Genomic analysis of bovine herpesvirus type 4 (BoHV-4) from Argentina: high genetic variability and novel phylogenetic groups.
Bovine herpesvirus 4 (BoHV-4) is a γ-herpesvirus that has been isolated both from apparently healthy animals and from cattle with a variety of clinical signs, including post-partum endometritis and abortion. BoHV-4 causes either a persistent or a latent infection in cells of the monocyte/macrophage lineage. Two groups of BoVH-4 strains have been defined based on their restriction patterns: the Movar-like strains (European prototype) and the DN 599-like strains (American prototype). The purpose of the present study was to genetically characterize wild type BoHV-4 strains isolated from vaginal discharges of aborted cows in Argentina. The virus was identified by isolation and nested PCR in all vaginal discharge samples from aborted cows, either as a sole agent or in association with other pathogens. Restriction enzyme profiling and phylogenetic analysis demonstrated that there is a high genetic variability among the studied field isolates. The existence of three groups of strains, which were designated as genotypes 1, 2 and 3, is described. Genotypes 1 and 2 possibly correspond to the Movar-like and DN 599-like groups, respectively, whereas Genotype 3 corresponds to a novel group. Two viral strains did not cluster into any of these three groups, indicating that other genotypes could be circulating in Argentina. These results suggest a complex epidemiological background for the Argentinean BoHV-4 strains, probably influenced by independent events of genetic drift. This hypothesis cannot be rejected based on the available data. However, there is no direct evidence supporting this possibility. Thus, it seems speculative to suggest that interspecific jumps are responsible for the observed phylogenetic grouping. On the other hand, our analyses suggest a geographical structure for the observed viral genotypes, since genotypes 1 and 2 included the European (Movar-like) and American (DN599-like) reference strains, respectively. Geographic dispersion is known to be a driver of herpes viruses diversification, and independent evolution in geographical isolated places ensures the emergence of particular mutations in each location due to genetic drift (Compans, 2007; Zong et al., 1999). Therefore, at this point, the genetic drift hypothesis is the one that requires less ad-hoc considerations and thus, to our understanding, is the one that fits to the findings from this study. The involvement of this genetic variability in the detection and pathogenesis of BoHV-4 remains to be investigated.


Acute disseminated encephalomyelitis and thrombocytopenia following Epstein-Barr virus infection.
Epstein-Barr Virus (EBV) causes a broad spectrum of disease in humans with several clinical syndromes and is ubiquitous, infecting more than 95% of the world's population. Central Nervous System (CNS) disease alone associated with Epstein-Barr virus rarely occurs in previously healthy individuals. Systemic viral illness in children and complications are rare, but may occur. In few cases, it is associated with a variety of CNS and hematological complications like acute disseminated encephalomyelitis, transverse myelitis, neuropsychiatric syndrome, GBS, autoimmune thrombocytopenia and hemolytic anemia and they usually respond to immunotherapy. We report previously healthy boy, who presented with left sided weakness, headache and thrombocytopenia following EBV infection. The thrombocytopenia was resistant to intravenous immunoglobulin and methylprednisolone but responded well to Rituximab.


Laparoscopic cholecystectomy. Analysis of results of first 322 cases.
From April 1994 to June 1995 322 laparoscopic cholecystectomies were performed in S. Camillo hospital Treviso, 294 in election and 28 in urgency. The technique and results of this experience are reported. No mortality was observed and morbility was 1.8%. Utility of gallstones in main biliary duct screening is discussed to select patients to subject to preoperative ERCP and the role of intraoperative cholangiography. Laparoscopic cholecystectomy is considered the choice option in cholelitiasis. In case of evidence of choledocholithiasis with anamnesis or preoperative tests, the tactics of choice in authors opinion is the sequential one, providing preoperative ERCP and laparoscopic cholecystectomy.


An increase of tissue cyclic AMP level by adenosine in the dog pancreas without stimulation of exocrine secretion.
In the vascularly isolated and self-hemoperfused dog pancreas, secretin (0.025 clinical units) and adenosine (1.0 mg) administered intra-arterially (i.a.) increased tissue cyclic AMP level by about 46% and 37%, respectively. Although the increases in the nucleotide were not statistically different, only secretin stimulated exocrine secretion. The increase in cyclic AMP induced by adenosine was significantly reversed by pretreatment with theophylline (0.3 mg, i.a.). These results suggest that the effect of adenosine on cyclic AMP formation occurs mainly in the non-exocrine system of the dog pancreas through A2/Ra-receptors.


The blood cells in NSCLC and the changes after RFA.
Lung cancer has attracted a lot of attention because of its high morbidity and mortality. The emergence of RFA provides a new treatment for unresectable NSCLC patients. In addition to killing in situ lung tumors, RFA also provides new immuno-activated antigens, for the treatment of lung cancer. It changes the tumor microenvironment and activates the entire immune system of patients. The peripheral blood cell count is easy to achieve and the blood cells are important in tumor immunity, which changes after RFA. On the one hand, the changes in blood cells identify the immune changes of NSCLC; on the other hand, it provides support and suspicion for the treatment of RFA.


Acute continuous moderate-intensity exercise, but not low-volume high-intensity interval exercise, attenuates postprandial suppression of circulating osteocalcin in young overweight and obese adults.
Bone remodeling markers (BRMs) are suppressed following the consumption of a meal. Our findings indicate that a single session of continuous moderate-intensity exercise, but not low-volume high-intensity interval exercise, performed 1 h after a meal attenuates the postprandial suppression of BRMs. Acute exercise transiently increases BRMs including osteocalcin (tOC) and the undercarboxylated form of osteocalcin (ucOC), a hormone that is implicated in glucose regulation. The effects of acute exercise and exercise-intensity on postprandial levels of tOC and ucOC are unknown. Twenty-seven adults that were overweight or obese (age 30 ± 1 years; BMI 30 ± 1 kg∙m-2; mean ± SEM) were randomly allocated to perform a single session of low-volume high-intensity interval exercise (LV-HIIE; nine females, five males) or continuous moderate-intensity exercise (CMIE; eightfemales, five males) 1 h after consumption of a standard breakfast. Serum tOC, ucOC, and ucOC/tOC were measured at baseline, 1 h, and 3 h after breakfast consumption on a rest day (no exercise) and the exercise day (exercise 1 h after breakfast). Compared to baseline, serum tOC and ucOC were suppressed 3 h after breakfast on the rest day (- 10 ± 1% and - 6 ± 2%, respectively; p < 0.05), whereas ucOC/tOC was elevated (2.5 ± 1%; p = 0.08). Compared to the rest day, CMIE attenuated the postprandial-induced suppression of tOC (rest day - 10 ± 2% versus CMIE - 5 ± 2%, p < 0.05) and ucOC (rest day - 6 ± 4% versus CMIE 11 ± 2%, p < 0.05), and increased postprandial ucOC/tOC (rest day 3 ± 2% versus CMIE 15 ± 1%, p < 0.05). In contrast, LV-HIIE did not alter postprandial tOC, ucOC, or ucOC/tOC (all p > 0.1). Acute CMIE, but not LV-HIIE, attenuates the postprandial-induced suppression of tOC and ucOC. CMIE may be an effective tool to control the circulating levels of BRMs following meal consumption in overweight/obese adults.


Full-length complementary DNA and the derived amino acid sequence of horse uteroglobin.
After its original description as a steroid-dependent protein in the rabbit uterus, uteroglobin became one of the best characterized proteins. However, detailed knowledge of its physiological role remains an enigma. In this study we investigate how its structure is phylogenetically conserved in the horse compared to other mammalian species. Northern blot analysis showed that in horses, the main expression of uteroglobin appears in lung, uterus, and prostate tissues. Western blot analysis demonstrated that the dimeric form of uteroglobin is found predominantly in biological compartments. Using a RACE-PCR technique, we cloned and sequenced the full-length cDNA (473 base pairs) that encodes equine uteroglobin. The nucleotide sequence was shown to characterize the primary structure of this protein. This enabled us to add equine uteroglobin to a comparative amino acid alignment of 8 other uteroglobin molecules, and finally, to unravel 14 evolutionary completely conserved amino acids. We summarize these results with a computer-based 3-D model of horse uteroglobin, and discuss new concepts on the physiological role of uteroglobin, in particular as a specific binding protein.


Synergistic Cytotoxicity of a Toxin Targeting the Epidermal Growth Factor Receptor and the Glycosylated Triterpenoid SO1861 in Prostate Cancer.
Treatment of advanced prostate cancer lacks specificity and curative intent. Therefore, the need for new targeted therapeutic approaches is high. In the present study, we generated the new targeted toxin EGF-PE24mutΔREDLK binding to the epidermal growth factor receptor (EGFR) on the surface of prostate cancer cells. It consists of the human epidermal growth factor (EGF) as binding domain and a de-immunized variant of Pseudomonas Exotoxin A (PE), called PE24mutΔREDLK, as toxin domain. The toxin domain contains a deletion of the C-terminal KDEL-like motif REDLK to prevent its transport from sorting endosomes via the KDEL receptor mediated pathway into the cytosol, where it can inhibit cellular protein biosynthesis and induce apoptosis. Indeed, REDLK deletion resulted in a strong decrease in cytotoxicity of the targeted toxin in prostate cancer cells compared to the parental targeted toxin EGF-PE24mut. However, addition of the plant glycosylated triterpenoid SO1861, which is known to mediate the release of biomolecules from endolysosomal compartments into the cytosol, resulted in an up to almost 7,000-fold enhanced synergistic cytotoxicity. Moreover, combination of PE24mutΔREDLK with SO1861 led to a cytotoxicity that was even 16- to 300-fold enhanced compared to that of EGF-PE24mut. Endolysosomal entrapment of the non-toxic targeted toxin EGF-PE24mutΔREDLK followed by activation through enhanced endosomal escape therefore represents a new promising approach for the future treatment of advanced prostate cancer with high efficacy and diminished side effects.


Socio-economic differences in the use of dairy fat in Russian and Finnish Karelia, 1994-2004.
To determine socio-economic differences from 1994 to 2004 in the use of butter and milk in Pitkäranta in the Republic of Karelia, Russia and North Karelia, Finland. Health behaviour surveys in Pitkäranta (n = 3,599) and North Karelia (n = 3,652) in 1994, 1996, 1998, 2000 and 2004. A clear overall decrease occurred in the use of butter in cooking in Pitkäranta from 1994 to 2004. In both areas, subjects with lower education used butter in cooking and drank fat-containing milk more often. Regarding butter on bread, the educational patterns in the two areas were opposite: in Pitkäranta, subjects with lower education used butter less often but in North Karelia, they used butter on bread more often. Practically, no changes in socio-economic differences from 1994 to 2004 were observed in either area. The socio-economic differences in the use of dairy fat were stable in both areas but larger in North Karelia than in Pitkäranta. Our results support earlier assumptions of a weaker role of education as a determinant of health in Russia than in the western societies.


