[Vitamin C and B1 allowance in various diseases of the gastrointestinal tract].
In 92 patients with various chronic affections of the gastro-intestinal tract vitamins C and B1 content in the blood and urine was measured. As controls 17 healthy persons were examined. Patients with gastro-intestinal affections were found to show a statistically significant fall of the vitamins C and B1 level in the blood and urine at all seasons of the year (winter, spring, summer and fall). But their decline is most intensive with exacerbation of the ailment and then the presence of the pain syndrome is attended by a steeper drop in the level of these vitamins. A course-wise parenteral administration of the vitamins C and B1 results in their statistically significant rise in the blood and urine, their level, however, remaining below that in healthy persons.


Glyceraldehyde 3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa.
Evidence is provided that the glycolytic enzyme glyceraldehyde 3-phosphate dehydrogenase is covalently linked to the fibrous sheath. The fibrous sheath is a typical structure of mammalian spermatozoa surrounding the axoneme in the principal piece of the flagellum. More than 90% of boar sperm glyceraldehyde 3-phosphate dehydrogenase activity is sedimented after cell disintegration by centrifugation. Detergents, different salt concentrations or short term incubation with chymotrypsin do not solubilize the enzyme, whereas digestion with trypsin or elastase does. Short term incubation with trypsin (15 minutes) even resulted in an activation of glyceraldehyde 3-phosphate dehydrogenase. Purification on phenyl-Sepharose yielded a homogeneous glyceraldehyde 3-phosphate dehydrogenase as judged from gel electrophoresis SDS-PAGE and native gradient PAGE. The molecular masses are 41.5 and 238 kDa, respectively, suggesting native glyceraldehyde 3-phosphate dehydrogenase to be a hexamer. Rabbit polyclonal antibodies raised to purified glyceraldehyde 3-phosphate dehydrogenase show a high specificity for mammalian spermatozoal glyceraldehyde 3-phosphate dehydrogenase, while other proteins of boar spermatozoa or the muscle glyceraldehyde 3-phosphate dehydrogenase are not labelled. Immunogold staining performed in a post-embedding procedure reveals the localization of glyceraldehyde 3-phosphate dehydrogenase along the fibrous sheath in spermatozoa of boar, bull, rat, stallion and man. Other structures such as the cell membrane, dense fibres, the axoneme or the mitochondria are free of label. During the process of sperm maturation, most of the cytoplasm of the sperm midpiece is removed as droplets during the passage through the epididymis. The labelling of this cytoplasm, in immature boar spermatozoa and in the droplets, indicates that glyceraldehyde 3-phosphate dehydrogenase is completely removed from the midpiece during sperm maturation in the epididymis. The inverse compartmentation of the glycolytic enzyme and mitochondria in the mammalian sperm flagella suggests that ATP-production in the principal piece mainly occurs by glycolysis and in the midpiece by respiration.


Spectroscopy of YO from first principles.
We report an ab initio study on the spectroscopy of the open-shell diatomic molecule yttrium oxide, YO. The study considers the six lowest doublet states, X2Σ+, A'2Δ, A2Π, B2Σ+, C2Π, D2Σ+, and a few higher-lying quartet states using high levels of electronic structure theory and accurate nuclear motion calculations. The coupled cluster singles, doubles, and perturbative triples, CCSD(T), and multireference configuration interaction (MRCI) methods are employed in conjunction with a relativistic pseudopotential on the yttrium atom and a series of correlation-consistent basis sets ranging in size from triple-ζ to quintuple-ζ quality. Core-valence correlation effects are taken into account and complete basis set limit extrapolation is performed for CCSD(T). Spin-orbit coupling is included through the use of both MRCI state-interaction with spin-orbit (SI-SO) approach and four-component relativistic equation-of-motion CCSD calculations. Using the ab initio data for bond lengths ranging from 1.0 to 2.5 Å, we compute 6 potential energy, 12 spin-orbit, 8 electronic angular momentum, 6 electric dipole moment and 12 transition dipole moment (4 parallel and 8 perpendicular) curves which provide a complete description of the spectroscopy of the system of six lowest doublet states. The Duo nuclear motion program is used to solve the coupled nuclear motion Schrödinger equation for these six electronic states. The spectra of 89Y16O simulated for different temperatures are compared with several available high resolution experimental studies; good agreement is found once minor adjustments are made to the electronic excitation energies.


Antagonism of ethanol and pentobarbital actions by benzodiazepine inverse agonists: neurochemical studies.
The benzodiazepine inverse agonist Ro 15-4513 has been shown to antagonize several behavioral effects of ethanol and to block the effects of ethanol on chloride flux across brain membranes. We used isolated mouse brain membranes to test whether Ro 15-4513 would reduce the effects of ethanol on membrane fluidity, voltage-dependent calcium channels, microsomal calcium release or binding of t-[35S]butylbicyclophosphorothionate. None of these actions of ethanol were altered by Ro 15-4513. The enhancement of gamma-aminobutyric acid (GABA)-activated chloride flux produced by ethanol or pentobarbital was antagonized partially by Ro 15-4513. Another inverse agonist, FG 7142, was more effective than Ro 15-4513 as an antagonist of ethanol actions on chloride flux. These results demonstrate that the ethanol antagonist action of Ro 15-4513 is specific for GABA-activated chloride flux and does not extend to other neurochemical actions of ethanol. The inverse agonist action (i.e., inhibition of GABA-activated chloride flux tested in the absence of ethanol) of Ro 15-4513 and FG 7142 was revealed by pretreatment of mice in vivo with ethanol. This raises the possibility that ethanol exposure increases the inverse agonist actions of Ro 15-4513 and related drugs and that these inverse agonist actions contribute to the ethanol antagonism observed in vivo and in vitro.


Living in the fast lane: rapid development of the locomotor muscle in immature harbor porpoises (Phocoena phocoena).
Cetaceans (dolphins and whales) are born into the aquatic environment and are immediately challenged by the demands of hypoxia and exercise. This should promote rapid development of the muscle biochemistry that supports diving, but previous research on two odontocete (toothed whales and dolphins) species showed protracted postnatal development for myoglobin content and buffering capacity. A minimum of 1 and 1.5 years were required for Fraser's (Lagenodelphis hosei) and bottlenose (Tursiops truncatus) dolphins to obtain mature myoglobin contents, respectively; this corresponded to their lengthy 2 and 2.5-year calving intervals (a proxy for the dependency period of cetacean calves). To further examine the correlation between the durations for muscle maturation and maternal dependency, we measured myoglobin content and buffering capacity in the main locomotor muscle (longissimus dorsi) of harbor porpoises (Phocoena phocoena), a species with a comparatively short calving interval (1.5 years). We found that at birth, porpoises had 51 and 69 % of adult levels for myoglobin and buffering capacity, respectively, demonstrating greater muscle maturity at birth than that found previously for neonatal bottlenose dolphins (10 and 65 %, respectively). Porpoises achieved adult levels for myoglobin and buffering capacity by 9-10 months and 2-3 years postpartum, respectively. This muscle maturation occurred at an earlier age than that found previously for the dolphin species. These results support the observation that variability in the duration for muscular development is associated with disparate life history patterns across odontocetes, suggesting that the pace of muscle maturation is not solely influenced by exposure to hypoxia and exercise. Though the mechanism that drives this variability remains unknown, nonetheless, these results highlight the importance of documenting the species-specific physiological development that limits diving capabilities and ultimately defines habitat utilization patterns across age classes.


Acute but not chronic macrophage recruitment in filarial infections in mice is dependent on C-C chemokine ligand 2.
Macrophages play an important role in the formation of granulomas and the clearance of Brugia pahangi infections in mice. However, the factors responsible for the recruitment of these cells to the site of infection are not known. In this study we examined the role of the C-C chemokine ligand 2 (CCL2; also known as macrophage chemotactic factor - MCP1) in macrophage recruitment in intraperitoneal infections with B. pahangi. We observed that CCL2 was expressed by peritoneal exudate cells and was present in the sera of wild-type mice. Serum levels of CCL2 peaked twice during the immune response, once during the early, acute phase and again during the late, chronic phase. To further elucidate the role of this chemokine in the anti-filarial immune response, we compared CCL2 deficient (CCL2(-/-)) mice to wild-type mice. We observed that macrophage recruitment was impaired only during the acute phase in the former. While macrophage recruitment was unaffected during the chronic phase, increased accumulation of B and T lymphocytes was seen in these mice. We further report that larval clearance and the in vitro adhesion of PECs to larvae were unimpaired in these mice.


Studies on the pathophysiology and genetic basis of migraine.
Migraine is a neurological disorder that affects the central nervous system causing painful attacks of headache. A genetic vulnerability and exposure to environmental triggers can influence the migraine phenotype. Migraine interferes in many facets of people's daily life including employment commitments and their ability to look after their families resulting in a reduced quality of life. Identification of the biological processes that underlie this relatively common affliction has been difficult because migraine does not have any clearly identifiable pathology or structural lesion detectable by current medical technology. Theories to explain the symptoms of migraine have focused on the physiological mechanisms involved in the various phases of headache and include the vascular and neurogenic theories. In relation to migraine pathophysiology the trigeminovascular system and cortical spreading depression have also been implicated with supporting evidence from imaging studies and animal models. The objective of current research is to better understand the pathways and mechanisms involved in causing pain and headache to be able to target interventions. The genetic component of migraine has been teased apart using linkage studies and both candidate gene and genome-wide association studies, in family and case-control cohorts. Genomic regions that increase individual risk to migraine have been identified in neurological, vascular and hormonal pathways. This review discusses knowledge of the pathophysiology and genetic basis of migraine with the latest scientific evidence from genetic studies.


Tetracene derivatives as potential red emitters for organic LEDs.
[structure: see text] As part of our program investigating the use of ethynylated acenes in organic electronics, we have prepared a series of functionalized tetracene derivatives in search of a material with red emission suitable for use in display technologies. A number of such compounds with functionalization on the alkyne and/or the acene ring were easily prepared in one or two steps from commercially available materials. Solution fluorescence quantum efficiencies were generally good for these derivatives, which possessed emission maxima spanning the range 540-637 nm. Simple light-emitting diodes fabricated from these compounds showed that one of the derivatives did exhibit red electroluminescence.


