was suggested by a progressive diminution of performance disrupting effects. In subjects pretreated with placebo, flumazenil increased subject-ratings of dizziness over preinjection ratings. Flumazenil produced an immediate reversal of lorazepam effects in subjects who were not tolerant to lorazepam (1- and 3-day pretreatment groups). Flumazenil did not precipitate withdrawal symptoms in subjects who received a single administration of lorazepam. Precipitated withdrawal symptoms were evident after 3 and 7 days of lorazepam pretreatment, and there was a tendency toward precipitated withdrawal symptoms (that included one panic attack) after 14 days of lorazepam pretreatment. Precipitated withdrawal was characterized by an elevation in subject-rated symptoms including dizziness, tenseness, tachycardia, perceptual disturbance and sweating. Symptoms were maximal immediately after injection, usually mild in severity and usually resolved within 1 hr. There was no evidence of precipitated withdrawal on psychomotor/behavioral performance or observer ratings. The present study provides the strongest human experimental evidence to date that flumazenil can precipitate withdrawal symptoms after a history of repeated benzodiazepine exposure.


Predictors of HIV-related stigmas among African American and Latino religious congregants.
To inform church-based stigma interventions by exploring dimensions of HIV stigma among African American and Latino religious congregants and determining how these are related to drug addiction and homosexuality stigmas and knowing someone HIV-positive. In-person, self-administered surveys of congregants 18+ years old across 2 African American and 3 Latino churches (n = 1,235, response rate 73%) in a western U.S. city with high HIV prevalence. Measures included 12 items that captured dimensions of HIV stigma, a 5-item scale that assessed attitudes toward people who are addicted to drugs, a 7-item scale assessing attitudes toward homosexuality, and questions regarding sociodemographics and previous communication about HIV. Of the survey participants, 63.8% were women, mean age was 40.2 years, and 34.4% were African American, 16.8% were U.S.-born Latinos, 16.0% were foreign-born, English-speaking Latinos, and 32.9% were foreign-born, Spanish-speaking Latinos. Exploratory and confirmatory factor analyses identified 4 dimensions of HIV stigma: discomfort interacting with people with HIV (4 items, α = .86), feelings of shame "if you had HIV" (3 items, α = .78), fears of rejection "if you had HIV" (3 items, α = .71), and feelings of blame toward people with HIV (2 items, α = .65). Across all dimensions, after controlling for sociodemographic characteristics and previous communication about HIV, knowing someone with HIV was associated with lower HIV stigma, and greater stigma concerning drug addiction and homosexuality were associated with higher HIV stigma. Congregation-based HIV stigma reduction interventions should consider incorporating contact with HIV-affected people. It may also be helpful to address attitudes toward drug addiction and sexual orientation. (PsycINFO Database Record


Interventions for tobacco use cessation in people living with HIV and AIDS.
Tobacco use is highly prevalent amongst people living with HIV/AIDS (PLWHA) and has a substantial impact on morbidity and mortality. To assess the effectiveness of interventions to motivate and assist tobacco use cessation for people living with HIV/AIDS (PLWHA), and to evaluate the risks of any harms associated with those interventions. We searched the Cochrane Tobacco Addiction Group's Specialised Register, Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, and PsycINFO in June 2015. We also searched EThOS, ProQuest, four clinical trial registries, reference lists of articles, and searched for conference abstracts using Web of Science and handsearched speciality conference databases. Controlled trials of behavioural or pharmacological interventions for tobacco cessation for PLWHA. Two review authors independently extracted all data using a standardised electronic data collection form. They extracted data on the nature of the intervention, participants, and proportion achieving abstinence and they contacted study authors to obtain missing information. We collected data on long-term (greater than or equal to six months) and short-term (less than six months) outcomes. Where appropriate, we performed meta-analysis and estimated the pooled effects using the Mantel-Haenszel fixed-effect method. Two authors independently assessed and reported the risk of bias according to prespecified criteria. We identified 14 studies relevant to this review, of which we included 12 in a meta-analysis (n = 2087). All studies provided an intervention combining behavioural support and pharmacotherapy, and in most studies this was compared to a less intensive control, typically comprising a brief behavioural intervention plus pharmacotherapy.There was moderate quality evidence from six studies for the long-term abstinence outcome, which showed no evidence of effect for more intense cessation interventions: (risk ratio (RR) 1.00, 95% confidence interval (CI) 0.72 to 1.39) with no evidence of heterogeneity (I(2) = 0%). The pooled long-term abstinence was 8% in both intervention and control conditions. There was very low quality evidence from 11 studies that more intense tobacco cessation interventions were effective in achieving short-term abstinence (RR 1.51, 95% CI 1.15 to 2.00); there was moderate heterogeneity (I(2) = 42%). Abstinence in the control group at short-term follow-up was 8% (n = 67/848) and in the intervention group was 13% (n = 118/937). The effect of tailoring the intervention for PLWHA was unclear. We further investigated the effect of intensity of behavioural intervention via number of sessions and total duration of contact. We failed to detect evidence of a difference in effect according to either measure of intensity, although there were few studies in each subgroup. It was not possible to perform the planned analysis of adverse events or HIV outcomes since these were not reported in more than one study. There is moderate quality evidence that combined tobacco cessation interventions provide similar outcomes to controls in PLWHA in the long-term. There is very low quality evidence that combined tobacco cessation interventions were effective in helping PLWHA achieve short-term abstinence. Despite this, tobacco cessation interventions should be offered to PLWHA, since even non-sustained periods of abstinence have proven benefits. Further large, well designed studies of cessation interventions for PLWHA are needed.


Degenerative Cervical Myelopathy in Higher-Aged Patients: How Do They Benefit from Surgery?
Degenerative cervical myelopathy (DCM) is the most common reason for spinal cord disease in elderly patients. This study analyzes the preoperative status and postoperative outcome of higher-aged patients in comparison to young and elderly patients in order to determine the benefit to those patients from DCM surgery. A retrospective analysis of the clinical data, radiological findings, and operative reports of 411 patients treated surgically between 2007 and 2016 suffering from DCM was performed. The preoperative and postoperative neurological functions were evaluated using the modified Japanese Orthopedic Association Score (mJOA Score), the postoperative mJOA Score improvement, the neurological recovery rate (NRR) of the mJOA Score, and the minimum clinically important difference (MCID). The Charlson Comorbidity Index (CCI) was used to evaluate the impact of comorbidities on the preoperative and postoperative mJOA Score. The comparisons were performed between the following age groups: G1: ≤50 years, G2: 51-70 years, and G3: >70 years. The preoperative and postoperative mJOA Score was significantly lower in G3 than in G2 and G1 (p < 0.0001). However, the mean mJOA Score's improvement did not differ significantly (p = 0.81) between those groups six months after surgery (G1: 1.99 ± 1.04, G2: 2.01 ± 1.04, G: 2.00 ± 0.91). Furthermore, the MCID showed a significant improvement in every age-group. The CCI was evaluated for each age-group, showing a statistically significant group effect (p < 0.0001). Analysis of variance revealed a significant group effect on the delay (weeks) between symptom onset and surgery (p = 0.003). The duration of the stay at the hospital did differ significantly between the age groups (p < 0.0001). Preoperative and postoperative mJOA Scores, but not the extent of postoperative improvement, are affected by the patients' age. Therefore, patients should be considered for DCM surgery regardless of their age.


