Evolution of C2H2-zinc finger genes and subfamilies in mammals: species-specific duplication and loss of clusters, genes and effector domains.
C2H2 zinc finger genes (C2H2-ZNF) constitute the largest class of transcription factors in humans and one of the largest gene families in mammals. Often arranged in clusters in the genome, these genes are thought to have undergone a massive expansion in vertebrates, primarily by tandem duplication. However, this view is based on limited datasets restricted to a single chromosome or a specific subset of genes belonging to the large KRAB domain-containing C2H2-ZNF subfamily. Here, we present the first comprehensive study of the evolution of the C2H2-ZNF family in mammals. We assembled the complete repertoire of human C2H2-ZNF genes (718 in total), about 70% of which are organized into 81 clusters across all chromosomes. Based on an analysis of their N-terminal effector domains, we identified two new C2H2-ZNF subfamilies encoding genes with a SET or a HOMEO domain. We searched for the syntenic counterparts of the human clusters in other mammals for which complete gene data are available: chimpanzee, mouse, rat and dog. Cross-species comparisons show a large variation in the numbers of C2H2-ZNF genes within homologous mammalian clusters, suggesting differential patterns of evolution. Phylogenetic analysis of selected clusters reveals that the disparity in C2H2-ZNF gene repertoires across mammals not only originates from differential gene duplication but also from gene loss. Further, we discovered variations among orthologs in the number of zinc finger motifs and association of the effector domains, the latter often undergoing sequence degeneration. Combined with phylogenetic studies, physical maps and an analysis of the exon-intron organization of genes from the SCAN and KRAB domains-containing subfamilies, this result suggests that the SCAN subfamily emerged first, followed by the SCAN-KRAB and finally by the KRAB subfamily. Our results are in agreement with the "birth and death hypothesis" for the evolution of C2H2-ZNF genes, but also show that this hypothesis alone cannot explain the considerable evolutionary variation within the subfamilies of these genes in mammals. We, therefore, propose a new model involving the interdependent evolution of C2H2-ZNF gene subfamilies.


Primary structure of the 5 S subunit of transcarboxylase as deduced from the genomic DNA sequence.
Transcarboxylase from Propionibacterium shermanii is a complex biotin-containing enzyme composed of 30 polypeptides of three different types. It is composed of six dimeric outer subunits associated with a central cylindrical hexameric subunit through 12 biotinyl subunits; three outer subunits on each face of the central hexamer. Each outer dimer is termed a 5 S subunit which associates with two biotinyl subunits. The enzyme catalyzes a two-step reaction in which methylmalonyl-CoA and pyruvate form propionyl-CoA and oxalacetate, the 5 S subunit specifically catalyzing one of these reactions. We report here the cloning, sequencing and expression of the monomer of the 5 S subunit. The gene was identified by matching amino acid sequences derived from isolated authentic 5 S peptides with the deduced sequence of an open reading frame present on a cloned P. shermanii genomic fragment known to contain the gene encoding the 1.3 S biotinyl subunit. The cloned 5 S gene encodes a protein of 519 amino acids, M(r) 57,793. The deduced sequence shows regions of extensive homology with that of pyruvate carboxylase and oxalacetate decarboxylase, two enzymes which catalyze the same or reverse reaction. A fragment was subcloned into pUC19 in an orientation such that the 5 S open reading frame could be expressed from the lac promoter of the vector. Crude extracts prepared from these cells contained an immunoreactive band on Western blots which co-migrated with authentic 5 S and were fully active in catalyzing the 5 S partial reaction. We conclude that we have cloned, sequenced and expressed the monomer of the 5 S subunit and that the expressed product is catalytically active.


Genetic variation in a metabolic signaling pathway and colon and rectal cancer risk: mTOR, PTEN, STK11, RPKAA1, PRKAG2, TSC1, TSC2, PI3K and Akt1.
Serine/threonine protein kinase 11 (STK11) and phosphatase tensin homolog deleted on chromosome 10 (PTEN) link insulin sensitivity and metabolic signaling to inflammation and other hormonal factors and colorectal cancer. We evaluate genetic variation in nine genes in a candidate pathway as follows: STK11 (3 tagSNPs), PTEN (9 tagSNPs), FRAP1 (mTOR) (4 tagSNPs), TSC1 (14 tagSNPs), TSC2 (8 tagSNPs), Akt1 (2 tagSNPs), PIK3CA (7 tagSNPs), PRKAA1 (13 tagSNPs) and PRKAG2 (68 tagSNPs) in two population-based case-control studies of colon (n = 1574 cases, 1940 controls) and rectal (n = 91 cases, 999 controls) cancer. FRAP1, PRKAA1, PRKAG2 and TSC2 genes were significantly associated with colon cancer; risk estimates ranged from 1.21 [95% confidence interval (CI) 1.05-1.38] for FRAP1rs1057079 for the AG/GG genotype to 1.51 (95% CI 1.09-2.09) for PRKAG2rs9648723 CC genotype. PIK3CA, PRKAG2, PTEN, STK11 and TSC1 were significantly associated with rectal cancer overall. The strongest association was observed for PIK3CA rs7651265 GG genotype (odds ratio 2.32 95% CI 1.02-5.30). FRAP1 was associated with microsatellite instability (MSI)+ colon tumors; PRKAA1, CpG island methylator phenotype (CIMP)+ and MSI+ colon tumors; PRKAG2 and KRAS2 colon tumors; TSC1 and CIMP+ and MSI+ colon tumors; TSC2 with MSI+ colon tumors; PIK3CA with KRAS2-mutated rectal tumors; PRKAG2 (rs6964824) with KRAS2- and TP53-mutated rectal tumors and with PRKAG2 (rs412396 and rs4725431) with CIMP+ rectal tumors. These data suggest that genetic variation in a predefined candidate pathway for colorectal cancer contributes to both colon and rectal cancer risk. Associations appear to be strongest for CIMP+ and MSI+ tumors.


Rested and stressed farmed Atlantic cod (Gadus morhua) chilled in ice or slurry and effects on quality.
The main objectives of this study were to investigate (1) whether rested harvest of farmed cod was better maintained by chilling with slurry rather than by traditional ice storage, (2) whether chilling with slurry would be a feasible chilling method to assure low core temperatures (≤0 °C) at packing of gutted fish, and (3) the effects of superchilling compared with traditional ice on selected quality parameters of cod during storage. In the experiment, seawater slurry at -2.0 ± 0.3 °C was used. Anesthetized (AQUI-S™), percussion stunned, and stressed cod chilled in slurry were compared. Cod stored on ice were used as reference group. The fish were evaluated at the day of slaughter, and after 7 and 14 d of storage according to handling stress (initial muscle pH, muscle twitches, rigor mortis), core temperatures, quality index method, microbial counts, weight changes, salt and water content, water distribution, pH, adenosine triphosphate-degradation products, K-value, water-holding capacity, fillet color, and texture. Chilling cod in slurry was more rapid than chilling in ice. Prechilling (1 d) of cod in slurry before subsequent ice storage resulted in lower quality 7 d postmortem compared with both ice and continuous slurry storage. The potential advantages of superchilling became more prominent after 14 d with lower microbiological activity, better maintenance of freshness (lower total quality index scores and lower K-values) compared with fish stored on ice. A drawback with slurry-stored fish was that cloudy eyes developed earlier, in addition to weight gain and salt uptake compared to ice-stored fish. Practical Application: Chilling is an essential operation in any fish-processing plant. This manuscript addresses different applications of slurry ice in the processing and storage of Atlantic cod. Cod quality was assessed after 7 and 14 d of iced and superchilled storage.


COVID-19 Pathogenesis: From Molecular Pathway to Vaccine Administration.
The Coronavirus 2 (SARS-CoV-2) infection is a global pandemic that has affected millions of people worldwide. The advent of vaccines has permitted some restitution. Aside from the respiratory complications of the infection, there is also a thrombotic risk attributed to both the disease and the vaccine. There are no reliable data for the risk of thromboembolism in SARS-CoV-2 infection in patients managed out of the hospital setting. A literature review was performed to identify the pathophysiological mechanism of thrombosis from the SARS-CoV-2 infection including the role of Angiotensin-Converting Enzyme receptors. The impact of the vaccine and likely mechanisms of thrombosis following vaccination were also clarified. Finally, the utility of the vaccines available against the multiple variants is also highlighted. The systemic response to SARS-CoV-2 infection is still relatively poorly understood, but several risk factors have been identified. The roll-out of the vaccines worldwide has also allowed the lifting of lockdown measures and a reduction in the spread of the disease. The experience of the SARS-CoV-2 infection, however, has highlighted the crucial role of epidemiological research and the need for ongoing studies within this field.


Cochlear reimplantation.
A small number of multichannel cochlear implant (CI) recipients require reimplantation. This study describes the causes of failure, surgical technique, and hearing outcomes in a consecutive series of 16 patients undergoing reimplantation of multichannel devices. We hypothesize that reimplantation is safe and that hearing results are at least as good as those measured following primary implantation. Retrospective analysis of consecutive clinical series. Chart analysis of 191 consecutive CI operations performed at the University of Miami Ear Institute between 1990 and 1997 revealed 16 patients who received a second multichannel device. All but one had a minimum follow-up of 1 year after reimplantation. Ten of these patients had initial implantation performed by us, and six were initially operated on elsewhere. Main outcomes of the initial procedure were compared with those of the reimplantation, including electrode insertion length, number of channels programmed, and audiometric results. In addition, cause of failure and relevant surgical findings at the second procedure are described. There were no surgical complications after reimplantation surgery. Device failure was the most frequent cause for reimplantation. Time between initial implantation and failure ranged from 0 to 46 months (mean, 22.4 mo; median, 23 mo). Common intraoperative findings include mastoid fibrosis, bone growth at the cochleostomy, and skin flap breakdown. Following reimplantation, mean length of insertion, number of channels actively programmed, and speech recognition scores were at least as good as findings before initial implant failure. CI reimplantation is safe and effective.


[How idiopathic is idiopathic scoliosis? Results of neurological studies with somatosensory evoked potentials (SSEP) in children and adolescents].
About 90% of all the scolioses are called "idiopathic". Various neurological diseases (for example poliomyelitis, etc.) are frequently accompanied by deformities of the spine. The so-called somatosensory evoked potentials are at our disposal being a very sensitive and a non-problematic neurological diagnostic tool. Formation of the question: Are there clinically non-detectable neurological changes demonstrable by use of evoked potentials in children with so-called idiopathic scoliosis? 45 non-operated patients suffering from scoliosis and 21 healthy children were examined clinically, neurologically, and by use of evoked potentials. 26/45 children with idiopathis scoliosis showed pathological evoked potentials (right-left-side-difference concerning latency and amplitudes of the potentials and generally delayed transmission from peripheral nerves to the somatosensory cortex), mostly concerning the lower extremities. 19/45 children showed normal evoked potentials (EP). There was no correlation between EP and direction respectively degree of the scoliosis. Neurological affections mostly located caudally of the cervical spine are to be discussed (for example protrusion of the intervertebral disc, dysrhaphic processes etc.).


