A comprehensive mathematical model of drug release kinetics from nano-liposomes, derived from optimization studies of cationic PEGylated liposomal doxorubicin formulations for drug-gene delivery.
This study focuses on the development of a universal mathematical model for drug release kinetics from liposomes to allow in silico prediction of optimal conditions for fine-tuned controlled drug release. As a prelude for combined siRNA-drug delivery, nanoliposome formulations were optimized using various mole percentages of a cationic lipid (1,2-dioleoyl-3-trimethylammonium-propane, DOTAP) in the presence or absence of 3 mol% distearoyl phosphoethanolamine, polyethylene glycol (PEG-2000mDSPE). Outcome parameters were particle size, zeta potential, entrapment efficiency, in vitro drug release, and tumor cell kill efficiency. The optimized formula (containing 20% DOTAP with 3% DSPE-mPEG(2000) was found to be stable for six months, with round-shaped particles without aggregate formation, an average diameter of 71 nm, a suitable positive charge, and 89% drug encapsulation efficiency (EE). The 41% drug release during 6 h confirmed controlled release. Furthermore, the release profiles as functions of pH and temperature were investigated and the kinetics of the drug release could adequately be fitted to Korsmeyer-Peppas' model by multiple regression analysis. The statistical parameters confirmed good conformity of final models. Functionality of the novel cationic liposome formulations (± DOX) was tested on osteosarcoma (OS) cell lines. Increased OS cell toxicity (1.3-fold) was observed by the DOX-loaded vs. the free DOX. A feasibility pilot showed that siRNA could be loaded efficiently as well. In conclusion, we have established a predictive mathematical model for the fine-tuning of controlled drug release from liposomal formulations, while creating functional drug-delivery liposomes with potential for siRNA co-delivery to increase specificity and efficacy.


Cancers of the maxillary sinus.
Maxillary sinus cancers are rare as compared with cancers of other sites. Because the onset is insidious, they are at an advanced stage at diagnosis and therefore present a therapeutic challenge for local control. Traditionally, treatment has been surgery or irradiation, or a combination of the two. Local control and survival, however, remains poor. Numerous chemotherapeutic agents have shown activity against squamous cell cancers of the head and neck. In several studies the use of either neoadjuvant or concurrent chemotherapy has improved local control and survival when compared with historical controls. Randomized studies are needed to confirm the benefit of chemotherapy over the standard forms of surgical and radiation therapy.


Desolvation and aggregation of sterically demanding alkali metal diarylphosphides.
The reaction between (Dipp)2PH and one equivalent of n-BuLi, PhCH2Na or PhCH2K in THF gives the complexes [(Dipp)2P]Li(THF)3 (2a), {[(Dipp)2P]Na(THF)2}2 (3a) and [(Dipp)2P]K(THF)4 (4a), respectively [Dipp = 2,6-iPr2C6H3]. Exposure of these compounds to vacuum yields the alternative solvates [(Dipp)2P]Li(THF)2 (2b), [(Dipp)2P]Na(THF)1.5 (3b), and [(Dipp)2P]K (4b), respectively; the alternative adduct [(Dipp)2P]Na(PMDETA) (3c) was prepared by treatment of 3a with PMDETA. Treatment of (Dipp)(Mes)PH or (Mes)2PH with one equivalent of n-BuLi in THF gives the complexes [(Dipp)(Mes)P]Li(THF)3 (7a) and [(Mes)2P]2Li2(THF)2(OEt2) (8a) after crystallisation from diethyl ether [Mes = 2,4,6-Me3C6H2]; crystallisation of 8a from hexane gives the alternative adduct [(Mes)2P]Li(THF)3 (8b). Exposure of 7a, 8a and 8b to vacuum leads to loss of coordinated solvent, yielding the solvates [(Dipp)(Mes)P]Li(THF)2 (7b) and [(Mes)2P]Li(THF) (8c). The solid-state structures of complexes 2a, 3a, 3c, 4a, 7a, 8a, and 8b have been determined by X-ray crystallography. Variable-temperature 31P{1H} and 7Li NMR spectroscopy indicates that 2b, 3b and 7b are subject to a monomer-dimer equilibrium in solution, where the monomeric forms are favoured at low temperature. In contrast, variable-temperature 31P{1H} and 7Li NMR spectroscopy suggests that 8c is subject to a dynamic equilibrium between a dimer and a cyclic trimer in solution, where the trimer is favoured at low temperatures.


In vivo imaging of chronic active lesions in multiple sclerosis.
New clinical activity in multiple sclerosis (MS) is often accompanied by acute inflammation which subsides. However, there is growing evidence that a substantial proportion of lesions remain active well beyond the acute phase. Chronic active lesions are most frequently found in progressive MS and are characterised by a border of inflammation associated with iron-enriched cells, leading to ongoing tissue injury. Identifying imaging markers for chronic active lesions in vivo are thus a major research goal. We reviewed the literature on imaging of chronic active lesion in MS, focussing on 'slowly expanding lesions' (SELs), detected by volumetric longitudinal magnetic resonance imaging (MRI) and 'rim-positive' lesions, identified by susceptibility iron-sensitive MRI. Both SELs and rim-positive lesions have been found to be prognostically relevant to future disability. Little is known about the co-occurrence of rims around SELs and their inter-relationship with other emerging techniques such as dynamic contrast enhancement (DCE) and positron emission tomography (PET).


The bovine herpesvirus-1 LR ORF2 is critical for this gene's ability to restore the high wild-type reactivation phenotype to a herpes simplex virus-1 LAT null mutant.
During neuronal latency of herpes simplex virus (HSV)-1, the latency-associated transcript (LAT) is the only viral gene readily detectable. LAT is required for the high-level reactivation phenotype in animal models. LAT's anti-apoptotic activity was recently demonstrated by our group and it was proposed that LAT's anti-apoptotic function is involved in enhancing the reactivation phenotype. Recently, using chimeric virus CJLAT, it was shown that the reactivation phenotype of LAT(-) mutant dLAT2903 can be restored to wild-type levels by inserting the bovine herpes virus (BHV)-1 latency-related (LR) gene into the LAT locus of this HSV-1 LAT deletion mutant. Although transcription of the LR gene, like LAT, inhibits apoptosis, LR appears to be multifunctional. To investigate whether the LR gene's anti-apoptotic function was responsible for restoring the high-reactivation phenotype, a mutated BHV-1 LR gene was inserted into the LAT locus of HSV-1 generating the chimeric virus CJLATmut. This mutation consists of three stop codons inserted just after the ATG of the first LR open reading frame (ORF2). In plasmids and in a BHV-1 mutant, this mutation eliminated the LR gene's anti-apoptotic activity, strongly suggesting that ORF2 encodes a protein responsible for LR's anti-apoptotic activity. Reactivation of the CJLATmut virus, in both rabbits and mice, was significantly lower than in wild-type McKrae virus (P=0.0001 and P=0.0003, respectively) and CJLAT virus, containing wild-type LR in place of LAT (P<0.0001) and was similar to LAT(-) dLAT2903 (P=0.8 and P=0.7, respectively). Thus, disruption of BHV-1 LR ORF2 eliminated the high-reactivation phenotype.


MS-Simulator: predicting y-ion intensities for peptides with two charges based on the intensity ratio of neighboring ions.
For the identification of peptides with tandem mass spectrometry (MS/MS), many software tools rely on the comparison between an experimental spectrum and a theoretically predicted spectrum. Consequently, the accurate prediction of the theoretical spectrum from a peptide sequence can potentially improve the peptide identification performance and is an important problem for mass spectrometry based proteomics. In this study a new approach, called MS-Simulator, is presented for predicting the y-ion intensities in the spectrum of a given peptide. The new approach focuses on the accurate prediction of the relative intensity ratio between every two adjacent y-ions. The theoretical spectrum can then be derived from these ratios. The prediction of a ratio is a closed-form equation that involves up to five consecutive amino acids nearby the two y-ions and the two peptide termini. Compared with another existing spectrum prediction tool MassAnalyzer, the new approach not only simplifies the computation, but also improves the prediction accuracy.


Cationic 5,10,15,20-tetrakis(N-methylpyridinium-4-yl)porphyrin fully intercalates at 5'-CG-3' steps of duplex DNA in solution.
The interaction of 5,10,15, 20-tetrakis(N-methylpyridinium-4-yl)porphyrin (T4MPyP(4+)) with the oligonucleotide DNA duplex [d(GCACGTGC)](2) was studied by two-dimensional (1)H NMR spectroscopy, optical absorbance, circular dichroism, and molecular dynamics simulation employing particle mesh Ewald methods. T4MPyP(4+) is one of the largest aromatic molecules for which intercalative binding to DNA has been proposed, although this has been called into question by recent X-ray crystallographic work [Lipscomb et al. (1996) Biochemistry 35, 2818-2823]. T4MPyP(4+) binding to [d(GCACGTGC)](2) produced a single set of (mostly) upfield-shifted DNA resonances in slow exchange with the resonances of the free DNA. Intra- and intermolecular NOEs observed in the complex showed that the porphyrin intercalates at the central 5'-CG-3' step of the DNA duplex without disrupting the flanking base pairs. Absorption and circular dichroism spectra of the complex also support intercalative binding. Molecular dynamics simulations (using explicit solvent and PME methods), carried out for fully and partially intercalated complexes, yielded stable trajectories and plausible structures, but only the symmetrical, fully intercalated model agreed with NOESY data. Stable hydrogen bonding was observed during 600 ps of MD simulation for the base pairs flanking the binding site.


Association between Schistosoma mansoni infection and access to improved water and sanitation facilities in Mwea, Kirinyaga County, Kenya.
Schistosomiasis remains a public health problem in Central Kenya despite concerted control efforts. Access to improved water and sanitation has been emphasized as important control measures. Few studies have assessed the association between access to improved water sources and sanitation facilities with Schistosoma mansoni infection in different environmental settings. This study assessed the association between S. mansoni infection and household access to improved water sources and sanitation facilities in Mwea, Kirinyaga County, Kenya. A cross sectional study was conducted between the months of August and October 2017. A total of 905 household heads from seven villages were interviewed and their stool samples screened for S. mansoni using the Kato Katz technique. Comparisons of demographic factors by S. mansoni infection were tested for significance using the chi-square test (χ2) or the Fisher exact test for categorical variables. Variables associated with S. mansoni infection were analyzed using univariable analysis and the strength of the association measured as odds ratio (OR) using mixed effects logistic regression at 95% CI, with values considered significant at p < 0.05. The overall prevalence of S. mansoni was, 23.1% (95% CI: 20.5-26.0%), with majority of the infections being of light intensity. Rurumi village had the highest prevalence at 33.3%, with Kirogo village having the least prevalence at 7.0%. Majority (84.1%) of the households lacked access to improved water sources but had access to improved sanitation facilities (75%). Households with access to piped water had the lowest S. mansoni infections. However, there was no significant association between S. mansoni infections with either the main source of water in the household (Odds Ratio (OR) =0.782 (95% CI: 0.497-1.229) p = 0.285 or sanitation facilities (OR = 1.018 (95% CI: 0.705-1.469) p = 0.926. Our study suggests that S. mansoni is still a public health problem among all age groups in Mwea irrigation scheme, Kirinyaga County, Central Kenya. Majority of the households lacks access to improved water sources but have access to improved sanitation facilities. This study recommends initiatives to ensure adequate provision of improved water sources, and the inclusion of the adult community in preventive chemotherapy programs.


