The role of diaphragmatic surgery during interval debulking after neoadjuvant chemotherapy: an analysis of 74 patients with advanced epithelial ovarian cancer.
The purpose of this retrospective study was to evaluate diaphragmatic surgery in achieving optimal cytoreductive results and its associated complications during interval debulking surgery in patients with advanced ovarian cancer. After retrospective review of medical records, diaphragmatic surgery was performed in 74 of 128 consecutive patients with advanced epithelial ovarian cancer who underwent interval debulking, between September 1993 and December 2007. Four different approaches were performed: coagulation (group 1), stripping (group 2), combination of stripping with coagulation (group 3), and diaphragm full-thickness resection including muscle with pleura (group 4). Cytoreductive outcome, morbidity, overall survival, and disease-free survival were analyzed. Two patients (2.7%) had International Federation of Gynecology and Obstetrics stage IIIB disease; 46 (62.16%), stage IIIC; and 26 (35.13%), stage IV. After 3 to 4 cycles of neoadjuvant platinum-based chemotherapy, the diaphragmatic disease was coagulated in 43 patients (58.10%) and was only stripped in 10 (13.51%); in 19 patients (25.67%), a combination of these techniques was applied; and in 2 (2.70%), the disease was resected, with the adjacent infiltrated part of the diaphragmatic muscle and the pleura above it. Debulking to no residual was achieved in 95%, 100%, 100%, and 50% for groups 1, 2, 3, and 4, respectively. The median disease-free survival was 15, 14, and 14 months, and the median overall survival was 34, 30, and 51 months for groups 1, 2, and 3, respectively, and were not reached for group 4. Minor and major complications were comparable among the groups. Pleural effusions were the most frequent associated complication, and chest tube placement (1.3%) or thoracocentesis (4%) were necessary for the relief of respiratory distress. The perioperative mortality rate was 0%. Diaphragmatic surgery during interval debulking enhances optimal cytoreduction rates and improves survival with acceptable and manageable morbidity. In patients with thick (>4 mm) or large (>1 cm) lesions, stripping the diaphragm or full-thickness resection of the diaphragmatic muscle is preferred.


Trends in serum citrulline and acute rejection among recipients of small bowel transplants.
A test for detecting acute cellular rejection (ACR) of small intestinal transplants (ITx) would be a major advance. Small preliminary studies suggest that serum citrulline levels correlate with ACR. The results for a group of 26 isolated intestinal and multivisceral transplant recipients are summarized here. Serum citrulline concentrations were determined by ion exchange chromatography and compared to biopsy-based grade of ACR. Other factors considered included patient and donor age and sex, ischemia time, and serum creatinine. Straight-line fits were employed to describe how each patient's citrulline levels changed over time. Estimated times to achieve normal citrulline (>or=30 micromol/L) ranged from 1 to 730 days posttransplant for 21 patients demonstrating increasing citrulline levels over time. Using stepwise linear regression, patients' ranks for time required to achieve normal citrulline levels were the only independent predictors of both maximum ACR (P <.0001) and average ACR (P =.0059) after 14 days posttransplant. The rate and direction of change in citrulline over time may be an indicator of the risk of acute rejection. We plan to further examine the use of citrulline as a marker for rejection in larger prospective studies.


Forecasting water quality variable using deep learning and weighted averaging ensemble models.
Water quality variables, including chlorophyll-a (Chl-a), play a pivotal role in comprehending and evaluating the condition of aquatic ecosystems. Chl-a, a pigment present in diverse aquatic organisms, notably algae and cyanobacteria, serves as a valuable indicator of water quality. Thus, the objectives of this study encompass: (1) the assessment of the predictive capabilities of four deep learning (DL) models - namely, recurrent neural network (RNN), long short-term memory (LSTM), gated recurrence unit (GRU), and temporal convolutional network (TCN) - in forecasting Chl-a concentrations; (2) the incorporation of these DL models into ensemble models (EMs) employing genetic algorithm (GA) and non-dominated sorting genetic algorithm (NSGA-II) to harness the strengths of each standalone model; and (3) the evaluation of the efficacy of the developed EMs. Utilizing data collected at 15-min intervals from Small Prespa Lake (SPL) in Greece, the models employed hourly Chl-a concentration lag times, extending up to 6 h, as models' inputs to forecast Chla (t+1). The proposed models underwent training on 70% of the dataset and were subsequently validated on the remaining 30%. Among the standalone DL models, the GRU model exhibited superior performance in Chl-a forecasting, surpassing the RNN, LSTM, and TCN models by 8%, 2%, and 2%, respectively. Furthermore, the integration of DL models through single-objective GA and multi-objective NSGA-II optimization algorithms yielded hybrid models adept at effectively forecasting both low and high Chl-a concentrations. The ensemble model based on NSGA-II outperformed standalone DL models as well as the GA-based model across a range of evaluation indices. For instance, considering the R-squared metric, the study's findings demonstrated that the EM-NSGA-II stands out with exceptional effectiveness compared to DL and EM-GA models, showcasing improvements of 14% (RNN), 8% (LSTM), 6% (GRU), 8% (TCN), and 3% (EM-GA) during the testing phase.