Scenarios for a post-COVID-19 world airline network.
The airline industry was severely hit by the COVID-19 crisis with an average demand decrease of about 64 % (IATA, April 2020), which triggered already several bankruptcies of airline companies all over the world. While the robustness of the world airline network (WAN) was mostly studied as a homogeneous network, we introduce a new tool for analyzing the impact of a company failure: the "airline company network" where two airlines are connected if they share at least one route segment. Using this tool, we observe that the failure of companies well connected with others has the largest impact on the connectivity of the WAN. We then explore how the global demand reduction affects airlines differently and provide an analysis of different scenarios if it stays low and does not come back to its pre-crisis level. Using traffic data from the Official Aviation Guide and simple assumptions about customer's airline choice strategies, we find that the local effective demand can be much lower than the average one, especially for companies that are not monopolistic and share their segments with larger companies. Even if the average demand comes back to 60 % of the total capacity, we find that between 46 % and 59 % of the companies could experience a reduction of more than 50 % of their traffic, depending on the type of competitive advantage that drives customer's airline choice. These results highlight how the complex competitive structure of the WAN weakens its robustness when facing such a large crisis.


Evaluation of a real-time polymerase chain reaction assay for the diagnosis of malaria in patients from Thailand.
We compared the diagnosis of malaria in 297 patients from Thailand by a real-time polymerase chain reaction (PCR) assay using the LightCycler with conventional microscopy using Giemsa-stained thick and thin blood films. The PCR assay can be completed in one hour and has the potential to detect and identify four species of Plasmodium in a single reaction by use of melting temperature curve analysis (however, we did not detect Plasmodium ovale in this study). Blood was collected, stored, and transported on IsoCode STIX, which provide a stable matrix for the archiving and rapid simple extraction of DNA. A genus-specific primer set corresponding to the 18S ribosomal RNA was used to amplify the target sequence. Fluorescence resonance energy technology hybridization probes were designed for P. falciparum over a region containing basepair mismatches, which allowed differentiation of the other Plasmodium species. The PCR results correlated with the microscopic results in 282 (95%) of 297 patient specimens. Most of these were single-species infections caused by P. vivax (150) and P. falciparum (120), along with 5 P. malariae, 2 mixed infections (P. falciparum and P. vivax), and 5 negative specimens. No negative microscopy specimens were positive by PCR (100% specificity for detection of any Plasmodium). The 15 discrepant results could not be resolved, but given the subjective nature of microscopy and the analytical objectivity of the PCR, the PCR results may be correct. The ability of the PCR method to detect mixed infections or to detect P. ovale could not be determined in this study. Within the limitations of initial equipment costs, this real-time PCR assay is a rapid, accurate, and efficient method for the specific diagnosis of malaria. It may have application in clinical laboratories, as well as in epidemiologic studies and antimalarial efficacy trials.


Water in ion channels and pores--simulation studies.
The microscopic properties of water in narrow pores are relevant to the function of ion channels and related membrane transport proteins. The emergence of several high-resolution structures allows one to perform molecular dynamics simulation studies of water in such pores. Simulations of bundles of parallel alpha-helical peptides (e.g. alamethicin) have enabled development of methodologies and concepts appropriate to such investigations. In the narrow channels formed by such bundles, water molecules exhibit reduced rotational and translation motion. This reduction in water mobility may be a general property of narrow pores. We have used simplified channel models to explore the role of hydrophobicity/hydrophilicity in the entry of water into pores. Narrow pores with a hydrophobic lining, although physically open, may not admit water molecules, acting as a 'hydrophobic gate' that prevents water and ion permeation. Such a gate can be opened either by widening the pore or making its lining more polar. Simulations have been used to explore the behaviour of water in GlpF, a member of the aquaporin family of water pores, and OmpA, a bacterial outer membrane protein. Preliminary results suggest that a continuous water wire is not formed within the amphipathic GlpF pore. Simulations of OmpA, in which polar residues line the channel, indicate that a small conformational change in one of the channel lining side chains may open the channel. In summary, comparison of the behaviour of water in different narrow transmembrane pores suggests that an amphipathic pore is ideal for water permeation, and that either a highly hydrophobic pore lining or a charged pore-lining region can act as a gate.


Circulating Metabolomic Signature in Generalized Pustular Psoriasis Blunts Monocyte Hyperinflammation by Triggering Amino Acid Response.
Generalized pustular psoriasis (GPP), the most grievous variant of psoriasis, is featured by dysregulated systemic inflammatory response. The cellular and molecular basis of GPP is poorly understood. Blood monocytes are key players of host defense and producers of inflammatory cytokines including IL-1β. How the immune response of monocytes is affected by metabolic internal environment in GPP remains unclear. Here, we performed a metabolomic and functional investigation of GPP serum and monocytes. We demonstrated a significant increase in IL-1β production from GPP monocytes. In GPP circulation, serum amyloid A (SAA), an acute-phase reactant, was dramatically increased, which induced the release of IL-1β from monocytes in a NLRP3-dependent manner. Using metabolomic analysis, we showed that GPP serum exhibited an amino acid starvation signature, with glycine, histidine, asparagine, methionine, threonine, lysine, valine, isoleucine, tryptophan, tyrosine, alanine, proline, taurine and cystathionine being markedly downregulated. In functional assay, under amino acid starvation condition, SAA-stimulated mature IL-1β secretion was suppressed. Mechanistically, at post-transcriptional level, amino acid starvation inhibited the SAA-mediated reactive oxygen species (ROS) formation and NLRP3 inflammasome activation. Moreover, the immune-modulatory effect of amino acid starvation was blocked by silencing general control nonderepressible 2 kinase (GCN2), suggesting the involvement of amino acid response (AAR) pathway. Collectively, our results suggested that decreased serum amino acids in GPP blunted the innate immune response in blood monocytes through AAR pathway, serving as a feedback mechanism preventing excessive inflammation in GPP.


Relevance of N6-methyladenosine regulators for transcriptome: Implications for development and the cardiovascular system.
N6-methyladenosine (m6A) is the most abundant and well-studied internal modification of messenger RNAs among the various RNA modifications in eukaryotic cells. Moreover, it is increasingly recognized to regulate non-coding RNAs. The dynamic and reversible nature of m6A is ensured by the precise and coordinated activity of specific proteins able to insert ("write"), bind ("read") or remove ("erase") the m6A modification from coding and non-coding RNA molecules. Mounting evidence suggests a pivotal role for m6A in prenatal and postnatal development and cardiovascular pathophysiology. In the present review we summarise and discuss the major functions played by m6A RNA methylation and its components particularly referring to the cardiovascular system. We present the methods used to study m6A and the most abundantly methylated RNA molecules. Finally, we highlight the possible involvement of the m6A mark in cardiovascular disease as well as the need for further studies to better describe the mechanisms of action and the potential therapeutic role of this RNA modification.