Isolation and characterization of human antigen-specific TCR alpha beta+ CD4(-)CD8- double-negative regulatory T cells.
Down-regulation of immune responses by regulatory T (Treg) cells is an important mechanism involved in the induction of tolerance to allo-antigens (Ags). Recently, a novel subset of Ag-specific T-cell receptor (TCR)alpha beta+ CD4(-)CD8- (double-negative [DN]) Treg cells has been found to be able to prevent the rejection of skin and heart allografts by specifically inhibiting the function of antigraft-specific CD8+ T cells. Here we demonstrate that peripheral DN Treg cells are present in humans, where they constitute about 1% of total CD3+ T cells, and consist of both naive and Ag-experienced cells. Similar to murine DN Treg cells, human DN Treg cells are able to acquire peptide-HLA-A2 complexes from antigen-presenting cells by cell contact-dependent mechanisms. Furthermore, such acquired peptide-HLA complexes appear to be functionally active, in that CD8+ T cells specific for the HLA-A2-restricted self-peptide, Melan-A, became sensitive to apoptosis by neighboring DN T cells after acquisition of Melan-A-HLA-A2 complexes and revealed a reduced proliferative response. These results demonstrate for the first time that a sizable population of peripheral DN Treg cells, which are able to suppress Ag-specific T cells, exists in humans. DN Treg cells may serve to limit clonal expansion of allo-Ag-specific T cells after transplantation.


A Japanese retrospective study of non-tuberculous mycobacterial infection in children, adolescents, and young adult patients with hematologic-oncologic diseases.
Non-tuberculous mycobacterial infection (NTM) is rare in healthy children, with lymphadenitis being the most common presentation. Immunocompromised populations are known to be at high risk, but the clinical picture of NTM infection in pediatric hematology/oncology patients is unclear. In this nationwide retrospective analysis of patients under the age of 40 treated in Japanese pediatric hematology/oncology departments who developed NTM infection between January 2010 and December 2020, 36 patients (21 patients with hematopoietic stem cell transplantation (HSCT) and 15 nontransplant patients) were identified. Post-transplant patients were infected with NTM at 24 sites, including the lungs (n = 12), skin and soft tissues (n = 6), bloodstream (n = 4), and others (n = 2). Nine of twelve patients with pulmonary NTM infection had a history of pulmonary graft-versus-host disease (GVHD), and rapid-growing mycobacteria (RGM) were isolated from five of them. In nontransplant patients, the primary diseases were acute lymphoblastic leukemia (ALL; n = 5), inborn errors of immunity (IEI; n = 6), and others (n = 4). All cases of ALL had bloodstream infections with RGM, whereas all cases of IEI were infected with slow-growing mycobacteria (SGM). In summary, three typical clinical scenarios for pediatric hematology/oncology patients have been established: RGM-induced pulmonary disease in patients with pulmonary GVHD, RGM bloodstream infection in patients with ALL, and SGM infection in patients with IEI. Our findings suggest that NTM must be regarded as a pathogen for infections in these high-risk patients, especially those with pulmonary GVHD, who may require active screening for NTM.


Borderline ovarian tumors: outcomes of fertility sparing surgery.
Borderline ovarian tumors(BOT) account for ten to 20 percent of all epitelial ovarian carcinomas and often occur in reproductive ages. The aim of this study was to evaluate the clinical and reproductive outcomes of patients who were diagnosed with BOT and underwent fertility sparing surgery. Patients younger than 40 years who underwent fertility sparing surgery for BOT from 2004 to 2012 were reviewed retrospectively and were evaluated according to the reproductive and clinical outcomes. Twenty-eight patients younger than 40 years with BOT underwent fertility sparing surgery. Median follow up time was 42 +/- 28.1 months. During the follow up period, two patients (7.1%) developed recurrence at 35 and 36 months, respectively. Five (17.9%) out of 28 patients became pregnant during the follow up period. Fertility sparing surgery should be the first choice for the treatment of BOT in patients who wish to preserve fertility.


A comparative study based on physical characteristics of suitable packing materials in biofiltration.
In the present work, 10 packing materials commonly used as support media in biofiltration are analysed and compared to evaluate their suitability according to physical characteristics. The nature of the packing material in biofilters is an important factor for the success in their construction and operation. Different packing materials have been used in biofiltration without a global agreement about which ones are the most adequate for biofiltration success. The materials studied were chosen according to previous works in the field of biofiltration including both organic and inorganic (or synthetic) materials. A set of nine different parameters were selected to cope with well-established factors, such as a material-specific surface area, pressure drop, nutrient supply, water retentivity, sorption capacity, and purchase cost. One ranking of packing materials was established for each parameter studied in order to define a relative suitability degree. Since biofiltration success generally depends on a combination of the ranked parameters, a procedure was defined to compare packing materials suitability under common situations in biofiltration. The selected scenarios, such as biofiltration of intermittent loads of pollutants and biofiltration of waste gases with low relative humidity, were investigated. The results indicate that, out of the packing materials studied, activated carbons were ranked top of several parameter rankings and were shown to be a significantly better packing material when parameters were combined to assess such selected scenarios.


Targeted and tailored diagnostic strategies in women with perimenopausal bleeding: advantages of the sonohysterographic approach.
Diagnosis and treatment of endometrial pathology nowadays ranges from clinical examination to transvaginal ultrasound (TVS), saline infusion sonohysterography (SIS), hysterosalpingography (HSG) and hysteroscopy (HYS). However, many gynaecologists prescribe blind endometrial biopsies, such as Vacuum ABRAsion (VABRA), as single strategy. The purpose of this work is to evaluate whether the procedure of VABRA should still be performed alone in perimenopausal women with abnormal uterine bleeding, compared to biopsies and samples obtained after a previous transvaginal sonohysterography. We retrospectively reviewed the records of 216 patients referred to our Ultrasonography and Day-Surgery Center between November 2005 and December 2006 with persistent premenopausal uterine bleeding, spotting or postmenopausal bleeding. One hundred and five out of 216 pts.(48.6%), defined as Group "A", underwent a sole endometrial sampling by VABRA; 111 out of 216 pts. (51.4%), Group "B", had a SIS first. Vabra showed a poor sensitivity in the diagnosis of polyps (19%) and submucosal myoma, with a negative predictive value of 73.4%. Likelihood ratio for test negative was 0.81, with an overall diagnostic accuracy of 75%. This study confirms that blind endometrial biopsies should no longer be performed as the only diagnostic strategy in perimenopausal women with abnormal uterine bleeding. On the other hand, a sonohysteroscopy-guided approach allows an accurate detection of focal lesions; nevertheless, it should not be forgotten that SIS is an ultrasound based procedure, and may provide further information on endometrial thickness, myomas, ovaries and pelvis.


Lysosomal proteolysis is the primary degradation pathway for cytosolic ferritin and cytosolic ferritin degradation is necessary for iron exit.
Cytosolic ferritins sequester and store iron, consequently protecting cells against iron-mediated free radical damage. However, the mechanisms of iron exit from the ferritin cage and reutilization are largely unknown. In a previous study, we found that mitochondrial ferritin (MtFt) expression led to a decrease in cytosolic ferritin. Here we showed that treatment with inhibitors of lysosomal proteases largely blocked cytosolic ferritin loss in both MtFt-expressing and wild-type cells. Moreover, cytosolic ferritin in cells treated with inhibitors of lysosomal proteases was found to store more iron than did cytosolic ferritins in untreated cells. The prevention of cytosolic ferritin degradation in MtFt-expressing cells significantly blocked iron mobilization from the protein cage induced by MtFt expression. These studies also showed that blockage of cytosolic ferritin loss by leupeptin resulted in decreased cytosolic ferritin synthesis and prolonged cytosolic ferritin stability, potentially resulting in diminished iron availability. Lastly, we found that proteasomes were responsible for cytosolic ferritin degradation in cells pretreated with ferric ammonium citrate. Thus, the current studies suggest that cytosolic ferritin degradation precedes the release of iron in MtFt-expressing cells; that MtFt-induced cytosolic ferritin decrease is partially preventable by lysosomal protease inhibitors; and that both lysosomal and proteasomal pathways may be involved in cytosolic ferritin degradation.


Chronic alcohol ingestion in rats alters lung metabolism, promotes lipid accumulation, and impairs alveolar macrophage functions.
Chronic alcoholism impairs pulmonary immune homeostasis and predisposes to inflammatory lung diseases, including infectious pneumonia and acute respiratory distress syndrome. Although alcoholism has been shown to alter hepatic metabolism, leading to lipid accumulation, hepatitis, and, eventually, cirrhosis, the effects of alcohol on pulmonary metabolism remain largely unknown. Because both the lung and the liver actively engage in lipid synthesis, we hypothesized that chronic alcoholism would impair pulmonary metabolic homeostasis in ways similar to its effects in the liver. We reasoned that perturbations in lipid metabolism might contribute to the impaired pulmonary immunity observed in people who chronically consume alcohol. We studied the metabolic consequences of chronic alcohol consumption in rat lungs in vivo and in alveolar epithelial type II cells and alveolar macrophages (AMs) in vitro. We found that chronic alcohol ingestion significantly alters lung metabolic homeostasis, inhibiting AMP-activated protein kinase, increasing lipid synthesis, and suppressing the expression of genes essential to metabolizing fatty acids (FAs). Furthermore, we show that these metabolic alterations promoted a lung phenotype that is reminiscent of alcoholic fatty liver and is characterized by marked accumulation of triglycerides and free FAs within distal airspaces, AMs, and, to a lesser extent, alveolar epithelial type II cells. We provide evidence that the metabolic alterations in alcohol-exposed rats are mechanistically linked to immune impairments in the alcoholic lung: the elevations in FAs alter AM phenotypes and suppress both phagocytic functions and agonist-induced inflammatory responses. In summary, our work demonstrates that chronic alcohol ingestion impairs lung metabolic homeostasis and promotes pulmonary immune dysfunction. These findings suggest that therapies aimed at reversing alcohol-related metabolic alterations might be effective for preventing and/or treating alcohol-related pulmonary disorders.