A Novel Biallelic STING1 Gene Variant Causing SAVI in Two Siblings.
STING-associated vasculopathy of infantile-onset (SAVI) is one of the newly identified types of interferonopathies. SAVI is caused by heterozygous gain-of-function mutations in the STING1. We herein report for the first time a homozygous variant in the STING1 gene in two siblings that resulted in constitutive activation of STING gene and the SAVI phenotype. Exome sequencing revealed a novel homozygous NM_198282.3: c.841C>T; p.(Arg281Trp) variant in exon 7 of the STING1 gene. The variant segregated in the family to be homozygous in all affected and either heterozygous or wild type in all healthy. Computational structural analysis of the mutants revealed changes in the STING protein structure/function. Elevated serum beta-interferon levels were observed in the patients compared to the control family members. Treatment with Janus kinase inhibitor (JAK-I) Ruxolitinib suppressed the inflammatory process, decreased beta-interferon levels, and stopped the progression of the disease.


Intrinsically disordered proteins in human diseases: introducing the D2 concept.
Intrinsically disordered proteins (IDPs) lack stable tertiary and/or secondary structures under physiological conditions in vitro. They are highly abundant in nature and their functional repertoire complements the functions of ordered proteins. IDPs are involved in regulation, signaling, and control, where binding to multiple partners and high-specificity/low-affinity interactions play a crucial role. Functions of IDPs are tuned via alternative splicing and posttranslational modifications. Intrinsic disorder is a unique structural feature that enables IDPs to participate in both one-to-many and many-to-one signaling. Numerous IDPs are associated with human diseases, including cancer, cardiovascular disease, amyloidoses, neurodegenerative diseases, and diabetes. Overall, intriguing interconnections among intrinsic disorder, cell signaling, and human diseases suggest that protein conformational diseases may result not only from protein misfolding, but also from misidentification, missignaling, and unnatural or nonnative folding. IDPs, such as alpha-synuclein, tau protein, p53, and BRCA1, are attractive targets for drugs modulating protein-protein interactions. From these and other examples, novel strategies for drug discovery based on IDPs have been developed. To summarize work in this area, we are introducing the D2 (disorder in disorders) concept.


Influence of thyroid status and growth hormone deficiency on ghrelin.
To assess whether some of the alterations in energy homeostasis present in thyroid function disorders and GH deficiency could be mediated by ghrelin. To assess the influence of thyroid status on ghrelin, adult male Sprague-Dawley rats were treated with vehicle (euthyroid), amino-triazole (hypothyroid) or l-thyroxine (hyperthyroid). The influence of GH on ghrelin was assessed in wild-type (control) and GH-deficient (dwarf) Lewis rats. Evaluation of gastric ghrelin mRNA expression in the stomach was carried out by Northern blot. Circulating levels of ghrelin were measured by radioimmunoassay. Hypothyroidism resulted in an increase in gastric ghrelin mRNA levels (euthyroid: 100+/-3.2% vs hypothyroid: 127.3+/-6.5%; P<0.01), being decreased in hyperthyroid rats (70+/-5.4%; P<0.01). In keeping with these results, circulating plasma ghrelin levels were increased in hypothyroid (euthyroid: 124+/-11 pg/ml vs hypothyroid: 262+/-39 pg/ml; P<0.01) and decreased in hyperthyroid rats (75+/-6 pg/ml; P<0.01). Using an experimental model of GH deficiency, namely the dwarf rat, we found a decrease in gastric ghrelin mRNA levels (controls: 100+/-6% vs dwarf: 66+/-5.5%; P<0.01) and circulating plasma ghrelin levels (controls: 124+/-12 pg/ml vs dwarf: 81+/-7 pg/ml; P<0.01). This study provides the first evidence that ghrelin gene expression is influenced by thyroid hormones and GH status and provides further evidence that ghrelin may play an important role in the alteration of energy homeostasis and body weight present in these pathophysiological states.


Evidence for age-related differences in heat acclimatisation responsiveness.
What is the central question of this study? Repeated heat exposure during the summer months can enhance heat loss in humans (seasonal heat acclimatisation), but does the magnitude of that enhancement differ between young and older adults when assessed during passive heat exposure? What is the main finding and its importance? While seasonal heat acclimatisation enhanced evaporative heat loss (i.e. sweating) in both young and older adults, those improvements led to a greater reduction in body heat storage in older adults. These outcomes indicate that heat acclimatisation may confer greater thermoregulatory benefits with increasing age. Repeated heat exposure throughout summer can enhance heat loss in humans (seasonal heat acclimatisation), although the effect of ageing on those improvements remains unclear. We therefore sought to assess thermoregulatory function in young and older adults during environmental heat exposure prior to and following seasonal heat acclimatisation, hypothesizing that the magnitude of adaptation would be greater in older relative to young adults. To achieve this, 14 young (19-27 years) and 10 older adults (55-72 years), who resided in a temperate humid-continental climate, completed a 3 h resting heat exposure (44°C, ∼30% relative humidity) in the winter-spring months as part of a larger investigation (pre-acclimatisation), before being re-evaluated using the same heat stress test following the summer months (post-acclimatisation). Whole-body dry and evaporative heat exchange, and metabolic rate were measured throughout using direct and indirect calorimetry (respectively), and used to quantify body heat storage (metabolic rate + dry heat gain - evaporative heat loss). Evaporative heat loss increased in both groups following acclimatisation, but those improvements led to a decrease in body heat storage in older (mean difference (95% CI); 213 (295, 131) kJ; P < 0.001), but not young adults (-25 (-94, 44) kJ; P = 0.458). Thus, body heat storage was greater in older compared to young adults before (222 (123, 314) kJ; P < 0.001), but not following acclimatisation (34 (-55, 123) kJ; P = 0.433). Although there is a need for larger and more controlled confirmatory studies, our findings indicate that seasonal heat acclimatisation may induce greater thermoregulatory adaptation in older compared to young adults.


A novel duplex qPCR assay for stepwise detection of multiple Perkinsea protistan infections of amphibian tissues.
Alveolate protists within the phylum Perkinsea have been found to infect amphibians across a broad taxonomic and geographic range. Phylogenetic analysis has suggested the existence of two clades of amphibian Perkinsea: a putatively non-pathogenic clade linked to asymptomatic infections of tadpoles in Africa, Europe and South America, and a putatively pathogenic clade linked to disease and mass mortality events of tadpoles in North America. Here, we describe the development of a duplex TaqMan qPCR assay to detect and discriminate between rDNA sequences from both clades of Perkinsea in amphibian tissues. The assay uses a single primer pair to target an 18S small subunit (SSU) ribosomal RNA (rRNA) gene region shared between the two clades, and two dual-labelled probes to target a region within this fragment that is diagnostic for each clade. This assay enables rapid screening for each of the two Perkinsea groups, allowing for detection, primarily of the phylogenetic group associated with disease outbreaks, and secondarily for the phylogenetic group with no current disease relationship identified. Incorporation of our novel qPCR assay into the routine surveillance of amphibian populations will allow for the assessment of the incidence of each protist clade, thereby providing an improved understanding of Perkinsea infection pervasiveness and a method to underpin future conservation planning.


High genomic plasticity and unique features of Xanthomonas translucens pv. graminis revealed through comparative analysis of complete genome sequences.
Xanthomonas translucens pv. graminis (Xtg) is a major bacterial pathogen of economically important forage grasses, causing severe yield losses. So far, genomic resources for this pathovar consisted mostly of draft genome sequences, and only one complete genome sequence was available, preventing comprehensive comparative genomic analyses. Such comparative analyses are essential in understanding the mechanisms involved in the virulence of pathogens and to identify virulence factors involved in pathogenicity. In this study, we produced high-quality, complete genome sequences of four strains of Xtg, complementing the recently obtained complete genome sequence of the Xtg pathotype strain. These genomic resources allowed for a comprehensive comparative analysis, which revealed a high genomic plasticity with many chromosomal rearrangements, although the strains were highly related. A high number of transposases were exclusively found in Xtg and corresponded to 413 to 457 insertion/excision transposable elements per strain. These mobile genetic elements are likely to be involved in the observed genomic plasticity and may play an important role in the adaptation of Xtg. The pathovar was found to lack a type IV secretion system, and it possessed the smallest set of type III effectors in the species. However, three XopE and XopX family effectors were found, while in the other pathovars of the species two or less were present. Additional genes that were specific to the pathovar were identified, including a unique set of minor pilins of the type IV pilus, 17 TonB-dependent receptors (TBDRs), and 11 plant cell wall degradative enzymes. These results suggest a high adaptability of Xtg, conferred by the abundance of mobile genetic elements, which could play a crucial role in pathogen adaptation. The large amount of such elements in Xtg compared to other pathovars of the species could, at least partially, explain its high virulence and broad host range. Conserved features that were specific to Xtg were identified, and further investigation will help to determine genes that are essential to pathogenicity and host adaptation of Xtg.


Celecoxib treatment alters the gene expression profile of normal colonic mucosa.
A clinical trial was recently conducted to evaluate the safety and efficacy of a selective inhibitor of cyclooxygenase-2 (celecoxib) in hereditary nonpolyposis colon cancer patients. In a randomized, placebo-controlled phase I/II multicenter trial, hereditary nonpolyposis colon cancer patients and gene carriers received either celecoxib at one of two doses or placebo. The goal was to evaluate the effects of these treatment arms on a number of endoscopic and tissue-based biomarker end points after 12 months of treatment. As part of this trial, we analyzed gene expression by cDNA array technology in normal descending (rectal) colonic mucosa of patients before and after treatment with celecoxib or placebo. We found that treatment of patients with celecoxib at recommended clinical doses (200 and 400 mg p.o. bid), in contrast to treatment with placebo, leads to changes in expression of >1,400 genes in the healthy colon, although in general, the magnitude of changes is <2-fold. Twenty-three of 25 pairs of colon biopsies taken before and after celecoxib treatment can be classified correctly by the pattern of gene expression in a leave-one-out cross-validation. Immune response, particularly T- and B-lymphocyte activation and early steps of inflammatory reaction, cell signaling and cell adhesion, response to stress, transforming growth factor-beta signaling, and regulation of apoptosis, are the main biological processes targeted by celecoxib as shown by overrepresentation analysis of the distribution of celecoxib-affected genes across Gene Ontology categories. Analysis of possible cumulative effects of celecoxib-induced changes in gene expression indicates that in healthy colon, celecoxib may suppress the immune response and early steps of inflammation, inhibit formation of focal contacts, and stimulate transforming growth factor-beta signaling.