Stability of the delta-endotoxin gene from Bacillus thuringiensis subsp. kurstaki in a recombinant strain of Clavibacter xyli subsp. cynodontis.
Deletion of chromosomally inserted gene sequences from Clavibacter xyli subsp. cynodontis, a xylem-inhabiting endophyte, was studied in vitro and in planta. We found that nonreplicating plasmid pCG610, which conferred resistance to kanamycin and tetracycline and contained segments of C. xyli subsp. cynodontis genomic DNA, integrated into a homologous sequence in the bacterial chromosome. In addition, pCG610 contains two copies of the gene encoding the CryIA(c) insecticidal protein of Bacillus thuringiensis subsp. kurstaki HD73. Using drug resistance phenotypes and specific DNA probes, we found that the loss of all three genes arose both in vitro under nonselective conditions and in planta. The resulting segregants are probably formed by recombination between the repeated DNA sequences flanking pCG610 that resulted from the integration event into the chromosome. Eventually, segregants predominated in the bacterial population. The loss of the integrated plasmid from C. xyli subsp. cynodontis revealed a possible approach for decreasing the environmental consequences of recombinant bacteria for agricultural use.


Growth factor-free, peptide-functionalized gelatin hydrogel promotes arteriogenesis and attenuates tissue damage in a murine model of critical limb ischemia.
Critical limb ischemia (CLI) occurs when blood flow is restricted through the arteries, resulting in ulcers, necrosis, and chronic wounds in the downstream extremities. The development of collateral arterioles (i.e. arteriogenesis), either by remodeling of pre-existing vascular networks or de novo growth of new vessels, can prevent or reverse ischemic damage, but it remains challenging to stimulate collateral arteriole development in a therapeutic context. Here, we show that a gelatin-based hydrogel, devoid of growth factors or encapsulated cells, promotes arteriogenesis and attenuates tissue damage in a murine CLI model. The gelatin hydrogel is functionalized with a peptide derived from the extracellular epitope of Type 1 cadherins. Mechanistically, these "GelCad" hydrogels promote arteriogenesis by recruiting smooth muscle cells to vessel structures in both ex vivo and in vivo assays. In a murine femoral artery ligation model of CLI, delivery of in situ crosslinking GelCad hydrogels was sufficient to restore limb perfusion and maintain tissue health for 14 days, whereas mice treated with gelatin hydrogels had extensive necrosis and autoamputated within 7 days. A small cohort of mice receiving the GelCad hydrogels were aged out to 5 months and exhibited no decline in tissue quality, indicating durability of the collateral arteriole networks. Overall, given the simplicity and off-the-shelf format of the GelCad hydrogel platform, we suggest it could have utility for CLI treatment and potentially other indications that would benefit from arteriole development.


Evaluation of compounding accuracy and aseptic techniques for intravenous admixtures.
Intravenous admixtures containing potassium collected from three hospital pharmacies were analyzed for compounding accuracy, sterility and pyrogenicity. The study was performed in two stages. During stage I, pharmacists and technicians were not informed of the study, but during stage II they were informed. In each stage 10 samples were collected from each person in the two personnel groups, analyzed and the results compared between the two personnel groups and the two stages. Results of the study showed that without monitoring (stage I) pharmacists had a higher mean percent error and contamination level than technicians. With monitoring, however, pharmacists showed a lower mean percent error and contamination level than technicians. Both personnel groups showed a decline in their mean percent error in the second stage, but there were still 83 (39.5%) errors in compounding accuracy greater than +/- 6%. No positive results with the Limulus test for pyrogens were obtained. It is recommended that a planned program of quality control be instituted for the preparation of i.v. admixtures by both pharmacists and technicians.


Evolution and development of the mammalian cerebral cortex.
Comparative developmental studies of the mammalian brain can identify key changes that can generate the diverse structures and functions of the brain. We have studied how the neocortex of early mammals became organized into functionally distinct areas, and how the current level of cortical cellular and laminar specialization arose from the simpler premammalian cortex. We demonstrate the neocortical organization in early mammals, which helps to elucidate how the large, complex human brain evolved from a long line of ancestors. The radial and tangential enlargement of the cortex was driven by changes in the patterns of cortical neurogenesis, including alterations in the proportions of distinct progenitor types. Some cortical cell populations travel to the cortex through tangential migration whereas others migrate radially. A number of recent studies have begun to characterize the chick, mouse and human and nonhuman primate cortical transcriptome to help us understand how gene expression relates to the development and anatomical and functional organization of the adult neocortex. Although all mammalian forms share the basic layout of cortical areas, the areal proportions and distributions are driven by distinct evolutionary pressures acting on sensory and motor experiences during the individual ontogenies.


Optimizing chemotherapy and targeted agent combinations in NSCLC.
Chemotherapy extends life and provides symptom palliation for patients with advanced non-small cell lung cancer (NSCLC). Numerous trials have been conducted that evaluate a variety of doublet regimens, but the majority of trials have found equal efficacy among the treatment arms. Indeed, a plateau appears to have been reached with respect to survival associated with traditional cytotoxic drug regimens. It was initially hoped that the addition of novel targeted agents to conventional chemotherapy would produce significant survival benefits for patients with advanced NSCLC; however, most trials have failed to show such a benefit. There is no survival benefit associated with adding erlotinib or gefitinib to a chemotherapy regimen, although there is a significant improvement in survival associated with erlotinib monotherapy in the second- and third-line advanced disease setting. In contrast, the results of E4599 clearly demonstrate that the addition of bevacizumab to paclitaxel-carboplatin chemotherapy extends survival in a select group of patients with non-squamous cell NSCLC. E4599 also represents a rational approach to drug development that could be modeled in other trials, namely, the use of a large, well designed, randomized trial prior to beginning a traditional phase II approach. This strategy can lead to the identification of subgroups most likely to benefit, as well as those that might experience increased toxicity, such as patients with squamous cell carcinoma treated with bevacizumab. Another approach to optimizing targeted therapy involves selecting a chemotherapy regimen with the greatest potential for synergy based on preclinical modeling. Because docetaxel has been shown to prolong survival in second-line treatment, a number of novel agents have been combined with docetaxel in order to improve efficacy. Alternatively, investigators have sought to combine novel agents with either carboplatin-paclitaxel or cisplatin-gemcitabine in first-line treatment. A number of trials are underway that combine these agents with inhibitors of the epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), and the proteasome, as well as COX2 inhibitors, and novel immunomodulators.


Omentum-wrapped scaffold with longitudinally oriented micro-channels promotes axonal regeneration and motor functional recovery in rats.
Tissue-engineered nerve scaffolds hold great potential in bridging large peripheral nerve defects. However, insufficient vascularization of nerve scaffolds limited neural tissues survival and regeneration, which hampered the successful implantation and clinical application of nerve scaffolds. The omentum possesses a high vascularization capacity and enhances regeneration and maturation of tissues and constructs to which it is applied. However, combined application of nerve scaffolds and omentum on axonal regeneration and functional recovery in the treatment of large peripheral nerve defects has rarely been investigated thus far. In the present study, an omentum-wrapped collagen-chitosan scaffold was used to bridge a 15-mm-long sciatic nerve defect in rats. Rats that received nerve autografts or scaffolds alone were served as positive control or negative control, respectively. The axonal regeneration and functional recovery were examined by a combination of walking track analysis, electrophysiological assessment, Fluoro-Gold (FG) retrograde tracing, as well as morphometric analyses to both regenerated nerves and target muscles. The results demonstrated that axonal regeneration and functional recovery were in the similar range between the omentum-wrapping group and the autograft group, which were significantly better than those in the scaffold alone group. Further investigation showed that the protein levels of vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were significantly higher in the omentum-wrapping group than those in the scaffold alone group in the early weeks after surgery. These findings indicate that the omentum-wrapped scaffold is capable of enhancing axonal regeneration and functional recovery, which might be served as a potent alternative to nerve autografts. The beneficial effect of omentum-wrapping on nerve regeneration might be related with the proteins produced by omentum.


Proteomic profiling of Mycobacterium tuberculosis identifies nutrient-starvation-responsive toxin-antitoxin systems.
In order to successfully enter the latent stage, Mycobacterium tuberculosis must adapt to conditions such as nutrient limitation and hypoxia. In vitro models that mimic latent infection are valuable tools for describing the changes in metabolism that occur when the bacterium exists in a non-growing form. We used two complementary proteomic approaches, label-free LC-MS/MS analysis and two-dimensional difference gel electrophoresis, to determine the proteome profile of extracellular proteins from M. tuberculosis cultured under nutrient starvation. Through the label-free LC-MS/MS analysis of fractionated samples, 1176 proteins were identified from culture filtrates of log phase and nutrient-starved cultures, and the protein levels of 230 proteins were increased in nutrient-starved culture filtrates, whereas those of 208 proteins were decreased. By means of Gene Ontology clustering analysis, significant differences in the overall metabolism during nutrient starvation were detected. Notably, members of the toxin-antitoxin systems were present in larger quantities in nutrient-starved cultures, supporting a role for these global modules as M. tuberculosis switches its metabolism into dormancy. Decreased abundance of proteins involved in amino acid and protein synthesis was apparent, as well as changes in the lipid metabolism. Further analysis of the dataset identified increased abundance of lipoproteins and decreased abundance of ESAT-6 family proteins. Results from the two-dimensional difference gel electrophoresis proteomics demonstrated overall agreement with the LC-MS/MS data and added complementary insights about protein degradation and modification.