Anticipatory activity of motor cortex neurons in relation to direction of an intended movement.
1. Monkeys were trained to 1) hold a handle in a central zone midway between "push" and "pull" while awaiting 2) an instruction telling them how to respond to a subsequent 3) perturbation, which triggered the instructed movement and was followed by 4) a reward if the movement was correct. 2. There were two sorts of instructions: push and pull. When the pull instruction had preceded the perturbation, the monkey responded to the perturbation by pulling, whereas after a push instruction, the monkey responded to the perturbation by pushing. 3. Recordings in pre- and postcentral sensorimotor cortex revealed instruction-induced changes of neuronal activity during the period intervening between the instruction and the perturbation-triggered movement. Effects of the instruction were differential depending on which of the two instructions was given, such differential responses to the instruction being detected in 61% of precentral pyramidal tract neurons (PTNs), 44% of precentral non-PTNs, and 11% of postcentral neurons. 4. Since motor cortex PTN axons end on alpha and gamma motoneurons and on interneurons of the spinal cord, changes of PTN activity with "intention" or "motor set" provide a mechanism for suprasegmental control and presetting of spinal cord reflex excitability specific to the nature of an impending movement.


Lysosomal Cholesterol Hydrolysis Couples Efferocytosis to Anti-Inflammatory Oxysterol Production.
Macrophages face a substantial amount of cholesterol after the ingestion of apoptotic cells, and the LIPA (lysosomal acid lipase) has a major role in hydrolyzing cholesteryl esters in the endocytic compartment. Here, we directly investigated the role of LIPA-mediated clearance of apoptotic cells both in vitro and in vivo. We show that LIPA inhibition causes a defective efferocytic response because of impaired generation of 25-hydroxycholesterol and 27-hydroxycholesterol. Reduced synthesis of 25-hydroxycholesterol after LIPA inhibition contributed to defective mitochondria-associated membrane leading to mitochondrial oxidative stress-induced NLRP3 (NOD-like receptor family, pyrin domain containing) inflammasome activation and caspase-1-dependent Rac1 (Ras-related C3 botulinum toxin substrate 1) degradation. A secondary event consisting of failure to appropriately activate liver X receptor-mediated pathways led to mitigation of cholesterol efflux and apoptotic cell clearance. In mice, LIPA inhibition caused defective clearance of apoptotic lymphocytes and stressed erythrocytes by hepatic and splenic macrophages, culminating in splenomegaly and splenic iron accumulation under hypercholesterolemia. Our findings position lysosomal cholesterol hydrolysis as a critical process that prevents metabolic inflammation by enabling efficient macrophage apoptotic cell clearance.


Striking a balance: regulation of transposable elements by Zfp281 and Mll2 in mouse embryonic stem cells.
Transposable elements (TEs) compose about 40% of the murine genome. Retrotransposition of active TEs such as LINE-1 (L1) tremendously impacts genetic diversification and genome stability. Therefore, transcription and transposition activities of retrotransposons are tightly controlled. Here, we show that the Krüppel-like zinc finger protein Zfp281 directly binds and suppresses a subset of retrotransposons, including the active young L1 repeat elements, in mouse embryonic stem (ES) cells. In addition, we find that Zfp281-regulated L1s are highly enriched for 5-hydroxymethylcytosine (5hmC) and H3K4me3. The COMPASS-like H3K4 methyltransferase Mll2 is the major H3K4me3 methylase at the Zfp281-regulated L1s and required for their proper expression. Our studies also reveal that Zfp281 functions partially through recruiting the L1 regulators DNA hydroxymethylase Tet1 and Sin3A, and restricting Mll2 at these active L1s, leading to their balanced expression. In summary, our data indicate an instrumental role of Zfp281 in suppressing the young active L1s in mouse ES cells.


A Novel Topical PPARγ Agonist Induces PPARγ Activity in Ulcerative Colitis Mucosa and Prevents and Reverses Inflammation in Induced Colitis Models.
Peroxisome proliferator-activated receptor-gamma (PPARγ) exerts anti-inflammatory effects and is therefore a potential target in ulcerative colitis (UC). A novel PPARγ agonist (AS002) developed for local action was evaluated ex vivo in biopsies from UC patients and in vivo in mice with low-grade dextran sodium sulfate (DSS)- and trinitrobenzene sulfonic acid (TNBS)-induced colitis. Colonic biopsies from UC patients (n = 18) and healthy controls (n = 6) were incubated with AS002 or rosiglitazone (positive control) to measure mRNA expression of the PPARγ-responsive gene ADIPOPHILIN and protein levels of UC-related cytokines (enzyme-linked immunosorbent assay). AS002 absorption was determined in the colonic mucosa of UC patients. DSS-colitis mice received PPARγ agonists or vehicle daily by intrarectal administration starting 2 days before induction of colitis (preventive) or from days 3 to 8 (curative). Myeloperoxidase (MPO) and cytokine levels in colonic mucosa were determined. In addition, AS002 effects were studied in TNBS colitis. AS002 displayed an absorption pattern of a lipophilic drug totally metabolized in the mucosa. AS002 and rosiglitazone increased ADIPOPHILIN mRNA expression (3-fold) and decreased TNF-α, IL-1β, and IL-13 levels in human UC biopsies. In DSS, in both preventive and curative treatment and in TNBS colitis, AS002 protected against macroscopic and histological damage and lowered MPO and TNF-α, IL-1β, and IL-13 levels. AS002 triggers anti-inflammatory PPARγ activity in the human colonic mucosa of UC patients and prevents and reverses colitis in mice. Our data suggest that AS002 has potential for topical maintenance treatment of UC, which warrants further studies in vivo in patients.


Genetic control of organ-reactive autoantibody production in mice by obesity (ob) diabetes (db) genes.
Inbred strains of mice exhibited genetic and sex-dependent differences in spontaneous production of organ-reactive autoantibodies detected by indirect immunofluorescence. Antitestis autoreactivity was found primarily in sera from C57BL/6J (B6) mice, whereas antigastric autoreactivity was common to both CBA/J and 129/J strains. Autoantibodies against islet cell cytoplasmic antigens (ICAs) were uniquely expressed by C57BL/KsJ (BKs) males. Introduction of the diabetes (db) mutation into these various inbred-strain backgrounds induced expression of ICA, with stronger induction observed in males. The stress imposed by the db or obesity (ob) mutation induced ICA in BKs mice at a higher frequency than in B6 mice; this differential sensitivity was somehow related to a gene linked to the H-2 complex because BKs.B6 H-2b congenic mice resembled B6 mice. The db3J mutation increased the expression of these autoantibodies in 129/J mice, which, like B6, were H-2b and therefore presumably possessed the same H-2-linked inducibility allele as BKs. Cytotoxic autoantibodies against islet cell surface antigens were only observed in C3HeB/FeJ db/db males, and their presence was correlated with beta-cell necrosis. It is concluded that db and/or ob genes appear to play an important role in the production of autoantibodies to islet cells, and sex-linked factor(s) may modify the phenotypic expression of the autoantibodies.


The Selective Nav1.7 Inhibitor, PF-05089771, Interacts Equivalently with Fast and Slow Inactivated Nav1.7 Channels.
Voltage-gated sodium (Nav) channel inhibitors are used clinically as analgesics and local anesthetics. However, the absence of Nav channel isoform selectivity of current treatment options can result in adverse cardiac and central nervous system side effects, limiting their therapeutic utility. Human hereditary gain- or loss-of-pain disorders have demonstrated an essential role of Nav1.7 sodium channels in the sensation of pain, thus making this channel an attractive target for new pain therapies. We previously identified a novel, state-dependent human Nav1.7 selective inhibitor (PF-05089771, IC50 = 11 nM) that interacts with the voltage-sensor domain (VSD) of domain IV. We further characterized the state-dependent interaction of PF-05089771 by systematically varying the voltage, frequency, and duration of conditioning prepulses to provide access to closed, open, and fast- or slow-inactivated states. The current study demonstrates that PF-05089771 exhibits a slow onset of block that is depolarization and concentration dependent, with a similarly slow recovery from block. Furthermore, the onset of block by PF-05089771 develops with similar rates using protocols that bias channels into predominantly fast- or slow-inactivated states, suggesting that channel inhibition is less dependent on the availability of a particular inactivated state than the relative time that the channel is depolarized. Taken together, the inhibitory profile of PF-05089771 suggests that a conformational change in the domain IV VSD after depolarization is necessary and sufficient to reveal a high-affinity binding site with which PF-05089771 interacts, stabilizing the channel in a nonconducting conformation from which recovery is slow.


[A case of occupational pulmonary siderosis: the pathogenetic and prognostic considerations].
Pulmonary siderosis is a disease which can be considered with the pneumoconioses of the so-called inert dust (either by deposit or by accumulation) and, from the earliest nosographic descriptions dating back to the thirties and forties, it has been substantially considered a benign form because of the scarce sclerogenic evolution and the preservation of discrete respiratory function. We present a case of pulmonary siderosis characterized on one hand by the importance and clarity of the cytopathological picture and on the other by the presence of changes, although modest, in lung function which confirm the most recent hypotheses in the literature on the potential evolution of pneumosiderotic diseases and their relatively less benign prognosis. From a preventive point of view the case draws attention to the need for greater control of environmental hygiene conditions in small factories. From an insurance and medico-legal point of view siderosis has recently been explicitly added to the occupational diseases which can be declared to the Italian state department dealing with occupational disease and disability payments (I.N.A.I.L.) (DPR of the 13.04.1994, section 47).


Tumor-associated macrophages mediate resistance of EGFR-TKIs in non-small cell lung cancer: mechanisms and prospects.
Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are the first-line standard treatment for advanced non-small cell lung cancer (NSCLC) with EGFR mutation. However, resistance to EGFR-TKIs is inevitable. Currently, most studies on the mechanism of EGFR-TKIs resistance mainly focus on the spontaneous resistance phenotype of NSCLC cells. Studies have shown that the tumor microenvironment (TME) also mediates EGFR-TKIs resistance in NSCLC. Tumor-associated macrophages (TAMs), one of the central immune cells in the TME of NSCLC, play an essential role in mediating EGFR-TKIs resistance. This study aims to comprehensively review the current mechanisms underlying TAM-mediated resistance to EGFR-TKIs and discuss the potential efficacy of combining EGFR-TKIs with targeted TAMs therapy. Combining EGFR-TKIs with TAMs targeting may improve the prognosis of NSCLC with EGFR mutation to some extent.