The German day-care study: multicomponent non-drug therapy for people with cognitive impairment in day-care centres supplemented with caregiver counselling (DeTaMAKS) - study protocol of a cluster-randomised controlled trial.
It is the wish of both people with cognitive impairment and their informal caregivers for the impaired person to live at home for as long as possible. This is also in line with economic arguments about health. The existing structure of day-care services for the elderly can be used to achieve this. Due to the current lack of empirical evidence in this field, most day-care centres do not offer a scientifically evaluated, structured intervention, but instead offer a mixture of individual activities whose efficacy has not yet been established. Informal caregivers of people with dementia use day-care centres primarily to relieve themselves of their care tasks and as a support service. The present study therefore investigates the effectiveness of a combination of a multicomponent activation therapy for people with mild cognitive impairment (MCI) or mild to moderate dementia at day-care centres and a brief telephone intervention for their informal caregivers. The study is conducted as a cluster-randomised intervention trial at 34 day-care centres in Germany with a 6-month treatment phase. The centres in the waitlist control group provide "care as usual". A power analysis indicated that 346 people should initially be included in the study. The primary endpoints of the study include the ability to perform activities of daily living (ADL) and cognitive capacities on the side of the day-care centre users and the subjectively perceived burden and well-being of the informal caregivers. The total duration of the study is 3 years, during which data are collected both by the psychometric testing of the people with cognitive impairment and by telephone interviews with informal caregivers. The project has three distinctive quality features. First, it is embedded in real care situations since the day-care services have already been established for this target group. Second, due to the large number of cases and the fact that the participating day-care centres are spread across the entire country, the results can be expected to be generalisable. Third, the interventions can be assumed to be implementable as they required only a one-day training event for the staff already working at the centres. ISRCTN16412551 (Registration date: 30 July 2014, registered retrospectively).


Hirudin and Decorsins of the North American Medicinal Leech Macrobdella decora: Gene Structure Reveals Homology to Hirudins and Hirudin-like Factors of Eurasian Medicinal Leeches.
Blood-sucking leeches, some of which are referred to as medicinal leeches, have caught attention not only because of their medical purposes, but also as study organisms to conduct research within fields as diverse as neurobiology, osmoregulation, ecology, and phylogeny. Of particular interest is the question whether hemophagy in leeches is of single origin or evolved independently several times. A key component in the saliva of hematophagous leeches is hirudin, a strong natural inhibitor of thrombin and hence the blood coagulation cascade. Multiple isoforms of hirudin have been described within and among several leech species and genera, often based on sequence data only. The identification of hirudin-like factors (HLFs) illustrated the necessity to underpin such predictions by functional tests. We overexpressed and purified the hirudin of the North American medicinal leech, Macrobdella decora, and proved its thrombin-inhibiting activity. In addition, analysis of the gene structure of both hirudin and some of the decorsins of M. decora clearly indicated conserved exon and intron positions when compared to genes of hirudins and HLFs of Eurasian medicinal leeches. Our data provide evidence for the incorporation of decorsins into the hirudin superfamily and support the concept of a single origin of blood feeding in jawed leeches.


Fractured occluder disc: a previously unrecognized complication of the Starr-Edwards disc prosthesis.
Fracture of the occluder disc of a low-profile Starr-Edwards prosthesis is a hitherto unrecognized complication. We describe a patient who presented with right heart failure and severe pulmonary hypertension 27 years after mitral valve replacement with a model 6520 caged-disc prosthesis. At surgery, there was a longitudinal split in the occluder disc, and organized thrombus was lodged between the split segments. This case offers a unique opportunity to study the long-term effects of wear on the polyethylene poppet and Stellite cage.


Nutrient Solution Flowing Environment Affects Metabolite Synthesis Inducing Root Thigmomorphogenesis of Lettuce (Lactuca sativa L.) in Hydroponics.
The principal difference between hydroponics and other substrate cultivation methods is the flowing liquid hydroponic cultivation substrate. Our previous studies have revealed that a suitable flowing environment of nutrient solution promoted root development and plant growth, while an excess flow environment was unfavorable for plants. To explain the thigmomorphogenetic response of excess flow-induced metabolic changes, six groups of lettuce (Lactuca sativa L.), including two flow conditions and three time periods, were grown. Compared with the plants without flow, the plants with flow showed decreased root fresh weight, total root length, root surface area, and root volume but increased average root diameter and root density. The roots with flow had more upregulated metabolites than those without flow, suggesting that the flow may trigger metabolic synthesis and activity. Seventy-nine common differential metabolites among six groups were screened, and enrichment analysis showed the most significant enrichment in the arginine biosynthesis pathway. Arginine was present in all the groups and exhibited greater concentrations in roots with flow than without flow. It can be speculated from the results that a high-flowing environment of nutrient solution promotes arginine synthesis, resulting in changes in root morphology. The findings provide insights on root thigmomorphogenesis affected by its growing conditions and help understand how plants respond to environmental mechanical forces.


Identification and molecular characterization of novel and divergent HoBi-like pestiviruses from naturally infected cattle in India.
HoBi-like pestiviruses have been sporadically reported from naturally infected cattle in South America, Asia and Europe. While the closely related bovine viral diarrhoea virus 1 (BVDV-1) and BVDV-2 have been reported from cattle in India, the prevalence and diversity of HoBi-like viruses have not yet been studied. Here we report the genetic diversity and molecular characteristics of HoBi-like viruses, through systematic surveillance in cattle (n=1049) from 21 dairy farms across India during 2012-2013. On the basis of real-time RT-PCR, virus isolation and nucleotide sequencing results, of the 20 pestivirus positive cattle, HoBi-like viruses were identified in 19 cattle from four farms in three states and BVDV-1b in one cattle. Phylogenetic analysis of 5'-UTR and N(pro) region identified the circulation of two lineages of HoBi-like viruses in India, that were distinct to those circulating globally, highlighting the independent evolution of at least three lineages of HoBi-like viruses globally. Antigenic differences were also evident between the two Indian lineages. In addition to revealing that HoBi-like virus may be more widespread in Indian cattle than previously reported, this study shows greater genetic divergence of HoBi-like viruses indicating a need for continued pestivirus surveillance in cattle.


A role for guanidino compounds in the brain.
Guanidino compounds of guanidinoethanesulfonic acid, guanidinoacetic acid, guanidinosuccinic acid, N-acetylarginine, beta-guanidinopropionic acid, creatinine, gamma-guanidinobutyric acid, arginine, guanidine, methylguanidine, homoarginine and alpha-guanidinoglutaric acid are present in the mammalian brain. These guanidino compounds except for arginine and guanidine induce seizures and convulsions in rat, rabbit and cat by intracisternal injection. Hirudonine, audonine, alpha-keto-delta-guanidinovaleric acid, N,N'-dibenzoylguanidine and phenylethylguanidine are also convulsants. Levels of creatinine, guanidinoethanesulfonic acid, creatinine, guanidinoacetic acid and methylguanidine in animal brain were changed at pre- and during convulsions induced by pentylentetrazol, amygdala kindling, iron-induced epileptogenesis and so on. These convulsions are thought to be due to depressed functions of serotonergic neurons and accumulated free radicals. Arginine is a substrate of nitric oxide production by nitric oxide synthase. alpha-Guanidinoglutaric acid is a generator of superoxide, hydroxyl radicals and nitric oxide, and induced C6 glial cell death. On the other hand, aminoguanidine is a free radical scavenger. Energy formation by creatine metabolism may inhibit apoptosis induced by pathogenesis. Free radical generation/ reaction and energy generation by guanidino compounds must be important key role in the brain.


Therapeutic Targeting of Nemo-like Kinase in Primary and Acquired Endocrine-resistant Breast Cancer.
Endocrine resistance remains a major clinical challenge in estrogen receptor (ER)-positive breast cancer. Despite the encouraging results from clinical trials for the drugs targeting known survival signaling, relapse is still inevitable. There is an unmet need to discover new drug targets in the unknown escape pathways. Here, we report Nemo-like kinase (NLK) as a new actionable kinase target that endows previously uncharacterized survival signaling in endocrine-resistant breast cancer. The effects of NLK inhibition on the viability of endocrine-resistant breast cancer cell lines were examined by MTS assay. The effect of VX-702 on NLK activity was verified by kinase assay. The modulation of ER and its coactivator, SRC-3, by NLK was examined by immunoprecipitation, kinase assay, luciferase assay, and RNA sequencing. The therapeutic effects of VX-702 and everolimus were tested on cell line- and patient-derived xenograft (PDX) tumor models. NLK overexpression endows reduced endocrine responsiveness and is associated with worse outcome of patients treated with tamoxifen. Mechanistically, NLK may function, at least in part, via enhancing the phosphorylation of ERα and its key coactivator, SRC-3, to modulate ERα transcriptional activity. Through interrogation of a kinase profiling database, we uncovered and verified a highly selective dual p38/NLK inhibitor, VX-702. Coadministration of VX-702 with the mTOR inhibitor, everolimus, demonstrated a significant therapeutic effect in cell line-derived xenograft and PDX tumor models of acquired or de novo endocrine resistance. Together, this study reveals the potential of therapeutic modulation of NLK for the management of the endocrine-resistant breast cancers with active NLK signaling.