Connexins may play a critical role in cigarette smoke-induced pulmonary hypertension.
Pulmonary hypertension (PH) is a chronic progressive disease characterized by pulmonary vasoconstriction and remodeling. It causes a gradual increase in pulmonary vascular resistance leading to right-sided heart failure, and may be fatal. Chronic exposure to cigarette smoke (CS) is an essential risk factor for PH group 3; however, smoking continues to be prevalent and smoking cessation is reported to be difficult. A majority of smokers exhibit PH, which leads to a concomitant increase in the risk of mortality. The current treatments for PH group 3 focus on vasodilation and long-term oxygen supplementation, and fail to stop or reverse PH-associated continuous vascular remodeling. Recent studies have suggested that pulmonary vascular endothelial dysfunction induced by CS exposure may be an initial event in the natural history of PH, which in turn may be associated with abnormal alterations in connexin (Cx) expression. The relationship between Cx and CS-induced PH development has not yet been directly investigated. Therefore, this review will describe the roles of CS and Cx in the development of PH and discuss the related downstream pathways. We also discuss the possible role of Cx in CS-induced PH. It is hoped that this review may provide new perspectives for early intervention.


"Ping-pong" gaze. Periodic alternating gaze deviation.
A clinicopathologic study of patient with periodic alternating gaze deviation is presented. The patient's eye moved rhythmically and conjugately from one extreme lateral position to the other. A 0.45-cm hemorrhage, found in the midline deep vermis, is thought to have disrupted the neural integrator that holds positions of gaze, resulting in periodic alternating gaze deviation. Three previous cases of periodic alternating gaze deviation are reviewed and illustrate the role of the cerebellum in the control of eye movements.


The use of nuclear transfer to produce transgenic pigs.
Manipulation of the pig genome has the potential to improve pig production and offers powerful biomedical applications. Genetic manipulation of mammals has been possible for over two decades, but the technology available has proven both difficult and inefficient. The development of new techniques to enhance efficiency and overcome the complications of random insertion is of importance. Nuclear transfer combined with homologous recombination provides a possible solution: precise genetic modifications in the pig genome may be induced via homologous recombination, and viable offspring can be produced by nuclear transfer using cultured transfected cell lines. The technique is still ineffective, but it is believed to have immense potential. One area that would benefit from the technology is that of xenotransplantation: transgenic pigs are expected to be available as organ donors in the foreseeable future.


5th JBF Symposium: 'Bridging for a Global Harmonization'.
Tower Hall Funabori, Tokyo, Japan, 6-7 March 2014 At the 5th Japan Bioanalysis Forum symposium, distinguished bioanalytical scientists from government and industry commentated on the bioanalytical method validation guidelines/guidance issued by their own regions, including the draft bioanalytical method validation guideline for ligand binding assay by the Japanese Ministry of Health, Labour and Welfare, which was finalized in April 2014. Additionally, the Japan Bioanalysis Forum Discussion Group, in which daily bioanalytical issues/interests were scientifically discussed, picked up five topics, and more than 200 conference attendees openly exchanged their views on a wide range of bioanalytical issues with discussion group members. This manuscript provides an overview of the highlights out of the symposium.


Absolute asymmetric synthesis of "chiral-at-metal" Grignard reagents and transfer of the chirality to carbon.
Two new six-coordinate Grignard reagents, cis-[(p-CH(3)C(6)H(4))MgBr(dme)(2)] (1) and cis-[MgCH(3)(thf)(dme)(2)]I (2), have been synthesized and their crystal structures have been determined. Both reagents are cis-octahedral and therefore chiral. They crystallize as conglomerates and racemize rapidly in solution. By utilizing these properties, the absolute asymmetric synthesis of specifically the Delta or the Lambda enantiomer was achieved for both Grignard reagents. Enantiopure 1 and 2 were then reacted with butyraldehyde or benzaldehyde to give the corresponding alcohol in up to 22 % enantiomeric excess. At -60 degrees C, the Grignard reagents crystallize as racemic phases instead of conglomerates. Consequently, the crystal structures of rac-cis-[(p-CH(3)C(6)H(4))MgBr(dme)(2)].DME (3) and rac-cis-[MgCH(3)(thf)(dme)(2)]I (4) could be determined.


Physical mapping of the murine casein locus reveals the gene order as alpha-beta-gamma-epsilon-kappa.
The murine casein locus has been characterized by long-range restriction mapping and the analysis of large fragment genomic clones. Cloned sequences from five mouse casein genes (alpha, beta, gamma, epsilon, kappa) were used to screen a murine (strain 129) genomic library in a bacterial artificial chromosome vector (BAC). Of the nine clones isolated, two contained three casein genes alpha, beta, gamma and gamma, epsilon, kappa, respectively. The following combinations were found in other clones: alpha + beta, beta + gamma; and gamma + epsilon. Thus, the gene order in the locus can be deduced to be alpha-beta-gamma-epsilon-kappa. This order was confirmed by restriction analysis of the clones. A contig map of the clones and flanking sequences has been established by characterizing seven BAC clones, which together span approximately 470 kb. Long-range restriction analysis of genomic DNA indicates that the murine casein locus is confined to a 250-kb partial Xho I fragment. The alpha and beta casein genes were shown to be arranged in a tail-to-tail orientation.


Alkyltelluro Substitution Improves the Radical-Trapping Capacity of Aromatic Amines.
The synthesis of a variety of aromatic amines carrying an ortho-alkyltelluro group is described. The new antioxidants quenched lipidperoxyl radicals much more efficiently than α-tocopherol and were regenerable by aqueous-phase N-acetylcysteine in a two-phase peroxidation system. The inhibition time for diaryl amine 9 b was four-fold longer than recorded with α-tocopherol. Thiol consumption in the aqueous phase was found to correlate inversely to the inhibition time and the availability of thiol is the limiting factor for the duration of antioxidant protection. The proposed mechanism for quenching of peroxyl radicals involves O-atom transfer from peroxyl to Te followed by H-atom transfer from amine to alkoxyl radical in a solvent cage.


Electrochemical sensor for organophosphate pesticides and nerve agents using zirconia nanoparticles as selective sorbents.
An electrochemical sensor for detection of organophosphate (OP) pesticides and nerve agents using zirconia (ZrO2) nanoparticles as selective sorbents is presented. Zirconia nanoparticles were electrodynamically deposited onto the polycrystalline gold electrode by cyclic voltammetry. Because of the strong affinity of zirconia for the phosphoric group, nitroaromatic OPs strongly bind to the ZrO2 nanoparticle surface. The electrochemical characterization and anodic stripping voltammetric performance of bound OPs were evaluated using cyclic voltammetric and square-wave voltammetric (SWV) analysis. SWV was used to monitor the amount of bound OPs and provide simple, fast, and facile quantitative methods for nitroaromatic OP compounds. The sensor surface can be regenerated by successively running SWV scanning. Operational parameters, including the amount of nanoparticles, adsorption time, and pH of the reaction medium have been optimized. The stripping voltammetric response is highly linear over the 5-100 ng/mL (ppb) methyl parathion range examined (2-min adsorption), with a detection limit of 3 ng/mL and good precision (RSD = 5.3%, n = 10). The detection limit was improved to 1 ng/mL by using 10-min adsorption time. The promising stripping voltammetric performances open new opportunities for fast, simple, and sensitive analysis of OPs in environmental and biological samples. These findings can lead to a widespread use of electrochemical sensors to detect OP contaminates.


[Relationship between plasma carbon monoxide and blood-brain barrier permeability in cirrhotic rats].
To explore the relationship between plasma levels of carbon monoxide and blood-brain permeability in cirrhotic rats. We measured mean arterial pressure (MAP), heart rate, plasma levels of carbon monoxide and amount of Evans blue in the brain tissue taken as the index of blood-brain permeability in cirrhotic rats (n=10) and controls (n=10). Cirrhotic rats showed significant increases in plasma carbon monoxide and amount of Evans blue in brain tissue compared with controls [(18.37 +/- 1.79) micromol/L,(18.52 +.- 1.39) ng/mg vs (10.27 +/- 1.21) micromol/L, (15.08 +/- 1.06) ng/mg; P< 0.01]. Carbon monoxide levels in cirrhotic rats correlated positively with blood-brain barrier permeability and inversely with MAP (r=0.72, P< 0.01; r= -0.67, P< 0.05). Carbon monoxide, involving in the occurrence of hypotension and the increase of blood-brain barrier permeability, may play a role in the development of hepatic encephalopathy.


Considerations in Orbital Reconstruction for the Oncologic Surgeon: Critical versus Optimal Objectives.
Background Orbital reconstruction following oncologic midface resection is uniquely challenging, and makes critical contributions to patient aesthetics, function, and identity. Approach is largely dependent on surgeon and patient preferences, and there exists no consensus on defect characterization. Objective The goal of the study is to provide a mental framework for the reconstructive oncologic surgeon to use as a foundation during his or her approach to the orbit. Design The design of the study is based on the review of current literature and expert opinion. Conclusions Critical versus optimal objectives must be set in orbital reconstruction, and a systematic approach should be followed. We approach orbital reconstruction by first deciding whether globe-sparing surgery is possible, or if orbital exenteration will be necessary. We then set critical and optimal objectives for our chosen pathway. Critical goals in globe-sparing reconstruction include maintaining orbital volume and preserving visual function, and an optimal goal includes preservation of the nasolacrimal system. Critical goals in orbital exenteration include obliterating the defect, sealing the skull base and nasal cavities, and allowing eye protection to be worn over the contralateral eye postoperatively. Optimal goals in exenteration include preparation for prosthetics, volume and bony replacement, eyelid-sparing technique, and consideration of postoperative radiation.


A DNA repair variant in POLQ (c.-1060A > G) is associated to hereditary breast cancer patients: a case-control study.
One of the hallmarks of cancer is the occurrence of high levels of chromosomal rearrangements as a result of inaccurate repair of double-strand breaks (DSB). Germline mutations in BRCA and RAD51 genes, involved in DSB repair, are strongly associated with hereditary breast cancer. Pol θ, a translesional DNA polymerase specialized in the replication of damaged DNA, has been also shown to contribute to DNA synthesis associated to DSB repair. It is noteworthy that POLQ is highly expressed in breast tumors and this expression is able to predict patient outcome. The objective of this study was to analyze genetic variants related to POLQ as new population biomarkers of risk in hereditary (HBC) and sporadic (SBC) breast cancer. We analyzed through case-control study nine SNPs of POLQ in hereditary (HBC) and sporadic (SBC) breast cancer patients using Taqman Real Time PCR assays. Polymorphisms were systematically identified through the NCBI database and are located within exons or promoter regions. We recruited 204 breast cancer patients (101 SBC and 103 HBC) and 212 unaffected controls residing in Southern Brazil. The rs581553 SNP located in the promoter region was strongly associated with HBC (c.-1060A > G; HBC GG = 15, Control TT = 8; OR = 5.67, CI95% = 2.26-14.20; p < 0.0001). Interestingly, 11 of 15 homozygotes for this polymorphism fulfilled criteria for Hereditary Breast and Ovarian Cancer (HBOC) syndrome. Furthermore, 12 of them developed bilateral breast cancer and one had a familial history of bilateral breast cancer. This polymorphism was also associated with bilateral breast cancer in 67 patients (OR = 9.86, CI95% = 3.81-25.54). There was no statistically significant difference of age at breast cancer diagnosis between SNP carriers and non-carriers. Considering that Pol θ is involved in DBS repair, our results suggest that this polymorphism may contribute to the etiology of HBC, particularly in patients with bilateral breast cancer.