Arterial reconstructive surgery for renovascular hypertension.
Many hypertensive patients with renal artery stenosis and abnormal findings on renin angiotensin assay can be cured or markedly improved by renal artery surgery. Because the mortality is low, we think that patients whose blood pressures are resistant to medical management should be examined carefully for renal artery stenosis as the cause of their hypertension. If such a lesion is found, many of these patients can expect to enjoy a good clinical response from renal artery reconstruction. Those patients who do not have satisfactory results from the surgical correction should undergo aggressive reevaluation to detect any correctable anatomic defects.


Assessment of left ventricular function and volumes for patients with dilated cardiomyopathy using gated myocardial perfusion SPECT and comparison with echocardiography.
Left ventricular function, volumes and regional wall motion provide valuable diagnostic information and are of long-term prognostic importance in patients with dilated cardiomyopathy (DCM). This study was designed to compare the effectiveness of two-dimensional echocardiography and gated single photon emission computed tomography (SPECT) to evaluate these parameters in patients with DCM. Gated SPECT and two-dimensional echocardiography were performed in 45 patients with DCM, and in 10 normal subjects as the control group. Patients were divided into two groups according to the aetiology of DCM: group I, ischaemic DCM (n=30); group II, non-ischaemic DCM (n=15). All patients and the control group underwent resting myocardial gated SPECT, 45 min after injection of 555 MBq of Tc-methoxyisobutyl-isonitrile (Tc-MIBI). Gated SPECT data, including left ventricular volumes and left ventricular ejection fraction (LVEF), were processed using an automated algorithm. Simpson's method was used to evaluate these parameters. Regional wall motion was evaluated using both modalities and scored using a 16-segment model with a five-point scoring system. Perfusion defects were expressed as a percentage of the whole myocardium planimetered by a bull's-eye polar map of composite non-gated SPECT. Myocardial perfusion was scored using a 16-segment model with a four-point scoring system. Mean perfusion defects and perfusion defect scores were 25+/-13% and 1.12+/-0.36 in group I and 4+/-8% and 0.76+/-0.26 in group II (P<0.01). The overall agreement between the two imaging modalities for the assessment of regional wall motion was 57% (403/720 segments: 269/480 segments in group I and 134/240 segments in group II). With gated SPECT, LVEF was 27+/-9%, the end-diastolic volume (EDV) was 212+/-71 ml and the end-systolic volume (ESV) was 160+/-67 ml. With echocardiography, these values were 29+/-8%, 197+/-56 ml and 139+/-47 ml, respectively. The correlation between gated SPECT and two-dimensional echocardiography was good (r=0.72, P<0.01) for the assessment of LVEF. The correlation was also good for EDV and ESV, but with wider limits of agreement (r= 0.71, P<0.01 and r=0.71, P<0.01, respectively) and with significantly higher values with gated SPECT (P<0.01). For patients with a perfusion defect of <20% or low myocardial perfusion scores, a higher correlation was found between the two methods for the assessment of LVEF, EDV and ESV. On the other hand, the correlation was lower for the assessment of wall motion. Gated SPECT and two-dimensional echocardiography correlate well for the assessment of left ventricular function and volumes. Gated SPECT has the advantage of providing information about left ventricular function, dimensions and perfusion.


Gene expression profiling of HCV genotype 3a initial liver fibrosis and cirrhosis patients using microarray.
Hepatitis C virus (HCV) causes liver fibrosis that may lead to liver cirrhosis or hepatocellular carcinoma (HCC), and may partially depend on infecting viral genotype. HCV genotype 3a is being more common in Asian population, especially Pakistan; the detail mechanism of infection still needs to be explored. In this study, we investigated and compared the gene expression profile between initial fibrosis stage and cirrhotic 3a genotype patients. Gene expression profiling of human liver tissues was performed containing more than 22000 known genes. Using Oparray protocol, preparation and hybridization of slides was carried out and followed by scanning with GeneTAC integrator 4.0 software. Normalization of the data was obtained using MIDAS software and Significant Microarray Analysis (SAM) was performed to obtain differentially expressed candidate genes. Out of 22000 genes studied, 219 differentially regulated genes found with P ≤ 0.05 between both groups; 107 among those were up-regulated and 112 were down-regulated. These genes were classified into 31 categories according to their biological functions. The main categories included: apoptosis, immune response, cell signaling, kinase activity, lipid metabolism, protein metabolism, protein modulation, metabolism, vision, cell structure, cytoskeleton, nervous system, protein metabolism, protein modulation, signal transduction, transcriptional regulation and transport activity. This is the first study on gene expression profiling in patients associated with genotype 3a using microarray analysis. These findings represent a broad portrait of genomic changes in early HCV associated fibrosis and cirrhosis. We hope that identified genes in this study will help in future to act as prognostic and diagnostic markers to differentiate fibrotic patients from cirrhotic ones.


Estimation of aerosol columnar size distribution and optical thickness from the angular distribution of radiance exiting the atmosphere: simulations.
We report the results of simulations in which an algorithm developed for estimation of aerosol optical properties from the angular distribution of radiance exiting the top of the atmosphere over the oceans [Appl. Opt. 33, 4042 (1994)] is combined with a technique for carrying out radiative transfer computations by synthesis of the radiance produced by individual components of the aerosol-size distribution [Appl. Opt. 33, 7088 (1994)], to estimate the aerosol-size distribution by retrieval of the total aerosol optical thickness and the mixing ratios for a set of candidate component aerosol-size distributions. The simulations suggest that in situations in which the true size-refractive-index distribution can actually be synthesized from a combination of the candidate components, excellent retrievals of the aerosol optical thickness and the component mixing ratios are possible. An exception is the presence of strongly absorbing aerosols. The angular distribution of radiance in a single spectral band does not appear to contain sufficient information to separate weakly from strongly absorbing aerosols. However, when two spectral bands are used in the algorithm, retrievals in the case of strongly absorbing aerosols are improved. When pseudodata were simulated with an aerosol-size distribution that differed in functional form from the candidate components, excellent retrievals were still obtained as long as the refractive indices of the actual aerosol model and the candidate components were similar. This underscores the importance of component candidates having realistic indices of refraction in the various size ranges for application of the method. The examples presented all focus on the multiangle imaging spectroradiometer; however, the results should be as valid for data obtained by the use of high-altitude airborne sensors.