Serum Anti-oligodendrocyte Autoantibodies in Patients With Multiple Sclerosis Detected by a Tissue-Based Immunofluorescence Assay.
Multiple sclerosis (MS), the most prevalent inflammatory disease of the central nervous system (CNS), is characterized by damaged to myelin sheaths and oligodendrocytes. Because MS patients have variable clinical courses and disease severities, it is important to identify biomarkers that predict disease activity and severity. In this study, we assessed the frequencies of serum autoantibodies against mature oligodendrocytes in MS patients using a tissue-based immunofluorescence assay (IFA) to determine whether anti-oligodendrocyte antibodies are associated with the clinical features of MS patients and whether they might be a biomarker to assess CNS tissue damage in MS patients. We assessed the binding of serum autoantibodies to mouse oligodendrocytes expressing Nogo-A, a reliable mature oligodendrocyte marker, by IFA with mouse brain and sera from 147 MS patients, comprising 103 relapsing-remitting MS (RRMS), 22 secondary progressive MS (SPMS), and 22 primary progressive MS (PPMS) patients, 38 neuromyelitis optica spectrum disorder (NMOSD) patients, 23 other inflammatory neurological disorder (OIND) patients, and 39 healthy controls (HCs). Western blotting (WB) was performed using extracted mouse cerebellum proteins and IgG from anti-oligodendrocyte antibody-positive MS patients. Tissue-based IFA showed that anti-oligodendrocyte antibodies were positive in 3/22 (13.6%) PPMS and 1/22 (4.5%) SPMS patients but not in RRMS, NMOSD, and OIND patients or HCs. WB demonstrated the target CNS proteins recognized by serum anti-oligodendrocyte antibodies were approximately 110 kDa and/or 150 kDa. Compared with anti-oligodendrocyte antibody-negative MS patients, MS patients with anti-oligodendrocyte antibodies were significantly older at the time of serum sampling, scored significantly higher on the Expanded Disability Status Scale and the Multiple Sclerosis Severity Score, and had a higher frequency of mental disturbance. Although the clinical significance of anti-oligodendrocyte antibodies is still unclear because of their low frequency, anti-oligodendrocyte autoantibodies are potential biomarkers for monitoring the disease pathology and progression in MS.


Affective-prosodic deficits in schizophrenia: comparison to patients with brain damage and relation to schizophrenic symptoms [corrected].
Although affective prosody seems to be a dominant and lateralised communication function of the right hemisphere, focal lesions of either hemisphere may cause problems with its modulation. When impairment occurs after brain damage, the profiles of affective-prosodic disturbances differ depending on the hemisphere injured. Patients with left brain damage (LBD) improve their performance whereas patients with right brain damage (RBD) do not when the verbal-articulatory demands of the test stimuli are reduced systematically. One of the major arguments for a right hemispheric contribution to schizophrenia has been the documentation of affective prosodic deficits under the assumption that these abnormalities reflect right hemispheric dysfunction. Thus, an essential question to resolve is whether the profile of affective prosodic disturbances in schizophrenia is similar to LBD or RBD, or represents a unique variation. Data were collected from four subject groups: 45 chronic, medication-stabilised, schizophrenic patients, 10 patients with focal LBD, nine patients with focal RBD, and 19 controls. All groups were tested on the aprosodia battery, which uses stimuli having incrementally reduced verbal-articulatory demands. Schizophrenic and aphasic symptoms were evaluated using standard assessment tools. For patients with impaired performance on the aprosodia battery, schizophrenic patients were statistically identical to patients with RBD and robustly different from those with LBD. Thirty eight schizophrenic patients (84.4%) were found to have some type of affective prosodic deficit with the predominant pattern indicating, at minimum, right posterior sylvian dysfunction (57.8%). When schizophrenic symptoms and aprosodic deficits were examined using a principal component analysis, affective comprehension and repetition loaded uniquely as separate factors. The profile of affective-prosodic deficits found in impaired schizophrenic patients is characteristic of RBD, supporting the concept that schizophrenia is a bihemispheric disease process. These deficits may also represent cardinal symptoms of schizophrenia as they are highly prevalent and, except for spontaneous affective prosody, are not associated statistically with traditional clusters of schizophrenic symptoms.


Prohibitin family members interact genetically with mitochondrial inheritance components in Saccharomyces cerevisiae.
Phb2p, a homolog of the tumor suppressor protein prohibitin, was identified in a genetic screen for suppressors of the loss of Mdm12p, a mitochondrial outer membrane protein required for normal mitochondrial morphology and inheritance in Saccharomyces cerevisiae. Phb2p and its homolog, prohibitin (Phb1p), were localized to the mitochondrial inner membrane and characterized as integral membrane proteins which depend on each other for their stability. In otherwise wild-type genetic backgrounds, null mutations in PHB1 and PHB2 did not confer any obvious phenotypes. However, loss of function of either PHB1 or PHB2 in cells with mitochondrial DNA deleted led to altered mitochondrial morphology, and phb1 or phb2 mutations were synthetically lethal when combined with a mutation in any of three mitochondrial inheritance components of the mitochondrial outer membrane, Mdm12p, Mdm10p, and Mmm1p. These results provide the first evidence of a role for prohibitin in mitochondrial inheritance and in the regulation of mitochondrial morphology.


Systematic acute and subchronic toxicity evaluation of polysaccharide-protein complex-functionalized selenium nanoparticles with anticancer potency.
Functionalized selenium nanoparticles (SeNPs) have demonstrated potential for applications in cancer chemotherapy, radio-sensitization, nephroprotection and drug delivery. However, their clinical application requires further systemic safety evaluation. Therefore, in this study, we examine the systematic acute and subchronic toxicity of polysaccharide-protein complex coated SeNPs (PTR-SeNPs). These particles exhibited a low oral acute toxicity (higher LD50) in SPF grade ICR mice and SD rats, and the evaluation of subchronic toxicity demonstrated that the no observed effect level (NOAEL) of the PTR-SeNPs was less than 200 μg Se per kg BW per day, which is about 30 times the tolerable upper intake levels of Se in the human body. In addition, we also found that, under a safe dose (0.75-7.5 mg kg-1), the oral administration of PTR-SeNPs dramatically inhibited the growth of cancer in a tumor-bearing nude mouse model, and the results of the histological analysis indicated that PTR-SeNPs did not significantly damage the major organs, including the liver, spleen, heart, kidneys and lungs. Moreover, the induction of caspase activation and mitochondrial dysfunction was the major anticancer action mechanism of PTR-SeNPs. Taken together, the results of this study provide a simple approach for the facile and large-scale manufacturing of SeNPs with reduced toxicity and enhanced anticancer activity through the regulation of the surface properties of SeNPs. Furthermore, this study generates evidence for the future exploration and translational application of these materials through oral administration in nanomedicine and nutritional sciences.


Micro- and macro-level correlates of adiposity in children.
Recently, studies using a social ecological perspective have identified important micro- and macro-level risk factors for excessive adiposity in youth. Although considerable research exists examining these relationships, few studies have applied a socioecological approach to simultaneously examine both micro- and macro-level factors in young children while objectively assessing adiposity via dual-energy x-ray absorptiometry (DXA). To examine race and sex differences in adiposity measured by DXA in a large sample of young children and to identify both micro- and macro-level correlates of adiposity. Cross-sectional. Elementary school children (N = 495) from the southeastern United States participated. Anthropometrics, percentage body fat via DXA, and psychosocial variables via questionnaire were assessed in the Fall of 2003. Community-level sociodemographic data and built-environment variables via geographic information system were collected in Spring 2009. Data analyses were completed in the Spring of 2010. Percentage body fat in white children was higher than in nonwhite children. Higher percentage body fat and poorer cardiovascular fitness were found in females compared with males. Percentage body fat was higher in children who had lower athletic competence and lived in neighborhoods with higher percentages of minority residents. This study provides preliminary support for the social-ecological model to explain variance in adiposity in children. Developers of health promotion programs for children living in minority neighborhoods should consider factors at multiple levels of the ecological model when designing and implementing programs.


Inhaled steroid/tobacco smoke particle interactions: a new light on steroid resistance.
Inhaled steroid resistance is an obstacle to asthma control in asthmatic smokers. The reasons of this phenomenon are not yet entirely understood. Interaction of drug particles with environmental tobacco smoke (ETS) could change the aerodynamic profile of the drug through the particle coagulation phenomenon. Aim of the present study was to examine whether steroid particles interact with smoke when delivered in the presence of ETS. Beclomethasone-hydrofluoralkane (BDP-HFA) pMDI particle profile was studied after a single actuation delivered in ambient air or in the presence of ETS in an experimental chamber using a light scattering Optical Particle Counter capable of measuring the concentrations of particle sized 0.3-1.0, 1.1-2.0, 2.1-3.0, 3.1-4.0, 4.1-5.0, and > 5.1 microm in diameter with a sampling time of one second. The number of drug particles delivered after a single actuation was measured as the difference between total particle number after drug delivery and background particle number. Two groups of experiments were carried out at different ambient background particle concentrations. Two-tail Student's t-test was used for statistical analysis. When delivered in ambient air, over 90% of BDP-HFA particles were found in the 0.3-1.0 microm size class, while particles sized 1.1-2.0 microm and 2.1-3.0 represented less than 6.6% and 2.8% of total particles, respectively. However, when delivered in the presence of ETS, drug particle profile was modified, with an impressive decrease of 0.3-1.0 microm particles, the most represented particles resulting those sized 1.1-2.0 microm (over 66.6% of total particles), and 2.1-3.0 microm particles accounting up to 31% of total particles. Our data suggest that particle interaction between inhaled BDP-HFA pMDI and ETS takes place in the first few seconds after drug delivery, with a decrease in smaller particles and a concurrent increase of larger particles. The resulting changes in aerosol particle profile might modify regional drug deposition with potential detriment to drug efficacy, and represent a new element of steroid resistance in smokers. Although the present study does not provide any functional or clinical assessment, it might be useful to advise smokers and non smokers with obstructive lung disease such as asthma or COPD, to avoid to act inhaled drugs in the presence of ETS in order to obtain the best therapeutic effect.


Inflammation and Arterial Stiffness in Chronic Kidney Disease: Findings From the CRIC Study.
Chronic kidney disease (CKD) and arterial stiffness are associated with increased cardiovascular morbidity and mortality. Inflammation is proposed to have a role in the development of arterial stiffness, and CKD is recognized as a proinflammatory state. Arterial stiffness is increased in CKD, and cross-sectional data has suggested a link between increased inflammatory markers in CKD and higher measures of arterial stiffness. However, no large scale investigations have examined the impact of inflammation on the progression of arterial stiffness in CKD. We performed baseline assessments of 5 inflammatory markers in 3,939 participants from the chronic renal insufficiency cohort (CRIC), along with serial measurements of arterial stiffness at 0, 2, and 4 years of follow-up. A total of 2,933 participants completed each of the follow-up stiffness measures. In cross-sectional analysis at enrollment, significant associations with at least 2 measures of stiffness were observed for fibrinogen, interleukin-6, high-sensitivity C-reactive protein, proteinuria, and composite inflammation score after adjustment for confounders. In longitudinal analyses, there were few meaningful correlations between baseline levels of inflammation and changes in metrics of arterial stiffness over time. In a large cohort of CKD participants, we observed multiple significant correlations between initial markers of inflammation and metrics of arterial stiffness, but baseline inflammation did not predict changes in arterial stiffness over time. While well-described biologic mechanisms provide the basis for our understanding of the cross-sectional results, continued efforts to design longitudinal studies are necessary to fully elucidate the relationship between chronic inflammation and arterial stiffening.