Drawing the line: the cultural cartography of utilization recommendations for mental health problems.
In the 1990s, sociologists began to rethink the failure of utilization models to explain whether and why individuals accessed formal treatment systems. This effort focused on reconceptualizing the underlying assumptions and processes that shaped utilization patterns. While we have built a better understanding of how social networks structure pathways to care and how disadvantaged sociocultural groups face substantial barriers to treatment, we have less understanding of the larger cultural context in which individuals recognize and respond to symptoms. Drawing from recent innovations in the sociology of culture, we develop the concept of "cultural mapping" to describe if and how individuals discriminate among different available sources of formal treatment. Using data from the 1996 Mental Health Module of the General Social Survey, we compare Americans' willingness to recommend providers in the general medical and specialty mental health sectors. The results indicate that, despite unrealistically high levels of endorsement, individuals do discriminate between providers based on their evaluation of the problem, underlying causes, and likely consequences. While perceived severity leads individuals to suggest any type of formal care, problems attributed to biological causes are directed to general or specialty medical providers (doctors, psychiatrists, and hospitals); those matching symptoms for schizophrenia or seen as eliciting violence are allocated to the specialty mental health sector (psychiatry, mental hospital); and those seen as being caused by stress are sent to nonmedical mental health providers (i.e., counselors). These findings help to explain inconsistencies in previous utilization studies, and they suggest the critical importance of maintaining a dialogue between medical sociology and the sociology of culture.


Pediatric head and neck lesions: assessment of vascularity by MR digital subtraction angiography.
Pediatric head and neck lesions can be difficult to characterize on clinical grounds alone. We investigated the use of dynamic MR digital subtraction angiography as a noninvasive adjunct for the assessment of the vascularity of these abnormalities. Twelve patients (age range, 2 days to 16 years) with known or suspected vascular abnormalities were studied. Routine MR imaging, time-of-flight MR angiography, and MR digital subtraction angiography were performed in all patients. The dynamic sequence was acquired in two planes at one frame per second by using a thick section (6-10 cm) selective radio-frequency spoiled fast gradient-echo sequence and an IV administered bolus of contrast material. The images were subtracted from a preliminary mask sequence and viewed as a video-inverted cine loop. In all cases, MR digital subtraction angiography was successfully performed. The technique showed the following: 1) slow flow lesions (two choroidal angiomas, eyelid hemangioma, and scalp venous malformation); 2) high flow lesions that were not always suspected by clinical examination alone (parotid hemangioma, scalp, occipital, and eyelid arteriovenous malformations plus a palatal teratoma); 3) a hypovascular tumor for which a biopsy could be safely performed (Burkitt lymphoma); and 4) a hypervascular tumor of the palate (cystic teratoma). Our early experience suggests that MR digital subtraction angiography can be reliably performed in children of all ages without complication. The technique provided a noninvasive assessment of the vascularity of each lesion that could not always have been predicted on the basis of clinical examination or routine MR imaging alone.


Loneliness and social isolation as risk factors for coronary heart disease and stroke: systematic review and meta-analysis of longitudinal observational studies.
The influence of social relationships on morbidity is widely accepted, but the size of the risk to cardiovascular health is unclear. We undertook a systematic review and meta-analysis to investigate the association between loneliness or social isolation and incident coronary heart disease (CHD) and stroke. Sixteen electronic databases were systematically searched for longitudinal studies set in high-income countries and published up until May 2015. Two independent reviewers screened studies for inclusion and extracted data. We assessed quality using a component approach and pooled data for analysis using random effects models. Of the 35 925 records retrieved, 23 papers met inclusion criteria for the narrative review. They reported data from 16 longitudinal datasets, for a total of 4628 CHD and 3002 stroke events recorded over follow-up periods ranging from 3 to 21 years. Reports of 11 CHD studies and 8 stroke studies provided data suitable for meta-analysis. Poor social relationships were associated with a 29% increase in risk of incident CHD (pooled relative risk: 1.29, 95% CI 1.04 to 1.59) and a 32% increase in risk of stroke (pooled relative risk: 1.32, 95% CI 1.04 to 1.68). Subgroup analyses did not identify any differences by gender. Our findings suggest that deficiencies in social relationships are associated with an increased risk of developing CHD and stroke. Future studies are needed to investigate whether interventions targeting loneliness and social isolation can help to prevent two of the leading causes of death and disability in high-income countries. CRD42014010225.


Hyperconnecitivity between dorsal attention and frontoparietal networks predicts treatment response in obsessive-compulsive disorder.
Obsessive-compulsive disorder (OCD) presented with repetitive obsessions and/or compulsions were associated with disrupted resting-state functional connectivity (rs-FC). To investigate the pharmacological treatment effect on rs-FC changes in OCD patients we conducted the seed-to-voxel FC analyses using dorsal attention network (DAN), default mode network (DMN), salience network (SN) and frontoparietal network (FPN) and basal ganglia seeds. Twenty-two healthy subjects and twenty-four unmedicated OCD patients underwent resting-state functional magnetic resonance imaging. Patients were rescanned after 12 weeks of escitalopram treatment. We found increased FC both within the DAN and between the DAN and the FPN which was ameliorated after medication and correlated significantly with the clinical improvement in obsession scores. We also observed an anticorrelation between the left caudate and the supplementary motor area in unmedicated OCD patients which also normalized with treatment. Results further showed treatment related normalization of orbitofrontal cortex hyperconnectivity with DMN and hypoconnectivity with DAN whereas aberrant FC between the SN and visual areas appears to be a medication effect. We suggest that DAN to FPN hyperconnectivity which is positively correlated with clinical improvement in obsession scores at pre-treatment stage in present study has a potential for being a neuroimaging marker to predict the treatment response in OCD.


Reducing diarrhea through the use of household-based ceramic water filters: a randomized, controlled trial in rural Bolivia.
Ceramic water filters have been identified as one of the most promising and accessible technologies for treating water at the household level. In a six-month trial, water filters were distributed randomly to half of the 50 participating households in a rural community in Bolivia; the remaining households continued to use customary water handling practices and served as controls. In four rounds of sampling following distribution of the filters, 100% of the 96 water samples from the filter households were free of thermotolerant coliforms compared with 15.5% of the control household samples. Diarrheal disease risk for individuals in intervention households was 70% lower than for controls (95% confidence interval [CI] = 53-80%; P < 0.001). For children less than five years old, the reduction in risk was 83% (95% CI = 51-94%; P < 0.001). These results show that affordable ceramic water filters enable low-income households to treat and maintain the microbiologic quality of their drinking water.


Pattern recognition receptors and their role in invasive aspergillosis.
Pattern recognition receptors (PRRs) are germline receptors that recognize conserved structures on microorganisms. Several PRRs have been identified in the recent years that are involved in the immune response against Aspergillus fumigatus. The role of PRRs in invasive pulmonary aspergillosis becomes especially apparent in the setting of an immunocompromised status of the host because of the redundancy of many PRRs in the host defense against A. fumigatus. Studies that investigated the PRRs and their effector pathways in invasive aspergillosis have led to a better understanding of the pathogenesis of invasive aspergillosis in immunocompromised patients. This knowledge may pave the way for novel diagnostic and immunomodulatory treatment strategies that are needed to overcome the high mortality associated with invasive A. fumigatus infection in immunocompromised patients.


A survey on robotic devices for upper limb rehabilitation.
The existing shortage of therapists and caregivers assisting physically disabled individuals at home is expected to increase and become serious problem in the near future. The patient population needing physical rehabilitation of the upper extremity is also constantly increasing. Robotic devices have the potential to address this problem as noted by the results of recent research studies. However, the availability of these devices in clinical settings is limited, leaving plenty of room for improvement. The purpose of this paper is to document a review of robotic devices for upper limb rehabilitation including those in developing phase in order to provide a comprehensive reference about existing solutions and facilitate the development of new and improved devices. In particular the following issues are discussed: application field, target group, type of assistance, mechanical design, control strategy and clinical evaluation. This paper also includes a comprehensive, tabulated comparison of technical solutions implemented in various systems.


Effective chemoimmunotherapy by co-delivery of doxorubicin and immune adjuvants in biodegradable nanoparticles.
Chemoimmunotherapy is an emerging combinatorial modality for the treatment of cancers resistant to common first-line therapies, such as chemotherapy and checkpoint blockade immunotherapy. We used biodegradable nanoparticles as delivery vehicles for local, slow and sustained release of doxorubicin, two immune adjuvants and one chemokine for the treatment of resistant solid tumors. Methods: Bio-compatible poly(lactic-co-glycolic acid)-PEG nanoparticles were synthesized in an oil/water emulsion, using a solvent evaporation-extraction method. The nanoparticles were loaded with a NIR-dye for theranostic purposes, doxorubicin cytostatic agent, poly (I:C) and R848 immune adjuvants and CCL20 chemokine. After physicochemical and in vitro characterization the nanoparticles therapeutic efficacy were carried-out on established, highly aggressive and treatment resistant TC-1 lung carcinoma and MC-38 colon adenocarcinoma models in vivo. Results: The yielded nanoparticles average size was 180 nm and -14 mV surface charge. The combined treatment with all compounds was significantly superior than separate compounds and the compounds nanoparticle encapsulation was required for effective tumor control in vivo. The mechanistic studies confirmed strong induction of circulating cancer specific T cells upon combined treatment in blood. Analysis of the tumor microenvironment revealed a significant increase of infiltrating leukocytes upon treatment. Conclusion: The multi-drug loaded nanoparticles mediated delivery of chemoimmunotherapy exhibited excellent therapeutic efficacy gain on two treatment resistant cancer models and is a potent candidate strategy to improve cancer therapy of solid tumors resistant to first-line therapies.


Evidence in vivo that the DEAD-box RNA helicase RhlB facilitates the degradation of ribosome-free mRNA by RNase E.
The RNA degradosome of Escherichia coli is a ribonucleolytic multienzyme complex containing RNase E, polynucleotide phosphorylase, RhlB, and enolase. Previous in vitro and in vivo work has shown that RhlB facilitates the exonucleolytic degradation of structured mRNA decay intermediates by polynucleotide phosphorylase in an ATPase-dependent reaction. Here, we show that deleting the gene encoding RhlB stabilizes a lacZ mRNA transcribed by bacteriophage T7 RNA polymerase. Deleting the gene encoding enolase has little if any effect. Other messages transcribed by T7 polymerase are also stabilized by DeltarhlB. The effect of point mutations inactivating RhlB is comparable with the effect of deleting the gene. Primer extension analysis of the lacZ message indicates that RhlB facilitates endoribonucleolytic cleavage by RNase E, demonstrating a functional interaction between the RNA helicase and the endoribonuclease. The possible physiological role of an RhlB-RNase E pathway and the mechanisms by which RhlB could facilitate RNase E cleavage are discussed.