Preparation and characterization of magnetic allylamine modified graphene oxide-poly(vinyl acetate-co-divinylbenzene) nanocomposite for vortex assisted magnetic solid phase extraction of some metal ions.
Magnetic allylamine modified graphene oxide-poly(vinyl acetate-co-divinylbenzene) (MGO-DVB-VA) was synthesized and used for magnetic solid phase extraction of Pb(II), Cd(II), Cu(II), Ni(II) and Co(II) prior to their determination by flame atomic absorption spectroscopy. The adsorbent surface functional group was characterized by using FT-IR and Raman spectroscopy. XRD pattern was used to determine the layers of GO. Surface morphology and elemental composition of the adsorbent were evaluated by using SEM and EDX analysis. Various parameters, effecting adsorption efficiency like initial solution pH, adsorbent dose, type and volume of eluent, volume of sample and diverse ions effects were optimized. The preconcentration factor (PF) is 40 for all the metals and the limits of detection for Pb, Cd, Cu, Ni and Co are in the range of 0.37-2.39 µg L(-1) and relative standard deviation below 3.1%. The method was validated by using the method for certified reference materials (Tobacco Leaves (INCT-OBTL-5), Tomato Leaves (1573a), Certified Water (SPS-ww2) and Certified Water (TMDA 64-2)). The method was successfully applied for natural water and food samples.


Breast-Conserving Surgery in Patients With Mammary Paget's Disease.
We aimed to analyze the association between Paget's disease (PD) and breast cancer (BC) subtypes and compare the effect of breast-conserving surgery (BCS) as a local treatment with mastectomy for PD. Data of patients with histologic type International Classification of Diseases-0-3 8540-8543 who were treated from 1973 to 2014 were retrieved from the Surveillance, Epidemiology, and End Results database of the National Cancer Institute. A chi-square test was used to identify differences in categorical data among different groups. Overall survival (OS) was analyzed using the Kaplan-Meier method, log-rank test, Cox proportional hazards models, sequential landmark analysis, and propensity score-matched analysis. The study cohort included 5398 patients. Triple-negative BC accounted for the fewest patients with PD-only (1/22, 4.54%), Paget's disease-ductal carcinoma in situ (PD-DCIS) (3/48, 6.25%), and Paget's disease-invading ductal carcinoma (PD-IDC) (23/352, 6.53%). According to the results of the log-rank test and Cox analysis, the 10-year OS rates were similar for the BCS and mastectomy subgroups among patients with PD-DCIS or PD-IDC. Furthermore, there were no significant differences in survival benefits among the different surgeries after propensity score matching. Landmark analyses for OS of patients with PD-DCIS or PD-IDC surviving more than 1, 3, and 5 y showed no significant differences in survival. There were statistical differences in 10-year OS rates for patients with PD-DCIS or PD-IDC who underwent radiation therapy, or not, following BCS (both, P < 0.001). For patients with PD-DCIS or PD-IDC, breast conservation therapy with lumpectomy and radiation is an effective local treatment strategy, compared with mastectomy.


Chimpanzee Adenovirus Vaccine Provides Multispecies Protection against Rift Valley Fever.
Rift Valley Fever virus (RVFV) causes recurrent outbreaks of acute life-threatening human and livestock illness in Africa and the Arabian Peninsula. No licensed vaccines are currently available for humans and those widely used in livestock have major safety concerns. A 'One Health' vaccine development approach, in which the same vaccine is co-developed for multiple susceptible species, is an attractive strategy for RVFV. Here, we utilized a replication-deficient chimpanzee adenovirus vaccine platform with an established human and livestock safety profile, ChAdOx1, to develop a vaccine for use against RVFV in both livestock and humans. We show that single-dose immunization with ChAdOx1-GnGc vaccine, encoding RVFV envelope glycoproteins, elicits high-titre RVFV-neutralizing antibody and provides solid protection against RVFV challenge in the most susceptible natural target species of the virus-sheep, goats and cattle. In addition we demonstrate induction of RVFV-neutralizing antibody by ChAdOx1-GnGc vaccination in dromedary camels, further illustrating the potency of replication-deficient chimpanzee adenovirus vaccine platforms. Thus, ChAdOx1-GnGc warrants evaluation in human clinical trials and could potentially address the unmet human and livestock vaccine needs.


Changes in deoxyribonucleic acid polymerase activities in synthesis of deoxyribonucleic acid during sporulation of Bacillus subtilis.
The deoxyribonucleic acid (DNA) polymerase activities in Bacillus subtilis strains Marburg 168 (thy-trp2) and D22, a DNA polymerase I-deficient mutant, were measured at various stages of sporulation. The DNA polymerase I activity, which had decreased after the exponential growth, began to increase at the early stage of sporulation, reached a maximum and then again decreased. The activity of neither DNA polymerase II nor III was observed to change so drastically as that of DNA polymerase I during sporulation. The incorporation of [3H]deoxythymidine 5'-triphosphate ([3H]dTTP) into Brij 58-treated permeable cells increased during sporulation. The stimulation of [3H]dTTP incorporation into the cells by irradiation with ultraviolet light was also observed to coincide with DNA polymerase I activity. In strain D22 the activities of DNA polymerase II and III were almost constant with time. Neither change of [3H]dTTP incorporation into Brij 58-treated cells nor stimulation of incorporation by irradiation with ultraviolet light was observed.


Finite element aided tracking of signal intensity changes in deforming intervertebral disc tissue.
Tracking of signal intensity changes in soft tissue over time is often hampered by deformation of the tissue. In this study a method is described that uses finite element modeling to compensate for tissue deformation. The method is applied to the quantification of fluid redistribution in an intervertebral disc that deforms under mechanical load. The clinical importance of this application emerges from the increased susceptibility of the intervertebral disc to damage after a period of mechanical loading. The study shows that the use of the finite element aided approach results in a detailed map of tissue MRI signal changes, where the distorting effects of tissue deformation are eliminated.


Candidal Prevalence in Diabetics and its Species Identification.
Candida species is a part of the normal mouth flora. Diabetes mellitus is a predisposing factor for the onset of oral candidiasis. The objective of this study was to estimate salivary glucose in patient with diabetes and healthy individuals, to determine total candidal counts as well as different candidal species in the saliva of patients with diabetes and nondiabetics. A sample size of 80 patients was taken. Out of 80 patients, 30 patients were uncontrolled diabetes patients (Group I), 30 patients were controlled diabetes patients (Group II), and 20 patients were healthy individuals (Group III). From all the salivary samples, salivary glucose estimation was done using the glucose oxidase-peroxidase method. Candidal colony-forming units (CFUs) were determined from all the salivary samples. CHROMagar Candida medium was used for the identification of various Candida species. One-way ANOVA was used to test for differences between the means of the three groups. Pearson's correlation coefficient test was used to evaluate the relationships between the variables. The mean salivary glucose levels were higher in Group I followed by Group II and Group III. The mean candidal CFU was significantly higher in Group I than in Group II and Group III. Candida glabrata was the predominant species found and more so in patients with diabetes and needs further study. Other common species isolated was C. albicans. Species identification can help in better treatment strategies and gain good control over the disease.