Calcium and phosphate effects on growth and alkaloid production in Coffea arabica: experimental results and mathematical model.
Plant, mammalian, and microbial cells are commonly immobilized in calcium alginate gels for the production of valuable secondary metabolites. However, calcium ions are known to inhibit growth in various types of cells, and calcium is an integral part of such gels. Therefore, an investigation was conducted to evaluate the effect of calcium on the growth and alkaloid production of a model cell-line, Coffea arabica, in suspension culture before, attempting to immobilize such cells in alginate. A kinetic model was then developed from the results to describe cell growth and alkaloid production and the mechanism by which calcium influences these variables. In addition, it was observed that there was a characteristic relationship between the concentration of calcium in the external medium and the concentration of extra cellular and intracellular phosphate. The intracellular phosphate level was, in turn, related to the production of alkaloids. Using these results, a dynamic mathematical model of cell growth and alkaloid production was developed based on the proposed roles of calcium and phosphate. The model showed satisfactory agreement with three sets of experiments at different calcium concentrations. A possible linkage between the calcium and phosphate results is postulated based on the limited solubility of calcium phosphate.


Revision of the Nasal Dorsum.
Revision of the dorsum in secondary rhinoplasty is challenging, regardless of the cause. Dorsal deformities should be evaluated for both aesthetic and functional. The authors briefly outline the presentation, etiology, pathogenesis, and treatment choices backed by evidence-based data, when applicable, for the following dorsal revision indications in secondary rhinoplasty: inverted-V and midvault narrowing, overresected dorsum, irregular dorsum, saddle nose, and pollybeak.


Evolutionary dynamics of HIV-1 and the control of AIDS.
Human immunodeficiency viruses (HIV) have exhibited an extraordinary capacity for genetic change, exploring new evolutionary space after each transmission to a new host. This presents a great challenge to the prevention and management of HIV-1 infection. At the same time, the relentless diversification of HIV-1, developing as it does under the constraints imposed by the human immune system and other selective forces, contains within it information useful for understanding HIV epidemiology and pathogenesis. Comparing the sheer mutational potential of HIV with actual data representing viral lineages that can survive selection suggests that HIV does not have unlimited capacity for change. Rather, clinical and bioinformatic data suggest that, even in the most diverse gene of the most highly variable organism, natural selection places severe limits on the portion of amino acid sequence space that ensures viability. This suggests some optimism for those attempting to identify sets of antigens that can generate effective humoral and cellular immune responses against HIV.


Effect of intraocular pressure on uveoscleral outflow following cyclodialysis in the monkey eye.
Cyclodialysis was performed in both eyes of five cynomolgus monkeys. Two days later the intraocular pressure (IOP) had fallen from 17.7 +/- 0.8 to 7.1 +/- 1.4 mm Hg (P less than 0.001). At that time, both eyes were perfused for 30 min with fluorescein-isothiocyanate (FITC) dextran (MW 70,000), one at 35 mm Hg and the other at 4 mm Hg. Four pairs of control eyes (without cyclodialysis) were perfused in the same manner. At 4 mm Hg, uveoscleral outflow was 0.02 +/- 0.02 microliter/min in control eyes and 0.05 +/- 0.04 microliter/min in eyes following cyclodialysis. However, at 35 mm Hg, uveoscleral outflow in eyes with cyclodialysis increased to 2.13 +/- 0.47 microliters/min compared to 0.32 +/- 0.10 microliter/min in control eyes. Thus the "facility" of uveoscleral outflow in control eyes is 0.01 microliter/min/mm Hg and in eyes following cyclodialysis is 0.07 microliter/min/mm Hg. It is concluded that cyclodialysis results in a pressure-dependent increase in uveoscleral outflow.


Sphingomimetic multiple sclerosis drug FTY720 activates vesicular synaptobrevin and augments neuroendocrine secretion.
Neurotransmission and secretion of hormones involve a sequence of protein/lipid interactions with lipid turnover impacting on vesicle trafficking and ultimately fusion of secretory vesicles with the plasma membrane. We previously demonstrated that sphingosine, a sphingolipid metabolite, promotes formation of the SNARE complex required for membrane fusion and also increases the rate of exocytosis in isolated nerve terminals, neuromuscular junctions, neuroendocrine cells and in hippocampal neurons. Recently a fungi-derived sphingosine homologue, FTY720, has been approved for treatment of multiple sclerosis. In its non-phosphorylated form FTY720 accumulates in the central nervous system, reaching high levels which could affect neuronal function. Considering close structural similarity of sphingosine and FTY720 we investigated whether FTY720 has an effect on regulated exocytosis. Our data demonstrate that FTY720 can activate vesicular synaptobrevin for SNARE complex formation and enhance exocytosis in neuroendocrine cells and neurons.


The effect of hyperthermia and melphalan on survival of human fibroblast strains and melanoma cell lines.
The survival of four human fibroblast strains and seven malignant melanoma cell lines was determined by the colony formation method following 4-hr treatment with various concentrations of melphalan, with or without simultaneous exposure to hyperthermia (42 degrees). The two amelanotic melanoma lines (MM127 and MM253) were 10 times more sensitive to melphalan at 36 degrees than were the four fibroblast strains, the five pigmented melanoma lines being of intermediate sensitivity. Sensitivity to melphalan was usually accompanied by sensitivity to heat, while combined treatment was not only synergistic in most lines but increased the differential between fibroblasts and melanoma cells. Survival studies carried out at 36 degrees, 40 degrees, 42 degrees, and 44 degrees, using human fetal lung fibroblasts and MM253 cells, showed that 42 degrees gave the greatest differential effect and allowed reasonable survival of the normal cells. Time-survival comparison of the same two lines demonstrated that there was no advantage in prolonging hyperthermia unless melphalan was used. A more convenient method for determination of survival was developed based on thymidine uptake of colonies grown in Linbro wells.


Protein Farnesyltransferase Catalyzes Unanticipated Farnesylation and Geranylgeranylation of Shortened Target Sequences.
Protein prenylation is a posttranslational modification involving the attachment of a C15 or C20 isoprenoid group to a cysteine residue near the C-terminus of the target substrate by protein farnesyltransferase (FTase) or protein geranylgeranyltransferase type I (GGTase-I), respectively. Both of these protein prenyltransferases recognize a C-terminal "CaaX" sequence in their protein substrates, but recent studies in yeast- and mammalian-based systems have demonstrated FTase can also accept sequences that diverge in length from the canonical four-amino acid motif, such as the recently reported five-amino acid C(x)3X motif. In this work, we further expand the substrate scope of FTase by demonstrating sequence-dependent farnesylation of shorter three-amino acid "Cxx" C-terminal sequences using both genetic and biochemical assays. Strikingly, biochemical assays utilizing purified mammalian FTase and Cxx substrates reveal prenyl donor promiscuity leading to both farnesylation and geranylgeranylation of these sequences. These findings expand the substrate pool of sequences that can be potentially prenylated, further refine our understanding of substrate recognition by FTase and GGTase-I, and suggest the possibility of a new class of prenylated proteins within proteomes.


Evaluation and development of deep neural networks for image super-resolution in optical microscopy.
Deep neural networks have enabled astonishing transformations from low-resolution (LR) to super-resolved images. However, whether, and under what imaging conditions, such deep-learning models outperform super-resolution (SR) microscopy is poorly explored. Here, using multimodality structured illumination microscopy (SIM), we first provide an extensive dataset of LR-SR image pairs and evaluate the deep-learning SR models in terms of structural complexity, signal-to-noise ratio and upscaling factor. Second, we devise the deep Fourier channel attention network (DFCAN), which leverages the frequency content difference across distinct features to learn precise hierarchical representations of high-frequency information about diverse biological structures. Third, we show that DFCAN's Fourier domain focalization enables robust reconstruction of SIM images under low signal-to-noise ratio conditions. We demonstrate that DFCAN achieves comparable image quality to SIM over a tenfold longer duration in multicolor live-cell imaging experiments, which reveal the detailed structures of mitochondrial cristae and nucleoids and the interaction dynamics of organelles and cytoskeleton.


Brief communication: Additional cases of maxillary canine-first premolar transposition in several prehistoric skeletal assemblages from the Santa Barbara Channel Islands of California.
This article identifies and discusses seven new cases of complete maxillary canine-premolar transposition in ancient populations from the Santa Barbara Channel region of California. A high frequency of this tooth transposition has been previously documented within a single prehistoric cemetery on one of the Channel Islands. A total of 966 crania representing 30 local sites and about 7,000 years of human occupation were examined, revealing an abnormally high prevalence of this transposition trait among islanders during the Early period of southern California prehistory ( approximately 5500-600 B.C.). One of the affected crania is from a cemetery more than 7,000-years-old and constitutes the earliest case of tooth transposition in humans so far reported. The results are consistent with findings by other studies that have indicated inbreeding among the early Channel Islands groups. Together with the normal transposition rates among mainland populations, the decreasing prevalence of maxillary canine-first premolar transposition among island populations across the Holocene suggests that inbreeding on the northern Channel Islands had all but ceased by the end of the first millennium B.C., most likely as a result of increased cross-channel migration and interaction.


Flow cytometric analysis of estrogen receptor expression in isolated nuclei and cells from mammary cancer tissues.
Cellular expression of receptors for the hormones estrogen and progesterone in human mammary tumors is of diagnostic and prognostic value. Ligand binding assays have been replaced by immunohistochemical analysis of receptor expression. However, both of these techniques are slow, and in the ligand-binding assay it is difficult to measure heterogeneity of receptor expression in individual cells. Flow cytometry has been used extensively for monitoring the expression of cellular receptors in hematopoietic tumors but has been of limited value in the analysis of mammary tumors, which are difficult to disaggregate into single cells for flow analysis. Hormone receptors have a predominant nuclear localization, and it is relatively easy to isolate nuclei from paraffin-embedded archival tissues for flow cytometric analysis of receptor expression. Thick sections from formalin-fixed paraffin-embedded archival mammary tumors were digested by different enzyme solutions for the isolation of single nuclei. Different fixatives were used to compare the results on subsequent staining of the nuclei for estrogen receptor (ER) expression. Double staining with propidium iodide and fluorescein isothiocyanate labeled secondary antibodies for ER expression was used for multiparametric analysis of ER and DNA content. Digestion of paraffin sections with low concentration of pepsin and detergents was ideal for isolation of single nuclei. Fixation with paraformaldehyde in the presence of Triton X-100 improved staining of the cells. Isolated nuclei had enhanced immunoreactivity compared with the whole cells, and subpopulations differing in reactivity could be identified in the nuclear fractions. Double staining of nuclei for ER expression and DNA content could allow for multiparametric analysis of these two important parameters. The procedures described can be used for processing of archival paraffin-embedded mammary tumors for monitoring of ER expression and aneuploidy. These two parameters have important diagnostic and prognostic significance in mammary tumors. Laser flow cytometry by providing multiparametric analysis can allow for correlation of these cellular markers with other important cellular and clinical parameters.