Heparin Enriched-WPI Coating on Ti6Al4V Increases Hydrophilicity and Improves Proliferation and Differentiation of Human Bone Marrow Stromal Cells.
Titanium alloy (Ti6Al4V) is one of the most prominent biomaterials for bone contact because of its ability to bear mechanical loading and resist corrosion. The success of Ti6Al4V implants depends on bone formation on the implant surface. Hence, implant coatings which promote adhesion, proliferation and differentiation of bone-forming cells are desirable. One coating strategy is by adsorption of biomacromolecules. In this study, Ti6Al4V substrates produced by additive manufacturing (AM) were coated with whey protein isolate (WPI) fibrils, obtained at pH 2, and heparin or tinzaparin (a low molecular weight heparin LMWH) in order to improve the proliferation and differentiation of bone-forming cells. WPI fibrils proved to be an excellent support for the growth of human bone marrow stromal cells (hBMSC). Indeed, WPI fibrils were resistant to sterilization and were stable during storage. This WPI-heparin-enriched coating, especially the LMWH, enhanced the differentiation of hBMSC by increasing tissue non-specific alkaline phosphatase (TNAP) activity. Finally, the coating increased the hydrophilicity of the material. The results confirmed that WPI fibrils are an excellent biomaterial which can be used for biomedical coatings, as they are easily modifiable and resistant to heat treatments. Indeed, the already known positive effect on osteogenic integration of WPI-only coated substrates has been further enhanced by a simple adsorption procedure.


New methods of removing debris and high-throughput counting of cyst nematode eggs extracted from field soil.
The soybean cyst nematode (SCN), Heterodera glycines, is the most damaging pathogen of soybeans in the United States. To assess the severity of nematode infestations in the field, SCN egg population densities are determined. Cysts (dead females) of the nematode must be extracted from soil samples and then ground to extract the eggs within. Sucrose centrifugation commonly is used to separate debris from suspensions of extracted nematode eggs. We present a method using OptiPrep as a density gradient medium with improved separation and recovery of extracted eggs compared to the sucrose centrifugation technique. Also, computerized methods were developed to automate the identification and counting of nematode eggs from the processed samples. In one approach, a high-resolution scanner was used to take static images of extracted eggs and debris on filter papers, and a deep learning network was trained to identify and count the eggs among the debris. In the second approach, a lensless imaging setup was developed using off-the-shelf components, and the processed egg samples were passed through a microfluidic flow chip made from double-sided adhesive tape. Holographic videos were recorded of the passing eggs and debris, and the videos were reconstructed and processed by custom software program to obtain egg counts. The performance of the software programs for egg counting was characterized with SCN-infested soil collected from two farms, and the results using these methods were compared with those obtained through manual counting.


The COVID-19 Pandemic Affects Seasonality, With Increasing Cases of New-Onset Type 1 Diabetes in Children, From the Worldwide SWEET Registry.
To analyze whether the coronavirus disease 2019 (COVID-19) pandemic increased the number of cases or impacted seasonality of new-onset type 1 diabetes (T1D) in large pediatric diabetes centers globally. We analyzed data on 17,280 cases of T1D diagnosed during 2018-2021 from 92 worldwide centers participating in the SWEET registry using hierarchic linear regression models. The average number of new-onset T1D cases per center adjusted for the total number of patients treated at the center per year and stratified by age-groups increased from 11.2 (95% CI 10.1-12.2) in 2018 to 21.7 (20.6-22.8) in 2021 for the youngest age-group, <6 years; from 13.1 (12.2-14.0) in 2018 to 26.7 (25.7-27.7) in 2021 for children ages 6 to <12 years; and from 12.2 (11.5-12.9) to 24.7 (24.0-25.5) for adolescents ages 12-18 years (all P < 0.001). These increases remained within the expected increase with the 95% CI of the regression line. However, in Europe and North America following the lockdown early in 2020, the typical seasonality of more cases during winter season was delayed, with a peak during the summer and autumn months. While the seasonal pattern in Europe returned to prepandemic times in 2021, this was not the case in North America. Compared with 2018-2019 (HbA1c 7.7%), higher average HbA1c levels (2020, 8.1%; 2021, 8.6%; P < 0.001) were present within the first year of T1D during the pandemic. The slope of the rise in pediatric new-onset T1D in SWEET centers remained unchanged during the COVID-19 pandemic, but a change in the seasonality at onset became apparent.


Autoimmune pancreatitis: the clinicopathological characteristics of the subtype with granulocytic epithelial lesions.
Autoimmune pancreatitis (AIP) has been established as a distinct form of chronic pancreatitis that is distinguishable from other types such as alcoholic, hereditary or obstructive chronic pancreatitis. AIP seems to be a global disease, since it has been reported in many different countries, especially from Japan, USA and Europe (Germany, Italy, United Kingdom). Typical histopathological findings in the pancreas in AIP include a periductal lymphoplasmacytic infiltration with fibrosis, causing narrowing of the involved ducts. The typical clinical features include presentation with obstructive jaundice/pancreatic mass and a dramatic response to steroids. However, while the reports from Japan describe uniform changes called lymphoplasmacytic sclerosing pancreatitis (LPSP) in the pancreas from AIP patients, the reports from Europe and USA distinguish two histopathologic patterns in AIP patients: one with the characteristics of LPSP and another with slightly different histological features, called idiopathic duct centric pancreatitis (IDCP) or AIP with granulocytic epithelial lesions (GELs). This article reviews the evidence that GEL-positive AIP or IDCP is a second type of AIP, distinct from LPSP, in regard to pancreatic pathology, immunology and epidemiology.


Rehabilitation of myositis ossificans in the brachialis muscle.
Theories on the etiology and management of myositis ossificans vary greatly. In this article, possible causative factors and treatment options are reviewed. We present a successful treatment regimen for myositis ossificans of the brachialis muscle in a 15-year-old football player. The treatment program emphasizes using joint mobilization and eccentric strengthening early in rehabilitation. The patient regained full elbow range of motion after nine weeks of this rehabilitation program. Follow-up at nine months revealed nearly complete maturation of the heterotopic bone formation, full pain-free range of motion, normal strength, and a safe return to athletic competition.


[A Case of Distal Bile Duct Cancer Occurring 38 Years after Cyst-to-Duct Anastomosis for a Congenital Biliary Dilatation].
A 41-year-old woman had undergone cyst-to-duct anastomosis for a congenital biliary dilatation when she was 3-yearold. After 38 years, she presented with a high fever, and laboratory findings showed elevation of liver and biliary enzyme levels. A CT scan showed a mass lesion in a choledochal cyst, and an adenocarcinoma was found in the anastomosis of the choledochal cyst and duodenum. We diagnosed distal bile duct cancer and performed SSPPD after neoadjuvant chemotherapy( gemcitabine plus cisplatin plus S-1). The resected tumor was pathologically diagnosed as papillary-infiltrating adenocarcinoma( ypT2N1M0, ypStage II B). Currently, 9 months after the second surgery, the patient is doing well without any signs of recurrence. The current case suggested the necessity of postoperative lifetime follow-up for patients with bile duct cancer who have undergone radical cyst excision for a congenital biliary dilatation.


Training current and future robotic surgeons simultaneously: initial experiences with safety and efficiency.
Robotic surgery is experiencing a rapidly-increasing presence in the field of general surgery. The adoption of any new technology carries the challenge of training current and future surgeons in a safe and effective manner. We report our experience with the initiation of a robotic general surgery program at an academic institution while simultaneously incorporating surgical trainees. The initial procedure performed was robotic-assisted cholecystectomy (RAC). Concurrent with the introduction of a robotic general surgical program, our institution implemented a progressive surgical trainee curriculum for all active residents and fellows. Immediately after being credentialed to perform RAC, attending surgeons began incorporating surgical trainees into robotic procedures. We retrospectively reviewed our first 50 RACs and compared them with our previous 50 standard laparoscopic cholecystectomies (SLC) to determine the impact of rapid integration of surgical trainees on developing technologies. Despite new technology and novice surgeons, there was no difference in mean operative time between the SLC and RAC groups (75.3 vs. 84.1 min, p = 0.077). Two patients in the robotic-assisted group required intraoperative conversion. Hospital length of stay was similar between groups, with the majority of patients leaving the same day. There were no postoperative complications in either group. A robotic general surgery program can be initiated while concurrently instructing surgical trainees on robotic surgery in a safe and efficient manner. We report our initial experience with the adoption of this rapidly advancing technology and describe our training model.


Evaluation of topography and vascularization of cervical paragangliomas by magnetic resonance imaging and color duplex sonography.
The management of cervical paragangliomas (PGs) depends on their specific type and their relation to adjacent vessels. The purpose of this study was to evaluate the potential of magnetic resonance imaging (MRI) and color duplex sonography (CDS) to classify PGs according to topography and vascularization. Sixteen patients harboring 22 PGs were studied retrospectively. With digital subtraction angiography as reference, the topographic relation of the tumors to the carotid arteries and the internal jugular vein and the patterns of vascularization were assessed. On MRI and CDS the typical feature of 15 carotid PGs was splaying of the carotid bifurcation, with the external carotid displaced anteriorly and the internal carotid artery and internal jugular vein located posteriorly. In five vagal PGs both modalities showed unidirectional anterior displacement of the external and internal carotid arteries. Two jugular PGs were found to extend within the lumen of the internal jugular vein. CDS completely depicted carotid body tumors but failed to delineate the high cervical portion of vagal and jugular PGs. MRI allowed us to assess the entire extent of all PGs. Nineteen lesions showed flow voids corresponding to abundant flow signal on CDS: three carotid body tumors appeared hypovascular on CDS and MRI. On CDS, intratumoral flow was directed cranially in carotid and inferiorly in vagal and jugular PGs. CDS and MRI are suitable for classification of cervical PGs as carotid, vagal or jugular PGs based on the topographic relation to the carotid arteries and internal jugular vein. Visualization of the intrinsic tumor vasculature proved an additional distinguishing criterion on CDS.