Risk factors for elementary school drinking: pubertal status, personality, and alcohol expectancies concurrently predict fifth grade alcohol consumption.
Little is known about the correlates and potential causes of very early drinking. The authors proposed this risk theory: (a) pubertal onset is associated with increased levels of positive urgency (the tendency to act rashly when experiencing intensely positive mood), negative urgency (the tendency to act rashly when distressed), and sensation seeking; (b) those traits predict increased endorsement of high-risk alcohol expectancies; (c) the expectancies predict drinker status among fifth graders; and (d) the apparent influence of positive urgency, negative urgency, and sensation seeking on drinker status is mediated by alcohol expectancies. The authors conducted a concurrent test of whether the relationships among these variables were consistent with the theory in a sample of 1,843 fifth grade students. In a well-fitting structural model, their hypotheses were supported. Drinker status among fifth graders is not just a function of context and factors external to children: it is predictable from a combination of pubertal status, personality characteristics, and learned alcohol expectancies.


Metazoan parasites of Hoplias aff. malabaricus, Trachelyopterus galeatus and Schizodon borellii (Osteichthyes) from the Protected Area and its main tributary, Brazil.
A study of the parasite-host interactions was conducted in the protected area popularly known as the "Refúgio Biológico de Santa Helena" and its tributary, the São Francisco Falso River. For each of the three host species, Schizodon borellii, Hoplias aff. malabaricus, and Trachelyopterus galeatus, 30 fish were collected from 2018 to 2019. A total of 2172 parasites were identified among the three host species. Among these, the Monogenea class had the highest number of species, with 26 taxa, followed by Copepoda with eight taxa, Digenea with six taxa, and Cestoda and Nematoda with one taxon each. Eleven new records of infection/infestation were found among the host species such as Urocleidoides paradoxus, Urocleidoides ramentacuminatus, Rhinoxenus arietinus, Mymarothecioides sp. (Monogenea), Ergasilus cf. bryconis, Lernaea devastatrix, and Brasergasilus sp. (Copepoda) parasitizing S. borellii. Trinigyrus sp., Vancleaveus sp. (Monogenea), Gamispinus diabolicus (Copepoda) present in T. galeatus, and Gamidactylus jaraquensis (Copepoda) present in H. aff. malabaricus. This study contributes to the record of parasite species occurrence in the vicinity of a protected area and helps fill gaps in the knowledge of fish parasitic fauna in the Neotropical region.


Thalamocortical synapses in the cat visual system in vivo are weak and unreliable.
The thalamocortical synapse of the visual system has been central to our understanding of sensory computations in the cortex. Although we have a fair understanding of the functional properties of the pre and post-synaptic populations, little is known about their synaptic properties, particularly in vivo. We used simultaneous recordings in LGN and V1 in cat in vivo to characterize the dynamic properties of thalamocortical synaptic transmission in monosynaptically connected LGN-V1 neurons. We found that thalamocortical synapses in vivo are unreliable, highly variable and exhibit short-term plasticity. Using biologically constrained models, we found that variable and unreliable synapses serve to increase cortical firing by means of increasing membrane fluctuations, similar to high conductance states. Thus, synaptic variability and unreliability, rather than acting as system noise, do serve a computational function. Our characterization of LGN-V1 synaptic properties constrains existing mathematical models, and mechanistic hypotheses, of a fundamental circuit in computational neuroscience.