Heteroplasmy of mouse mtDNA is genetically unstable and results in altered behavior and cognition.
Maternal inheritance of mtDNA is the rule in most animals, but the reasons for this pattern remain unclear. To investigate the consequence of overriding uniparental inheritance, we generated mice containing an admixture (heteroplasmy) of NZB and 129S6 mtDNAs in the presence of a congenic C57BL/6J nuclear background. Analysis of the segregation of the two mtDNAs across subsequent maternal generations revealed that proportion of NZB mtDNA was preferentially reduced. Ultimately, this segregation process produced NZB-129 heteroplasmic mice and their NZB or 129 mtDNA homoplasmic counterparts. Phenotypic comparison of these three mtDNA lines demonstrated that the NZB-129 heteroplasmic mice, but neither homoplasmic counterpart, had reduced activity, food intake, respiratory exchange ratio; accentuated stress response; and cognitive impairment. Therefore, admixture of two normal but different mouse mtDNAs can be genetically unstable and can produce adverse physiological effects, factors that may explain the advantage of uniparental inheritance of mtDNA.


Saving babies' lives (SBL) - a programme to reduce neonatal mortality in rural Cambodia: study protocol for a stepped-wedge cluster-randomised trial.
Neonatal mortality remains unacceptably high. Many studies successful at reducing neonatal mortality have failed to realise similar gains at scale. Effective implementation and scale-up of interventions designed to tackle neonatal mortality is a global health priority. Multifaceted programmes targeting the continuum of neonatal care, with sustainability and scalability built into the design, can provide practical insights to solve this challenge. Cambodia has amongst the highest neonatal mortality rates in South-East Asia, with rural areas particularly affected. The primary objective of this study is the design, implementation, and assessment of the Saving Babies' Lives programme, a package of interventions designed to reduce neonatal mortality in rural Cambodia. This study is a five-year stepped-wedge cluster-randomised trial conducted in a rural Cambodian province with an estimated annual delivery rate of 6615. The study is designed to implement and evaluate the Saving Babies' Lives programme, which is the intervention. The Saving Babies' Lives programme is an iterative package of neonatal interventions spanning the continuum of care and integrating into the existing health system. The Saving Babies' Lives programme comprises two major components: participatory learning and action with community health workers, and capacity building of primary care facilities involving facility-based mentorship. Standard government service continues in control arms. Data collection covering the whole study area includes surveillance of all pregnancies, verbal and social autopsies, and quality of care surveys. Mixed methods data collection supports iteration of the complex intervention, and facilitates impact, outcome, process and economic evaluation. Our study uses a robust study design to evaluate and develop a holistic, innovative, contextually relevant and sustainable programme that can be scaled-up to reduce neonatal mortality. ClinicalTrials.gov: NCT04663620 . Registered on 11th December 2020, retrospectively registered.


Secondhand smoke exposure in public places in Guatemala: comparison with other Latin American countries.
To measure secondhand smoke levels in workplaces in Guatemala and to compare exposure to levels in other Latin American cities. Exposure was estimated by passive sampling of vapor phase nicotine using a filter badge. Filters were placed in 1 hospital, 1 school, 2 universities, 1 government building, the airport, and 10 restaurants/bars. In total, 103 filters were deployed (plus 7 duplicates and 10 blanks). Nicotine (microg/m(3)) was measured by gas chromatography. Medians [interquartile ranges (IQR)] of nicotine concentrations were reported and compared with other Latin American cities. A survey about attitudes for smoke-free workplaces was distributed among employees. Nicotine was detected in most (68%) locations surveyed (including workplaces where smoking is banned). The highest levels were found in bars [median, 4.58 microg/m(3) (IQR, 1.71-6.44)] and restaurants [median, 0.56 microg/m(3) (IQR, 0.46-0.71)]. Nicotine concentrations in bars and restaurants were 710 and 114 times higher, respectively, compared with hospital concentrations after adjustment for smoking ban signs, type of ventilation, and volume of the area. Support for smoke-free environments was high, except in bar/restaurant and airport workers. Airborne nicotine levels in Guatemala were similar to those found in other Latin American cities. In Guatemala, exposure to secondhand smoke is highly prevalent. Workers in bars and restaurants are disproportionately exposed to secondhand smoke compared with other workers. There is an urgent need for complete smoke-free legislation and for educating workers about the benefits of smoke-free workplaces.


Methods for fabricating microarrays of motile bacteria.
Motile bacterial cell microarrays were fabricated by attaching Escherichia coli K-12 cells onto predesigned 16-mercaptohexadecanoic acid patterned microarrays, which were covalently functionalized with E. coli antibodies or poly-L-lysine. By utilizing 11-mercaptoundecyl-penta(ethylene glycol) or 11-mercapto-1-undecanol as passivating molecules, nonspecific binding of E. coli was significantly reduced. Microcontact printing and dip-pen nanolithography were used to prepare microarrays for bacterial adhesion, which was studied by optical fluorescence and atomic force microscopy. These data indicate that single motile E. coli can be attached to predesigned line or dot features and binding can occur via the cell body or the flagella of bacteria. Adherent bacteria are viable (remain alive and motile after adhesion to patterned surface features) for more than four hours. Individual motile bacterial cells can be placed onto predesigned surface features that are at least 1.3 microm in diameter or larger. The importance of controlling the adhesion of single bacterial cell to a surface is discussed with regard to biomotor design.


Acceptance and commitment therapy as guided self-help for psychological distress and positive mental health: a randomized controlled trial.
In order to reduce the high prevalence of depression, early interventions for people at risk of depression are warranted. This study evaluated the effectiveness of an early guided self-help programme based on acceptance and commitment therapy (ACT) for reducing depressive symptomatology. Participants with mild to moderate depressive symptomatology were recruited from the general population and randomized to the self-help programme with extensive email support (n=125), the self-help programme with minimal email support (n=125) or to a waiting list control group (n=126). Participants completed measures before and after the intervention to assess depression, anxiety, fatigue, experiential avoidance, positive mental health and mindfulness. Participants in the experimental conditions also completed these measures at a 3-month follow-up. In the experimental conditions significant reductions in depression, anxiety, fatigue, experiential avoidance and improvements in positive mental health and mindfulness were found, compared with the waiting list condition (effect sizes Cohen's d=0.51-1.00). These effects were sustained at the 3-month follow-up. There were no significant differences between the experimental conditions on the outcome measures. The ACT-based self-help programme with minimal email support is effective for people with mild to moderate depressive symptomatology.


Estimating the value of tropical coastal wetland habitats to fisheries: Caveats and assumptions.
Habitat valuation can provide an objective basis for the prioritisation of conservation and restoration actions. The attribution of fisheries production to particular habitat units is an important measure of value, but is difficult to estimate. Using the case study of habitat use by juvenile banana prawns in a tropical estuary, we assessed the potential to produce valid value estimates at two spatio-conceptual scales: estuary reach and whole estuary. Additionally, we also explore the potential to produce meaningful value estimates at the scale of whole estuary contribution to the offshore fisheries stock. A diversity of potential and actual sources of error and logical problems means that quantification at any scale is at best of uncertain validity and produces estimates that are likely to produce unreliable results if treated as quantitative inputs to production models. Estimates for the whole estuary were the most viable, although still requiring substantial assumptions that may or may not be reasonable in particular situations. Estimates for individual habitats required the unreasonable assumption of limited prawn movement, while estimates of contribution of an estuary to the fishery required difficult-to-obtain and usually unavailable information. Because low occupancy habitats can have trophic value, we also used stable isotope analysis to assess the importance of mangroves and saltmarshes to prawn nutrition. No particular habitat was of critical trophic importance, again suggesting that the habitat-production link is most usefully assessed at the whole-of-estuary scale. While valuable and required to support targeted ecosystem management and protection and restoration efforts, valid estimates of the contribution of particular units to fisheries are likely to be unachievable in many situations.