Outcomes of patients with metastatic pancreatic cancer who progress on first restaging imaging.
Objective responses to first-line systemic chemotherapy in metastatic pancreatic cancer patients are seen in less than one third of cases. Unfortunately, a significant amount will have disease progression (PD) on their first restaging imaging. With patients' short life expectancy, it is crucial for clinicians to be prudent when deciding whom and when to treat. Our study aimed to evaluate outcomes of patients that progressed on their first restaging imaging on 1st line therapy. We retrospectively analyzed patients diagnosed between 2010-2017 whose first restaging imaging demonstrated PD. The primary outcome was overall survival (OS) from metastatic diagnosis date to death. Patients who were lost to follow-up were excluded. Out of 262 total patients reviewed, 98 patients (37%) were included. Sixty-five (66%) received 2nd line therapy, and 33 (34%) did not. Reasons patients did not pursue 2nd line therapy were performance status (PS) decline, organ dysfunction, or patient choice for alternative therapy. Median ages for patients who did and did not receive 2nd line therapy were 61 and 67, respectively (P<0.001). More patients had a poor PS at the time of initial diagnosis in the non-2nd line therapy group (7.5% vs. 31.0%, P=0.021). Median OS for those receiving 2nd line therapy was 9 months (95% CI: 7-11 months) compared to 4 months (95% CI: 3-5 months) for those not receiving 2nd-line therapy (P<0.001). Although likely biased due to better performance status and younger age, our patients who progressed rapidly on 1st line therapy showed an OS benefit if they received 2nd line therapy. These results suggest that patients maintaining a good PS after immediate progression on 1st line therapy should be offered 2nd line therapy.


Identification of a novel hypocholesterolemic protein, major royal jelly protein 1, derived from royal jelly.
Royal jelly (RJ) intake lowers serum cholesterol levels in animals and humans, but the active component in RJ that lowers serum cholesterol level and its molecular mechanism are unclear. In this study, we set out to identify the bile acid-binding protein contained in RJ, because dietary bile acid-binding proteins including soybean protein and its peptide are effective in ameliorating hypercholesterolemia. Using a cholic acid-conjugated column, we separated some bile acid-binding proteins from RJ and identified the major RJ protein 1 (MRJP1), MRJP2, and MRJP3 as novel bile acid-binding proteins from RJ, based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Purified MRJP1, which is the most abundant protein of the bile acid-binding proteins in RJ, exhibited taurocholate-binding activity in vitro. The micellar solubility of cholesterol was significantly decreased in the presence of MRJP1 compared with casein in vitro. Liver bile acids levels were significantly increased, and cholesterol 7α-hydroxylase (CYP7A1) mRNA and protein tended to increase by MRJP1 feeding compared with the control. CYP7A1 mRNA and protein levels were significantly increased by MRJP1 tryptic hydrolysate treatment compared with that of casein tryptic hydrolysate in hepatocytes. MRJP1 hypocholesterolemic effect has been investigated in rats. The cholesterol-lowering action induced by MRJP1 occurs because MRJP1 interacts with bile acids induces a significant increase in fecal bile acids excretion and a tendency to increase in fecal cholesterol excretion and also enhances the hepatic cholesterol catabolism. We have identified, for the first time, a novel hypocholesterolemic protein, MRJP1, in RJ. Interestingly, MRJP1 exhibits greater hypocholesterolemic activity than the medicine β-sitosterol in rats.


Long-range correlations and complex regulation of pacing in long-distance road racing.
To analyze time-domain, spectral, and fractal properties of speed regulation during half-marathon racing. In 21 male experienced runners, high-resolution data on speed (V), stride frequency (SF), and stride length (SL) were assessed during half-marathon competition (21,098 m). Performance times, time- and frequency-domain variability, spectral-scaling exponent (beta), and fractal dimension (FD) of V, SF, and SL were analyzed. V of 3.65 ± 0.41 m/s, SF of 1.41 ± 0.05 Hz, and SL of 2.58 ± 0.25 m occurred with higher (P < .05) individual variability in V and SL than in SF. Beta and FD were always 1.04-1.88 and 1.56-1.99, respectively. Beta and FD differed (P < .05) in SF and SL compared with V and were correlated in V and SL (r = .91, P < .05). Spectral peaks of V, SF, and SL occurred at wavelengths of 3-35 min, and those of V and SL were interrelated (r = .56, P < .05). Mean SF and mean SL were significantly correlated with performance (r = .59 and r = .95, P < .05). SL accounted for 84% ± 6% and SF for 16% ± 6% of speed variability. The observed nonrandom fluctuations in V, SF, and SL correspond to nonstationary fractional Brownian motion with inherent long-range correlations. This indicates a similar complex regulation process in experienced runners that is primarily mediated via SL.


Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis.
The Arabidopsis genome contains approximately 200 genes that encode proteins with similarity to the nucleotide binding site and other domains characteristic of plant resistance proteins. Through a reiterative process of sequence analysis and reannotation, we identified 149 NBS-LRR-encoding genes in the Arabidopsis (ecotype Columbia) genomic sequence. Fifty-six of these genes were corrected from earlier annotations. At least 12 are predicted to be pseudogenes. As described previously, two distinct groups of sequences were identified: those that encoded an N-terminal domain with Toll/Interleukin-1 Receptor homology (TIR-NBS-LRR, or TNL), and those that encoded an N-terminal coiled-coil motif (CC-NBS-LRR, or CNL). The encoded proteins are distinct from the 58 predicted adapter proteins in the previously described TIR-X, TIR-NBS, and CC-NBS groups. Classification based on protein domains, intron positions, sequence conservation, and genome distribution defined four subgroups of CNL proteins, eight subgroups of TNL proteins, and a pair of divergent NL proteins that lack a defined N-terminal motif. CNL proteins generally were encoded in single exons, although two subclasses were identified that contained introns in unique positions. TNL proteins were encoded in modular exons, with conserved intron positions separating distinct protein domains. Conserved motifs were identified in the LRRs of both CNL and TNL proteins. In contrast to CNL proteins, TNL proteins contained large and variable C-terminal domains. The extant distribution and diversity of the NBS-LRR sequences has been generated by extensive duplication and ectopic rearrangements that involved segmental duplications as well as microscale events. The observed diversity of these NBS-LRR proteins indicates the variety of recognition molecules available in an individual genotype to detect diverse biotic challenges.


The cost effectiveness of tamoxifen in the prevention of breast cancer.
In the National Surgical Adjuvant Breast and Bowel Project P-1 Breast Cancer Prevention Trial (BCPT), women considered to be at high risk for developing breast cancer who received tamoxifen experienced 49% and 50% reductions in the risk of developing invasive and noninvasive breast cancer, respectively, compared with women receiving placebo. Although the BCPT addressed the clinical benefits of tamoxifen, this study sought to assess its cost effectiveness in the prevention of breast cancer in women at increased risk for developing the disease. Women were considered to be at an increased risk if they were: 1) 60 years of age or older, 2) age 35 to 59 years with a history of lobular carcinoma in situ, or 3) age 35 to 59 years with additional risk factors that made their 5-year predicted breast cancer risk at least as great as that of women 60 years of age. A decision-analysis model was used to estimate the incremental cost effectiveness of using tamoxifen compared with no intervention as preventive therapy in age-group defined cohorts of women who were at high risk for developing breast cancer. The analysis used data on the benefits and risks of tamoxifen as observed in the BCPT. In a subgroup analysis, tamoxifen's cost effectiveness was also evaluated in women who had had a hysterectomy, because of evidence that suggested an increased risk of endometrial cancer in women receiving tamoxifen. Under conservative assumptions from a base-case analysis, the incremental cost effectiveness of tamoxifen is $41,372 per life-year gained for women age 35 to 49 years, whereas for women age 50 to 59 years and 60 to 69 years, these values are $68,349 and $74,981, respectively. For women with a previous hysterectomy, tamoxifen's cost effectiveness is $46,060 per life-year gained. A strategy of using tamoxifen in high-risk women to prevent breast cancer in high-risk women may be cost effective, particularly in the 35-to-49 year-old age group and in those of any age who have had a hysterectomy.


Interlaboratory agreement among results of human papillomavirus type 16 enzyme-linked immunosorbent assays.
Serological assays for measuring antibodies to human papillomavirus type 16 (HPV-16) virus-like particles (VLPs) have become important epidemiologic tools in recent years. However, the interlaboratory replicability of these assays has not been assessed. In this investigation, three laboratories tested a panel of specimens obtained from two different groups: 265 subjects in a vulvar cancer case-control study and 107 healthy volunteer blood donors. Each laboratory used an enzyme-linked immunosorbent assay (ELISA), but no attempt was made to standardize assay procedures among the three laboratories. The data showed good day-to-day intralaboratory replicability in laboratory 1 (correlation coefficient, > or = 0.88) and good intra-assay variability in laboratory 3 (correlation coefficient, > or = 0.93). Interlaboratory correlations, likewise, ranged between 0.61 and 0.80 in both case-control study subjects and healthy blood donors, indicating that ELISA optical density (OD) values between laboratories were linearly related regardless of the population. Kappa coefficients (kappa), based on each laboratory's categorical interpretation of its results (as positive or negative), showed good agreement (kappa, > 0.6) in case-control study subjects and moderate agreement (kappa, > or = 0.4) in blood donors, a population that had few strongly positive sera. When OD values near seropositive cutoffs were treated as indeterminates, there was little discordance between laboratories in either population. The data suggest that each laboratory measured the same humoral immune response and that their HPV-16 VLP ELISAs performed similarly (Pearson correlations). Interlaboratory differences, however, probably due to reagents and procedures, were considerably greater than intralaboratory day-to-day variability. Interlaboratory agreement in determining seropositivity (kappa) could be improved by sharing positive and negative serum controls and by treating marginal results as indeterminate. As part of continuing cooperation to improve interlaboratory agreement, we are preparing bulk serum control specimens to be shared and made available to interested researchers.


Improved Parameterization for Combined Isotopomer Analysis of Diatomic Spectra and Its Application to HF and DF.
A new way of representing vibration-rotation term values for multiple isotopomers of a given electronic state of a diatomic molecule is presented which resolves problems associated with the way the conventional combined isotopomer expansion represents the atomic mass-dependent JWKB and Born-Oppenheimer breakdown correction terms. Its application to infrared and microwave data for HF and DF yields new Dunham expansion coefficients and Born-Oppenheimer breakdown correction terms for this species. This procedure is implemented in a generally available computer program for fitting to various types of data involving one or several electronic states of multiple isotopomers of a diatomic molecule. Copyright 1999 Academic Press.