Nonlinear chaotic component extraction for postural stability analysis.
This paper proposes a nonlinear analysis of the human postural steadiness system. The analyzed signal is the displacement of the centre of pressure (COP) collected from a force plate used for measuring postural sway. Instead of analyzing the classical nonlinear parameters on the whole signal, the proposed method consists of analyzing the nonlinear dynamics of the intrinsic mode functions (IMF) of the COP signal. Based on the computation of the IMFs Lyapunov exponents, it is shown that pre-processing the COP signal with the Empirical Mode Decomposition allows an efficient extraction of its chaotic component.


Untranslated regions from C4 amaranth AhRbcS1 mRNAs confer translational enhancement and preferential bundle sheath cell expression in transgenic C4 Flaveria bidentis.
Many aspects of photosynthetic gene expression are posttranscriptionally regulated in C4 plants. To determine if RbcS mRNA untranslated regions (UTRs) in themselves could confer any characteristic C4 expression patterns, 5'- and 3'-UTRs of AhRbcS1 mRNA from the C4 dicot amaranth were linked to a gusA reporter gene. These were constitutively transcribed from a cauliflower mosaic virus promoter and assayed for posttranscriptional expression patterns in transgenic lines of the C4 dicot Flaveria bidentis. Three characteristic C4 expression patterns were conferred by heterologous AhRbcS1 UTRs in transgenic F. bidentis. First, the AhRbcS1 UTRs conferred strong translational enhancement of gusA expression, relative to control constructs lacking these UTRs. Second, while the UTRs did not appear to confer tissue-specific expression when analyzed by beta-glucuronidase activity assays, differences in gusA mRNA accumulation were observed in leaves, stems, and roots. Third, the AhRbcS1 UTRs conferred preferential gusA expression (enzyme activity and gusA mRNA accumulation) in leaf bundle sheath cells. AhRbcS1 UTR-mediated translational enhancement was also observed in transgenic C3 plants (tobacco [Nicotiana tabacum]) and in in vitro translation extracts. These mRNAs appear to be translated with different efficiencies in C4 versus C3 plants, indicating that processes determining overall translational efficiency may vary between these two categories of higher plants. Our findings suggest that the AhRbcS1 5'-UTR functions as a strong translational enhancer in leaves and other tissues, and may work synergistically with the 3'-UTR to modulate overall levels of Rubisco gene expression in different tissues and cell types of C4 plants.


Impaired endothelium-mediated relaxation in coronary arteries from insulin-resistant rats.
The insulin resistance syndrome is associated with atherosclerosis and cardiovascular events; however, the underlying mechanism of vascular dysfunction is unknown. The purpose of the current study was to assess endothelium- and smooth-muscle-mediated vasodilation in isolated coronary arteries from insulin-resistant rats and to determine whether insulin resistance alters the activity of the specific endothelium-derived relaxing factors. Male Sprague-Dawley rats were randomized to insulin resistance or control. Insulin resistance was induced by a fructose-rich diet. After 4 weeks of diet, coronary arteries were removed and vascular function was assessed in vitro using videomicroscopy. Acetylcholine (10(-9)-3 x 10(-5) M)- or sodium-nitroprusside (10(-9)-3 x 10(-4) M)-induced relaxations were determined. To evaluate the role of the specific endothelium-derived relaxing factors, several inhibitors were used, including N-nitro-L-arginine (LNNA), charybdotoxin/apamin (CTX/apamin), and indomethacin. Studies with nitroprusside showed that smooth-muscle-dependent relaxation did not differ between insulin resistance and control groups. In contrast, maximal relaxation (E(max)) to acetylcholine was decreased in the insulin resistance group (56 +/- 7%) versus control (93 +/- 3%). LNNA pretreatment further impaired E(max) in the IR group from 56 +/- 7 to 17 +/- 2% (p < 0.01). In control, E(max) was only slightly impaired by LNNA (93 +/- 3 to 63 +/- 6%; p < 0.05). The addition of CTX/apamin also decreased relaxation in the control group (93 +/- 3 to 47 +/- 7%; p < 0.05), whereas relaxation in insulin-resistant rats was not affected (45 +/- 5% with CTX/apamin vs. 56 +/- 7% with acetylcholine alone, NS). Pretreatment with indomethacin did not affect relaxation in either group, while pretreatment with the combination of LNNA and CTX/ apamin completely abolished relaxation in both groups. Endothelium-dependent relaxation is impaired in small coronary arteries from insulin-resistant rats. The mechanism of this defect is related to a decrease in an endothelium-dependent, nitric oxide/prostanoid-independent relaxing factor or endothelium-derived hyperpolarizing factor.