End-stage renal disease: can dietary protein restriction prevent it?
The hypothesis that dietary protein restriction slows the rate of progression of chronic renal disease has been tested in four large clinical trials, the largest and most elegant of which was the Modification of Diet in Renal Disease (MDRD) study conducted recently in the United States. During these trials the majority of patients with nondiabetic renal disease had slow progression, and no overall benefit from dietary intervention was demonstrated in three of four studies. However, MDRD is the first trial to suggest that protein restriction has a two-stage effect on renal function. During the first 4 months, glomerular filtration rate (GFR) fell more rapidly in patients with protein restriction than in patients with normal protein intake, probably because of a hemodynamic effect on the renal microcirculation. After this 4-month period, the rate of decline in GFR was slower in patients on the low protein diet. This suggests but does not prove that low protein diet may slow progression of renal disease in the long run. MDRD also showed that strict control of hypertension slows progression in patients with heavy proteinuria.


Second extracellular protease mediating maturation of Vibrio mimicus hemolysin.
Vibrio mimicus is a bacterium that causes gastroenteritis in humans. This pathogen produces an enterotoxic hemolysin called V. mimicus hemolysin (VMH), which is secreted extracellularly as an inactive 80-kDa protoxin and converted to a 66-kDa mature toxin through cleavage between Arg151 and Ser152. The 56-kDa serine protease termed V. mimicus trypsin-like protease (VmtA) is known to mediate this maturating process. However, some strains including strain ES-20 does not possess the vmtA gene. In the present study, the vmtA-negative strains were found to have a replaced gene that encodes a 43-kDa (403 aa) precursor of a serine protease designated by VmtX (V. mimicus trypsin-like protease X). To examine whether VmtX is also involved in the maturation of VMH, VmtX was isolated from the culture supernatant of V. mimicus strain NRE-20, a metalloprotease-negative mutant constructed from strain ES-20. Concretely, the culture supernatant was fractionated with 70% saturated ammonium sulfate and subjected to affinity column chromatography using a HiTrap Benzamidine FF column. The analysis of the N-terminal amino acid sequences of the proteins in the obtained VmtX preparation indicated that the 39-kDa protein was active VmtX consisting of 371 aa (Ile33-Ser403). The VmtX preparation was found to activate pro-VMH through generation of the 66-kDa protein. Additionally, treatment of the VmtX preparation with serine protease inhibitors, such as leupeptin and phenylmethylsulfonyl fluoride, significantly suppressed the activities to hydrolyze the specific peptide substrate and to synthesize the 66-kDa toxin. These findings indicate that VmtX is the second protease that mediats the maturation of VMH.


Mannose-binding lectin polymorphisms and the risk of sepsis: evidence from a meta-analysis.
Several studies have evaluated the association between mannose-binding lectin (MBL) polymorphisms and sepsis. However, the results are inconclusive and conflicting. To better understand the roles of MBL polymorphisms in sepsis, we conducted a comprehensive meta-analysis. All relevant studies were searched from PubMed, EMBASE and Web of Knowledge databases, with the last report up to 7 May 2013. Twenty-nine studies addressing four MBL polymorphisms (-550G/C, -221G/C, structure variant A/O, Gly54Asp) were analysed for susceptibility to sepsis and one study for sepsis-related mortality. Overall, significant associations between structure variant A/O and susceptibility to sepsis were observed for AO + OO vs. AA [odds ratio (OR) 1·27, 95% confidence interval (CI) 1·05-1·52, P = 0·01] and O vs. A (OR 1·19, 95% CI 1·02-1·40, P = 0·03). In subgroup analysis based on age group, increased risk was found in the paediatric group in the dominant model (OR 1·72, 95% CI 1·16-2·56, P = 0·007). Moreover, there was a slight association between the +54A/B polymorphism and susceptibility to sepsis in Caucasians (recessive model: OR 10·64, 95% CI 1·24-91·65, P = 0·03). However, no association was observed for -550G/C and -221G/C polymorphisms both overall and in subgroup analysis. For sepsis-related mortality, only one study suggested AO/OO was associated with in-hospital mortality in pneumococcal sepsis patients after controlling for confounding variables. Our meta-analysis indicated that MBL structure variants might be associated with susceptibility to sepsis but further studies with a large sample size should be conducted to confirm these findings.


Substrate specificity of retinyl ester hydrolase activity in retinal pigment epithelium.
In the eye, hydrolysis of stored retinyl esters is catalyzed by retinyl ester hydrolase (REH) activities in retinal pigment epithelium (RPE) membranes. In the present study, biochemical analyses were conducted to determine the substrate specificity of these activities. Specific activities determined for hydrolysis of various retinol isomers of retinyl palmitate (9-cis-, 11-cis-, 13-cis-, and all-trans-retinyl palmitates) indicated that 11-cis-retinyl palmitate is preferentially hydrolyzed (1.7 nmol/min/mg) compared to the other isomers (0.1-0.3 nmol/min/mg). Examination of the specificity of REH activity for 11-cis-retinyl esters of varied acyl chain length (-myristate, -palmitate, and -stearate) and degree of saturation (-oleate and -linoleate) further demonstrated that palmitate is the preferred fatty acyl moiety. Notably, retinyl esters possessing chain lengths which more closely approximate that of the palmitate ester exhibited higher rates of hydrolysis. Similar results were obtained in retinyl ester-plasma membrane fusion studies in which hydrolysis took place within the membrane domain rather than at the lipid-water interface. REH substrate specificity was further assessed in competition studies in which 11-cis-retinyl palmitate hydrolysis was monitored in the presence of 13-cis-, 9-cis-, or all-trans-retinyl palmitate. Results show that addition of these retinyl palmitate isomers does not affect the rate of hydrolysis of 11-cis-retinyl palmitate. However, the hydrolytic rates associated with other retinyl palmitate isomers were significantly reduced in the presence of 11-cis-retinyl palmitate. Finally, cholesterol ester hydrolase activity was found to be distinct from the observed 11-cis-REH activity and the presence of cholesterol oleate did not affect the rate of 11-cis-retinyl palmitate hydrolysis. Collectively, these data support the hypothesis that a distinct, membrane-associated, 11-cis-retinyl palmitate-specific retinyl ester hydrolase activity exists in the retinal pigment epithelium.


CD56-positive acute lymphoblastic leukemia.
We report a patient with lung cancer who developed CD56-positive acute lymphoblastic leukemia. He was referred to our hospital for thrombocytopenia. Atypical cells were found in the blood and the bone marrow. These cells were immunophenotypically positive for CD3epsilon, CD56, and terminal deoxynucleotidyl transferase, and negative for surface CD3, CD4, CD19, CD33, and myeloperoxidase. A small proportion of leukemic cells express CD13. There were no rearrangements of T-cell receptor (TCR)-beta, TCR-gamma, or immunoglobulin heavy chain. No Epstein-Barr virus was detected. Systemic examination did not detect any tumors other than pulmonary adenocarcinoma, and the patient was diagnosed as having acute natural killer (NK) cell leukemia. Chemotherapy was effective, and he achieved complete remission. The course of the disease was complicated by a lung abscess, and the patient died 3 months after the diagnosis. We considered that the diagnosis was blastic NK cell lymphoma/leukemia subtype. However, it actually was myeloid/NK cell precursor leukemia subtype that weakly expressed CD13.


Vertebral hydatid disease.
Hydatid disease affecting the bones is a rare condition, its incidence ranging from 0.5-2% of all cases of hydatidosis. The bones most often involved are the vertebrae (44%). A 22-year-old man presented to the hospital with low back pain. A CT scan revealed intracanal and paravertebral cystic lesions at the level of L5-S1 with destruction of the corpus and lamina, and sacral foramen invasion. An operation was performed and histopathological findings confirmed the diagnosis. In countries where hydatidosis is endemic, the possibility of the disease must be borne in mind.


Contribution of circulating maternal lipids to fetal tissues in the guinea pig.
Using a constant infusion of [1-14C] palmitic acid into 7 late pregnant guinea-pigs the contribution of circulating maternal lipid to the fetuses was assessed. Within the mother plasma radioactivity was found mainly in free fatty acid with only a small amount circulating as triacylglycerol. In contrast in the fetal plasma [14C] was distributed as one-quarter free fatty acid and three-quarters triacylglycerol, with total [14C] levels being four-fold higher than in the maternal plasma after 160 min of infusion. [14C] Triacylglycerol was found in the fetal liver within 10 min of commencing infusion and levels rose to 12 times the levels of [14C] triacylglycerol in the fetal plasma (per/g or ml) by 100-160 min. [14C] Distribution into the lipid moieties of fetal tissues was found to change with time. By 100-160 min the distribution was: liver, triacylglycerol 83%, phospholipid 7%, free fatty acid 4%; fat, triacylglycerol 80%; heart, triacylglycerol 38%, phospholipid 33%, free fatty acid 9%; kidney, triacylglycerol 15%, phospholipid 58%, free fatty acid 10%; muscle, triacylglycerol 70%, phospholipid 10%, free fatty acid 10%; lung, phospholipid 75%; brain, phospholipid 80%. It was concluded that free fatty acid transferred from the mother contributes substantially to fetal lipids in late gestation in this species.


Interorganizational Sensemaking of the Transition Toward a Circular Value Chain.
The transition toward the circular economy requires stakeholders to collaborate along value chains. Yet, such collaborations are considerably challenging. Given the paradigmatic change, stakeholders face high levels of uncertainty and also need to align on a common way forward. We extend research on interorganizational sensemaking and the circular economy by exploring the process of interorganizational alignment in a European consortium of over 150 companies representing the value chain for flexible packaging with the objective to transform the value chain from linear to circular. We find that the interorganizational sensemaking process unfolds across three levels-organization, value chain, and ecosystem-which provide different reference frames for the process. We provide insights into how these frames, power dynamics, and identity considerations influence this process. Our findings highlight the importance of considering interdependencies between stakeholders and a collective reconceptualization of the established value chain to successfully transition toward a circular one.