Genetic diversity analysis of French goat populations reveals selective sweeps involved in their differentiation.
After domestication 11 000 years ago in Asia Minor, the goat followed human migration to Europe and Asia. It was then introduced in Africa and is now raised all over the world. In this study, we exploited a dataset composed of 54 000 SNPs (Illumina goat DNA chip) to analyze the genetic diversity of 223 individuals belonging to eight French breeds (Alpine, Angora, Corse, Fossés, Poitevine, Provençale, Pyrénées and Saanen). Analyses carried out included individual-based approaches (principal component analysis and population structure) and population-based approaches (phylogenetic tree constructions). The results of the genetic diversity analyses revealed that French breeds are clearly differentiated, in particular, the Angora breed that originates from south west Asia. The Provençale breed shows a very original genetic pattern that could be the result of ancient admixture. Then, selection signatures were detected by identifying regions of outlying genetic differentiation between populations. Five genomic regions were detected under selection on chromosomes 5, 6, 11, 13 and 20, revealing mainly soft selective sweeps and a few hard selective sweeps and highlighting candidate genes that had been selected for during the evolutionary history of these breeds. Among them, two coat coloration genes (ADAMTS20 and ASIP) and one gene related to milk composition (CSN1S1) were involved.


The spatial distribution and physico-chemical characteristic of microplastics in the sediment and cockle (Anadara granosa) from the coastal waters of East Java, Indonesia, and the health hazards associated with cockle consumption.
This study evaluated microplastic (MP) abundances and physico-chemical characteristics in sediments and Anadara granosa along the East Java coast and their health implications. Fibers (74 %) dominated sediment MPs at south coast, while fragments (49-61 %) dominated north coast. Fiber (43-52 %) is the predominant MP in cockle tissues in all locations. Most MP in sediments (31-47 %) and cockle tissues (41-49 %) is black. The majority of microplastics (100-1500 μm) are found in sediment (73-90 %), and cockles (77-79 %). Very weak correlations found between the amount of MP and the length of the cockle shell. However, Spearman correlation shows that as the amount of MP in sediment increases, so does the amount of MP in cockle tissue. Each year, individuals of varying ages consume an average of 20,800 to 156,000 MP items. Cockles contain plasticizer components and microplastic polymers which are classified from II to V regarding of hazard levels, with V being the most hazardous.


Cutaneous laser therapy.
The carbon dioxide laser is useful for vaporizing lesions and applying incisions, the argon laser coagulates superficial vascular lesions and the neodymium-YAG laser is used for large vascular and more deeply situated lesions. Many patients with port-wine stains have been treated with excellent to poor results, major problems consisting of incomplete color removal and hypertrophic scarring (occurring in 4% to 23% of cases). While results are imperfect, patients are satisfied with the improvement in 86% of cases. Lasers have been used with good results for treating patients with strawberry angioma of infancy, pyogenic granuloma, telangiectasia of the face, decorative tattoos, genital condylomata and warts. The results of laser treatment of essential telangiectasia of the lower extremities have generally been poor. The CO(2) laser has been effective in excising small lesions and elevating skin flaps.


[In vitro systems for the primary search for biologically active substances with carcinostatic action].
Improvement and development of new methods for primary screening of antitumour drugs in vitro are based on the data on the points open to injury in the tumour cell metabolism and on the evidence of the known carcinostatic drug mechanism. Further development of the primary screening methods should proceed, probably, in two main directions. Further improvement of the methods for cloning tumour cells in the semifluid nutrient media is of great interest. On the whole the creation of new test systems is a significant trend of the scientific screening.


[Barium sulfate and water-soluble contrast medium--a suitable mixture for contrast radiography in acute small bowel obstruction].
Experiences with a mixture of two different contrast media for radiography in acute small bowel obstruction are reported. Due to different physical and chemical properties, barium sulfate and a water-soluble, high-osmolar contrast medium provided high contrast density in all segments of the small bowel and a shortening of the transit time to the colon. This method proved to be effective and accurate in predicting which patients with suspected small bowel obstruction required surgical intervention. No complications caused by the contrast media were observed intraoperatively or in the postoperative period.


Patterns of failure after the reduced volume approach for elective nodal irradiation in nasopharyngeal carcinoma.
To evaluate the patterns of nodal failure after radiotherapy (RT) with the reduced volume approach for elective neck nodal irradiation (ENI) in nasopharyngeal carcinoma (NPC). Fifty-six NPC patients who underwent definitive chemoradiotherapy with the reduced volume approach for ENI were reviewed. The ENI included retropharyngeal and level II lymph nodes, and only encompassed the echelon inferior to the involved level to eliminate the entire neck irradiation. Patients received either moderate hypofractionated intensity-modulated RT for a total of 72.6 Gy (49.5 Gy to elective nodal areas) or a conventional fractionated three-dimensional conformal RT for a total of 68.4-72 Gy (39.6-45 Gy to elective nodal areas). Patterns of failure, locoregional control, and survival were analyzed. The median follow-up was 38 months (range, 3 to 80 months). The out-of-field nodal failure when omitting ENI was none. Three patients developed neck recurrences (one in-field recurrence in the 72.6 Gy irradiated nodal area and two in the elective irradiated region of 39.6 Gy). Overall disease failure at any site developed in 11 patients (19.6%). Among these, there were six local failures (10.7%), three regional failures (5.4%), and five distant metastases (8.9%). The 3-year locoregional control rate was 87.1%, and the distant failure-free rate was 90.4%; disease-free survival and overall survival at 3 years was 80% and 86.8%, respectively. No patient developed nodal failure in the omitted ENI site. Our investigation has demonstrated that the reduced volume approach for ENI appears to be a safe treatment approach in NPC.


Modeling civil violence: an agent-based computational approach.
This article presents an agent-based computational model of civil violence. Two variants of the civil violence model are presented. In the first a central authority seeks to suppress decentralized rebellion. In the second a central authority seeks to suppress communal violence between two warring ethnic groups.


Iptakalim, opener of K(ATP), reverses the enhanced expression of genes encoding K(ATP) subunits in spontaneously hypertensive rats.
ATP-sensitive potassium channels (K(ATP)) are thought to be targets for antihypertensive drugs that are potassium channel openers. In this study, the expression of genes encoding the K(ATP) subunits, SUR2, Kir6.1 and Kir6.2, was detected in tissues from Wistar-Kyoto rats (WKY), spontaneously hypertensive rats (SHR), and SHR undergoing long-term treatment with iptakalim, a novel antihypertensive drug that acts via K(ATP). The transcript levels for SUR2, Kir6.1 and Kir6.2 in the heart, aortic smooth muscle, and tail artery smooth muscle were determined by reverse transcription-polymerase chain reaction (RT-PCR). In general, Kir6.2 and SUR2 were more highly represented in all SHR tissues compared with those of WKY, and transcripts of Kir6.2 were significantly higher in tail artery smooth muscle from SHR. Following long-term treatment with iptakalim, mRNA levels of Kir6.2 and SUR2 were reduced significantly in all tissues compared with those of untreated SHR. Kir6.1 expression was not significantly different between SHR and WKY, and was unaffected by iptakalim treatment. These results indicate that the expression of the K(ATP) subunits genes, SUR2 and Kir6.2, are closely associated with hypertensive pathological states, and the effect of iptakalim on K(ATP) mRNA levels may explain, in part, the effects of iptakalim in reversing vascular and cardiac remodeling. Furthermore, changes in Kir6.2 mRNA levels suggest that Kir6.x, as well as SUR, is responsible for drug binding.


Hemorrhage after ischemic stroke - relation to age and previous hemorrhage in a nationwide cohort of 58,868 patients.
In randomized controlled trials of secondary prevention after stroke, the risk of hemorrhage varies between 1% and 5% per year in patients with antithrombotic therapy, i.e. anticoagulants and antiplatelets. To explore the rate and the risk of hemorrhage after stroke in a nationwide cohort. We identified 58 868 first ever ischemic stroke patients in the Swedish Stroke Register during 2001 to 2005 (=index stroke) and followed them by record linkage to the National Patient Register. Rates of hemorrhage and hazard ratios, for comparisons of rates between subgroups, were calculated. Of the 58 586 ischemic stroke patients identified, 5527 (9·4%) had a history of hemorrhage. During follow-up (mean 2·0 years), 2876 patients endured a hemorrhage, giving an average hemorrhage rate of 2·6 (95% confidence interval 2·5-2·7) per 100 person-years. After index stroke, 11% of the patients were discharged with anticoagulants, and 79% with antiplatelets. Given the differences in baseline characteristics, the hemorrhage rates (per 100 person-years) were 2·5 (95% confidence interval 2·2-2·8), 2·4 (95% confidence interval 2·3-2·5), and 3·8 (95% confidence interval 3·5-4·2) in patients prescribed anticoagulants, antiplatelets, and no antithrombotics, respectively. There was an increased risk of hemorrhage in patients ≥75 years compared with those <75 years (hazard ratio = 1·61, 95% confidence interval 1·49-1·73) and in patients with previous hemorrhages compared with those without (hazard ratio = 1·82, 95% confidence interval 1·64-2·02). When antithrombotics were used in large-scale clinical practice, the observed rates of hemorrhage were similar with anticoagulant therapy but increased with antiplatelet therapy compared with rates reported in randomized controlled trials. Old age and previous hemorrhage were associated with an increased risk of hemorrhage after an ischemic stroke.


The role of bariatric surgery in the treatment of type 2 diabetes: current evidence and clinical guidelines.
The incidence of type 2 diabetes (T2D) continues to rise worldwide. The management of T2D is challenging and therefore amenable to multimodality treatment options. Many published observations of obese individuals with T2D that have undergone bariatric surgery consistently demonstrate remarkable improvement and short-term remission of T2D. Recently published randomized trials confirm these findings and demonstrate significantly improved glycemic control following bariatric procedures, especially Roux-en-Y gastric bypass and laparoscopic sleeve gastrectomy. However, the question of long-term remission remains uncertain. Clinicians may consider the use of bariatric surgery as a treatment option for certain obese patients with T2D who have failed intensive lifestyle intervention and conventional pharmacotherapy.


Slow and fast light via SBS in optical fibers for short pulses and broadband pump.
Slow-light effect via stimulated Brillouin scattering (SBS) in single-mode optical fibers was considered for short probe pulses of nanosecond duration relevant to Gb/s data streams. Unlike recent estimations of delay versus pump based on steady-state small-signal approximation we have used numerical solution of three-wave equations describing SBS for a realistic fiber length. Both regimes of small signal and pump depletion (gain saturation) were considered. The physical origin of Stokes pulse distortion is revealed which is related to excitation of long-living acoustic field behind the pulse and prevents effective delay control by pump power increase at cw pumping. We have shown different slope of the gain-dependent delay for different pulse durations. Spectrally broadened pumping by multiple cw components, frequency-modulated pump and pulse train were studied for short pulses which allow to obtain large delay and suppress pulse distortion. In the pump-depletion regime of pumping by pulse train, both pulse delay and distortion decrease with increasing pump, and the pulse achieves advancement.