Analysis of performance of root-form endosseous implants placed in the maxillary sinus.
To evaluate the long-term effect on metal-bone interface of the perforation of the maxillary antral floor by root-form titanium implants, a study was undertaken in five mature Macaca fascicularis monkeys. One root-form titanium implant was placed bilaterally in the maxillary sinus perforating the sinus 5 mm and a second implant was placed completely in bone tissue. The protruding implant in the right antrum was grafted with autogenous cancellous bone and porous bone mineral (Bio-Oss) and the contralateral antral implant protruding in the antrum was left without grafting. The bilateral implants were used to support "free-standing" fixed prostheses, which were in full function for 14 months, during which time there was excellent clinical function and no mobility of the implants. Histomorphometric analysis of specimens revealed that there was no difference in the trabecular pattern or the amount of calcified matrix vs. marrow vascular spaces along the titanium implant interface. New reparative bone partially restored the osseous surface along the implant on the ungrafted side. Implants in the grafted antrum demonstrated bone regeneration over the implant surface ranging from 2 cm to complete regeneration. Histomorphometric analysis revealed the peri-implant bone to contain some particles of the Bio-Oss, although most of the implant bone interface exhibited only vital bone and marrow vascular spaces. The study indicates that the protrusion of implants maximally into the sinus floor does not necessarily require a bone graft to produce functional abutments for fixed prostheses and that implants can function well with 5 mm of protrusion into the antrum with excellent bone-metal interface.


Cytotoxic lymphocytes in the lungs of mice infected with respiratory syncytial virus.
Mice infected with respiratory syncytial virus (RSV) developed cytotoxic lymphocytes in the lungs, which lysed RSV-infected, but not uninfected cells. Cytotoxic activity was greatest 7 to 9 days after infection, was virus-specific, MHC-restricted and abolished by treatment of lymphocytes with anti-Thy 1.2 or with anti-Lyt 2.2 sera and complement. There was a close temporal relationship between the appearance of these cytotoxic lymphocytes in the lung and clearance of virus. In contrast, RSV persisted in the lungs of athymic (nude) mice and such animals failed to develop RSV-specific cytotoxic lymphocytes. Thus, cytotoxic T-cells may have an important role in recovery from RSV infection.


Nucleotide modifications enable rational design of TLR7-selective ligands by blocking RNase cleavage.
Toll-like receptors 7 (TLR7) and 8 (TLR8) each sense single-stranded RNA (ssRNA), but their activation results in different immune activation profiles. Attempts to selectively target either TLR7 or TLR8 have been hindered by their high degree of homology. However, recent studies revealed that TLR7 and TLR8 bind different ligands resulting from the processing of ssRNA by endolysosomal RNases. We demonstrate that by introducing precise 2' sugar-modified bases into oligoribonucleotides (ORNs) containing known TLR7 and TLR8 binding motifs, we could prevent RNase-mediated degradation into the monomeric uridine required for TLR8 activation while preserving TLR7 activation. Furthermore, a novel, optimized protocol for CRISPR-Cas9 knockout in primary human plasmacytoid dendritic cells showed that TLR7 activation is dependent on RNase processing of ORNs and revealed a previously undescribed role for RNase 6 in degrading ORNs into TLR ligands. Finally, 2' sugar-modified ORNs demonstrated robust innate immune activation in mice. Altogether, we identified a strategy for creating tunable TLR7-selective agonists.