How to Effectively Decrease Patient Co-Payments of High-Cost Drugs Through Innovation: Lessons From the Karmanos Specialty Pharmacy.
High-cost drugs impose a financial burden on patients with cancer. Karmanos Specialty Pharmacy (KSP) developed a process to automate financial assistance (FA) applications to decrease patient drug cost. We evaluate the outcomes of this program on cost to patients and payers. This is an observational, retrospective study of the KSP claims data set from January to December 2019, accessed by 13 statewide cancer centers within Michigan. Drug cost of patients, payers, FA (funds to lower patient drug cost), and types of FA were obtained. A subset analysis was performed to determine drug delivery times. In 2019, 869 prescriptions and 1,722 prescription fills were provided to 463 patients through KSP. The total cost of drug claims was approximately $10 million US dollars (USD) among Medicare patients (58%), approximately $3.4 million USD for privately insured patients (20%), and approximately $3.7 million USD for Medicaid patients (22%). Twenty-seven percent of patients (22% of all prescription fills) required additional FA with initial total co-payment claims of $335,216 USD. $280,988 USD of FA was obtained, which substantially lowered total patient costs by 81%. $250,818 USD of FA obtained was from foundation grants (327 fills), and $21,441 USD from manufacturer co-pay cards (47 fills). An additional $12,260 USD (12 fills) from a Karmanos Patient Assistance Fund was used. There was high dependence on foundation grant assistance among Medicare patients (33% of claims). In a subset analysis, the median time from prescription written to delivery to the patient was < 7 days (0-56 days). Twenty-seven percent of patients (22% of prescriptions fills) in 2019 required additional FA for high-cost drugs. KSP substantially reduced patient cost by implementing an efficient process using additional pharmacy assistants to obtain FA.


Androgen receptor gene CAG repeat polymorphism and breast cancer risk in Iranian women: a case-control study.
We investigated the association between polymorphic expansion of trinucleotide CAG repeats in androgen receptor (AR) gene and breast cancer risk among Iranian women in a matched case-control study. There was a strong overall association between per CAG repeat increments in average repeat length and the risk of the malignancy [OR=3.56; 95% CI, 2.80-5.29]. Women carrying one or two alleles with [CAG]n repeat ≥22 units were at increased risk of breast cancer [OR=2.03; 95% CI, 1.56-2.6]. The risk was significantly increased in homozygous longer repeats, versus homozygous alleles <22. We observed reduced risk of developing the tumor in positive familial breast cancer subjects carrying repeats ≥22 and 23. Homozygosity for the longer [CAG]n repeats may be linked to the increased breast cancer risk. In contrast to previous reports, longer AR [CAG]n repeat alleles may decline the risk among women with a familial breast cancer.


Methicillin and Vancomycin Resistant Staphylococcus aureus and Associated Factors from Surgical Ward Inpatients at Debre Markos Referral Hospital, Northwest Ethiopia.
Staphylococcus aureus is a well-known superbug and leading causes of wound infections. The clinical epidemiology of methicillin resistant S. aureus (MRSA) and vancomycin resistant S. aureus (VRSA) is not well documented in Ethiopia. The aim of this study was to determine the proportion of MRSA, VRSA and associated factors from surgical inpatients in Debre Markos Referral Hospital (DMRH), Northwest Ethiopia. A facility-based cross-sectional study was conducted from February to April 2020 at DMRH. A structured questionnaire was used to gather demographic and clinical data. Wound swab was collected from inpatients and then inoculated on blood agar and mannitol salt agar. The presence of MRSA and VRSA was determined using the cefoxitin (30 μg) antibiotic disk diffusion and vancomycin E-test methods, respectively. The data were analyzed using SPSS 20. Data were analyzed using bivariate and multivariate logistic regression analysis. A P-value <0.05 was considered statistically significant. A total of 242 wound cases were enrolled and the majority of them were males 172 (71.1%). Among the total enrolled cases, S. aureus was identified from 71 (29.3%) of the admitted patients. The proportion of MRSA was 32 (13.22%) and that of VRSA was 4.1%. The proportion of vancomycin intermediate S. aureus (VISA) was gauged at 4.5%. Hospital stay over 72 hrs, wound depth, current antibiotic use, and previous history of wound infection showed statistically significant association with MRSA. On the contrary, VRSA did not showed any significant association against the analyzed variables. High proportions of S. aureus isolates became MRSA; resistant to all β-lactam antimicrobial agents excluding newer cephalosporin. In addition, the proportion of VRSA/VISA was also high. Multiple variables demonstrated significant associations with MRSA. Hence, intervention measures for MRSA risk groups must be in place. Furthermore, hospital infection control and an antibiotic stewardship program should be strengthened.


Contrasting Strategies for Sucrose Utilization in a Floral Yeast Clade.
Yeast species in the Wickerhamiella and Starmerella genera (W/S clade) thrive in the sugar-rich floral niche. We have previously shown that species belonging to this clade harbor an unparalleled number of genes of bacterial origin, among which is the SUC2 gene, encoding a sucrose-hydrolyzing enzyme. In this study, we used complementary in silico and experimental approaches to examine sucrose utilization in a broader cohort of species representing extant diversity in the W/S clade. Distinct strategies and modes of sucrose assimilation were unveiled, involving either extracellular sucrose hydrolysis through secreted bacterial Suc2 or intracellular assimilation using broad-substrate-range α-glucoside/H+ symporters and α-glucosidases. The intracellular pathway is encoded in two types of gene clusters reminiscent of the MAL clusters in Saccharomyces cerevisiae, where they are involved in maltose utilization. The genes composing each of the two types of MAL clusters found in the W/S clade have disparate evolutionary histories, suggesting that they formed de novo. Both transporters and glucosidases were shown to be functional and additionally involved in the metabolization of other disaccharides, such as maltose and melezitose. In one Wickerhamiella species lacking the α-glucoside transporter, maltose assimilation is accomplished extracellularly, an attribute which has been rarely observed in fungi. Sucrose assimilation in Wickerhamiella generally escaped both glucose repression and the need for an activator and is thus essentially constitutive, which is consistent with the abundance of both glucose and sucrose in the floral niche. The notable plasticity associated with disaccharide utilization in the W/S clade is discussed in the context of ecological implications and energy metabolism. IMPORTANCE Microbes usually have flexible metabolic capabilities and are able to use different compounds to meet their needs. The yeasts belonging to the Wickerhamiella and Starmerella genera (forming the so-called W/S clade) are usually found in flowers or insects that visit flowers and are known for having acquired many genes from bacteria by a process called horizontal gene transfer. One such gene, dubbed SUC2, is used to assimilate sucrose, which is one of the most abundant sugars in floral nectar. Here, we show that different lineages within the W/S clade used different solutions for sucrose utilization that dispensed SUC2 and differed in their energy requirements, in their capacity to scavenge small amounts of sucrose from the environment, and in the potential for sharing this resource with other microbial species. We posit that this plasticity is possibly dictated by adaptation to the specific requirements of each species.


Relationships between lymphocyte counts and treatment-related toxicities and clinical responses in patients with solid tumors treated with PD-1 checkpoint inhibitors.
The relationships between absolute lymphocyte counts (ALC), drug- related toxicities, and clinical responses remain unclear in cancer patients treated with PD-1 (programmed cell death 1) inhibitors. We performed a retrospective review of 167 adult solid tumor patients treated with nivolumab or pembrolizumab at a single institution between January 2015 and November 2016. Patients with an ALC >2000 at baseline had an increased risk of irAE (OR 1.996, p<0.05) on multivariate analysis. In a multivariate proportional hazards model, a shorter time to progression was noted in patients who were lymphopenic at baseline (HR 1.45 (p<0.05)) and at three months (HR 2.01 (p<0.05)). Patients with baseline lymphopenia and persistent lymphopenia at month 3 had a shorter time to progression compared to those who had baseline lymphopenia but recovered with ALC > 1000 at 3 months (HR 2.76, p<0.05). Prior radiation therapy was the characteristic most strongly associated with lymphopenia at 3 months (OR 2.24, p<0.001). These data suggest that patients with higher baseline lymphocyte counts have a greater risk for irAE, whereas patients with lymphopenia at baseline and persistent lymphopenia while on therapy have a shorter time to progression on these agents. These associations require further validation in additional patient cohorts.


Significance of the suspensory and collateral ligaments in lesser metatarsal neck surgery.
In this study the authors have endeavored to define more closely the distinct nature of the collateral and suspensory ligaments of the lesser metatarsophalangeal joints and to propose a mechanism through which the ligaments confer stability to the metatarsal head. The lesser metatarsal neck dorsal V-osteotomy is one surgical procedure that has the potential of disrupting the intracapsular ligaments, depending on the placement of the "V" cut. The angulation of the "V" cut redirects the capital fragment in the sagittal plane while maintaining stability in the transverse and frontal planes. The authors contend that the sagittal stability of the free capital fragment after a V-osteotomy is maintained, as well, by the attachment of the newly defined suspensory ligaments and the collateral ligaments of the metatarsophalangeal joint. Disruption of these ligaments, such as occurs when placing the apex of the V-osteotomy proximally, serves to increase the instability within the metatarsal head, ultimately leading to a potential increase in number of complications.


[Monitoring calcium in outer hair cells with confocal microscopy and fluorescence ratios of fluo-3 and fura-red].
Calcium distribution and mobilization during mechanical stimulation in outer hair cells of the guinea pig were monitored using laser scanning confocal microscopy and co-loaded fluo-3 and fura-red fluorescent probes. Spatial calcium gradients were revealed among various subcellular areas. The ratios of the fluorescence intensity of fluo-3 and fura-red were 1.71 +/- 0.85, 1.61 +/- 0.75, 1.47 +/- 0.65 and 1.39 +/- 0.66 for the cytoplasm, the cytoplasmic membrane, the cuticular plate and the nucleus respectively, indicating that free calcium ion concentrations are the highest in the cytoplasm and the lowest in the nucleus. While the calcium concentration remained relatively constant under resting conditions, it increased during mechanical stimulation. The results show that confocal ratio imaging of fluo-3 and fura-red enables us to determine more accurately the subcellular calcium distribution and that the calcium ions make a contribution to the mechanic-electrical transduction in hair cells.


A convenient synthesis of beta-D-galactosyl disaccharide derivatives using the beta-D-galactosidase from Bacillus circulans.
beta-D-Gal-(1-->4)-beta-D-GlcNAc-OC6H4NO2-p (p-nitrophenyl N-acetyl-beta-lactosaminide) and beta-D-Gal-(1-->6)-beta-D-GlcNAc-OC6H4NO2-p (p-nitrophenyl N-acetyl-beta-isolactosaminide) were regioselectively synthesized from lactose and p-nitrophenyl 2-acetamido-2-deoxy-glucopyranoside, employing transglycosylation by the beta-D-galactosidase from Bacillus circulans and by controlling the concentration of organic solvent in the reaction system. The (1-->4)-linked disaccharide was formed exclusively when the concentration of organic solvent was high, whereas the (1-->6)-linked isomer was produced with a low concentration. Further utilization of the transglycosylation by the enzyme led to the regioselective formation of beta-D-Gal-(1-->4)-D-GalNAc and beta-D-Gal-(1-->4)-beta-D-GalNAc-OC6H4NO2-p. With the enzyme, beta-D-galactosyl transfer occurred preferentially at the O-4 position of GlcNAc and GalNAc, regardless of the configuration of the hydroxyl group.