Vaccinating a billion people against COVID-19: India's quest for systems leadership in exceptional times.
In January 2021, India launched the world's most extensive vaccination campaign against COVID-19. It is estimated that India would need to vaccinate over a billion people to achieve herd immunity. Even though the Indian Government focuses on improving and delivering its vaccination programme, significant challenges still exist. This paper aims to discuss current challenges to scale up India's vaccination campaign and addresses strategies for achieving this. The paper is based on a review of secondary sources, including journal articles from scholarly and grey literature and information available in the public domain. The search focused explicitly on the COVID-19 scenario, vaccination programme, public health management and systems leadership in the Indian health care system. The analysis revealed that various factors have disrupted India's vaccination campaign, including shortage of vaccine doses, mandatory prior online registration, lack of infrastructure, safety concerns for older people, untrained workforce and absence of a solid public health framework. Furthermore, India appears to have struggled to reduce tensions and instill trust in its ability to effectively manage the COVID-19 pandemic and vaccination programme due to a lack of cooperation between union government, state governments and other stakeholders, namely, policymakers, hospitals, industry and community. The findings indicate that scaling up India's anti-COVID vaccination programme would require system-level leadership strategies that work within the country's limited resources. Deeper reforms in vaccine development, storage, delivery, training and regulatory frameworks are also needed to extend the world's largest anti-COVID-19 vaccination campaign.


[In vitro observation of ciliary activity of the tracheal membrane under a common light microscope].
To establish a new method for in vitro observation of ciliary activity of the tracheal membrane in rabbit under a common light microscope. Nine healthy adult rabbits were used. Two equal sized cervical trachea flaps were removed after the rabbits were anesthetized by urethane (4 mg/kg). The removed trachea flap was randomly assigned to the ephedrine (Eph) and DMEM control group, respectively. The observed trachea flap was placed in a small flat-bottomed glass container with its membranous side upward in DMEM culture medium solution (DMEM control group) or in 0.5% ephedrine solution prepared with DMEM (Eph group). One drop of 1% methylthioninium stained autologous blood cells was added into the glass container as the tracer, and the trachea flap was observed under a common light microscope (400 x). The latter was attached with a digital camera linking to an image manipulation system, the computed dynamic image analyser. The velocity of the tracer cell movement worked as the indicator for ciliary activity and was automatically determined by the digital image manipulation system. The variation coefficient (VC) was used as the indicator of the cell movement velocity. There was no significant difference among the VC at different time points in the DMEM control group. VC of the Eph group decreased regularly with the time point. A thin layer of flowing fluid was found on the surface of the tracheal mucociliary blanket which is driven by the activity of the mucociliary system. The new method of using the tracer to evaluate the ciliary activity of mammalia tracheal membrane in vitro is reliable and stable. It is practical and valuable in the in vitro observation and evaluation of ciliary activity of the tracheal membrane.


Synthesis of health promotion concepts in children's palliative care.
Palliative care improves the health of children with a life-limiting condition and appears to draw implicitly on concepts shared with a model of health promotion. However, to date there has been no scrutiny about how this relationship may shape understanding about children's palliative care. To explore the influence of health promoting concepts on children's palliative care models, policies and guidelines. Health and social care databases were searched for policies, models and guidelines published between 2000-2018. Additional searches of professional national and international healthcare websites, children's palliative care charities and UK and Ireland government websites were conducted. A best fit framework synthesis was used. A total of 55 policies and guidelines were reviewed for the framework synthesis. Eight themes were generated: (1) health promoting children's palliative care policy and guidelines; (2) planning ahead; (3) creating a supportive environment; (4) enabling coping and independence; (5) reorienting children's palliative care sectors; (6) the lengthening trajectory of need for support; (7) strengthening community engagement in children's palliative care; and (8) quality of life and value-based ideologies. The best fit framework synthesis confirmed a conceptual relationship between children's palliative care and health promotion. This is captured in a new model that will extend professionals' understanding.


[The characteristics of the prevalence of vertebrogenic lumbosacral radiculalgias among cotton and tobacco growers in Uzbekistan].
An epidemiological study of the prevalence of vertebrogenic lumbosacral radiculalgias (VLSR) was made among 910 cotton growers and 311 tobacco growers of Samarkand Province of Uzbekistan. Out of the 1221 persons aged 18 to 70, VLSR were revealed in 223. The incidence of the disease among cotton growers was 6.3 and among tobacco growers, it amounted to 7.1 per 100 workers. A definite correlation was established between the disease and the character of work and specificity of the working postures: a long stay in a bent position aggravated by the pressure of the apron strap weighing 8-10 kg on the lumbar part of the spine. A correlation was also established between the incidence of VLSR and the length of service.


Changes in the microflora of man during long-term confinement.
Extended space missions may change the human normal microflora, including the intestinal flora. The bacterial composition of the intestinal microflora is an important factor in natural resistance to infection. During the year-long medico-engineering experiment human intestinal microflora was thoroughly studied. It was necessary to collect more detailed information concerning the composition in healthy people. 83 microbiological tests of faeces of 27 healthy male subjects were carried out. It was found that spore-less anaerobic bacteria predominated in the fecal microflora, reaching 90% of total micro-organisms. Aerobic micro-organisms were less than 6% of total micro-organisms. Long-term enclosure of men was shown to induce changes in the intestinal microflora which involved a sharp reduction of different microbial groups, and occasionally complete elimination of certain representatives of the intestinal flora. The composition tended to be simplified. A study of the isolated cultures of Cl. perfringens demonstrated an increased amount of strains with toxicogenic properties. Staphylococci were isolated from the test subjects during their long-term enclosure. The pharynx and nose of the three test subjects were examined and 1445 staphylococcal strains were isolated, of which 745 strains (51.5%) were coagulase producers. In 745 strains phage-type, coagulase, toxicity, hemolysins, lecitinase, fibrinolysin and DNA-ase were determined. Phage-typing of the isolated staphylococci revealed a microbial exchange between the men. Staphylococcal strains of the same phage-type (29/52) isolated during the experiment showed an increase of the toxin titre (from 1:40-1:80 to 1:640-1:1280) and development of additional pathogenic properties that were not found previously. An increase in beta-hemolysin and fibrinolysin production and an acceleration of DNA-ase activity. Experiments on mice demonstrated increased virulence of the staphylococcal strains isolated by the end of the experiment. The data are discussed from the point of view of the immunological responses of the human body.


Association of proliferative activity and size in acoustic neuroma: implications for timing of surgery.
Acoustic neuroma is the most frequent benign tumor of the cerebellopontine angle, and surgery is still the most common form of treatment. To gain better insight into the dysregulated mechanisms causing growth of acoustic neuroma, the authors studied the proliferative activity of 34 consecutive samples by analyzing immunohistochemical staining with Ki-67 and proliferating cell nuclear antigen (PCNA), and apoptosis based on the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling. Data from these analyses were correlated with clinical parameters (that is, tumor size, duration of symptoms, and patient age). Apoptotic cells were found in none of the tumors. Proliferation measured on staining with Ki-67 and PCNA correlated with tumor size, but not with patient age or duration of symptoms. The authors demonstrated that tumors 18 mm or smaller in diameter have lower proliferation indices and growth rates, compared with tumors larger than 18 mm with high proliferative indices and growth rates. Additionally, they observed that these more aggressive, larger tumors occur mostly in patients younger than 50 years of age. Patients with tumors larger than 18 mm in diameter and who are younger than 50 years of age sustain an enhanced risk for fast-growing tumors because of these lesions' enhanced proliferative activity. For these patients the authors recommend active therapy.


Chronic administration of losartan reverses cardiovascular changes in hypertensive fructose-fed rats.
The cluster of risk factors including hyperinsulinemia, insulin resistance, hypertriglyceridemia and hypertension has been called syndrome X. Several evidences link the insulin resistance syndrome with endothelial dysfunction. Since the participation of the renin-angiotensin system (RAS) in this pathology is still unclear, the present study examined the effect of chronic administration of an angiotensin AT1 receptor antagonist, losartan (L), on endothelial nitric oxide synthase (eNOS) activity in aortic endothelium and cardiac tissue, and on the proliferation of primary cultured aortic smooth muscle cells (SMC), obtained from fructose-fed rats (FFR), an experimental model of syndrome X Male Wistar rats were used: Control, FFR and FFR+L (n = 8 in each group). After 8 weeks, tissue samples were obtained and 10% fetal calf serum (FCS) proliferative effect was examined in SMC by 3H-thymidine incorporation and cell counting. The eNOS activity was estimated in aortic endothelial lining and cardiac homogenates by conversion of 3H-arginine into 3H-citrulline. FFR aortic SMC showed a significantly increased 10% FCS-induced 3H-thymidine incorporation and cell number compared to controls. FFR aortic and cardiac eNOS activities were significantly decreased. Chronic treatment with L decreased systolic blood pressure,reverted cardiac hypertrophy, abolished the increased SMC proliferation and restoredeNOS activity. These data confirm that changes in SMC proliferation and endothelial dysfunction at different levels of the cardiovascular system are involved in syndrome "X", and that AT1 receptor blocking can revert those changes, suggesting an important role of the RAS, possibly mediated by AT2 receptors and kinins, in the physiopathological mechanisms of this model.