Incidence of facial pain in the general population.
Facial pain has a considerable impact on quality of life. Accurate incidence estimates in the general population are scant. The aim was therefore to estimate the incidence rate (IR) of trigeminal neuralgia (TGN), postherpetic neuralgia (PHN), cluster headache (CH), occipital neuralgia (ON), local neuralgia (LoN), atypical facial pain (AFP), glossopharyngeal neuralgia (GPN) and paroxysmal hemicrania (PH) in the Netherlands. In the population-based Integrated Primary Care Information (IPCI) medical record database potential facial pain cases were identified from codes and narratives. Two medical doctors reviewed medical records, questionnaires from general practitioners and specialist letters using criteria of the International Association for the Study of Pain. A pain specialist arbitrated if necessary and a random sample of all cases was evaluated by a neurologist. The date of onset was defined as date of first specific symptoms. The IR was calculated per 100,000PY. Three hundred and sixty-two incident cases were ascertained. The overall IR [95% confidence interval] was 38.7 [34.9-42.9]. It was more common among women compared to men. Trigeminal neuralgia and cluster headache were the most common forms among the studied diseases. Paroxysmal hemicrania and glossopharyngeal neuralgia were among the rarer syndromes. The IR increased with age for all diseases except CH and ON, peaking in the 4th and 7th decade, respectively. Postherpetic neuralgia, CH and LoN were more common in men than women. From this we can conclude that facial pain is relatively rare, although more common than estimated previously based on hospital data.


Seroadaptive behaviours among men who have sex with men in San Francisco: the situation in 2008.
To assess changes in seroadaptive behaviours among men who have sex with men (MSM) in San Francisco over the past 4 years. 461 MSM were recruited in 2008 as the second wave of the US National HIV Behavioural Surveillance (NHBS) survey in San Francisco. Participants were classified into patterns of seroadaptive behaviours based on reported sexual practices (ie, episodes of insertive and receptive anal sex), condom use, HIV serostatus and partners' serostatus for up to five partners in the preceding 6 months. The prevalence of seroadaptive behaviours was compared with the first wave of NHBS, which used identical methods in 2004. In 2008, 33.7% of HIV-negative and 18.9% of HIV-positive MSM used condoms 100% of the time; nearly half (48.0%) of HIV-negative MSM and two-thirds (66.7%) of HIV-positive MSM had unprotected anal intercourse (UAI). Collectively, seroadaptive behaviours comprised the most common form of risk management; 40.5% of HIV-negative MSM and 51.1% of HIV-positive MSM engaged in some form of seroadaptation, the most common being 'pure serosorting' (all UAI with same serostatus partners) reported by 27.5% of HIV-negative MSM and 22.2% of HIV-positive MSM. None of these behaviours were significantly different from their corresponding measures in 2004. Seroadaptation continues to describe the prevailing form of sexual risk management for MSM in San Francisco, suggesting that these behaviours are not novel and require careful measurement to gauge the true potential for the spread of HIV, and nuanced prevention messages to reduce risk.


[Alternative foods: a critical analysis of a proposal for nutritional intervention]
The nationwide use of a 'Multimixture' a formula based on alternative foods such as rice and/or wheat bran, sesame and squash seeds, cassava, beet and carrot leaves, several indigenous leafy vegetables, and ground egg shells has been proposed by the National Institute of Food and Nutrition (INAN) as an official solution to fight hunger among poor Brazilians. The fragile nutritional state of the target population may make technical or ethical questions appear purely academic, yet nutritional, toxicological, and practical feasibility considerations appear to warrant a revision of the INAN proposal. While the Multimixture approach may prove valid as a temporary solution in cases of extreme poverty, it is not universally applicable for the intended use, failing to take into account the age and nutritional status of the subjects or duration of the intervention.


Traditional antimalarial phytotherapy remedies used by the South Coast community, Kenya.
This study was conducted to document herbal medicines used in the treatment of malaria as well as the existing knowledge, attitudes and practices related to malaria recognition, control and treatment in South Coast, Kenya. Data was collected using semistructured questionnaires and interviews. A focused group discussion held with the community members, one in each of the study villages supplemented the interview and questionnaire survey. The respondents were found to have a good understanding of malaria and could distinguish it from other fever types. They were also aware that malaria was spread by mosquitoes. Malaria prevalence was high, and affected individuals an average of four times a year. Community members avoided mosquito bites by using mosquito nets, clearing bushes around their homesteads and burning plant parts to generate smoke. They prevented and treated malaria by taking decoctions or concoctions of traditional herbal remedies. Forty plant species in thirty-five genera distributed in twenty-four families were used as antimalarials in the study area. Five plant species, namely; Heeria insignis Del. (Anacardiaceae), Rottboelia exaltata L.F (Gramineae), Pentanisia ouranogyne S. Moore (Rubiaceae), Agathisanthenum globosum (A. Rich) Hiern (Rubiaceae), and Grewia trichocarpa Hochst ex A. Rich (Tiliaceae) are documented for the first time in South Coast, Kenya, for the treatment of malaria. The plants documented in the current study are a potential source for new bioactive compounds of therapeutic value in malaria treatment. The results provide data for further pharmacological and toxicological studies and development of commercial antimalarial phytotherapy products.


Perioperative hyperglycemia is a strong correlate of postoperative infection in type II diabetic patients after coronary artery bypass grafting.
This study was planned to assess the relationship of perioperative glycemic control to the subsequent risk of infectious complications and to compare early clinical outcomes of coronary artery bypass surgery in diabetics with nondiabetics in a single center. A total of 1090 adults who underwent coronary artery surgery in a five year period were included in a retrospective cohort study based on available chart review. Of 1090 patients, 400 had type II diabetes mellitus. Intraoperative and postoperative blood glucose levels in diabetic group were manipulated by means of a continuous insulin infusion. Data of pre- and postoperative blood glucose levels were evaluated with respect to postoperative infection risk for diabetics. Risks of early mortality, cerebrovascular accident, and postoperative infection in diabetic patients were compared with the nondiabetic group. High preoperative mean glucose levels were the main risk factor for the development of postoperative infection (p = 0.012 and p = 0.028 for the mean glucose levels 1 and 2 days before operation, respectively). For diabetic group, of 400 patients 20 (5%) were diagnosed to have postoperative infection (superficial sternal wound in 3 (0.75%), donor site infection in 4 (1%), mediastinitis in 5 (1.25%), urinary tract infection in 6 (1.5%), and lung infection in 2 (0.5%) patients). The diabetic group had significantly higher prevalence of mediastinitis, donor site infection, urinary tract infection and total infection (p values were 0.048, 0.013, 0.009, and 0.044, respectively). Early mortality was higher among diabetics than in nondiabetics (1.73% vs 3%, p = 0.048) but the risk of cerebrovascular accident in diabetics was not greater than in nondiabetics in early period. In patients with diabetes who undergo coronary artery bypass surgery, preoperative hyperglycemia is an independent predictor of short-term infectious complications and total length of stay in hospital.