The G-protein-coupled bile acid receptor Gpbar1 (TGR5) suppresses gastric cancer cell proliferation and migration through antagonizing STAT3 signaling pathway.
Gpbar1 (TGR5), a membrane-bound bile acid receptor, is well known for its roles in regulation of energy homeostasis and glucose metabolism. Here we show that TGR5 is a suppressor of gastric cancer cell proliferation and migration through antagonizing STAT3 signaling pathway. We firstly show that TGR5 activation greatly inhibited proliferation and migration of human gastric cancer cells and strongly induced gastric cancer cell apoptosis. We then found that TGR5 activation antagonized STAT3 signaling pathway through suppressing the phosphorylation of STAT3 and its transcription activity induced by lipopolysaccharide (LPS) or interleukin-6. TGR5 overexpression with ligand treatment inhibited gene expression mediated by STAT3. It suggests that TGR5 antagonizes gastric cancer proliferation and migration at least in part by inhibiting STAT3 signaling. These findings identify TGR5 as a suppressor of gastric cancer cell proliferation and migration that may serve as an attractive therapeutic tool for human gastric cancer.


Sulfur dioxide inhalation stimulated mitochondrial biogenesis in rat brains.
Sulfur dioxide (SO(2)) is a common environmental pollutant. Mitochondria play essential roles in energy metabolism, generation of reactive oxygen species, and regulation of apoptosis in response to neuronal brain injury. It is of interest to observe the effect of SO(2) on mitochondrial function in brain. In the present study, male Wistar rats were housed in exposure chambers and treated with 3.5, 7 and 14mg/m(3) SO(2) for 4h/day for 30days, while control rats were exposed to filtered air in the same condition. Mitochondrial membrane potential (MMP) was assessed in cerebral mitochondria using the lipophilic cationic probe JC-1. The amount of ATP was measured by the luciferinluciferase method. Analyses of mitochondrial replication and transcription were performed by real time PCR. The protein levels were detected using Western blotting. Our results showed that cerebral mtDNA content was markedly increased in rats after SO(2) exposure. Paralleling the change in mtDNA content, MMP, ATP content, MDA level, CO1 & 4 and ATP6 & 8 expression, and cytochrome c oxidase activity were increased in rat cortex after SO(2) inhalation. Moreover, mitochondrial biogenesis was accompanied by increased expression of NRF1 and TFAM, whereas PGC-1α was not changed. We report for the first time increased mitochondrial biogenesis in brain of rats exposed to SO(2), which might be an adaptive response to mitochondrial depletion by oxidant damage.


Adolescent but not adult ethanol binge drinking modulates cocaine withdrawal symptoms in mice.
Ethanol (EtOH) binge drinking is an increasingly common behavior among teenagers that induces long-lasting neurobehavioral alterations in adulthood. An early history of EtOH abuse during adolescence is highly correlated with cocaine addiction in adulthood. Abstinence of cocaine abuse can cause psychiatric symptoms, such as anxiety, psychosis, depression, and cognitive impairments. This study assessed the consequences of adolescent exposure to EtOH on the behavioral alterations promoted by cocaine withdrawal in adulthood. We pretreated juvenile (34-47 days old) or adult (68-81 days old) mice with EtOH (1.25 g/kg) following a binge-drinking pattern. Then, after a three-week period without drug delivery, they were subjected to a chronic cocaine treatment in adulthood and tested under cocaine withdrawal by the ensuing paradigms: open field, elevated plus maze, prepulse inhibition, tail suspension test, and object recognition. Another set of mice were treated with the same EtOH binge-drinking procedure during adolescence and were tested immediately afterwards under the same behavioral paradigms. Adolescent EtOH pretreatment undermined the anxiogenic effects observed after cocaine abstinence, reduced prepulse inhibition, and increased immobility scores in the tail suspension test following cocaine withdrawal. Moreover, the memory deficits evoked by these substances when given separately were enhanced in cocaine-withdrawn mice exposed to EtOH during adolescence. EtOH binge drinking during adolescence also induced anxiety, depressive symptoms, and memory impairments when measured immediately afterwards. In contrast, neither EtOH nor cocaine alone or in combination altered any of these behaviors when given in adulthood. EtOH binge drinking induces short- and long-term behavioral alterations and modulates cocaine withdrawal symptoms when given in adolescent mice.


Detection of zonal flow spectra in DIII-D by a dual-channel Doppler backscattering system.
Doppler backscattering (DBS) has been successfully used to measure the E x B flow velocity and local intermediate wavenumber density fluctuation levels in the DIII-D tokamak. Depending on the launch angle and the frequency of the probing beam, the signal backscattered from the plasma cut-off layer is sensitive to density fluctuations at a specific perpendicular wavenumber (1 < or = k(perpendicular rho(s)) < or = 4). Due to the localization and high time resolution for poloidal flow measurements, DBS is well suited to detect stationary and time-dependent shear flows [zonal flows (ZFs)]. We present a novel scheme to measure ZF spectra using a dual-channel DBS system capable of simultaneously probing two minor radii separated by a distance of 0.2 cm < Delta r < 3 cm. Frequency spectra of geodesic acoustic modes and low frequency ZFs (f or = 10 kHz) have been obtained for 0.6 < r/a < 0.95.


[Ocular Behçet].
Our study aimed to determine the clinical, therapeutic and prognostic features of ocular involvement in patients with Behçet's disease treated in our Department of ophthalmology. We conducted a retrospective data collection from medical records of 20 patients treated at the military hospital in Laayoune. All patients underwent complete ophthalmological examination and fluorescein angiography if necessary. OCT exam was performed in two patients. Ten patients had anterior uveitis, complicated in one case by ocular hypertonia; two patients had intermediate uveitis; eight patients had posterior segment involvement complicated in one case by intravitreal hemorrhage. Behcet's Disease (BD) is an systemic idiopathic inflammatory disease currently classified within primary non-necrotizing vasculitis. Ocular involvement is common and severe in Behçet's disease, with the potential to compromise the visual prognosis. Behcet's disease is common in Morocco. It can compromise patient's visual prognosis making the collaboration between ophthalmologists and internists particularly important.


Electric properties tomography: Biochemical, physical and technical background, evaluation and clinical applications.
Electric properties tomography (EPT) derives the patient's electric properties, i.e. conductivity and permittivity, using standard magnetic resonance (MR) systems and standard MR sequences. Thus, EPT does not apply externally mounted electrodes, currents or radiofrequency (RF) probes, as is the case in competing techniques. EPT is quantitative MR, i.e. it yields absolute values of conductivity and permittivity. This review summarizes the physical equations underlying EPT, the corresponding basic and advanced reconstruction techniques and practical numerical aspects to realize these reconstruction techniques. MR sequences which map the field information required for EPT are outlined, and experiments to validate EPT in phantom and in vivo studies are described. Furthermore, the review describes the clinical findings which have been obtained with EPT so far, and attempts to understand the physiologic background of these findings.


Emotional Well-Being Following a Later Life Career Change: The Roles of Agency and Resources.
More individuals are attempting career changes in later life, as an increasing number of people face precarious retirement prospects. Although many of these older job seekers eventually find new livelihoods, little is known about their emotional well-being subsequent to these changes. Using the 2014 American Institute for Economic Research Older Worker Survey, we evaluate the contributions of demographic characteristics, agency, and resources when estimating three measures of emotional well-being following a successful later life career change ( n = 337). We found that having financial resources during the career transition was associated with all three positive emotional outcomes, while family support and intentionality were also associated with positive emotions. Conversely, prior job prestige and purposeful job training had no relationships with subsequent well-being. These results suggest that later life career changes, despite their challenges, often result in positive emotional outlooks-for those who have the resources to support them.


The synaptic vesicle cycle.
The ins and outs of the synaptic vesicle cycle are being examined in increasing detail with diverse investigative tools in a variety of cell types, particularly those with large granules. The cycle begins with the opening of a fusion pore that connects the vesicle lumen to the extracellular fluid. Sensitive electrophysiological techniques reveal the often-stuttering behavior of single pores in non-neuronal cells, through which small molecules trickle until the fusion pore expands and the remaining contents erupt from the vesicle. The granule membranes are then retrieved by multiple processes that appear to act in parallel and that are distinguished from each other kinetically and ultrastructurally. Following endocytosis, synaptic vesicles are then shuttled back into the vesicle pool, where they briefly mix with other vesicles, become immobilized, and remain gelled with their neighbors, even while moving en masse again to the presynaptic membrane as a prelude for another round of exocytosis.


Stress response of Porphyromonas gingivalis.
The heat shock response of Porphyromonas gingivalis was examined by 1- and 2-dimensional polyacrylamide gel electrophoresis after metabolic labeling with [14C]-amino acids. When P. gingivalis cells were shifted from 37 degrees C to 42 degrees C, elevated synthesis of 4 proteins with the apparent molecular weight of 92, 80, 74, and 62 kDa was observed, and synthesis of a 50-kDa protein decreased during heat shock. The 74- and 62-kDa proteins were identified as homologs of Escherichia coli DnaK and GroEL respectively by Western immunoblotting. On 2-dimensional Western blots, 2 forms of DnaK and 4 forms of GroEL were identified due to their slightly different isolelectric points. dnaK and groEL gene homologs were identified in several P. gingivalis strains and some other black-pigmented oral species. DnaK and GroEL homolog proteins were induced when P. gingivalis cells were challenged by ethanol. These proteins were not induced by oxidative stress or change in pH.


Religious activity improves life satisfaction for some physicians and older patients.
To assess religious perceptions and activities of physicians and older patients and to determine whether religious activities are associated with life satisfaction. A cross-sectional survey of practicing Virginia internists and psychiatrists and hospitalized or institutionalized (nursing home) older adults. One hundred randomly sampled practicing physicians and 55 hospitalized or institutionalized older patients. A mailed survey was used for the physicians and a structured interview for the patients. All subjects provided information pertaining to demographics and life satisfaction using the Life Satisfaction Index (LSI-B). For physicians or patients who engaged in any religious activity, the Intrinsic/Extrinsic Religiosity (I/E-R) scale was used. Of the 100 physicians (49 internists and 51 psychiatrists) who answered the survey (50% response rate), 75% used religious activity as a coping resource (39% somewhat, 36% definitely). There was a positive correlation between intrinsic religious activity (e.g., prayer, Bible reading) and life satisfaction (r = .293, P = .042). Of the 55 patients interviewed, 47 (86%) used religion as a coping resource, and intrinsic religious activity was positively associated with life satisfaction (r = .843, P < .001). Even after controlling for age, gender, health, and marital status, intrinsic religious activity remained a predictor of higher life satisfaction. Intrinsic religious activity is associated positively with life satisfaction in physicians and ill older adults.