The molecular determinants of neurosteroid binding in the GABA(A) receptor.
Neurosteroids positively modulate GABA-A receptor (GABAAR) channel activity by binding to a transmembrane domain intersubunit site. Understanding the interactions in this site that determine neurosteroid binding and its effect is essential for the design of neurosteroid-based therapeutics. Using photo-affinity labeling and an ELIC-α1GABAAR chimera, we investigated the impact of mutations (Q242L, Q242W and W246L) within the intersubunit site on neurosteroid binding. These mutations, which abolish the thermal stabilizing effect of allopregnanolone on the chimera, reduce neither photolabeling within the intersubunit site nor competitive prevention of labeling by allopregnanolone. Instead, these mutations change the orientation of neurosteroid photolabeling. Molecular docking of allopregnanolone in WT and Q242W receptors confirms that the mutation favors re-orientation of allopregnanolone within the binding pocket. Collectively, the data indicate that mutations at Gln242 or Trp246 that eliminate neurosteroid effects do not eliminate neurosteroid binding within the intersubunit site, but significantly alter the preferred orientation of the neurosteroid within the site. The interactions formed by Gln242 and Trp246 within this pocket play a vital role in determining the orientation of the neurosteroid that may be necessary for its functional effect.


[Primary structure of gene H of cattle plague virus strain K].
Synthesis, cDNA cloning, and identification of H gene nucleotide sequence of rinderpest virus (RPV) K strain are carried out. Analysis of the identified nucleotide sequence has revealed the single open reading frame encoding a protein consisting of 609 amino acids with molecular weight of 68 kDa. The mean nucleotide homology between H genes of K, Kabete O and L strains in 88.0%, the mean amino acid homology of the corresponding proteins is 88.2%. RPV K strain hemagglutinin contains 5 potential glycosylation sites. The position of all 13 cystein bases is identical to positions in H proteins of RPV Kabete O and L strains. Studies of the hydrophobic profile of the compared proteins have shown 2 potential transmembrane fragments.


Studies on the degradation pathway of iron-sulfur centers during unfolding of a hyperstable ferredoxin: cluster dissociation, iron release and protein stability.
The ferredoxin from the thermoacidophile Acidianus ambivalens is a representative of the archaeal family of di-cluster [3Fe-4S][4Fe-4S] ferredoxins. Previous studies have shown that these ferredoxins are intrinsically very stable and led to the suggestion that upon protein unfolding the iron-sulfur clusters degraded via linear three-iron sulfur center species, with 610 and 520 nm absorption bands, resembling those observed in purple aconitase. In this work, a kinetic and spectroscopic investigation on the alkaline chemical denaturation of the protein was performed in an attempt to elucidate the degradation pathway of the iron-sulfur centers in respect to protein unfolding events. For this purpose we investigated cluster dissociation, iron release and protein unfolding by complementary biophysical techniques. We found that shortly after initial protein unfolding, iron release proceeds monophasically at a rate comparable to that of cluster degradation, and that no typical EPR features of linear three-iron sulfur centers are observed. Further, it was observed that EDTA prevents formation of the transient bands and that sulfide significantly enhances its intensity and lifetime, even after protein unfolding. Altogether, our data suggest that iron sulfides, which are formed from the release of iron and sulfide resulting from cluster degradation during protein unfolding in alkaline conditions, are in fact responsible for the observed intermediate spectral species, thus disproving the hypothesis suggesting the presence of a linear three-iron center intermediate. Kinetic studies monitored by visible, fluorescence and UV second-derivative spectroscopies have elicited that upon initial perturbation of the tertiary structure the iron-sulfur centers start decomposing and that the presence of EDTA accelerates the process. Also, the presence of EDTA lowers the observed melting temperature in thermal ramp experiments and the midpoint denaturant concentration in equilibrium chemical unfolding experiments, further suggesting that the clusters also play a structural role in the maintenance of the conformation of the folded state.


[Pleiotrophic effect of statins (3-hydroxy-3-methylglutaryl-coenzyme a reductase inhibitors)].
The development of statins improved the therapy of hypercholesterolemia and atherosclerotic disease tremendously. The beneficial effects of statins were clearly demonstrated in large scale primary and secondary prevention studies. In addition to the reduction of plasma cholesterol, inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase also results in the depletion of intermediates of cholesterol biosynthesis that are important for cellular integrity. The so called pleiotrophic effects of statins and probably also their adverse events can be attributed to the inhibition of synthesis of these intermediates. The review article describes the pathogenesis of atherosclerosis, the pharmacokinetic and pharmacodynamik of statins, and their pleiotrophic effects concerning endothelial function, LDL (low density lipoprotein) oxidation, macrophages, smooth muscle cell proliferation, atherosclerotic plaque, platelets, thrombosis, proinflammatory factors, haemorheology, hypertension, venous thrombosis, bone metabolism, stroke, and the possible influence on the prevention of Alzheimer's disease.


Pump-controlled directional light emission from random lasers.
The angular emission pattern of a random laser is typically very irregular and difficult to tune. Here we show by detailed numerical calculations that one can overcome the lack of control over this emission pattern by actively shaping the spatial pump distribution. We demonstrate, in particular, how to obtain customized pump profiles to achieve highly directional emission. Going beyond the regime of strongly scattering media where localized modes with a given directionality can simply be selected by the pump, we present an optimization-based approach which shapes extended lasing modes in the weakly scattering regime according to any predetermined emission pattern.


Influenza A/subtype and B/lineage effectiveness estimates for the 2011-2012 trivalent vaccine: cross-season and cross-lineage protection with unchanged vaccine.
We estimate vaccine effectiveness (VE) against both influenza A/subtypes and B/lineages in Canada for the 2011-2012 trivalent inactivated influenza vaccine (TIV) with components entirely unchanged from the 2010-2011 TIV and in the context of phenotypic and genotypic characterization of circulating viruses. In a test-negative case-control study VE was estimated as [1-(adjusted)OddsRatio] × 100 for RT-PCR-confirmed influenza in vaccinated vs nonvaccinated participants. Viruses were characterized by hemagglutination inhibition (HI) and sequencing of antigenic sites of the hemagglutinin (HA) gene. There were 1507 participants. VE against A(H1N1)pdm09 was 80% (95% confidence interval [CI], 52%-92%): circulating viruses were HI-characterized as vaccine-matched and bore just 2 aminoacid (AA) differences from vaccine. VE against A/H3N2 was 51% (95% CI, 10%-73%): circulating viruses were HI-characterized as vaccine-related but bore ≥11AA differences from vaccine. VE against influenza B was 51% (95% CI, 26%-67%) in total: 71% (95% CI, 40%-86%) for lineage-matched B/Victoria and 27% (95% CI, -21% to 56%) for lineage-mismatched B/Yamagata. For both influenza A and B types, VE was similar among recipients of either 2010-2011 or 2011-2012 TIV alone, higher when vaccinated both seasons. Phenotypic and genotypic characterization of circulating and vaccine viruses enhances understanding of TIV performance, shown in 2011-2012 to be substantial against well-conserved A(H1N1)pdm09 and lineage-matched influenza B, suboptimal against genetic-variants of A/H3N2, and further reduced against lineage-mismatched influenza B. With unchanged vaccine components, protection may extend beyond a single season.


Anterior debridement and fusion followed by posterior pedicle screw fixation in pyogenic spondylodiscitis: autologous iliac bone strut versus cage.
An anterior approach for debridement and fusion with autologous bone graft has been recommended as the gold standard for surgical treatment of pyogenic spondylodiscitis. The use of anterior foreign body implants at the site of active infection is still a challenging procedure for spine surgeons. Several authors have recently introduced anterior grafting with titanium mesh cages instead of autologous bone strut in the treatment of spondylodiscitis. The authors present their experience of anterior fusion with 3 types of cages followed by posterior pedicle screw fixation. They also compare their results with the use of autologous iliac bone strut. The authors retrospectively reviewed the cases of 60 patients with pyogenic spondylodiscitis treated by anterior debridement between January 2003 and April 2005. Fusion using either cages or iliac bone struts was performed during the same course of anesthesia followed by posterior fixation. Twenty-three patients underwent fusion with autologous iliac bone strut, and 37 patients underwent fusion with 1 of the 3 types of cages. The infections resolved in all patients, as noted by normalization of their erythrocyte sedimentation rates and C-reactive protein levels. Patients in both groups were evaluated in terms of their preoperative and postoperative clinical and imaging findings. Single-stage anterior debridement and cage fusion followed by posterior pedicle screw fixation can be effective in the treatment of pyogenic spondylodiscitis. There was no difference in clinical and imaging outcomes between the strut group and cage group except for the subsidence rate. The subsidence rate was higher in the strut group than in the cage group. The duration until subsidence was also shorter in the strut group than in the cage group.


Pediatric indications for deep brain stimulation.
Based on the success of deep brain stimulation (DBS) in the treatment of adult disorders, it is reasonable to assume that the application of DBS in the pediatric population is an emerging area worthy of study. The purpose of this paper is to outline the current movement disorder indications for DBS in the pediatric population, and to describe areas of investigation, including possible medically refractory psychiatric indications. We performed a structured review of the English language literature from 1990 to 2011 related to studies of DBS in pediatrics using Medline and PubMed search results. Twenty-four reports of DBS in the pediatric population were found. Based on published data on the use of DBS for pediatric indications, there is a spectrum of clinical evidence for the use of DBS to treat different disorders. Dystonia, a disease associated with a low rate of remission and significant disability, is routinely treated with DBS and is currently the most promising pediatric application of DBS. We caution the application of DBS to conditions associated with a high remission rate later in adulthood, like obsessive-compulsive disorder and Tourette's syndrome. Moreover, epilepsy and obesity are currently being investigated as indications for DBS in the adult population; however, both are associated with significant morbidity in pediatrics. While currently dystonia is the most promising application of DBS in the pediatric population, multiple conditions currently being investigated in adults also afflict children and adolescents, and thus warrant further research.