Pseudosynapsis and decreased stringency of meiotic repair pathway choice on the hemizygous sex chromosome of Caenorhabditis elegans males.
During meiosis, accurate chromosome segregation relies on homology to mediate chromosome pairing, synapsis, and crossover recombination. Crossovers are dependent upon formation and repair of double-strand breaks (DSBs) by homologous recombination (HR). In males of many species, sex chromosomes are largely hemizygous, yet DSBs are induced along nonhomologous regions. Here we analyzed the genetic requirements for meiotic DSB repair on the completely hemizygous X chromosome of Caenorhabditis elegans males. Our data reveal that the kinetics of DSB formation, chromosome pairing, and synapsis are tightly linked in the male germ line. Moreover, DSB induction on the X is concomitant with a brief period of pseudosynapsis that may allow X sister chromatids to masquerade as homologs. Consistent with this, neither meiotic kleisins nor the SMC-5/6 complex are essential for DSB repair on the X. Furthermore, early processing of X DSBs is dependent on the CtIP/Sae2 homolog COM-1, suggesting that as with paired chromosomes, HR is the preferred pathway. In contrast, the X chromosome is refractory to feedback mechanisms that ensure crossover formation on autosomes. Surprisingly, neither RAD-54 nor BRC-2 are essential for DSB repair on the X, suggesting that unlike autosomes, the X is competent for repair in the absence of HR. When both RAD-54 and the structure-specific nuclease XPF-1 are abrogated, X DSBs persist, suggesting that single-strand annealing is engaged in the absence of HR. Our findings indicate that alteration in sister chromatid interactions and flexibility in DSB repair pathway choice accommodate hemizygosity on sex chromosomes.


Interferon-beta modulates protein synthesis in the central nervous system.
Interferon-beta (IFN-beta), acting canonically via the modulation of transcription, affects neocortical pyramidal neurons. By use of 2-D differential gel electrophoresis and subsequent mass spectrometry we identified IFN-beta regulated proteins in the central nervous system. These proteins are involved in cytoskeleton assembly, protein transport and nucleotide metabolism and, as such, serve regenerative and protective functions. Electrophysiologically, IFN-beta mediated protein synthesis is essential for part of the excitatory neuronal effects, as revealed under blockade of protein biosynthesis. This study presents novel effects of IFN-beta in the central nervous system and begins to unravel the mechanism behind the known excitability changes in neurons.


[Risk assessment of adverse environmental factors affecting the health of the Russian population by the Russian State Sanitary and Epidemiological Surveillance].
In the article there are adduced the practical aspects of realization of a concept of risk evaluation of the adverse environment factors influencing the health of the Russian population by the State Service of Sanitary and Epidemiological Surveillance. Epidemiological data on the influence of the most spread free air contaminants is summarized and a matter of monitoring food products safety is considered. There are outlined arrangements for the successful application of the evaluation of risk of the different environment factors influencing the populations health to practical activities of the State Service of Sanitary and Epidemiological Surveillance of Russia.


COVID-19 and lung cancer.
COVID-19 pandemic had affected health services around the world, also reducing the diagnosis of lung cancer. On the other hand, examination of surgical specimens in patients with lung cancer and SARS-CoV-2 gave the opportunity to evidence early histologic features related to this emerging pandemic. Different prioritization of health organizations during COVID-19 pandemic resulted in a significant decline of lung cancer screening (up to 56%), delayed diagnosis (up to 30-40%) and higher advanced stage, with some exceptions (i.e., Canada). Increased use of stereotactic radiation treatments in stage I-IIA have been noticed in better-organized health systems. Surgical specimens performed for lung cancer in patients with incipient SARS-CoV-2 permitted to appreciate early histologic findings of COVID-19 with hyperplastic pneumocytes with/without fibrin exudate, alveolar macrophages/myeloid cells, perivascular T-lymphocytic infiltrate and lack of hyaline membrane. While the COVID-19 pandemic has declined the rate of lung cancer diagnosis worldwide, some institutions have significantly limited detrimental effects. Histology related to early SARS-CoV-2 infection in surgical samples for lung cancer revealed specific histologic changes.


Enhancing orthographic knowledge helps spelling production in eight-year-old Chinese children at risk for dyslexia.
We investigated the effects of enhancing orthographic knowledge on the spelling of Chinese characters and words in 131 eight-year-old Chinese children at risk for dyslexia. The traditional approach (37 children) emphasizing memory and repeated writing was the control condition. The analytic and synthetic approach (ASA, 33 children) stressed insight into character structure. The integrated analytic and synthetic approach added to ASA self-correction and metacognitive activities (INA, 61 children). The children were first asked to write down as many words as possible associated with pictures of home, school, and community; the correctly written words formed the baseline information. The children were then instructed by their classroom teachers in six especially designed short texts and assessed in eight measurable bujian or radical tasks subserving three constructs: morpheme completion, bujian analysis and synthesis and bujian compounding. Multivariate analyses of variance showed that the children in the INA condition outperformed those in the other conditions in three of the measurable bujian tasks. A confirmatory factor analysis verified the stability of the eight tasks and their clustering into three constructs. From these results, we tentatively propose a "bujian sensitivity hypothesis" as a means of helping young Chinese children at risk for spelling disorders.


Nanofluidic digital PCR for the quantification of Norovirus for water quality assessment.
Sensitive detection of water- and foodborne enteric viruses is extremely relevant, especially due to the low concentrations in which they are found. Accurate and sensitive detection of Norovirus, the primary responsible for water- and foodborne outbreaks, is of particular importance. Quantification of Norovirus is commonly performed by quantitative RT-PCR (RT-qPCR). In recent years a new platform was developed, digital PCR, that quantifies without the need for a standard curve thus decreasing the errors associated with its utilization. The platform developed by LifeTechnologies, QuantStudio 3D Digital PCR is amongst the least studied digital platform and although it allows the direct detection of DNA targets it requires a two-step RT-PCR for the detection of RNA targets. In this work we developed a new protocol able to detect Norovirus using a one-step digital PCR reaction (RT-dPCR). The performance of the newly developed one-step digital PCR was compared to RT-qPCR for the detection of Norovirus genogroup I and genogroup II. The sensitivity of RT-dPCR was identical to that of RT-qPCR, and the quantitative data determined by both methods were not significantly different for most samples. This one-step absolute quantification approach is a useful tool to minimize the time spent currently using this particular platform to amplify viral RNA and to standardize quantification of enteric viruses in food and environmental samples. This study proved the usefulness of the newly developed RT-dPCR protocol for a sensitive and accurate detection of low-copy targets.


Integrated bioinformatics analysis reveals CDK1 and PLK1 as potential therapeutic targets of lung adenocarcinoma.
This study is to identify potential biomarkers and therapeutic targets for lung adenocarcinoma (LUAD).GSE6044 and GSE118370 raw data from the Gene Expression Omnibus database were normalized with Robust Multichip Average. After merging these two datasets, the combat function of sva packages was used to eliminate batch effects. Then, limma packages were used to filtrate differentially expressed genes. We constructed protein-protein interaction relationships using STRING database and hub genes were identified based on connectivity degrees. The cBioportal database was used to explore the alterations of the hub genes. The promoter methylation of cyclin dependent kinase 1 (CDK1) and polo-like Kinase 1 (PLK1) and their association with tumor immune infiltration in patients with LUAD were investigated using DiseaseMeth version 2.0 and TIMER databases. The Cancer Genome Atlas-LUAD dataset was used to perform gene set enrichment analysis.We identified 10 hub genes, which were upregulated in LUAD, among which 8 were successfully verified in the Cancer Genome Atlas and Oncomine databases. Kaplan-Meier analysis indicated that the expressions of CDK1 and PLK1 in LUAD patients were associated with overall survival and disease-free survival. The methylation levels in the promoter regions of these 2 genes in LUAD patients were lower than those in normal lung tissues. Their expressions in LUAD were associated with tumor stages and relative abundance of tumor infiltrating immune cells, such as B cells, CD4+ T cells, and macrophages. Moreover, cell cycle, DNA replication, homologous recombination, mismatch repair, P53 signaling pathway, and small cell lung cancer signaling were significantly enriched in CDK1 and PLK1 high expression phenotype.CDK1 and PLK1 may be used as potential biomarkers and therapeutic targets for LUAD.


Glutathione-mediated detoxification mechanisms of human placenta.
Glutathione peroxidase and GSH S-transferases play significant physiological roles in the detoxification mechanisms. The present study indicates that only selenium-dependent GSH-peroxidase I is present in human placenta. GSH-peroxidase II (non-selenium GSH-peroxidase) is totally absent from this tissue. The placental GSH-peroxidase has a relative molecular mass of 85 000 and is a tetramer of equal-size subunits. It is cyanide sensitive and its properties are similar to those of human erythrocyte GSH-peroxidase. In contrast to human liver, which has seven GSH S-transferases, the placenta has only one species of GSH S-transferase. This enzyme (pI 4.5, relative molecular mass 45 000, subunit size 22 500) has a different amino acid composition from that of liver and erythrocyte GSH S-transferases. From immunological studies, the placental GSH S-transferase appears also to have a genetic origin distinct from that of liver, erythrocyte and kidney GSH S-transferases. The placental GSH S-transferase does not express GSH-peroxidase activity. The overall substrate specificities of placental GSH S-transferase are also narrow, compared with liver enzymes. The present study indicates that, although these two GSH-linked detoxification mechanisms are operative in placenta, they appear to be less efficient than the similar mechanisms operating in liver.


Effect of carbidopa on prolactin, growth hormone and cortisol secretion in man.
Serum prolactin, growth hormone and cortisol levels were examined in normal volunteers following administration of carbidopa, a peripheral decarboxylase inhibitor. 24-hour urinary tryptamine levels dropped markedly indicating that inhibition of decarboxylase was effective. Prolactin levels rose while growth hormone and cortisol remained unchanged. Since the tuberoinfundibular dopamine nerve terminals lie outside the blood brain barrier, this study suggests that these neurons are involved in prolactin but not in growth hormone or cortisol regulation. Findings are compatible with two alternate hypotheses--either that dopamine is a physiologic prolactin inhibiting factor (PIF) or that tuberoinfundibular dopamine neurons regulate the release of PIF.


[Retinal detachment: temporary internal tamponade with silicone oil after vitrectomy. Anatomic and functional results].
Forty-seven patients with retinal detachment treated by vitrectomy and silicone oil injection benefited from semi-premature ablation of the oil: after a mean of 5 months for supple retinal detachments and 6 1/2 months for those associated with vitreoretinal proliferation. Ablation was performed after obtaining a solid chorioretinal scar by cryopexy or endodiathermy, usually combined with laser panphotocoagulation. The technique used for ablation is described. After follow up for from 9 months to 3 1/2 years (mean 18 months) two-thirds of the patients had flat retinas. Recurrences developed usually during the first month. An explanation for these relapses is proposed.