Anogenital Ulcers: An Unusual Manifestation of Invasive Aspergillosis.
Aspergillus spp. is one of the most ubiquitous fungi but generally does not cause disease in immunocompetent patients. It is the second most frequent agent of opportunistic fungal infections, after Candida albicans, with a rise in its incidence on recent years. Invasive fungal diseases represent an important cause of morbidity and mortality. Its origin can be primary, in relation to a cutaneous injury, or secondary, by extension from contiguous tissues, or by hematogenous spread, usually in the context of pulmonary aspergillosis. In this report, we describe the case of an elderly woman with invasive aspergillosis that manifested with anogenital and skin ulcers, with unfavorable outcome, despite intensive therapy and intravenous antifungals.


Perspectives in the reconstruction of paediatric facial paralysis.
In this review article, aimed at those likely to encounter but relatively unfamiliar with paediatric facial paralysis, we summarize reconstructive options in its surgical management. The article looks at static and dynamic reconstructions available to address the problems encountered in the different anatomical regions of paralyzed face. Although multiple surgical options are available for each anatomical region, decision-making should be taken in a multidisciplinary environment, and take into account the individual requirements of the family and patients.


Identification of kazinol Q, a natural product from Formosan plants, as an inhibitor of DNA methyltransferase.
DNA methylation plays a pivotal role in the epigenetic regulation of the transcription of a number of cancer-related genes, thereby representing an important target for cancer prevention and treatment. In our search for DNA methyltransferase (DNMT) inhibitors from Formosan plants, by screening against a library consisting of 12 structurally distinct natural products, we identified kazinol Q {4-[6-(1,1-dimethyl-allyl)-7-hydroxy-chroman-2-yl]-3,6-bis-(3-methyl-but-2-enyl)-benzene-1,2-diol} as an inhibitor of recombinant DNMT1 with IC50 of 7 μM. The effect of kazinol Q on DNMT inhibition was validated by its ability to reactivate the expression of a DNA methylation-silenced gene, E-cadherin, in MDA-MB-231 breast cancer cells. Moreover, kazinol Q suppressed the proliferation of MCF-7 breast and LNCaP prostate cancer cells, in part, through apoptosis induction. The role of DNMT1 inhibition in mediating kazinol Q's antiproliferative effect was supported by the protective effect of ectopic expression of DNMT1 on kazinol Q-induced cell death. Molecular modeling analysis suggests that kazinol Q inhibited DNMT activity by competing with cytosine binding, a mechanism similar to that described for (-)-epigallocatechin-3-gallate (EGCG). Relative to EGCG, kazinol Q exhibits several desirable features for drug development, including chemical stability and increased hydrophobicity, and might have therapeutic relevance to cancer treatment.


Antimicrobial resistance of Shigella isolated in New York City in 1973.
One hundred and two Shigella sonnei and fourteen S. flexneri strains isolated from individual patients in New York City hospitals were investigated for antibiotic resistance. The S. sonnei showed 60% resistance to ampicillin and 58% to tetracycline and streptomycin. S. flexneri showed no ampicillin resistance but 50% resistance to tetracycline. There were marked differences in resistance of S. sonnei between hospitals which were not explainable by ethnic or socio-economic differences in the patient populations.


Epithelial ingrowth after femtosecond laser-assisted in situ keratomileusis.
To describe the incidence and clinical features of epithelial ingrowth after femtosecond laser-assisted in situ keratomileusis (LASIK). All eyes with clinically significant epithelial ingrowth were identified from patients who underwent primary femtosecond laser-assisted LASIK and their subset of patients who underwent enhancement procedures from January 2005 to March 2007 at Kudret Eye Hospital. Clinical course in terms of severity, type of epithelial ingrowth, time of appearance, effect on visual acuity, and treatment was described for each patient. Two eyes of 6415 eyes after primary procedure and 2 eyes of 108 eyes after enhancement procedure developed clinically significant epithelial ingrowth. All eyes were treated with surgical removal of epithelial ingrowth. The epithelial ingrowths in all the patients were seen as solitary islands of cells, with a size ranging between 1 and 3 mm, beneath the flap with no connection with the surface epithelium. There was recurrence of ingrowth in 1 eye. Visually significant epithelial ingrowth after femtosecond laser-assisted LASIK is an infrequent complication. The lower incidence of epithelial ingrowth after femtosecond LASIK surgery compared with mechanical microkeratome-assisted LASIK may be attributed to the anatomy of the femtosecond laser-created side cut in contrast to that created with a mechanical microkeratome and the creation of less peripheral trauma at the time of flap creation.


Sedentary Occupation Workers Who Meet the Physical Activity Recommendations Have a Reduced Risk for Metabolic Syndrome: A Cross-Sectional Study.
We tested the hypothesis that sedentary occupation workers who meet the physical activity recommendations present a lower risk for metabolic syndrome (MetS) than their nonactive counterparts. A cross-sectional study involving 502 sedentary occupation workers. Physical activity level was self-reported. MetS was defined by International Diabetes Federation criteria. The active group showed lower odds for MetS [odds ratio (OR) 0.52, 95% confidence interval (95% CI) 0.27 to 0.98], abdominal obesity (OR 0.36, 95% CI 0.16 to 0.82), elevated blood pressure (OR 0.47, 95% CI 0.26 to 0.84), and reduced high-density lipoprotein cholesterol (OR 0.54, 95% CI 0.31 to 0.93) than the sedentary group after adjustments for age, time in job, body mass index, and tobacco use. Sedentary occupation workers who meet the physical activity recommendations have a reduced risk for MetS.


Human herpesvirus-8 (HHV-8) sero-detection and HIV association in Kaposi's sarcoma (KS), non-KS tumors and non-neoplastic conditions.
The association of the human herpesvirus-8/Kaposi's sarcoma (KS)-associated herpesvirus (HHV-8/KSHV) serology with various malignancies in Tanzania is not currently well established while previous studies were based on either PCR or immunofluorescence assays [IFA] but not with a sensitive enzyme-linked immunosorbent assay (ELISA). Selected archival diagnostic biopsies (n = 184) and sera from indigenous patients with KS (n = 120), non-KS tumors (n = 24) and non-neoplastic lesions (n = 40) at Muhimbili National Hospital (MNH), Tanzania, were evaluated by diagnostic histopathology, immunohistology [anti-HHV-8 latency-associated nuclear antigen (LANA)] and serology for HIV (ELISA) and HHV-8 (IFA and ELISA). About 66.3% (n = 122) cases including AIDS-associated Kaposi's sarcoma (AKS) (n = 93), reactive conditions (n = 28) and only one non-KS tumour were HIV positive. Endemic KS (EKS) patients were mostly males (96.3%, 26/27) who were less (69.9%, 65/93) predominant in AIDS-associated (AKS). A high (89%) percentage of patients with anti-HHV-8 antibodies was found in the cohort including the HIV positive (92%) cases, males (81.2%), KS patients (93%), non-KS tumors (92%), and reactive conditions (75%). All HHV-8 seronegative KS cases were nodular stage whereas both sera and corresponding biopsies from early stage KS were HHV-8+. Assay sensitivity, positive predictive value (PPV) and specificity were 98.6%, 93.5% and 16.7% for IFA and 93.5%, 98.6% and 50.0% for ELISA respectively. HHV-8 seroprevalence at MNH appears high as expected among AKS cases and males but also in non-KS patients. ELISA showed a combination of high HHV-8 sensitivity as well as higher PPV and specificity than IFA which however, showed higher sensitivity. The apparent stage-dependent, inverted serum HHV-8 immunoreactivity supports a notion of viral immune-segregation during KS development. Routine HHV-8 screening should be considered particularly in patients at risk of KS and for selection of blood/organ donations.


Involvement of rapid nucleotide synthesis in recovery from phosphate starvation of Catharanthus roseus cells.
Growth of suspension-cultured Catharanthus roseus cells ceased during phosphate starvation, but the cells grew again upon addition of Pi even after long-term starvation. The metabolic fate of [(33)P]Pi was studied in 1-week-old stationary phase cells in ordinary culture and in 1- or 2-week-old Pi-starved cells. Immediately after administration, the most heavily labelled organic compounds are nucleotides, followed by sugar phosphates. Two weeks Pi starvation slowed down the speed of incorporation of (33)P into nucleotides. The RNA, protein, and free nucleotide content all decreased gradually during Pi starvation; however, these compounds, especially nucleotides, increased markedly in the 24 h after addition of Pi. These responses are found in all cells examined, although the total amounts of these compounds were lower in the long-term Pi-deficient cells. Of the nucleotides, a marked increase was observed in nucleoside triphosphates and UDP-glucose. The transcript level of phosphate transporter and the activities of acid phosphatase, 5'- and 3'-nucleotidase, and adenosine nucleosidase were all reduced by the addition of Pi. In contrast, the activities of adenine phosphoribosyltransferase, nicotinate phosphoribosyltransferase, and nicotinamidase, which are salvage enzymes of purine and pyridine nucleotides, were markedly increased in the Pi-fed cells. Little or no increase was observed in adenosine kinase. In the light of these results, the possible involvement of net nucleotide synthesis in the initial metabolic events of recovery from Pi deficiency are discussed.


Generation of Newly Discovered Resistance Gene mcr-1 Knockout in Escherichia coli Using the CRISPR/Cas9 System.
The mcr-1 gene is a new "superbug" gene discoverd in China in 2016 that makes bacteria highly resistant to the last-resort class of antibiotics. The mcr-1 gene raised serious concern about its possible global dissemination and spread. Here, we report a potential anti-resistant strategy using the CRISPR/Cas9-mediated approach that can efficiently induce mcr-1 gene knockout in Escherichia coli. Our findings suggested that using the CRISPR/Cas9 system to knock out the resistance gene mcr-1 might be a potential anti-resistant strategy. Bovine myeloid antimicrobial peptide-27 could help deliver plasmid pCas::mcr targeting specific DNA sequences of the mcr-1 gene into microbial populations.


[Secretory immunoglobulin A (S-IgA)/immunoglobulin A (IgA) ratio in cervico-vaginal lavage fluids].
Investigation of the immunoglobulins in the vaginal lavage fluids are semiquantitatively. The aim of the present study was to complete the semiquantitative S-IgA estimations with an "S-IgA/IgA" ratio. An antiserum against the secretory component (Anti-SC) and an S-IgA standard as well as an antiserum against the immunoglobulin A(Anti-IgA) and a serum IgA standard were used. The ratio was proved in vaginal lavage fluids from 5 fertile and 2 infertile women in the course of the menstrual cycle on days 8th, 11th, 14th, 18th and 24th. Progesterone, IgG and IgA were estimated simultaneously in serum samples. According to the "S-IgA/IgA" ratio patients were divided in 2 groups with values higher than 1 on the one side and with values lower than 1 on the other side. In cases with high "S-IgA/IgA" ratio an increased local antibody production and secretion is supposed.