Virulence of Meloidogyne incognita to expression of N gene in pepper.
Four pepper genotypes classified as resistant and four pepper genotypes classified as susceptible to several avirulent populations of M. incognita were compared for their reactions against a population of Meloidogyne incognita (Chitwood) Kofoid and White which had been shown to be virulent to resistant bell pepper (Capsicum annuum) in preliminary tests. The virulent population of M. incognita originated from a commercial bell pepper field in California. The resistant pepper genotypes used in all experiments were the Capsicum annuum cultivars Charleston Belle, Carolina Wonder, and Carolina Cayenne, and the C. chinense cultigen PA-426. The susceptible pepper genotypes used in the experiments were the C. annuum cultivars Keystone Resistant Giant, Yolo Wonder B, California Wonder, and the C. chinense cultigen PA-350. Root gall indices (GI) were ≥ 3.0 for all genotypes in both tests except for PA-426 (GI=2.57) in test 1 and 'Carolina Cayenne' (GI=2.83) in test 2. Numbers of eggs per gram fresh root weight ranged from 20,635 to 141,319 and reproductive indices ranged from 1.20 to 27.2 for the pepper genotypes in both tests, indicating that all eight pepper genotypes tested were susceptible to the M. incognita population used in these tests. The M. incognita population used in these studies overcame resistance conferred by the N gene in all resistant genotypes of both C. annuum and C. chinense.


Lipid peroxidation in electroporated hepatocytes occurs much more readily than does hydroxyl-radical formation.
1. Rat hepatocytes suspended in 0.25 M-sucrose were electropermeabilized. This completely disrupted their plasma-membrane permeability barrier. 2. The endoplasmic reticulum in electroporated hepatocytes appeared morphologically preserved and maintained its permeability barrier as evidenced by electron-microscopic examination and latency measurements on luminal reticular enzymes. 3. Upon aerobic incubation with an NADPH-generating system and iron/ADP, porated hepatocytes peroxidized their membrane lipids at rates similar to those of matched microsomal preparations. 4. When hepatocytes were incubated with iron/EDTA and azide, radical formation detectable with dimethyl sulphoxide (DMSO) was only 10-20% that shown by microsomes. Omitting azide abolished hepatocyte reactivity with DMSO completely. Effects of hydroxyl-radical (.OH) scavengers and of added catalase suggest that the radical detected by DMSO is .OH. 5. Cytosolic inhibitor(s) from hepatocytes seemed to be a major factor limiting .OH formation. These were macromolecular, but showed a degree of heat-stability. Dialysis largely abolished inhibition, but this could be restored again by adding GSH. 6. Since .OH formation in hepatocytes seems to be much more stringently prevented than lipid peroxidation, free-radical damage originating from intracellular redox systems seems more likely to take the form of lipid peroxidation.


Reducing the amount of blood transfused: a systematic review of behavioral interventions to change physicians' transfusion practices.
Transfusion services have used various techniques to reduce blood product utilization. Given the potential adverse effects of transfusions and the resources consumed in implementing strategies to reduce transfusions, there is a need to understand their effectiveness. Therefore, we performed a systematic review of the literature to examine the effectiveness of behavioral interventions to reduce blood product utilization. We identified all relevant articles through the use of electronic searches of MEDLINE and EMBASE, as well as hand searches of review articles and personal files. The electronic searches included articles published between January 1966 and May 2003. The searches included the terms blood transfusion, plasma exchange, guidelines, education, practice patterns, and professional practice. The outcomes of interest were the number of units transfused and the proportion of patients who received transfusions. Nineteen studies examining the effectiveness of single (guidelines, prospective audits, retrospective audits, and reminders) or multifaceted interventions in reducing red blood cell, platelet, plasma, cryoprecipitate, and albumin transfusions met the inclusion criteria. Eighteen studies demonstrated a relative reduction in the number of units given (range, 9%-77%) or the proportion of patients receiving transfusions (range, 17%-79%). The reported reductions were qualitatively similar for the different blood products studied. No particular intervention or combination of interventions appeared more effective in reducing utilization. Behavioral interventions, including simple interventions, appear to be effective in changing physician transfusion practices and reducing blood utilization. Appropriately designed clinical trials are still needed to determine the relative effectiveness of different interventions to change practices.


Pemphigus Vulgaris and Foliaceus IgG Autoantibodies Directly Block Heterophilic Transinteraction between Desmoglein and Desmocollin.
Anti-desmoglein (Dsg) 1 and Dsg3 IgG autoantibodies in pemphigus foliaceus and pemphigus vulgaris cause blisters through loss of desmosomal adhesion. It is controversial whether blister formation is due to direct inhibition of Dsg, intracellular signaling events causing desmosome destabilization, or both. Recent studies show that heterophilic binding between Dsg and desmocollin (Dsc) is the fundamental adhesive unit of desmosomes. To eliminate cellular contributions to potential pathogenicity of pemphigus antibodies, bead assays coated with recombinant Dsg1, Dsc1, Dsg3, or Dsc3 ectodomains were developed. A mixture of Dsg beads and Dsc beads formed large aggregates, confirming that the heterophilic binding is dominant. The pathogenic anti-Dsg1 and anti-Dsg3 mAbs, which bind the transadhesive interface, blocked the aggregation of Dsg1/Dsc1 and Dsg3/Dsc3 beads, respectively, whereas nonpathogenic mAbs did not. All sera tested from eight patients with pemphigus foliaceus and eight patients with mucosal pemphigus vulgaris with active disease inhibited the adhesion of Dsg1/Dsc1 and Dsg3/Dsc3 beads, respectively. When paired sera obtained from seven patients with pemphigus foliaceus and six patients with pemphigus vulgaris in active disease and remission were compared, the former inhibited aggregation better than the latter. These findings strongly suggest that steric hindrance of heterophilic transinteraction between Dsg and Dsc is important for disease pathology in both pemphigus foliaceus and pemphigus vulgaris.


Evaluation of a high-pathogenicity H5N1 avian influenza A virus isolated from duck meat.
The introduction of an influenza A virus possessing a novel hemagglutinin (HA) into an immunologically naive human population has the potential to cause severe disease and death. Such was the case in 1997 in Hong Kong, where H5N1 influenza was transmitted to humans from infected poultry. Because H5N1 viruses are still isolated from domestic poultry in southern China, there needs to be continued surveillance of poultry and characterization of virus subtypes and variants. This study provides molecular characterization and evaluation of pathogenesis of a recent H5N1 virus isolated from duck meat that had been imported to South Korea from China. The HA gene of A/Duck/Anyang/AVL-1/01 (H5N1) isolate was found to be closely related to the Hong Kong/97 H5N1 viruses. This virus also contained multiple basic amino acids adjacent to the cleavage site between HA1 and HA2, characteristic of high-pathogenicity avian influenza viruses (HPAI). The pathogenesis of this virus was characterized in chickens, ducks, and mice. The DK/Anyang/AVL-1/01 isolate replicated well in all species and resulted in 100% and 22% lethality for chickens and mice, respectively. No clinical signs of disease were observed in DK/Anyang/AVL-1/01-inoculated ducks, but high titers of infectious virus could be detected in multiple tissues and oropharyngeal swabs. The presence of an H5N1 influenza virus in ducks bearing a HA gene that is highly similar to those of the pathogenic 1997 human/poultry H5N1 viruses raises the possibility of reintroduction of HPAI to chickens and humans.


[Properties of phosphorylase kinase activated by subtilisin].
The activation of phosphorylase kinase during limited proteolysis by subtilisin was studied. It was shown that phosphorylase kinase undergoes rapid activation and its activity remains unchanged throughout a prolonged incubation. Electrophoresis in the presence of Na-SDS revealed a rapid decomposition of the alpha-subunit and a gradual disappearance of the beta-subunit; the protein molecule was shown to be composed of the degradation products of alpha- and beta-subunits with different molecular weights and unchanged proteolysis of the gamma-subunit. The phosphorylase kinase hydrolysate was separated using chromatography on a cellulose phosphate column. The active protein fraction contains a new form of phosphorylase kinase with a low molecular weight (approximately 80 000) which is insensitive to Ca2+. The subtilisin-activated phosphorylase kinase does not affect the activity of phosphodiesterase from cyclic nucleotides.


Factors Governing Selectivity of Dopamine Receptor Binding Compounds for D2R and D3R Subtypes.
Targeting the D3 dopamine receptor (D3R) is a promising pharmacotherapeutic strategy for the treatment of many disorders. The structure of the D3R is similar to the D2 dopamine receptor (D2R), especially in the transmembrane spanning regions that form the orthosteric binding site, making it difficult to identify D3R selective pharmacotherapeutic agents. Here, we examine the molecular basis for the high affinity D3R binding and D3R vs D2R binding selectivity of substituted phenylpiperazine thiopheneamides. We show that removing the thiophenearylamide portion of the ligand consistently decreases the affinity of these ligands at D3R, while not affecting their affinity at the D2R. Our long (>10 μs) molecular dynamics simulations demonstrated that both dopamine receptor subtypes adopt two major conformations that we refer to as closed or open conformations, with D3R sampling the open conformation more frequently than D2R. The binding of ligands with conjoined orthosteric-allosteric binding moieties causes the closed conformation to populate more often in the trajectories. Also, significant differences were observed in the extracellular loops (ECL) of these two receptor subtypes leading to the identification of several residues that contribute differently to the ligand binding for the two receptors that could potentially contribute to ligand binding selectivity. Our observations also suggest that the displacement of ordered water in the binding pocket of D3R contributes to the affinity of the compounds containing an allosteric binding motif. These studies provide a better understanding of how a bitopic mode of engagement can determine ligands that bind selectively to D2 and D3 dopamine receptor subtypes.


Maculopapular skin eruptions associated with Covid-19: A systematic review.
In this systematic review, we anticipated in summarizing clinical features, histopathological hallmarks, and possible pathology behind the maculopapular skin eruptions occurring in Covid-19 patients. A literature search was executed using MEDLINE/PubMed and Embase databases for articles published till 20 November 2020. All eligible articles including observational studies, case reports, and case series reporting the maculopapular skin lesion in Covid-19 patients were included. Data were obtained for 354 Covid-19 patients presenting with maculopapular lesions from 40 studies. The mean age of these patients was 53 years, and with 42% of them being male. These maculopapular lesions differed considerably in terms of distribution and appearance, ranging from diffuse erythematous maculopapular lesions to scattered erythematous macules coalescing into papules to maculopapular lesions in plaques. The mean duration of the lesion was 8 days. These lesions were frequently localized on trunks and extremities. Superficial perivascular dermatitis with lymphocytic infiltrate was a histopathological hallmark of these lesions. As these skin lesions may have a possible association with diagnosis, management, prognosis, and severity of the disease, all health practitioners need to be well acquainted with these Covid-19 skin lesions. Also, in the middle of this worldwide pandemic, early identification of this eruption may help manage this infection's further spread.