Learning Intention-Aware Policies in Deep Reinforcement Learning.
Deep reinforcement learning (DRL) provides an agent with an optimal policy so as to maximize the cumulative rewards. The policy defined in DRL mainly depends on the state, historical memory, and policy model parameters. However, we humans usually take actions according to our own intentions, such as moving fast or slow, besides the elements included in the traditional policy models. In order to make the action-choosing mechanism more similar to humans and make the agent to select actions that incorporate intentions, we propose an intention-aware policy learning method in this letter To formalize this process, we first define an intention-aware policy by incorporating the intention information into the policy model, which is learned by maximizing the cumulative rewards with the mutual information (MI) between the intention and the action. Then we derive an approximation of the MI objective that can be optimized efficiently. Finally, we demonstrate the effectiveness of the intention-aware policy in the classical MuJoCo control task and the multigoal continuous chain walking task.


Advances in Nanomaterials for Sustainable Gas Separation and Storage: Focus on Clathrate Hydrates.
ConspectusClathrate hydrates, also known as gas hydrates, are a type of inclusion compound formed in highly developed nanoporous lattice spaces created by water molecules, where gas molecules such as CO2, H2, CH4, and other low-molecular-weight liquid molecules are trapped. The nanoporous cage formed by water molecules serves as the "host", while the trapped gas or low-molecular-weight liquid molecules such as tetrahydrofuran act as "guests". Early on, clathrate hydrates drew attention as a potential replacement for conventional natural gas due to their natural gas hydrate form, which contains natural gases as guests and exists in permafrost or sea floors. Recently, based on the unique physicochemical properties of clathrate hydrates, efforts are being made to utilize synthetic clathrate hydrates in various separation processes such as post- and pre-combustion CO2 capture, H2 storage, natural gas storage and transportation, wastewater desalination, and more. While it is undeniable that clathrate hydrates are based on principles that are beneficial for the separation and storage of gas molecules, there are several challenges that must be addressed for their practical application. These challenges include (i) the limitation of gas storage capacity due to the confined size of nanoporous cages, (ii) the relatively high-pressure and low-temperature thermodynamic storage conditions typically required for clathrate hydrate formation, and (iii) slow formation kinetics and low gas hydrate conversion, which are also essential issues that need to be resolved for the meaningful implementation of clathrate hydrates. In this Account, we aim to introduce recent noteworthy research findings, including those from our research team, focusing on addressing these challenges. We explored the untapped potential of clathrate hydrates by bridging the gap between macroscopic and microscopic properties. This has led to breakthroughs in sustainable gas separation and storage applications. By revealing the hidden nature of these hydrates, we have effectively mitigated their inherent limitations, setting the stage for more feasible and efficient H2 storage solutions through the introduction of hydrogen-natural gas blends to clathrate hydrates. Additionally, we have demonstrated the tuning effect on all naturally formed hydrate structures, offering new insights into their underlying properties and macroscopic behavior. Furthermore, our research has proposed a highly efficient hydrate-based pre-combustion CO2 capture approach that leverages porous media with appropriate wettability and considers the implications of microstructure properties. This emphasizes the crucial connection between nano-structure and macroscopic properties, underscoring the significance of understanding their interplay for economic feasibility. We believe that our efforts to unveil the hidden nature of gas hydrates provide strategies to address challenges and lay the groundwork for practical applications.


[Consequences of the COVID-19 pandemic for surgical service].
The COVID-19 pandemic has a serious impact on surgical service, emergency and especially elective surgical care. Many hospitals were re-designated as COVID hospitals due to resource constraints and large number of COVID-19 patients requiring hospitalization. This led to cancellation or postponement of scheduled surgeries. In addition, restrictions in elective surgery were associated with the risk of infection in surgical patients. Various protocols and guidelines recommended non-surgical or outpatient treatment if possible. During the pandemic, postoperative morbidity and mortality in emergency surgery increased significantly. The same is true for elective surgeries in 7-8 weeks after previous coronavirus infection. The authors analyze the issues of organization, priorities for restoration of elective surgery and criteria for patient selection.


Outcomes of Parathyroidectomy in Patients with Primary Hyperparathyroidism: A Systematic Review and Meta-analysis.
Parathyroidectomy is a definitive treatment for primary hyperparathyroidism. Patients contemplating this intervention will benefit from knowledge regarding the expected outcomes and potential risks of the currently available surgical options. To appraise and summarize the available evidence regarding benefits and harms of minimally invasive parathyroidectomy (MIP) and bilateral neck exploration (BNE). A comprehensive search of multiple databases (MEDLINE, EMBASE, and Scopus) from each database's inception to September 2014 was performed. Eligible studies evaluated patients with primary hyperparathyroidism undergoing MIP or BNE. Reviewers working independently and in duplicate extracted data and assessed the risk of bias. We identified 82 observational studies and 6 randomized trials at moderate risk of bias. Most of them reported outcomes after MIP (n = 71). Using random-effects models to pool results across studies, the cure rate was 98 % (95 % CI 97-98 %, I (2) = 10 %) with BNE and 97 % (95 % CI 96-98 %, I (2) = 86 %) with MIP. Hypocalcemia occurred in 14 % (95 % CI 10-17 % I (2) = 93 %) of the BNE cases and in 2.3 % (95 % CI 1.6-3.1 %, I (2) = 87 %) with MIP (P < 0.001). There was a statistically significant lower risk of laryngeal nerve injury with MIP (0.3 %) than with BNE (0.9 %), but similar risk of infection (0.5 vs. 0.5 %) and mortality (0.1 vs. 0.5 %). The available evidence, mostly observational, is at moderate risk of bias, and limited by indirect comparisons and inconsistency for some outcomes (cure rate, hypocalcemia). MIP and BNE are both effective surgical techniques for the treatment of primary hyperparathyroidism. The safety profile of MIP appears superior to BNE (lower rate of hypocalcemia and recurrent laryngeal nerve injury).


A case-study on the accuracy of mass balances for xenobiotics in full-scale wastewater treatment plants.
Removal efficiencies of micropollutants in wastewater treatment plants (WWTPs) are usually evaluated from mass balance calculations using a small number of observations drawn from short sampling campaigns. Since micropollutant loads can vary greatly in both influent and effluent and reactor tanks exhibit specific hydraulic residence times, these short-term approaches are particularly prone to yield erroneous removal values. A detailed investigation of micropollutant transit times at full-scale and on how this affects mass balancing results was still lacking. The present study used hydraulic residence time distributions to scrutinize the match of influent loads to effluent loads of 10 polar micropollutants with different influent dynamics in a full-scale WWTP. Prior hydraulic modeling indicated that a load sampled over one day in the effluent is composed of influent load fractions of five preceding days. Results showed that the error of the mass balance can be reduced with increasing influent sampling duration. The approach presented leads to a more reliable estimation of the removal efficiencies of those micropollutants which can be constantly detected in influents, such as pharmaceuticals, but provides no advantage for pesticides due to their sporadic occurrence. The mismatch between sampled influent and effluent loads was identified as a major error source and an explanation was provided for the occurrence of negative mass balances regularly reported. This study indicates that the accurate determination of global removal values is only feasible in full-scale investigations with sampling durations much longer than 1 day. In any case, the uncertainty of these values needs to be reported when used in removal assessment, model selection or validation.


AOA-Based Three-Dimensional Multi-Target Localization in Industrial WSNs for LOS Conditions.
High-precision and fast relative positioning of a large number of mobile sensor nodes (MSNs) is crucial for smart industrial wireless sensor networks (SIWSNs). However, positioning multiple targets simultaneously in three-dimensional (3D) space has been less explored. In this paper, we propose a new approach, called Angle-of-Arrival (AOA) based Three-dimensional Multi-target Localization (ATML). The approach utilizes two anchor nodes (ANs) with antenna arrays to receive the spread spectrum signals broadcast by MSNs. We design a multi-target single-input-multiple-output (MT-SIMO) signal transmission scheme and a simple iterative maximum likelihood estimator (MLE) to estimate the 2D AOAs of multiple MSNs simultaneously. We further adopt the skew line theorem of 3D geometry to mitigate the AOA estimation errors in determining locations. We have conducted extensive simulations and also developed a testbed of the proposed ATML. The numerical and field experiment results have verified that the proposed ATML can locate multiple MSNs simultaneously with high accuracy and efficiency by exploiting the spread spectrum gain and antenna array gain. The ATML scheme does not require extra hardware or synchronization among nodes, and has good capability in mitigating interference and multipath effect in complicated industrial environments.


Amino acid sequences around the disulphide bridges and methionine residues of porcine pepsin.
1. The amino acid sequences around three disulphide bridges and four methionine residues of porcine pepsin were studied by using diagonal electrophoresis methods. 2. Two of the three disulphide bridges were in small loops of five and six residues. The sequence around one of the two half-cystine residues of the third disulphide bridge had a large number of acidic residues. 3. The sequence of a tetrapeptide containing phosphoserine was also determined. 4. Four unique methionine-containing sequences were constructed. The information is sufficient for the determination of the overlaps in the cyanogen bromide fragments of pepsin. 5. The usefulness of diagonal methods in the study of protein structure, the relative positions of cystinyl and methionyl residues in porcine pepsin and the homology between pepsin and rennin are discussed.


Structure of the C-terminal domain of lettuce necrotic yellows virus phosphoprotein.
Lettuce necrotic yellows virus (LNYV) is a prototype of the plant-adapted cytorhabdoviruses. Through a meta-prediction of disorder, we localized a folded C-terminal domain in the amino acid sequence of its phosphoprotein. This domain consists of an autonomous folding unit that is monomeric in solution. Its structure, solved by X-ray crystallography, reveals a lollipop-shaped structure comprising five helices. The structure is different from that of the corresponding domains of other Rhabdoviridae, Filoviridae, and Paramyxovirinae; only the overall topology of the polypeptide chain seems to be conserved, suggesting that this domain evolved under weak selective pressure and varied in size by the acquisition or loss of functional modules.