Data Model Requirements for a Digital Cognitive Aid for Anesthesia to Support Intraoperative Crisis Management.
The aim of this study is to define data model requirements supporting the development of a digital cognitive aid (CA) for intraoperative crisis management in anesthesia, including medical emergency text modules (text elements) and branches or loops within emergency instructions (control structures) as well as their properties, data types, and value ranges. The analysis process comprised three steps: reviewing the structure of paper-based CAs to identify common text elements and control structures, identifying requirements derived from content, design, and purpose of a digital CA, and validating requirements by loading exemplary emergency checklist data into the resulting prototype data model. The analysis of paper-based CAs identified 19 general text elements and two control structures. Aggregating these elements and analyzing the content, design and purpose of a digital CA revealed 20 relevant data model requirements. These included checklist tags to enable different search options, structured checklist action steps (items) in groups and subgroups, and additional information on each item. Checklist and Item were identified as two main classes of the prototype data model. A data object built according to this model was successfully integrated into a digital CA prototype. To enable consistent design and interactivity with the content, presentation of critical medical information in a digital CA for crisis management requires a uniform structure. So far it has not been investigated which requirements need to be met by a data model for this purpose. The results of this study define the requirements and structure that enable the presentation of critical medical information. Further research is needed to develop a comprehensive data model for a digital CA for crisis management in anesthesia, including supplementation of requirements resulting from simulation studies and feasibility analyses regarding existing data models. This model may also be a useful template for developing data models for CAs in other medical domains.


Chronic mild stress leads to anxiety-like behavior and decreased p70 S6K1 activity in the hippocampus of male mice.
Major affective disorders are highly prevalent, however, current treatments are limited in their effectiveness due to a lack of understanding of underlying molecular mechanisms. Recent studies have shown that reduced activity of p70 S6 kinase 1 (S6K1), a downstream target of the mechanistic target of rapamycin complex 1 (mTORC1), is linked to anxiety-like behavior in both humans and rodents. The purpose of this study was to investigate the relationship between S6K1 and anxiety-like behavior following chronic mild stress (CMS) and drug-induced inhibition of S6K1. Following CMS, anxiety-like behavior was evaluated using an open field (OF) and elevated plus maze (EPM) in adult male C57/Bl6 mice. After behavior analysis, samples of the hippocampus were harvested for quantification of S6K1, S6 ribosomal protein, glycogen synthase kinase-3 β (GSK3β), and beta tubulin via western blot. Our results demonstrate that CMS mice exhibit anxiety-like behavior in the OF and EPM and reduced activity of S6K1 in the hippocampus (HPC). We measured phosphorylation levels of GSK3β and found that GSK3β phosphorylation was also reduced following CMS compared to control mice. Furthermore, pharmacological inhibition of S6K1 with PF-4708671 in male mice was sufficient to produce anxiety-like behavior in the OF and EPM. These results further support the significant role of S6K1 in the pathogenesis of anxiety and affective disorders.


Structural framework for the understanding spectroscopic and functional signatures of the cyanobacterial Orange Carotenoid Protein families.
The Orange Carotenoid Protein (OCP) is a unique photoreceptor crucial for cyanobacterial photoprotection. Best studied Synechocystis sp. PCC 6803 OCP belongs to the large OCP1 family. Downregulated by the Fluorescence Recovery Protein (FRP) in low-light, high-light-activated OCP1 binds to the phycobilisomes and performs non-photochemical quenching. Recently discovered families OCP2 and OCP3 remain structurally and functionally underexplored, and no systematic comparative studies have ever been conducted. Here we present two first crystal structures of OCP2 from morphoecophysiologically different cyanobacteria and provide their comprehensive structural, spectroscopic and functional comparison with OCP1, the recently described OCP3 and all-OCP ancestor. Structures enable correlation of spectroscopic signatures with the effective number of hydrogen and discovered here chalcogen bonds anchoring the ketocarotenoid in OCP, as well as with the rotation of the echinenone's β-ionone ring in the CTD. Structural data also helped rationalize the observed differences in OCP/FRP and OCP/phycobilisome functional interactions. These data are expected to foster OCP research and applications in optogenetics, targeted carotenoid delivery and cyanobacterial biomass engineering.


Revealing the Angiopathy of Lacrimal Gland Lesion in Type 2 Diabetes.
For a better understanding of diabetic angiopathy (DA), the potential biomarkers in lacrimal DA and its potential mechanism, we evaluated the morphological and hemodynamic alterations of lacrimal glands (LGs) in patients with type 2 diabetes and healthy counterparts by color Doppler flow imaging (CDFI). We further established a type 2 diabetic mice model and performed hematoxylin-eosin (HE) staining, immunofluorescence staining of CD31, RNA-sequencing analysis, and connectivity map (CMap) analysis. We found atrophy and ischemia in patients with type 2 diabetes and mice models. Furthermore, we identified 846 differentially expressed genes (DEGs) between type 2 diabetes mellitus (T2DM) and vehicle mice by RNA-seq. The gene ontology (GO) analysis indicated significant enrichment of immune system process, regulation of blood circulation, apoptotic, regulation of secretion, regulation of blood vessel diameter, and so on. The molecular complex detection (MCODE) showed 17 genes were involved in the most significant module, and 6/17 genes were involved in vascular disorders. CytoHubba revealed the top 10 hub genes of DEGs, and four hub genes (App, F5, Fgg, and Gas6) related to vascular regulation were identified repeatedly by MCODE and cytoHubba. GeneMANIA analysis demonstrated functions of the four hub genes above and their associated molecules were primarily related to the regulation of circulation and coagulation. CMap analysis found several small molecular compounds to reverse the altered DEGs, including disulfiram, bumetanide, genistein, and so on. Our outputs could empower the novel potential targets to treat lacrimal angiopathy, diabetes dry eye, and other diabetes-related diseases.


Adipocytokines and achievement of low disease activity in rheumatoid arthritis.
To determine if adipocytokines are independently associated with the achievement of low disease activity (LDA) over long-term follow-up in a large rheumatoid arthritis (RA) registry. This cohort study evaluated adults with RA from the Veteran's Affairs RA Registry. Adipocytokines (adiponectin, leptin, and fibroblast growth factor [FGF]-21) and inflammatory cytokines were measured as part of a multi-analyte panel on banked serum from enrollment. Covariates were derived from medical record, biorepository, and registry databases. Multivariable Cox proportional hazard models evaluated associations between adipocytokines and rates of 1) DAS28 LDA and remission, 2) individual Boolean remission criteria and 3) initiation of a new bDMARD or tsDMARD. There were 1,276 participants with a DAS28 >3.2 at enrollment. Of these, 827 achieved LDA and 598 achieved remission over 2,287 and 4,096 person-years, respectively. Patients in the highest quartile of adiponectin had lower rates LDA before and after adjustment [aHR Q4: 0.68 (0.53,0.87) p<0.001]. Those in the highest quartile of leptin and FGF-21 also had lower rates of LDA. Higher quartiles of adipocytokines were also associated with lower rates of achieving a low patient/evaluator global scores and low tender joint counts. Among 1,236 biologic-naïve participants, values above the median for adiponectin [HR: 1.67 (1.23,1.26) p = 0.001] and FGF-21 [HR: 1.27 (1.09,1.47) p = 0.002] were associated with a greater likelihood of initiating a b/tsDMARD. Adipocytokines may serve as prognostic biomarkers of a more severe RA disease course. Additional study is needed to determine whether adipocytokines are phenotypic markers or whether they actively promote disease progression.


Dichloridobis(pyridine-2-thiol-ato-κ²N,S)tin(IV): a new polymorph.
The title compound, [SnCl₂(C₅H₄NS)₂], is the product of reaction of 2,2'-dipyridyl disulfide with tin tetra-chloride. The Sn(IV) atom adopts a distorted octa-hedral geometry, with the two bidentate pyridine-2-thiol-ate ligands forming two planar four-membered chelate rings. The two Sn-Cl, two Sn-N and two Sn-S bonds are in cis, cis and trans configurations, respectively. The crystal grown from acetonitrile represents a new monoclinic polymorph in space group C2/c with the mol-ecule having twofold rotational symmetry, the Sn(IV) atom lying on the twofold axis. The mol-ecular structure of the monoclinic polymorph is very close to that of the triclinic polymorph studied previously in space group P-1, the mol-ecule occupying a general position [Masaki & Matsunami (1976 ▶). Bull. Chem. Soc. Jpn, 49, 3274-3279; Masaki et al. (1978 ▶). Bull. Chem. Soc. Jpn, 51, 3298-3301]. Apparently, the formation of the two polymorphs is determined by the different systems of inter-molecular inter-actions. In the crystal of the monoclinic polymorph, mol-ecules are bound into ribbons along the c axis by C-H⋯Cl hydrogen bonds, whereas in the crystal of the triclinic polymorph, mol-ecules form chains along the a axis by attractive S⋯S inter-actions. The crystal studied was a pseudo-merohedral twin; the refined BASF value is 0.221 (1).


Human adenovirus type 5 vectors expressing rabies glycoprotein.
The prevalence of wildlife rabies throughout the world and the continued spread of this disease in North America highlights the need for oral vaccines which may be used safely and effectively to vaccinate a number of species that are reservoirs or vectors of rabies. We have previously shown that AdRG1, a replication competent recombinant human adenovirus type 5 (Ad5) expressing a rabies glycoprotein (RG), can induce immunity to rabies in rodent, canine, and skunk model systems. To improve the Ad5 vector system as a potential oral vaccine, we have constructed additional Ad5 recombinant vectors and compared RG expression in cell culture and immunogenicity in animals. Two new replication competent vectors are compared. AdRG1.3, which carries RG with accompanying SV40 poly A addition sequences within an E3 deletion, and AdRG4, which has RG in the E3 deletion but under the control of an exogenous Ad2 major late promoter, both express higher levels of RG in permissive cell culture than did AdRG1 and both elicit high levels of serum anti-rabies antibodies by parenteral or oral routes in animals. AdRG1.3 may be a more effective vaccine vector in species which are non-permissive for the replication of human Ad5.