The Impact of Nonequilibrium Conditions in Lung Surfactant: Structure and Composition Gradients in Multilamellar Films.
The lipid-protein mixture that covers the lung alveoli, lung surfactant, ensures mechanical robustness and controls gas transport during breathing. Lung surfactant is located at an interface between water-rich tissue and humid, but not fully saturated, air. The resulting humidity difference places the lung surfactant film out of thermodynamic equilibrium, which triggers the buildup of a water gradient. Here, we present a millifluidic method to assemble multilamellar interfacial films from vesicular dispersions of a clinical lung surfactant extract used in replacement therapy. Using small-angle X-ray scattering, infrared, Raman, and optical microscopies, we show that the interfacial film consists of several coexisting lamellar phases displaying a substantial variation in water swelling. This complex phase behavior contrasts to observations made under equilibrium conditions. We demonstrate that this disparity stems from additional lipid and protein gradients originating from differences in their transport properties. Supplementing the extract with cholesterol, to levels similar to the endogenous lung surfactant, dispels this complexity. We observed a homogeneous multilayer structure consisting of a single lamellar phase exhibiting negligible variations in swelling in the water gradient. Our results demonstrate the necessity of considering nonequilibrium thermodynamic conditions to study the structure of lung surfactant multilayer films, which is not accessible in bulk or monolayer studies. Our reconstitution methodology also opens avenues for lung surfactant pharmaceuticals and the understanding of composition, structure, and property relationships at biological air-liquid interfaces.


Astroglial glutamate transporter deficiency increases synaptic excitability and leads to pathological repetitive behaviors in mice.
An increase in the ratio of cellular excitation to inhibition (E/I ratio) has been proposed to underlie the pathogenesis of neuropsychiatric disorders, such as autism spectrum disorders (ASD), obsessive-compulsive disorder (OCD), and Tourette's syndrome (TS). A proper E/I ratio is achieved via factors expressed in neuron and glia. In astrocytes, the glutamate transporter GLT1 is critical for regulating an E/I ratio. However, the role of GLT1 dysfunction in the pathogenesis of neuropsychiatric disorders remains unknown because mice with a complete deficiency of GLT1 exhibited seizures and premature death. Here, we show that astrocyte-specific GLT1 inducible knockout (GLAST(CreERT2/+)/GLT1(flox/flox), iKO) mice exhibit pathological repetitive behaviors including excessive and injurious levels of self-grooming and tic-like head shakes. Electrophysiological studies reveal that excitatory transmission at corticostriatal synapse is normal in a basal state but is increased after repetitive stimulation. Furthermore, treatment with an N-methyl-D-aspartate (NMDA) receptor antagonist memantine ameliorated the pathological repetitive behaviors in iKO mice. These results suggest that astroglial GLT1 has a critical role in controlling the synaptic efficacy at corticostriatal synapses and its dysfunction causes pathological repetitive behaviors.


Living on the edge: A continental-scale assessment of forest vulnerability to drought.
Globally, forests are facing an increasing risk of mass tree mortality events associated with extreme droughts and higher temperatures. Hydraulic dysfunction is considered a key mechanism of drought-triggered dieback. By leveraging the climate breadth of the Australian landscape and a national network of research sites (Terrestrial Ecosystem Research Network), we conducted a continental-scale study of physiological and hydraulic traits of 33 native tree species from contrasting environments to disentangle the complexities of plant response to drought across communities. We found strong relationships between key plant hydraulic traits and site aridity. Leaf turgor loss point and xylem embolism resistance were correlated with minimum water potential experienced by each species. Across the data set, there was a strong coordination between hydraulic traits, including those linked to hydraulic safety, stomatal regulation and the cost of carbon investment into woody tissue. These results illustrate that aridity has acted as a strong selective pressure, shaping hydraulic traits of tree species across the Australian landscape. Hydraulic safety margins were constrained across sites, with species from wetter sites tending to have smaller safety margin compared with species at drier sites, suggesting trees are operating close to their hydraulic thresholds and forest biomes across the spectrum may be susceptible to shifts in climate that result in the intensification of drought.


Effects of chemical, biological, and physical aging as well as soil addition on the sorption of pyrene to activated carbon and biochar.
In this study, the suitability of biochar and activated carbon (AC) for contaminated soil remediation is investigated by determining the sorption of pyrene to both materials in the presence and absence of soil and before as well as after aging. Biochar and AC were aged either alone or mixed with soil via exposure to (a) nutrients and microorganisms (biological), (b) 60 and 110 °C (chemical), and (c) freeze-thaw cycles (physical). Before and after aging, the pH, elemental composition, cation exchange capacity (CEC), microporous SA, and sorption isotherms of pyrene were quantified. Aging at 110 °C altered the physicochemical properties of all materials to the greatest extent (for example, pH increased by up to three units and CEC by up to 50% for biochar). Logarithmic K(Fr) values ranged from 7.80 to 8.21 (ng kg(-1))(ng L(-1))(-nF) for AC and 5.22 to 6.21 (ng kg(-1))(ng L(-1))(-nF) for biochar after the various aging regimes. Grinding biochar to a smaller particle size did not significantly affect the sorption of d(10) pyrene, implying that sorption processes operate on the subparticle scale. Chemical aging decreased the sorption of pyrene to the greatest extent (up to 1.8 log unit for the biochar+soil). The sorption to AC was affected more by the presence of soil than the sorption to biochar was. Our results suggest that AC and biochar have a high sorption capacity for pyrene that is maintained both in the presence of soil and during harsh aging. Both materials could therefore be considered in contaminated land remediation.


Production of Pseudomonas aeruginosa Rhamnolipid Biosurfactants in Heterologous Hosts.
The high-level production of rhamnolipid biosurfactants is a unique feature of Pseudomonas aeruginosa and is strictly regulated in response to environmental conditions. The final step in rhamnolipid biosynthesis is catalyzed by the rhlAB genes encoding a rhamnosyltransferase. The expression of the cloned rhlAB genes was studied in heterologous hosts, either under the control of the rhlR and rhlI rhamnolipid regulatory elements or under the control of the tac promoter. A recombinant P. fluorescens strain harboring multiple plasmid-encoded copies of the rhamnolipid gene cluster produced rhamnolipids (0.25 g liter(sup-1)) when grown under nitrogen-limiting conditions. The highest yields (0.6 g liter(sup-1)) and productivities (24 mg liter(sup-1) h(sup-1)) were obtained in a recombinant Pseudomonas putida strain, KT2442, harboring promoterless rhlAB genes fused to the tac promoter on a plasmid. Active rhamnosyltransferase was synthesized, but no rhamnolipids were produced, by recombinant Escherichia coli upon induction of rhlAB gene expression.