A Research Roadmap: Connected Health as an Enabler of Cancer Patient Support.
The evidence that quality of life is a positive variable for the survival of cancer patients has prompted the interest of the health and pharmaceutical industry in considering that variable as a final clinical outcome. Sustained improvements in cancer care in recent years have resulted in increased numbers of people living with and beyond cancer, with increased attention being placed on improving quality of life for those individuals. Connected Health provides the foundations for the transformation of cancer care into a patient-centric model, focused on providing fully connected, personalized support and therapy for the unique needs of each patient. Connected Health creates an opportunity to overcome barriers to health care support among patients diagnosed with chronic conditions. This paper provides an overview of important areas for the foundations of the creation of a new Connected Health paradigm in cancer care. Here we discuss the capabilities of mobile and wearable technologies; we also discuss pervasive and persuasive strategies and device systems to provide multidisciplinary and inclusive approaches for cancer patients for mental well-being, physical activity promotion, and rehabilitation. Several examples already show that there is enthusiasm in strengthening the possibilities offered by Connected Health in persuasive and pervasive technology in cancer care. Developments harnessing the Internet of Things, personalization, patient-centered design, and artificial intelligence help to monitor and assess the health status of cancer patients. Furthermore, this paper analyses the data infrastructure ecosystem for Connected Health and its semantic interoperability with the Connected Health economy ecosystem and its associated barriers. Interoperability is essential when developing Connected Health solutions that integrate with health systems and electronic health records. Given the exponential business growth of the Connected Health economy, there is an urgent need to develop mHealth (mobile health) exponentially, making it both an attractive and challenging market. In conclusion, there is a need for user-centered and multidisciplinary standards of practice to the design, development, evaluation, and implementation of Connected Health interventions in cancer care to ensure their acceptability, practicality, feasibility, effectiveness, affordability, safety, and equity.


Salmonella typhimurium encodes a putative iron transport system within the centisome 63 pathogenicity island.
Upon entry into the host, Salmonella enterica strains are presumed to encounter an iron-restricted environment. Consequently, these bacteria have evolved a variety of often-redundant high-affinity acquisition systems to obtain iron in this restricted environment. We have identified an iron transport system that is encoded within the centisome 63 pathogenicity island of Salmonella typhimurium. The nucleotide composition of this locus is significantly different from that of the rest of this pathogenicity island, suggesting a different ancestry and a mosaic structure for this region of the S. typhimurium chromosome. This locus, designated sit, consists of four open reading frames which encode polypeptides with extensive homology to the yfe ABC iron transport system of Yersinia pestis, as well as other ABC transporters. The sitA gene encodes a putative periplasmic binding protein, sitB encodes an ATP-binding protein, and sitC and sitD encode two putative permeases (integral membrane proteins). This operon is capable of complementing the growth defect of the enterobactin-deficient Escherichia coli strain SAB11 in iron-restricted minimal medium. Transcription of the sit operon is repressed under iron-rich growth conditions in a fur-dependent manner. Introduction of a sitBCD deletion into wild-type S. typhimurium resulted in no apparent growth defect in either nutrient-rich or minimal medium and no measurable virulence phenotype. These results further support the existence of redundant iron uptake systems in S. enterica.


Isolation, Purification, and Characterization of Five Active Diketopiperazine Derivatives from Endophytic Streptomyces SUK 25 with Antimicrobial and Cytotoxic Activities.
In our search for new sources of bioactive secondary metabolites from Streptomyces sp., the ethyl acetate extracts from endophytic Streptomyces SUK 25 afforded five active diketopiperazine (DKP) compounds. The aim of this study was to characterize the bioactive compounds isolated from endophytic Streptomyces SUK 25 and evaluate their bioactivity against multiple drug resistance (MDR) bacteria such as Enterococcus raffinosus, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and Enterobacter spp., and their cytotoxic activities against the human hepatoma (HepaRG) cell line. The production of secondary metabolites by this strain was optimized through Thornton's medium. Isolation, purification, and identification of the bioactive compounds were carried out using high-performance liquid chromatography, high-resolution mass liquid chromatography-mass spectrometry, Fourier transform infrared spectroscopy, and nuclear magnetic resonance, and cryopreserved HepaRG cells were selected to test the cytotoxicity. The results showed that endophytic Streptomyces SUK 25 produces four active DKP compounds and an acetamide derivative, which were elucidated as cyclo-(L-Val-L-Pro), cyclo-(L-Leu-L-Pro), cyclo-(L-Phe-L-Pro), cyclo-(L-Val-L-Phe), and N-(7-hydroxy-6-methyl-octyl)-acetamide. These active compounds exhibited activity against methicillin-resistant S. aureus ATCC 43300 and Enterococcus raffinosus, with low toxicity against human hepatoma HepaRG cells. Endophytic Streptomyces SUK 25 has the ability to produce DKP derivatives biologically active against some MDR bacteria with relatively low toxicity against HepaRG cells line.


Possibilities for IPM Strategies in European Laying Hen Farms for Improved Control of the Poultry Red Mite (Dermanyssus gallinae): Details and State of Affairs.
The Poultry Red Mite (PRM), Dermanyssus gallinae, is a major threat to the poultry industry worldwide, causing serious problems to animal health and welfare, and huge economic losses. Controlling PRM infestations is very challenging. Conventionally, D. gallinae is treated with synthetic acaricides, but the particular lifestyle of the mite (most of the time spent off the host) makes the efficacy of acaracide sprays often unsatisfactory, as sprays reach only a small part of the population. Moreover, many acaricides have been unlicensed due to human consumer and safety regulations and mites have become resistant to them. A promising course of action is Integrated Pest Management (IPM), which is sustainable for animals, humans and the environment. It combines eight different steps, in which prevention of introduction and monitoring of the pest are key. Further, it focusses on non-chemical treatments, with chemicals only being used as a last resort. Whereas IPM is already widely applied in horticulture, its application is still in its infancy to control D. gallinae in layer houses. This review presents the currently-available possibilities for control of D. gallinae in layer houses for each of the eight IPM steps, including monitoring techniques, established and emerging non-chemical treatments, and the strategic use of chemicals. As such, it provides a needed baseline for future development of specific IPM strategies, which will allow efficient and sustainable control of D. gallinae in poultry farms.


Endolysosomal N-glycan processing is critical to attain the most active form of the enzyme acid alpha-glucosidase.
Acid alpha-glucosidase (GAA) is a lysosomal glycogen-catabolizing enzyme, the deficiency of which leads to Pompe disease. Pompe disease can be treated with systemic recombinant human GAA (rhGAA) enzyme replacement therapy (ERT), but the current standard of care exhibits poor uptake in skeletal muscles, limiting its clinical efficacy. Furthermore, it is unclear how the specific cellular processing steps of GAA after delivery to lysosomes impact its efficacy. GAA undergoes both proteolytic cleavage and glycan trimming within the endolysosomal pathway, yielding an enzyme that is more efficient in hydrolyzing its natural substrate, glycogen. Here, we developed a tool kit of modified rhGAAs that allowed us to dissect the individual contributions of glycan trimming and proteolysis on maturation-associated increases in glycogen hydrolysis using in vitro and in cellulo enzyme processing, glycopeptide analysis by MS, and high-pH anion-exchange chromatography with pulsed amperometric detection for enzyme kinetics. Chemical modifications of terminal sialic acids on N-glycans blocked sialidase activity in vitro and in cellulo, thereby preventing downstream glycan trimming without affecting proteolysis. This sialidase-resistant rhGAA displayed only partial activation after endolysosomal processing, as evidenced by reduced catalytic efficiency. We also generated enzymatically deglycosylated rhGAA that was shown to be partially activated despite not undergoing proteolytic processing. Taken together, these data suggest that an optimal rhGAA ERT would require both N-glycan and proteolytic processing to attain the most efficient enzyme for glycogen hydrolysis and treatment of Pompe disease. Future studies should examine the amenability of next-generation ERTs to both types of cellular processing.


The preventive effect of oxytocin to Cisplatin-induced neurotoxicity: an experimental rat model.
Peripheral neurotoxicity is a frequent dose-limiting side effect of the chemotherapeutic agent cisplatin. This study was conducted to investigate the preventive effect of oxytocin (OT) on cisplatin-induced neurotoxicity in rats. Forty-four adult female rats were included in the study. Thirty-six rats were administered intraperitoneally (i.p.) single dose cisplatin 10 mg/kg and divided in to 3 groups. The first group (n=12) received saline i.p., whereas the second group (n=12) and the third group (n=12) were injected with 80 µg/kg and 160 µg/kg OT, respectively, for 10 days. The remaining 8 rats served as the control group. Electromyography (EMG) studies were recorded and blood samples were collected for the measurement of plasma lipid peroxidation (malondialdehyde; MDA), tumor necrosis factor (TNF)-α, and glutathione (GSH) levels. EMG findings revealed that compound muscle action potential amplitude was significantly decreased and distal latency was prolonged in the nontreated cisplatin-injected rats compared with the control group (P<0.005). Also, nontreated cisplatin-injected rats showed significantly higher TNF-α and MDA levels and lower GSH level than control group. The administration of OT significantly ameliorated the EMG alterations, suppressed oxidative stress and inflammatory parameters, and increased antioxidative capacity. We suggest that oxytocin may have beneficial effects against cisplatin-induced neurotoxicity.


Evaluation of Functional Spray Coatings for Mitigating the Uptake of Volatile Phenols by Pinot Noir Wine Grapes via Blocking, Absorption, and/or Adsorption.
Spray coatings have shown promising potential in preventing the uptake of smoke phenols from wildfires into wine grapes. Three cellulose nanofiber-based coatings with low methoxyl pectin or varying concentrations of chitosan were made into films and their potential for blocking, absorption, or adsorption of phenols (guaiacol, m-cresol, and syringol) was evaluated using a custom-built smoke diffusion box. The coatings were also applied to Pinot noir grapes in a vineyard. GC-MS analysis for smoke phenols from headspace gases of diffusion study and extractions of films indicated that chitosan-based films can block guaiacol and syringol, and all films are able to capture m-cresol. The type of coating and application time in a vineyard did not affect (P < 0.05) physicochemical properties, size, and weight of the berries, whereas chitosan-based coatings resulted in a higher anthocyanin content of berries. This study provided new information about the key mechanisms (i.e., blocking phenols) of coatings to mitigate smoke phenol uptake in wine grapes.