Metabolic control in newly diagnosed type 1 diabetic children. Effect of continuous subcutaneous infusion.
15 insulin-dependent diabetic children at onset were randomly allocated to one of two different therapeutical protocols: continuous subcutaneous insulin infusion (CSII) and intensified conventional insulin treatment with three daily insulin injections (CIT). Both treatments were performed for 10 days; the initial insulin dose was 1.5 U/kg/day and thereafter the insulin dosage was modified in order to obtain a satisfactory control. Near-normal blood glucose levels were obtained after 24 h in the CSII group, and after 3 days in the CIT group. All subjects underwent 1 year of follow-up. HbA1 levels and insulin requirements decreased similarly in the two groups; C-peptide secretion did not increase significantly in both groups. A clear advantage of CSII cannot be assumed, and the usefulness of this therapeutical approach needs to be confirmed by further investigations.


A Rare Variant Identified Within the GluN2B C-Terminus in a Patient with Autism Affects NMDA Receptor Surface Expression and Spine Density.
NMDA receptors (NMDARs) are ionotropic glutamate receptors that are crucial for neuronal development and higher cognitive processes. NMDAR dysfunction is involved in a variety of neurological and psychiatric diseases; however, the mechanistic link between the human pathology and NMDAR dysfunction is poorly understood. Rare missense variants within NMDAR subunits have been identified in numerous patients with mental or neurological disorders. We specifically focused on the GluN2B NMDAR subunit, which is highly expressed in the hippocampus and cortex throughout development. We analyzed several variants located in the GluN2B C terminus and found that three variants in patients with autism (S1415L) or schizophrenia (L1424F and S1452F) (S1413L, L1422F, and S1450F in rodents, respectively) displayed impaired binding to membrane-associated guanylate kinase (MAGUK) proteins. In addition, we observed a deficit in surface expression for GluN2B S1413L. Furthermore, there were fewer dendritic spines in GluN2B S1413L-expressing neurons. Importantly, synaptic NMDAR currents in neurons transfected with GluN2B S1413L in GluN2A/B-deficient mouse brain slices revealed only partial rescue of synaptic current amplitude. Functional properties of GluN2B S1413L in recombinant systems revealed no change in receptor properties, consistent with synaptic defects being the result of reduced trafficking and targeting of GluN2B S1413L to the synapse. Therefore, we find that GluN2B S1413L displays deficits in NMDAR trafficking, synaptic currents, and spine density, raising the possibility that this mutation may contribute to the phenotype in this autism patient. More broadly, our research demonstrates that the targeted study of certain residues in NMDARs based on rare variants identified in patients is a powerful approach to studying receptor function.SIGNIFICANCE STATEMENT We have used a "bedside-to-bench" approach to investigate the functional regulation of NMDA receptors (NMDARs). Using information from deep sequencing of patients with neurological or psychiatric disorders, we investigated missense variants identified in the intracellular C-terminal domain of the GluN2B NMDAR subunit. We found several variants that displayed altered properties. In particular, one variant identified in a patient with autism, human GluN2B S1415L, displayed reduced surface expression and binding to PSD-95. Furthermore expression of GluN2B S1415L (S1413L in mouse) showed a deficit in rescue of synaptic NMDAR currents and fewer dendritic spines, consistent with other reports of spine abnormalities being associated with autism. More broadly, we demonstrate that using patient data is an effective approach to probing the structure/function relationship of NMDARs.


Characterization of microsatellite loci in fulvous fruit bat Rousettus leschenaulti.
Rousettus leschenaulti is an abundant species in many countries of South-East Asia, including south China. We isolated seven microsatellite loci in R. leschenaulti from genomic DNA enriched for CA repeats with the enriched library method. A total of 56 samples from a population in the Guangxi Province of China were tested with these microsatellite markers. The polymorphism ranged from seven to 16 alleles, and the observed heterozygosity was 84-94%. It is the first time microsatellite markers were characterized from R. leschenaulti, and these markers can be an important tool for analysing population structure and genotypic diversity.


Hepatitis E virus antibodies in HIV-infected patients and blood donors from western Poland: A preliminary report.
Hepatitis E virus (HEV) infection is an emerging problem in industrialized countries, including Europe. Little data exists on HEV seroprevalence in Poland. The aim of this study was to assess the prevalence of anti-HEV IgG antibodies in Polish patients infected with the human immunodeficiency virus (HIV) and blood donors. Two hundred and ten individuals (n = 105 of HIV-infected patients and n = 105 of ageand sex-matched blood donors from the same area; 178 men and 32 women), aged 18-50 (median age: 38 years), were tested for the presence of anti-HEV IgG antibodies with the EUROIMMUN Anti-Hepatitis E Virus (HEV) ELISA (IgG) tests (Lübeck, Germany). Additionally, some simple clinical and laboratory data was collected. The overall anti-HEV IgG prevalence was 2.4% (5/210). One HIV-positive patient (0.95%) and 4 blood donors (3.8%) were seropositive (p = 0.1745). All the HEV-exposed individuals were men with a history of travel abroad and no icteric disease in the past. Exposure to HEV infection among Polish HIV patients and blood donors seems to be uncommon. Data on this issue is scarce and conflicting for HIV-infected individuals. Further investigations applying different serological tests and concomitant HEV RNA testing are needed to reliably assess the risk and practical impact of HEV infection in Poland.


Restriction fragment length polymorphisms at the growth hormone gene in pigs.
Pigs from four Danish and two Swedish populations were examined for restriction fragment length polymorphism (RFLP) at the growth hormone (GH) gene. Polymorphism was detected with the restriction enzymes DraI and TaqI. A comparison of the Danish populations showed significant differences among their allelic frequencies.


NLRP3 and CARD8 Polymorphisms Influence Higher Disease Activity in Rheumatoid Arthritis.
The activation of NLRP3-inflammasome may contribute to inflammatory processes in rheumatoid arthritis (RA). Functional polymorphisms in the genes coding for its components NLRP3 and CARD8 were associated with a proinflammatory phenotype. Our aim was to investigate the influence of these polymorphisms on RA susceptibility and disease activity at the time of diagnosis and after six months of treatment. A group of 128 RA patients treated with methotrexate and 122 healthy controls were genotyped for NLRP3 rs35829419 (p. Q705K) and CARD8 rs2043211 (p. C10X) polymorphisms. RA susceptibility was not influenced by the investigated polymorphisms or their interaction. The investigated polymorphisms explained 8% of variability in DAS28 at the time of diagnosis. Carriers of NLRP3 rs35829419 or CARD8 rs2043211 polymorphisms had significantly higher DAS28 at the time of diagnosis (p=0.003; p=0.022; respectively). Polymorphic CARD8 rs2043211 TT genotype was also associated with higher DAS28 after six months of treatment (p=0.033). Genetic variability of inflammasome components may contribute to higher disease activity at the time of diagnosis and after 6 months of methotrexate treatment in RA patients. Better understanding of the immunological mechanisms behind a more active course of RA may suggest novel treatment approaches in a subset of patients with a proinflammatory phenotype.


Neural Processing of Threat Cues in Young Children With Attention-Deficit/Hyperactivity Symptoms.
A growing literature indicates that attention deficit/hyperactivity disorder (ADHD) involves difficulty processing threat-related emotion faces. This deficit is especially important to understand in young children, as threat emotion processing is related to the development of social skills and related behavioral regulation. Therefore, the current study aimed to better understand the neural basis of this processing in young children with ADHD symptoms. Forty-seven children between 4 and 7 years of age were included in the analysis, 28 typical developing and 19 with clinically significant levels of ADHD hyperactive/impulsive symptoms. Participants completed a passive affective face-viewing task. Event-related potentials were assessed for each emotion, and parental report of child behavior and emotion regulation abilities was assessed. Children with ADHD symptoms showed altered N170 modulation in response to specific emotion faces, such that the N170 was less negative in response to fearful compared to neutral faces, whereas typically developing children showed the opposite pattern. Groups did not differ in reactivity to anger or non-threat-related emotion faces. The N170 difference in fearful compared to neutral faces correlated with reported behavior, such that less fear reactivity predicted fewer prosocial behaviors. Abnormalities in the underlying neural systems for fear processing in young children with ADHD symptoms may play an important role in social and behavioral deficits within this population.


Asymptotic current-voltage relations for currents exceeding the diffusion limit.
We consider the one-dimensional transport of ions into a perm-selective solid. Direct attempts to evaluate the current-voltage characteristics for currents exceeding the diffusion limit are frustrated by the appearance of nonconverging integrals. We describe how to overcome this obstacle using a regularization scheme.


Retrospective comparison of nab-paclitaxel plus ramucirumab and paclitaxel plus ramucirumab as second-line treatment for advanced gastric cancer focusing on peritoneal metastasis.
Background Ramucirumab (RAM) plus solvent-based (sb)-paclitaxel (PTX) is the standard second-line chemotherapy for advanced gastric cancer (AGC). The subset analysis of the ABSOLUTE trial, which confirmed non-inferiority of weekly nanoparticle albumin-bound (nab)-PTX to weekly sb-PTX, suggested that nab-PTX might have better efficacy than sb-PTX in patients with peritoneal metastasis. We retrospectively evaluated the efficacy and safety of RAM plus sb-PTX and nab-PTX in patients with peritoneal metastasis of AGC. Methods AGC patients who received RAM plus sb-PTX or nab-PTX as second-line chemotherapy from June 2015 to February 2019 were included in the study. Overall survival (OS), progression-free survival (PFS), response rate, and safety were assessed. Results A total of 128 patients were included in this study (93 in the RAM plus sb-PTX group and 35 in the RAM plus nab-PTX group). PFS was 4.1 months in the RAM plus sb-PTX group and 4.6 months in the RAM plus nab-PTX group (HR 0.90; 95%CI 0.58-1.41, p = 0.643). OS was 8.9 months in the RAM plus sb-PTX group and 11.4 months in the RAM plus nab-PTX group (HR 0.95; 95%CI 0.56-1.62, p = 0.847). A total of 62 and 31 patients had peritoneal metastasis in the RAM plus sb-PTX and the RAM plus nab-PTX groups, respectively. RAM plus nab-PTX showed a slightly longer survival compared to RAM plus sb-PTX in patients with peritoneal metastasis (PFS 5.8 vs 3.5 months, HR 0.66; 95% CI 0.40-1.10, p = 0.109). Conclusion This study suggests that RAM plus nab-PTX might be a more effective treatment for peritoneal metastasis of AGC.