Treponema pallidum recombinant protein Tp47 activates NOD-like receptor family protein 3 inflammasomes in macrophages via glycolysis.
Glycolysis is a key pathway in cellular glucose metabolism for energy supply and regulates immune cell activation. Whether glycolysis is involved in the activation of NOD-like receptor family protein 3 (NLRP3) inflammasomes during Treponema pallidum (T. pallidum) infection is unclear. In this study, the effect of T. pallidum membrane protein Tp47 on NLRP3 inflammasome activation in rabbit peritoneal macrophages was analysed and the role of glycolysis in NLRP3 inflammasome activation was explored. The results showed that Tp47 promoted NLRP3, caspase-1, and IL-1β mRNA expression in macrophages, enhanced glycolysis and glycolytic capacity of macrophage, and promoted the production of macrophage glycolytic metabolites citrate, phosphoenolpyruvate, and lactate. The M2 pyruvate kinase (PKM2) inhibitor shikonin down-regulated the Tp47-promoted NLRP3, caspase-1, and IL-1β mRNA expression in macrophages, and suppressed the Tp47-enhanced glycolysis and glycolytic capacity. Similarly, si-PKM2 significantly inhibited Tp47-promoted NLRP3, caspase-1, and IL-1β mRNA expression and the Tp47-enhanced glycolysis and glycolytic capacity in macrophages. In conclusion, Tp47 activated NLRP3 inflammasomes via PKM2-dependent glycolysis and provided a new perspective on the effect of T. pallidum infection on host macrophages, which would contribute to the understanding of the infection mechanism and host immune mechanism of T. pallidum.


Painting of fourth and chromosome-wide regulation of the 4th chromosome in Drosophila melanogaster.
Drosophila melanogaster exhibits two expression-regulating systems that target whole, specific chromosomes: the dosage compensation system whereby the male-specific lethal complex doubles transcription of genes on the male X-chromosome and the chromosome 4-specific protein Painting of fourth, POF. POF is the first example of an autosome-specific protein and its presence raises the question of the universality of chromosome-specific regulation. Here we show that POF and heterochromatin protein 1 (HP1) are involved in the global regulation of the 4th chromosome. Contrary to previous conclusions, Pof is not essential for survival of diplo-4th karyotype flies. However, Pof is essential for survival of haplo-4th individuals and expression of chromosome 4 genes in diplo-4th individuals is decreased in the absence of Pof. Mapping of POF using chromatin immunoprecipitation suggested that it binds within genes. Furthermore, we show that POF binding is dependent on heterochromatin and that POF and HP1 bind interdependently to the 4th chromosome. We propose a balancing mechanism involving POF and HP1 that provides a feedback system for fine-tuning expression status of genes on the 4th chromosome.


How is inflammatory bowel disease managed in Spanish gastroenterology departments? The results of the GESTIONA-EII survey.
Not all national health centers include specialized units or clinicians devoted to inflammatory bowel disease. The goal of the survey was to gain an insight into the management of this disease within Spanish gastroenterology departments via a survey among their members. An online survey was conducted in February and March 2015, among SEPD members (2017 clinician members), who were split into three categories: heads of department, general gastroenterologists, and experts in this disease. The results of the last two surveys are reported, including demography-related questions and specific questions on the strategies and resources available for the care of these patients. A total of 166 responses were received (response rate 8.19%), excluding those from heads of department (previously published). Sixty gastroenterologists considered themselves experts in inflammatory bowel disease, and 106 non-experts in it, the latter being either general gastroenterologists or specialists in other areas, mainly endoscopy. Twenty-eight percent of non-expert gastroenterologists said their hospitals had specific units, with a monographic clinic in 46%. However, 26% reported that they were treating affected patients themselves. Experts in inflammatory bowel disease reported that their institute had resources to support their work, but there was a lack of surgeons with expertise in this condition, particularly in county hospitals. At least, within SEPD members, 2 out of 3 experts in inflammatory bowel disease seem to have the resources available for their work (nurses, day unit, telephone line, database, referrals, joint sessions). Although there is room for improvement (email to contact patients, devoted surgeon, absence of referral protocols), and 2 out of 3 are concerned about pharmacy costs. Since a substantial number of patients remain treated by general practitioners, rapid referral programs might be helpful in this setting.


Precision Reengineering of the Oral Microbiome for Caries Management.
Technological advancements have revolutionized our understanding of the complexity and importance of the human microbiome. This progress has also emphasized the need for precision therapeutics, as it has underscored the dilemmas, such as dysbiosis and increasing antibiotic resistance, associated with current, broad-spectrum treatment modalities. Dental caries remains the most common chronic disease worldwide, accompanied by a tremendous financial and social burden, despite widespread and efficacious fluoride and hygienic regimens. Over the past several decades, various precision approaches to combat dental caries, including vaccines, probiotics, and antimicrobial compounds, have been pursued. Despite the distinct overall conceptual strengths of each approach, for various reasons, there are currently no approved precision antibiotic therapeutics to prevent dental caries. Specifically targeted antimicrobial peptides (STAMPs) are synthetic molecules that combine the antibiotic moiety of a traditional antimicrobial peptide with a targeting domain to provide specificity against a particular organism. Conjoining the killing domain from the antimicrobial, novispirin G10, and a targeting domain derived from the Streptococcus mutans pheromone, CSP, the STAMP C16G2 was designed to provide targeted killing of S. mutans, widely considered the keystone species in dental caries pathogenesis. C16G2 was able to selectively eliminate S. mutans from complex ecosystems while leaving closely related, yet health-associated, oral species unharmed. This remodeling of the dental plaque community is expected to have significant advantages compared to conventional broad-spectrum mouthwashes, as the intact, surviving community is apt to prevent reinfection by pathogens. Following successful phase I clinical trials that evaluated the safety and basic microbiology of C16G2 treatments, the phase II trials of several C16G2 formulations are currently in progress. C16G2 represents an exciting advance in precision therapeutics, and the STAMP platform provides vast opportunities for both the development of additional therapeutics and the overall study of microbial ecology.


Young retrotransposons and non-B DNA structures promote the establishment of dominant rye centromere in the 1RS.1BL fused centromere.
The fine centromere structure in Robertsonian wheat-rye translocation chromosomes exhibits variation among different translocation genotypes. Within extensively employed wheat-rye 1RS.1BL translocation lines in wheat breeding, their translocated chromosomes frequently display fused centromere. Nevertheless, the mechanism governing the functionality of the fused centromere in 1RS.1BL translocated chromosomes remains to be clarified. In this study, we investigated the fine centromere structure of the 1RS.1BL translocated chromosome through a combination of cytological and genomics methods. We found that only the rye-derived centromere exhibits functional activity, whether in breeding applications or artificially synthesized translocation chromosomes. The active rye-derived centromere had higher proportion of young full-length long terminal repeat retrotransposons (flLTR-RTs) and more stable non-B DNA structures, which may be beneficial toward transcription of centromeric repeats and CENH3 loading to maintain the activity of rye centromeres. High levels of DNA methylation and H3K9me2 were found in the inactive wheat-derived centromeres, suggesting that it may play a crucial role in maintaining the inactive status of the wheat centromere. Our works elucidate the fine structure of 1RS.1BL translocations and the potential mechanism of centromere inactivation in the fused centromere, contributing knowledge to the application of fused centromere in wheat breeding formation of new wheat-rye translocation lines.


Survival after In-Hospital Cardiopulmonary Resuscitation in a Major Referral Center during 2001-2008.
Despite efforts to save more people suffering from in-hospital cardiac arrest, rates of survival after in-hospital cardiopulmonary resuscitation (CPR) are no better today than they were more than a decade ago. This study was undertaken to assess the demographics, clinical parameters and outcomes of patients undergoing CPR by the code blue team at our center during 2001 to 2008. Data were collected retrospectively from adult patients (n=2262) who underwent CPR. Clinical outcomes of interest were survival at the end of CPR and survival at discharge from the hospital. Factors associated with survival were evaluated using binomial and tests. Of the patients included (n=2262), 741 patients (32.8%) had successful CPR. The number of male patients requiring CPR was more than females in need of the procedure. The majority of patients requiring CPR were older than 60 years (56.4±17.9). The number of successful CPR cases in long-day shift (7:00 to 19:00) was more than that in the night shift (19:00 to 7:00). Furthermore, 413 (18.4%) cases were resuscitated on holidays and 1849 (81.7%) on the working days. The duration of CPR was 10 min or less in 710 (31.4%) cases. Cardiopulmonary resuscitations which lasted less than 10 minutes were associated with better outcomes. The findings of the present study indicate that some manageable factors including the duration of CPR, working shift, working day (holiday or non-holiday) could affect the CPR outcomes. The findings might also be taken as evidence to suggest that the allocation of more personnel in each shift especially in night shifts and holidays, planning to increase the personnel's CPR skills, and decreasing the waste time would result in the improvement of CPR outcome.


Phylogeny of Mycoplasma bovis isolates from Hungary based on multi locus sequence typing and multiple-locus variable-number tandem repeat analysis.
Mycoplasma bovis is an important pathogen causing pneumonia, mastitis and arthritis in cattle worldwide. As this agent is primarily transmitted by direct contact and spread through animal movements, efficient genotyping systems are essential for the monitoring of the disease and for epidemiological investigations. The aim of this study was to compare and evaluate the multi locus sequence typing (MLST) and the multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) through the genetic characterization of M. bovis isolates from Hungary. Thirty one Hungarian M. bovis isolates grouped into two clades by MLST. Two strains had the same sequence type (ST) as reference strain PG45, while the other twenty nine Hungarian isolates formed a novel clade comprising five subclades. Isolates originating from the same herds had the same STs except for one case. The same isolates formed two main clades and several subclades and branches by MLVA. One clade contained the reference strain PG45 and three isolates, while the other main clade comprised the rest of the strains. Within-herd strain divergence was also detected by MLVA. Little congruence was found between the results of the two typing systems. MLST is generally considered an intermediate scale typing method and it was found to be discriminatory among the Hungarian M. bovis isolates. MLVA proved to be an appropriate fine scale typing tool for M. bovis as this method was able to distinguish closely related strains isolated from the same farm. We recommend the combined use of the two methods for the genotyping of M. bovis isolates. Strains have to be characterized first by MLST followed by the fine scale typing of identical STs with MLVA.


Ototoxicity of organic solvents - from scientific evidence to health policy.
The scientific workshop, organized under the 6th European Framework Programme, the Marie Curie Host Fellowship for the Transfer of Knowledge "NoiseHear" Project, by the Nofer Institute of Occupational Medicine (Łódź, Poland, 15-16 November 2006), gathered world specialists in noise, chemicals, and ototoxicity, including hearing researchers, toxicologists, otolaryngologists, audiologists and occupational health physicians.The workshop examined the evidence and the links between isolated exposure to organic solvents, combined exposure to noise and solvents, and effects on the auditory system. Its main purpose was to review the key scientific evidence to gather the necessary knowledge for developing adequate occupational health policies. This paper summarizes the workshop sessions and subsequent discussions.