The pathogenesis of a North American H5N2 clade 184.108.40.206 group A highly pathogenic avian influenza virus in surf scoters (Melanitta perspicillata).
Aquatic waterfowl, particularly those in the order Anseriformes and Charadriiformes, are the ecological reservoir of avian influenza viruses (AIVs). Dabbling ducks play a recognized role in the maintenance and transmission of AIVs. Furthermore, the pathogenesis of highly pathogenic AIV (HPAIV) in dabbling ducks is well characterized. In contrast, the role of diving ducks in HPAIV maintenance and transmission remains unclear. In this study, the pathogenesis of a North American A/Goose/1/Guangdong/96-lineage clade 126.96.36.199 group A H5N2 HPAIV, A/Northern pintail/Washington/40964/2014, in diving sea ducks (surf scoters, Melanitta perspicillata) was characterized. Intrachoanal inoculation of surf scoters with A/Northern pintail/Washington/40964/2014 (H5N2) HPAIV induced mild transient clinical disease whilst concomitantly shedding high virus titers for up to 10 days post-inoculation (dpi), particularly from the oropharyngeal route. Virus shedding, albeit at low levels, continued to be detected up to 14 dpi. Two aged ducks that succumbed to HPAIV infection had pathological evidence for co-infection with duck enteritis virus, which was confirmed by molecular approaches. Abundant HPAIV antigen was observed in visceral and central nervous system organs and was associated with histopathological lesions. Collectively, surf scoters, are susceptible to HPAIV infection and excrete high titers of HPAIV from the respiratory and cloacal tracts whilst being asymptomatic. The susceptibility of diving sea ducks to H5 HPAIV highlights the need for additional research and surveillance to further understand the contribution of diving ducks to HPAIV ecology.


Speciation of protein-bound trace elements by gel electrophoresis and atomic spectrometry.
The metabolism of trace elements, in particular their binding to proteins in biological systems is of great importance in biochemical, toxicological, and pharmacological studies. As a result there has been a sustained interest over the last two decades in the speciation of protein-bound metals. Various analytical approaches have been employed, combining efficient separation of metalloproteins by liquid chromatography or electrophoresis with high-sensitivity elemental detection. Slab-gel electrophoresis (GE) is a key platform for high-resolution protein separation, and has been combined with autoradiography and various atomic spectrometric techniques for in-gel determination of protein-bound metals. Recently, the combination of GE with state-of-the-art inductively coupled plasma-mass spectrometry (ICP-MS), particularly when linked to laser ablation (LA) for direct gel interrogation, has opened up new opportunities for rapid characterization of metalloproteins. The use of GE and atomic spectrometry for the speciation of protein-bound trace elements is reviewed in this paper. Technical requirements for gel electrophoresis/atomic spectrometric measurement are considered in terms of method compatibilities, detection capability and potential usefulness. The literature is also surveyed to illustrate current status and future trends.


Inherent Oxidative Stress in the Lewis Rat Is Associated with Resistance to Toxoplasmosis.
The course of Toxoplasma gondii infection in rats closely resembles that in humans. However, compared to the Brown Norway (BN) rat, the Lewis (LEW) rat is extremely resistant to T. gondii infection. Thus, we performed RNA sequencing analysis of the LEW rat versus the BN rat, with or without T. gondii infection, in order to unravel molecular factors directing robust and rapid early T. gondii-killing mechanisms in the LEW rat. We found that compared to the uninfected BN rat, the uninfected LEW rat has inherently higher transcript levels of cytochrome enzymes (Cyp2d3, Cyp2d5, and Cybrd1, which catalyze generation of reactive oxygen species [ROS]), with concomitant higher levels of ROS. Interestingly, despite having higher levels of ROS, the LEW rat had lower transcript levels for antioxidant enzymes (lactoperoxidase, microsomal glutathione S-transferase 2 and 3, glutathione S-transferase peroxidase kappa 1, and glutathione peroxidase) than the BN rat, suggesting that the LEW rat maintains cellular oxidative stress that it tolerates. Corroboratively, we found that scavenging of superoxide anion by Mn(III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) decreased the refractoriness of LEW rat peritoneal cells to T. gondii infection, resulting in proliferation of parasites in LEW rat peritoneal cells which, in turn, led to augmented cell death in the infected cells. Together, our results indicate that the LEW rat maintains inherent cellular oxidative stress that contributes to resistance to invading T. gondii, and they thus unveil new avenues for developing therapeutic agents targeting induction of host cell oxidative stress as a mechanism for killing T. gondii.


Genetic Architecture of Circulating Very-Long-Chain (C24:0 and C22:0) Ceramide Concentrations.
Total ceramide concentrations are linked with increased insulin resistance and cardiac dysfunction. However, recent studies have demonstrated that plasma concentrations of specific very-long-chain fatty ceramides (C24:0 and C22:0) are associated with a reduced incidence of coronary heart disease and all-cause mortality. We hypothesized that specific genetic loci are associated with plasma C22:0 and C24:0 concentrations. Heritability and genome-wide association studies of plasma C24:0 and C22:0 ceramide concentrations were performed among 2,217 participants in the Framingham Heart Study Offspring Cohort, adjusting for cardiovascular risk factor covariates and cardiovascular drug treatment. The multivariable-adjusted heritability for C22:0 and C24:0 ceramides was 0.42 (standard error [SE], 0.07; p=1.8E-9) and 0.25 (SE, 0.08; p=0.00025), respectively. Nineteen single nucleotide polymorphisms (SNPs), all on chromosome 20, significantly associated with C22:0 concentrations; the closest gene to these variants was SPTLC3. The lead SNP (rs4814175) significantly associated with 3% lower plasma C22:0 concentrations (p=2.83E-11). Nine SNPs, all on chromosome 20 and close to SPTLC3, were significantly associated with C24:0 ceramide concentrations. All 9 were also significantly related to plasma C22:0 levels. The lead SNP (rs168622) was significantly associated with 10% lower plasma C24:0 ceramide concentrations (p=9.94E-09). SNPs near the SPTLC3 gene, which encodes serine palmitoyltransferase long chain base subunit 3 (SPTLC3; part of the enzyme that catalyzes the rate-limiting step of de novo sphingolipid synthesis) were associated with plasma C22:0 and C24:0 ceramide concentrations. These results are biologically plausible and suggest that SPTLC3 may be a potential therapeutic target for C24:0 and C22:0 ceramide modulation.


Sarcoid optic disc edema and optociliary shunts.
We report the case of a patient with sarcoid granulomatous optic disc edema with optociliary shunts, normal visual acuity, and negative orbit computed tomographic studies. A literature review disclosed four cases of optociliary veins associated with sarcoid optic neuropathy, all the patients having very poor vision. Optociliary shunts may not carry a poor visual prognosis in optic nerve sarcoidosis.


Handling missing responses in generalized linear mixed model without specifying missing mechanism.
In longitudinal studies, missingness of data is often unavoidable. Valid estimators from the generalized linear mixed model usually rely on the correct specification of the missing data mechanism. An incorrectly specified missing mechanism may lead to a biased estimator. In this article, we propose a class of unbiased estimating equations using pairwise conditional technique to deal with the generalized linear mixed model under benign non-ignorable missingness where specification of the missing model is not needed. We show that the proposed estimator is consistent and asymptotically normal under certain conditions. Simulation results and an example using longitudinal course of neuropsychological data are also shown.


Addiction in Singapore: changing patterns and evolving challenges.
Data from recent studies have shown an increase in substance use and addictive behaviours in Singapore. However, training and services still lag behind this trend, which means the provision of adequate treatment is lacking in spite of good evidence that treatment approaches for addiction disorders are effective. There is a need to train more manpower for this purpose, especially in primary healthcare. Another challenge is to ramp up services as quickly as possible to manage the growing number of addicts. Research should also focus on special populations, while public health education and national policies can be improved, particularly to curb binge drinking. We need to be open to the use of harm-reduction therapies and substitution treatments, as addiction is a chronic relapsing disorder with significant health and psychosocial morbidity.


Nationwide age references for sitting height, leg length, and sitting height/height ratio, and their diagnostic value for disproportionate growth disorders.
To obtain age references for sitting height (SH), leg length (LL), and SH/H ratio in the Netherlands; to evaluate how SH standard deviation score (SDS), LL SDS, SH/H SDS, and SH/LL SDS are related to height SDS; and to study the usefulness of height corrected SH/H cut-off lines to detect Marfan syndrome and hypochondroplasia. Cross-sectional data on height and sitting height were collected from 14,500 children of Dutch origin in the age range 0-21 years. Reference SD charts were constructed by the LMS method. Correlations were analysed in three age groups. SH/H data from patients with Marfan syndrome and genetically confirmed hypochondroplasia were compared with height corrected SH/H references. A positive association was observed between H SDS, SH SDS, and LL SDS in all age groups. There was a negative correlation between SH/H SDS and height SDS. In short children with a height SDS <-2 SDS, a cut-off limit of +2.5 SD leads to a more acceptable percentage of false positive results. In exceptionally tall children, a cut-off limit of -2.2 SDS can be used. Alternatively, a nomogram of SH/H SDS versus H SDS can be helpful. The sensitivity of the height corrected cut-off lines for hypochondroplasia was 80% and for Marfan syndrome only 30%. In exceptionally short or tall children, the dependency of the SH/H ratio (SDS) on height SDS has to be taken into consideration in the evaluation of body proportions. The sensitivity of the cut-off lines for hypochondroplasia is fair.


Locoregional management of hepatocellular carcinoma. Surgical and ablation therapies.
The selection of an appropriate treatment strategy for patients with HCC depends on careful tumor staging and assessment of the underlying liver disease (Fig. 5). All patients with localized HCC (involvement of one single lobe, no vascular invasion or extrahepatic disease) should be evaluated for the potentially curative therapeutic options of partial hepatectomy or OLT. Candidates for partial hepatectomy must have no liver disease or Child's A cirrhosis, normal portal pressure, and normal serum bilirubin. For patients not meeting these criteria, OLT should be considered if there is a solitary lesion smaller than 5 cm in diameter or fewer than three lesions smaller than 3 cm. Local ablative therapies such as PEI, RFA, and TACE offer palliation for patients for whom surgical approaches are contraindicated. Percutaneous alcohol injection and RFA are minimally invasive and can be used on an outpatient basis, usually for tumor nodules smaller than 3 cm. When these therapies are used for small tumors, the survival rates can be similar to those achieved by partial hepatectomy. Transcatheter [figure: see text] arterial chemoembolization may be used as an interim treatment for patients waiting for OLT. Although TACE is often used for the palliation of large tumors, significant survival benefits have not yet been demonstrated for this indication.