Telomerase activity and serum levels of p53 protein as prognostic factors of survival in patients with advanced non-small cell lung cancer.
Evaluation of relationships between survival time of patients with advanced, non-resectable non-small cell lung cancer (NSCLC) and telomerase activity in aspirates, collected from primary lung tumours, and serum p53 protein levels. The study group consisted of 52 patients with advanced (stage IIIB and IV) non-small cell lung cancer. In all of them, transthoracic fine-needle biopsy (TFNB) of focal pulmonary lesion was performed. The aspirates were subjected to telomerase activity by the PCR-ELISA PLUS method and serum levels of p53 protein were determined by the ELISA method. Additionally, clinical advancement of cancer and the time period of survival were assessed in the studied group. Kaplan-Meyer method and Cox analysis were used for statistical evaluation of survival prognosis. Increased telomerase activity was observed in 42 (81%) of the patients with non-resectable non-small cell lung cancer. Elevated concentrations of serum p53 protein were found in 28 (54%) of the participants. The following death rates were noted during the entire study period: twenty-three (23) (62%), out of 37 patients with increased telomerase activity, 7 (47%), out of 15 without detectable telomerase activity in primary lung tumour, 16 (57%), out of 28 subjects with increased serum levels of p53 protein and 14 (58%), out of 24 with no increased serum levels of p53. A significant relationship was observed in Cox hazard analysis between the time of survival and telomerase activity, while no such relationship was observed between the survival time period and serum p53 protein levels or sex, age, primary lung tumour size, lymph node status or development of distant metastases. Telomerase activity in advanced primary non-small cell lung cancer is a better predictor of patients' survival than serum levels of p53 protein. The assessment of telomerase activity supplements in the prognosis of survival in the course of non-resectable NSCLC.


Bird orientation: a method of study.
A psychophysical technique has been applied to the study of bird orientation. The technique enables precise determinations of the accuracy with which a bird can orient itself. Data indicate that a pigeon can detect angular displacement as little as 3.4 degrees.


Continuous cell suspension processing using magnetically stabilized fluidized beds.
The behavior of a cell suspension in a continuous magnetically stabilized fluidized bed (MSFB) was investigated both experimentally and theoretically. The low, constant pressure drop and fluidity of the solids phase in the MSFB allowed a continuous countercurrent separator to be constructed. The magnetic field eliminated all motion of the solids phase (nickel spheres) and produced a device similar to a packed-bed depth filter. Yeast cells were used as the suspended solids and the performance of the MSFB filter was assessed as a function of the bed height, solids velocity, cell concentration, and liquid composition. Removal rates could be adjusted by controlling the cell/support interaction and were found to be as high as 99%. A mathematical model was used to aid in understanding this filtration and was found to agree qualitatively with all experimental observations. Comparison of the model with the data suggests that both cell/cell binding and cell shadowing are occurring.


Synthesis and biological evaluation of nonpeptide mimetics of omega-conotoxin GVIA.
A benzothiazole-derived compound (4a) designed to mimic the C(alpha)-C(beta) bond vectors and terminal functionalities of Lys2, Tyr13 and Arg17 in omega-conotoxin GVIA was synthesised, together with analogues (4b-d), which had each side-chain mimic systematically truncated or eliminated. The affinity of these compounds for rat brain N-type and P/Q-type voltage gated calcium channels (VGCCs) was determined. In terms of N-type channel affinity and selectivity, two of these compounds (4a and 4d) were found to be highly promising, first generation mimetics of omega-conotoxin. The fully functionalised mimetic (4a) showed low microM binding affinity to N-type VGCCs (IC(50)=1.9 microM) and greater than 20-fold selectivity for this channel sub-type over P/Q-type VGCCs, whereas the mimetic in which the guanidine-type side chain was truncated back to an amine (4d, IC(50)= 4.1 microM) showed a greater than 25-fold selectivity for the N-type channel.


Association of Adiponectin with High-Sensitivity C-Reactive Protein and Clinical Outcomes in Peritoneal Dialysis Patients: A 3.5-Year Follow-Up Study.
Adiponectin (ADPN), one of most abundant fat-derived biologically active substances, plays an important role in anti-atherosclerotic process. There are conflicting results about the impact of ADPN on cardiovascular (CV) outcomes and mortality, particularly in patients undergoing peritoneal dialysis (PD). Moreover, the relationship between ADPN and inflammatory mediators has been seldom explored in this population. Therefore, we examined the relationship between ADPN and longitudinal high-sensitivity C-reactive protein (hs-CRP) changes and investigated whether ADPN or hs-CRP levels could predict CV outcomes and mortality in prevalent PD patients after comprehensive adjustment of possible confounders. In this prospective cohort study, 78 PD patients were enrolled and followed from February 2009 to August 2012. During follow-up, CV events and all-cause mortality were recorded. The mean baseline ADPN value was 29.46±18.01 μg/ml and duration of PD treatment was 37.76±36.96 months. In multiple linear regression analysis, plasma ADPN levels positively correlated with high-density lipoprotein and negatively associated with hs-CRP, body mass index, D4/D0 glucose, triglyceride, and duration of PD treatment. After stratified by genders, the inverse association between baseline ADPN and hs-CRP was more significant in the female group. The hs-CRP levels were followed up annually and remained significantly lower in the high ADPN group in the first 2 years. Patients were then stratified into two groups according to the median ADPN value (23.8 μg/ml). The results of Kaplan-Meier survival analysis demonstrated less CV events and better survival in high ADPN group. On multivariate Cox regression analysis, only ADPN level (HR: 0.93, 95% CI: 0.88-0.98, p = 0.02), age and history of CV diseases were independent risk factors for future CV events. Furthermore, hs-CRP (HR: 1.11, 95% CI:1.001-1.22, p = 0.04) was identified as independent predictor of all-cause mortality. Serum hs-CRP levels were consistently lower in the high ADPN group during 2-year follow-up. We also demonstrated the importance of ADPN and hs-CRP in predicting CV events and all-cause mortality in PD population during 3.5-year follow-up.


Telomeric site position heterogeneity in macronuclear DNA of Paramecium primaurelia.
In Paramecium primaurelia, the macronuclear gene encoding the G surface protein is located near a telomere. In this study, multiple copies of this telomere have been isolated and the subtelomeric and telomeric regions of some of them have been sequenced. The telomeric sequences consist of tandem repeats of the hexanucleotides C4A2 or C3A3. We show that the location where these repeats are added, which we call the telomeric site, is variable within a 0.6-0.8-kb region. These results are discussed in relation with the formation of macronuclear DNA.


RAF inhibitors that evade paradoxical MAPK pathway activation.
Oncogenic activation of BRAF fuels cancer growth by constitutively promoting RAS-independent mitogen-activated protein kinase (MAPK) pathway signalling. Accordingly, RAF inhibitors have brought substantially improved personalized treatment of metastatic melanoma. However, these targeted agents have also revealed an unexpected consequence: stimulated growth of certain cancers. Structurally diverse ATP-competitive RAF inhibitors can either inhibit or paradoxically activate the MAPK pathway, depending whether activation is by BRAF mutation or by an upstream event, such as RAS mutation or receptor tyrosine kinase activation. Here we have identified next-generation RAF inhibitors (dubbed 'paradox breakers') that suppress mutant BRAF cells without activating the MAPK pathway in cells bearing upstream activation. In cells that express the same HRAS mutation prevalent in squamous tumours from patients treated with RAF inhibitors, the first-generation RAF inhibitor vemurafenib stimulated in vitro and in vivo growth and induced expression of MAPK pathway response genes; by contrast the paradox breakers PLX7904 and PLX8394 had no effect. Paradox breakers also overcame several known mechanisms of resistance to first-generation RAF inhibitors. Dissociating MAPK pathway inhibition from paradoxical activation might yield both improved safety and more durable efficacy than first-generation RAF inhibitors, a concept currently undergoing human clinical evaluation with PLX8394.


Extended vomer flaps in cleft palate repair: a preliminary report.
It is the author's belief that poor healing due to deficient mesenchyme at the junction of the hard and soft palates probably constitutes the essential reason for some failures of palatoplasty. In order to provide extra tissue to reinforce the above-mentioned region and to facilitate efficient velar lengthening, the use of an extension of the vomer flap detached from the roof of the pharynx is proposed. Such extended vomer flaps have been applied successfully both on the oral and nasal sides in 50 patients. The necessity of further and longer observations is obvious.


Multivariate classification of smokers and nonsmokers using SVM-RFE on structural MRI images.
Voxel-based morphometry (VBM) studies have revealed gray matter alterations in smokers, but this type of analysis has poor predictive value for individual cases, which limits its applicability in clinical diagnoses and treatment. A predictive model would essentially embody a complex biomarker that could be used to evaluate treatment efficacy. In this study, we applied VBM along with a multivariate classification method consisting of a support vector machine with recursive feature elimination to discriminate smokers from nonsmokers using their structural MRI data. Mean gray matter volumes in 1,024 cerebral cortical regions of interest created using a subparcellated version of the Automated Anatomical Labeling template were calculated from 60 smokers and 60 nonsmokers, and served as input features to the classification procedure. The classifier achieved the highest accuracy of 69.6% when taking the 139 highest ranked features via 10-fold cross-validation. Critically, these features were later validated on an independent testing set that consisted of 28 smokers and 28 nonsmokers, yielding a 64.04% accuracy level (binomial P = 0.01). Following classification, exploratory post hoc regression analyses were performed, which revealed that gray matter volumes in the putamen, hippocampus, prefrontal cortex, cingulate cortex, caudate, thalamus, pre-/postcentral gyrus, precuneus, and the parahippocampal gyrus, were inversely related to smoking behavioral characteristics. These results not only indicate that smoking related gray matter alterations can provide predictive power for group membership, but also suggest that machine learning techniques can reveal underlying smoking-related neurobiology.


Utilization of an Escherichia coli mutant for carbon-13 enrichment of tRNA for NMR studies.
The enrichment of tRNA at specific sites with carbon-13 has been accomplished in vivo using a mutant of Escherichia coli. A relaxed strain of E. coli auxotrophic for methionine was grown in a specifically defined medium supplemented with either [14C] or [13C]-methyl labeled methionine. Cells were collected at the end of the log-phase of growth and tRNA was extracted. Analysis of the radioactivity of the [14C]-labeled tRNA established an incorporation ratio of three labeled carbons per tRNA molecule. Incorporation of the [14C]-label in vivo was confined to the methylation of nucleotides as determined by thin layer chromatography of nucleotides resulting from a ribonuclease digestion of [14C]-labeled tRNA. The carbon-13 NMR spectrum of [13C]-enriched tRNA indicated a similar degree of incorporation into the methylated nucleotides by the substantial enhancement of [13C]-methyl NMR signals only. Assignment of signals has been made for the methyl groups of ribothymidine and N7-methylguanosine in E. coli tRNA.


Changes in microbial community during biohydrogen production using gamma irradiated sludge as inoculum.
The changes in microbial community structures during fermentative hydrogen production process were investigated by analyzing 16S rDNA gene sequences using gamma irradiated sludge as inoculum. The experimental results showed that the microbial community structure of untreated sludge was very rich in diversity. After gamma irradiation, lots of species were inhibited, and species with high survival rates under radiation conditions became dominant. After fermentation, Clostridium butyrium and a sequence closely related to Clostridium perfringens ATCC 13124(T) (CP000246) became predominant, which were all common hydrogen producers. Microbial distribution analysis indicated that gamma irradiation was a good pretreatment method for enriching hydrogen-producing strains from digested sludge.