Comparison of Concussion Sideline Screening Measures Across Varying Exertion Levels Within Simulated Games.
Currently, there is no gold standard to evaluate the effect of varying game-like exertion states on Sport Concussion Assessment Tool 3rd Edition (SCAT-3) outcomes. Baseline assessments may occur before, during, or after physical activity, while postinjury evaluations predominantly occur following physical activity. Thus, clinicians may be comparing postinjury evaluations completed following exertion to baseline evaluations completed following varying levels of rest or exertion, which may not be a valid method for clinical decision making. To determine the effect of various physical exertion levels on sideline concussion assessment outcomes and reliability. Within-subjects, repeated measures. Field. Physically active participants (N = 36) who regularly participate in basketball activity. Subjects participated in 2 simulated basketball games, completing a symptom checklist, Standardized Assessment of Concussion, and Balance Error Scoring System before game play, during halftime, and at the completion of each simulated game. Pulse rate was assessed as a proxy of physical exertion. Total symptom, Standardized Assessment of Concussion, and Balance Error Scoring System scores. Physical exertion did not significantly predict symptom, Standardized Assessment of Concussion, or Balance Error Scoring System scores, although a trend toward higher symptom scores was observed for females (ß = 0.03, P = .09). All assessments had poor to moderate reliability across sessions (.15 < interclass correlation coefficient [2,1] < .60). Low- to moderate-intensity physical activity did not have a significant effect on clinical concussion sideline assessments; however, the low test-retest reliability observed prevents strong conclusions on these relationships. The poor overall reliability does not allow for clear recommendations for what state of baseline physical exertion (ie, rested or exerted) provides optimal data to make postinjury clinical decisions, although baseline concussion assessments completed at rest have the most valid and conservative normative values for injury comparison.


Total mechanical energy of a ventricle model and cardiac oxygen consumption.
Mechanical energy (ENG) required by a time-varying elastance model of the ventricle was compared with oxygen consumption per beat (VO2) of the canine left ventricle contracting under a variety of loading conditions. ENG needed for this model to increase its elastance during systole is shown to be equal to the sum of the potential energy built in the elastance during systole plus the external mechanical stroke work. This ENG is equivalent to the area (PVA) bounded by the end-systolic and end-diastolic P-V curves and the systolic limb of the P-V loop trajectory in the P-V plane. There was a high correlation (r = 0.89) between VO2s documented in the literature and PVAs assessed by the author from the accompanying P-V data from both isovolumic and ejecting contractions in 11 hearts. A linear regression analysis yielded an empirical equation: VO2 (ml O2/beat) = a . PVA (mmHg . ml/beat) + b, where a = 1.37 X 10(-5) and b = 0.027, which can be used to predict VO2 from PVA. A preliminary experimental study in my laboratory confirmed the validity of this empirical equation.


Purification and properties of the MalT protein, the transcription activator of the Escherichia coli maltose regulon.
Expression of the Escherichia coli maltose regulon is controlled by MalT, a transcriptional activator (Mr = 102,288) encoded by the malT gene. Activation of transcription depends on the presence of the inducer, maltotriose. Using an in vitro transcription/translation assay to monitor the protein, we have purified MalT in native form from MalT-overproducing bacteria. The purified protein is able to promote transcription from different MalT-controlled promoters in well-defined in vitro systems. Maltotriose and the MalT protein suffice to stimulate initiation of transcription at malPp by the E. coli RNA polymerase holoenzyme. In contrast, both MalT protein and cAMP receptor protein are required with their respective effectors, maltotriose and cyclic AMP, for activation of malEp. These data are in agreement with in vivo observations. In addition, we present evidence that MalT is an ATP-binding protein, a result suggesting that ATP may play a role in transcription initiation.


Measuring the rate of change of voice fundamental frequency in fluent speech during mental depression.
A method of measuring the rate of change of fundamental frequency has been developed in an effort to find acoustic voice parameters that could be useful in psychiatric research. A minicomputer program was used to extract seven parameters from the fundamental frequency contour of tape-recorded speech samples: (1) the average rate of change of the fundamental frequency and (2) its standard deviation, (3) the absolute rate of fundamental frequency change, (4) the total reading time, (5) the percent pause time of the total reading time, (6) the mean, and (7) the standard deviation of the fundamental frequency distribution. The method is demonstrated on (a) a material consisting of synthetic speech and (b) voice recordings of depressed patients who were examined during depression and after improvement.


The male biological clock is ticking: a review of the literature.
The term biological clock is usually used by physicians and psychologists to refer to the declining fertility, increasing risk of fetal birth defects and alterations to hormone levels experienced by women as they age. Female fecundity declines slowly after the age of 30 years and more rapidly after 40 and is considered the main limiting factor in treating infertility. However, there are several scientific reports, chapters in books and review articles suggesting that men may also have a biological clock. The aim of our study was to conduct a review of the literature, based on the Medical Literature Analysis and Retrieval System Online (Medline), to evaluate the male biological clock. After adjustments for other factors, the data demonstrate that the likelihood that a fertile couple will take more than 12 months to conceive nearly doubles from 8% when the man is < 25 years old to 15% when he is > 35 years old. Thus, paternal age is a further factor to be taken into account when deciding on the prognosis for infertile couples. Also, increasing male age is associated with a significant decline in fertility (five times longer to achieve pregnancy at the age of 45 years). Patients and their physicians therefore need to understand the effects of the male biological clock on sexual and reproductive health, in that it leads to erectile dysfunction and male infertility, as well as its potential implications for important medical conditions such as diabetes and cardiovascular diseases.


The tumour-suppressor protein ASPP1 is nuclear in human germ cells and can modulate ratios of CD44 exon V5 spliced isoforms in vivo.
The ASPP1 (Apoptosis Stimulating Protein of p53) protein is an important tumour-suppressor. We have detected a novel protein interaction between the human ASPP1 (hASPP1) protein and the predominantly nuclear adaptor protein SAM68. In the human testis, full-length endogenous hASPP1 protein is located in the nucleus like SAM68, predominantly within meiotic and postmeiotic cells. Mouse ASPP1 (mASPP1) protein is mainly expressed in the brain and testis. The interaction with nuclear SAM68 is likely to be restricted to human germ cells, since endogenous mASPP1 protein is exclusively cytoplasmic. The C-terminal region of hASPP1 efficiently targeted a fused GFP molecule to the nucleus, whereas the N-terminus of hASPP1 targeted GFP to the cytoplasm. In the context of the full-length molecule this cytoplasmic targeting sequence is dominant in HEK293 and Saos-2 cells, since full-length hASPP1-GFP is almost exclusively cytoplasmic. Despite its predominantly cytoplasmic location, we show that ASPP1-GFP expression in HEK293 cells can regulate the ratio of alternative spliced isoforms derived from a pre-mRNA regulated downstream of cytoplasmic signalling pathways, and our data suggest that ASPP1 may operate in this case downstream or parallel to RAS signalling pathways.