Reproductive performance of Arab mares in the Kingdom of Saudi Arabia.
The aim of this study was to evaluate the reproductive performance of the Arab mares in the Kingdom of Saudi Arabia (KSA). Managers of 26 studfarms containing 1014 Arab mares at five districts were asked to complete a questionnaire on the reproductive efficiency of their mares. The mean age at first mating was 3.06 ± 0.5 years. Mares were mated equally in winter and all over the year. Natural mating was mostly used (76.9%). The means of estrus duration and estrus interval were 6.58 ± 1 days and 19.57 ± 1.8 days, respectively. The means of first service and over- all pregnancy rates and the number of cycles/pregnancy were 52.71 ± 20.2%, 83.72 ± 15.0%, and 1.46 ± 0.3, respectively, with no effect of the postpartum interval, district, or season. The average pregnancy duration was 335.5 ± 10.2 days. It was shorter in mares mated by day 9 postpartum than in those mated later (p < 0.05). The ratio of stallion/mares ranged from 1:4 to 1:24, with no correlation with the pregnancy rates or the number of cycles/pregnancy. Photoperiodic regulation and changes in the hypothalamic-hypophyseal axis of mares are usually associated with the seaso- nal reproductive activity of mares in the temperate but not in subtropical areas. Short duration of gestation for mares mated by day 9 after foaling may be related to the nutritional status of dams. The obtained data represent the first record of the reproductive performance of Arab mares in the KSA, which could be used to anticipate their performance under different managements. The Arab mares in the KSA are not seasonal and can reproduce efficiently all over the year. District and season did not influence the fertility.


Unraveling the swine genome: implications for human health.
The pig was first used in biomedical research in ancient Greece and over the past few decades has quickly grown into an important biomedical research tool. Pigs have genetic and physiological traits similar to humans, which make them one of the most useful and versatile animal models. Owing to these similarities, data generated from porcine models are more likely to lead to viable human treatments than those from murine work. In addition, the similarity in size and physiology to humans allows pigs to be used for many experimental approaches not feasible in mice. Research areas that employ pigs range from neonatal development to translational models for cancer therapy. Increasing numbers of porcine models are being developed since the release of the swine genome sequence, and the development of additional porcine genomic and epigenetic resources will further their use in biomedical research.


Quantitative Analysis of Differentiation Activity for Mouse Embryonic Stem Cells by Deep Learning for Cell Center Detection using Three-Dimensional Confocal Fluorescence Microscopy Images.
Accurate single cell segmentation provides means to monitor the behavior of single cell within a population of cells. Time-lapse fluorescence images are used to reveal heterogeneous nature of single mouse embryonic stem cell (ESC) colony and monitor fluctuations of the cell states. Automatic quantification of speed and status shifts of the ESCs depends on accurate single cell segmentation that is used to calculate the 3D center of every cell and track this cell for the quantification. This study proposes a new 3D U-net to accurately detect center of each single cell in 3D confocal images. The dimension of input 3D images to the U-net is flexible so that multiple center detections from different image directions can be implemented simultaneously to improve the center detection accuracy. This study showed that our method can improve accuracy for cell center detection and thus the quantification for ESC speeds and status shifts.


[Analysis of human anti-rabies post exposure treatment in the great area of Sao Paulo, Brazil].
The treatment of human beings with risk of exposure to rabies virus infection still presents some cases of undesirable post treatment reactions. The knowledge resulting from the regular analysis of the epidemiological features associated with these practices would minimize the occurrence of these troubles. The records of the treatment of 8,758 human anti-rabies cases performed in Osasco City, S. Paulo, Brazil between 1984 and 1994 with Fuenzalida & Palacios vaccine and/or anti-rabies serum produced in equines were analysed by Epi Info program. It was found that the higher risk of exposure was for male children of from five to nine years of age. The most frequent scene of aggression was the victim's own house. Dogs were responsible for most of the accidents (62.8%); the majority of dogs and cats involved had not been immunized against rabies (51.0% of dogs and 73.2% of cats). Most people aged nine or under were attacked in the head (36.6%), followed by the upper limbs (35.1%); above this age the part of the body most frequently hurt was the upper (45.8%) and lower limbs (43. 7%); 26.5% of patients had already been vaccinated against rabies at least once in their lifetime; 90.7% of them sought medical care within five days, anti-rabies treatment was prescribed for 41.9% of them. Post vaccinal reactions were observed in 0.25% of the patients 0.3% of which were of neurological; 80.1% of the dogs and 58.4% of the cats involved coere kept under observation. The months of March, July, August and September presented the highest aggression rates.


Studies in the use of magnetic microspheres for immunoaffinity extraction of paralytic shellfish poisoning toxins from shellfish.
Paralytic shellfish poisoning (PSP) is a potentially fatal human health condition caused by the consumption of shellfish containing high levels of PSP toxins. Toxin extraction from shellfish and from algal cultures for use as standards and analysis by alternative analytical monitoring methods to the mouse bioassay is extensive and laborious. This study investigated whether a selected MAb antibody could be coupled to a novel form of magnetic microsphere (hollow glass magnetic microspheres, brand name Ferrospheres-N) and whether these coated microspheres could be utilized in the extraction of low concentrations of the PSP toxin, STX, from potential extraction buffers and spiked mussel extracts. The feasibility of utilizing a mass of 25 mg of Ferrospheres-N, as a simple extraction procedure for STX from spiked sodium acetate buffer, spiked PBS buffer and spiked mussel extracts was determined. The effects of a range of toxin concentrations (20-300 ng/mL), incubation times and temperature on the capability of the immuno-capture of the STX from the spiked mussel extracts were investigated. Finally, the coated microspheres were tested to determine their efficiency at extracting PSP toxins from naturally contaminated mussel samples. Toxin recovery after each experiment was determined by HPLC analysis. This study on using a highly novel immunoaffinity based extraction procedure, using STX as a model, has indicated that it could be a convenient alternative to conventional extraction procedures used in toxin purification prior to sample analysis.


From general discrimination to segmented inequality: Migration and inequality in urban China.
Internal migration in China during the last three decades, the largest in human history, offers a rare opportunity to understand inequalities in the making. Using data spanning 10years from China's largest metropolis, Shanghai, this study assesses how enduring state institutions interplay with the spread of market forces to shape income inequality between migrants and native urban workers. Though the wages of both Chinese migrants and urban workers rose considerably, economic restructuring during the decade under study resulted in diminished privileges for urbanites and subsequently increased collision between migrants and urban workers in the private sectors. These shifts, rather than substantially reducing inequality, have led to an evolving form of inequality, from an initial general blatant discrimination against migrants across the board, to a new and more subtle form of inequality characterized by substantial segmented discrimination against migrants within economic sectors, with the degree of inequality varying from sector to sector. We discuss how this changing inequality reflects complementary rather than competing roles of the state and market institutions in inequality creation and maintenance.


Recent advances in our understanding of the allograft response.
Organ transplantation is a life-saving treatment for end-stage organ failure. However, despite advances in immunosuppression, donor matching, tissue typing, and organ preservation, many organs are still lost each year to rejection. Ultimately, tolerance in the absence of immunosuppression is the goal, and although this seldom occurs spontaneously, a deeper understanding of alloimmunity may provide avenues for future therapies which aid in its establishment. Here, we highlight the recent key advances in our understanding of the allograft response. On the innate side, recent work has highlighted the previously unrecognised role of innate lymphoid cells as well as natural killer cells in promoting the alloresponse. The two major routes of allorecognition have recently been joined by a third newly identified pathway, semi-direct allorecognition, which is proving to be a key active pathway in transplantation. Through this review, we detail these newly defined areas in the allograft response and highlight areas for potential future therapeutic intervention.


In vivo priming by DNA injection occurs predominantly by antigen transfer.
DNA vaccines can stimulate both humoral and cytolytic immune responses. Although bone marrow-derived elements present the expressed Ag, the mechanisms for acquiring immunogenic peptides have yet to be fully elucidated. APCs may become directly transfected by plasmid DNA or process extracellular proteins produced by other transfected cells. Using a transactivating plasmid system and bone marrow chimeras, we show that both mechanisms appear to be involved; however, the bulk of the immune response is dependent on expression of Ag by nonlymphoid tissues and transfer to APCs. These in vivo studies are the first to define the role of transfected nonlymphoid cells in generating Ag for presentation by bone marrow-derived APCs after needle injection with plasmid DNA.


The tumor suppressor semaphorin 3B triggers a prometastatic program mediated by interleukin 8 and the tumor microenvironment.
Semaphorins are a large family of evolutionarily conserved morphogenetic molecules originally identified for their repelling role in axonal guidance. Intriguingly, semaphorins have recently been implicated in cancer progression (Neufeld, G., T. Lange, A. Varshavsky, and O. Kessler. 2007. Adv. Exp. Med. Biol. 600:118-131). In particular, semaphorin 3B (SEMA3B) is considered a putative tumor suppressor, and yet we found that it is expressed at high levels in many invasive and metastatic human cancers. By investigating experimental tumor models, we confirmed that SEMA3B expression inhibited tumor growth, whereas metastatic dissemination was surprisingly increased. We found that SEMA3B induced the production of interleukin (IL) 8 by tumor cells by activating the p38-mitogen-activated protein kinase pathway in a neuropilin 1-dependent manner. Silencing the expression of endogenous SEMA3B in tumor cells impaired IL-8 transcription. The release of IL-8, in turn, induced the recruitment of tumor-associated macrophages and metastatic dissemination to the lung, which could be rescued by blocking IL-8 with neutralizing antibodies. In conclusion, we report that SEMA3B exerts unexpected functions in cancer progression by fostering a prometastatic environment through elevated IL-8 secretion and recruitment of macrophages coupled to the suppression of tumor growth.


A combination of platelet features allows detection of early-stage cancer.
Detection of early-stage cancer significantly improves patient survival. As platelets play an important role in cancer progression, we aimed to investigate whether platelets can be used for the discovery of early-stage cancer. Patients with lung (n = 86) or head of pancreas (n = 42) cancer were included, as were healthy sex- and age-matched controls (n = 92). Blood was collected before initiation of treatment. Platelet count, volume and activation status were quantified in whole blood. Next, concentrations of vascular endothelial growth factor, platelet-derived growth factor, platelet factor 4, thrombospondin-1 and connective tissue-activating peptide III were measured in both platelets and plasma. Using the results, two multivariable diagnostic models were developed and internally validated. Multiple platelet features, including platelet count, volume and protein content, were significantly changed in lung and head of pancreas cancer patients. However, the pattern of changes differed between both groups. The diagnostic model developed for lung cancer discriminated very well between patients and controls (AUC = 88.7%). Addition of smoking as a variable significantly increased the AUC of the model to 94.5%. The diagnostic model for head of pancreas cancer also performed well (AUC = 82.7%). Both models were internally validated, resulting in optimism-corrected AUC's of 86.8% and 80.8%, respectively. In patients with lung or head of pancreas cancer, several platelet characteristics are changed compared to healthy sex- and age-matched controls. A cancer type-specific combination of these platelet features can be used to discriminate between patients with early-stage cancer and healthy individuals.