Physical inactivity in Parkinson's disease.
Patients with Parkinson's disease (PD) are likely to become physically inactive, because of their motor, mental, and emotional symptoms. However, specific studies on physical activity in PD are scarce, and results are conflicting. Here, we quantified daily physical activities in a large cohort of PD patients and another large cohort of matched controls. Moreover, we investigated the influence of disease-related factors on daily physical activities in PD patients. Daily physical activity data of PD patients (n = 699) were collected in the ParkinsonNet trial and of controls (n = 1,959) in the Longitudinal Aging Study Amsterdam (LASA); data were determined using the LAPAQ, a validated physical activity questionnaire. In addition, variables that may affect daily physical activities in PD were recorded, including motor symptoms, depression, disability in daily life, and comorbidity. Patients were physically less active; a reduction of 29% compared to controls (95% CI, 10-44%). Multivariate regression analyses demonstrated that greater disease severity, gait impairment, and greater disability in daily living were associated with less daily physical activity in PD (R (2) = 24%). In this large study, we show that PD patients are about one-third less active compared to controls. While disease severity, gait, and disability in daily living predicted part of the inactivity, a portion of the variance remained unexplained, suggesting that additional determinants may also affect daily physical activities in PD. Because physical inactivity has many adverse consequences, work is needed to develop safe and enjoyable exercise programs for patients with PD.


Glutathione Peroxidase-like Antioxidant Activity of 1,3-Benzoselenazoles: Synthesis and In Silico Molecular Docking Studies as Pancreatic Lipase Inhibitors.
The synthesis of 1,3-benzoselenazoles was achieved by the reaction of corresponding bis[3-amino-N-(p-tolyl)benzamide-2-yl] diselenide, bis[3-amino-N-(4-methoxyphenyl)benzamide-2-yl] diselenide, and bis[3-amino-N-(4-(dimethylamino)phenyl) benzamide-2-yl] diselenide with aryl aldehydes. The 1,3-benzoselenazoles continued to exist as planar molecules due to the presence of secondary Se···O interactions as revealed by the single-crystal X-ray analysis. The presence of secondary Se···O interactions in 1,3-benzoselenazoles was confirmed using natural bond orbital (NBO) and atoms in molecules (AIM) calculations. Nucleus-independent chemical shift (NICS) values suggested the presence of aromatic character in a five-membered benzoselenazole heterocyclic ring. The glutathione peroxidase (GPx)-like antioxidant activity of all 1,3-benzoselenazoles was assessed using a thiophenol assay, exhibiting greater antioxidant activity than Ph2Se2 used as a reference. The most active catalyst carrying a strong electron-donating group (-NMe2) at the ortho-position to the benzoselenazole ring was further investigated at different concentrations of thiophenol, H2O2, and 1,3-benzoselenazoles as catalyst for determining their catalytic parameters. Moreover, the potential applications of all 1,3-benzoselenazoles against pancreatic lipase (PL) have been identified using in silico interactions between the active sites of the 1LPB protein as evaluated using a molecular docking study.


Computed tomography of the brain, chest, and abdomen in the preoperative assessment of non-small cell lung cancer.
The benefit to be gained from carrying out computed tomography of brain and abdomen in addition to the chest has been evaluated retrospectively in 114 consecutive patients with non-small cell lung cancer who, on the basis of history, clinical examination, chest radiography, and bronchoscopy had been considered potentially operable. Computed tomography of the chest showed potentially inoperable tumour in 37 patients, of whom 25 had tumour confined to the chest. Three patients were shown to have malignant disease within the mediastinum and abdomen; five within the mediastinum and brain; and four within the mediastinum, abdomen, and brain. Computed tomography of the abdomen disclosed deposits in nine patients, but in only two were the abnormalities restricted to the abdomen. Computed tomography of the brain showed metastases in 10 patients, of whom only one had metastatic disease confined to the brain. Thus three patients had isolated deposits in the abdomen and brain. In 12 patients the identification of metastases in the abdomen and brain removed the need for mediastinoscopy. Preoperative computed tomography of the abdomen and brain detected occult metastases in 15 patients (13%) in this study. In three patients the extrathoracic abnormality proved the only contraindication to surgery, but in the other 12 it provided valuable corroborative evidence of incurability and facilitated the assessment of the mediastinal abnormality.


Summary of track C: epidemiology and public health.
To review Track C on epidemiology and public health. Recent trends in the descriptive epidemiology are reported, for example, the rapid spread of HIV in certain Asian countries and the more precise insights in the spread thanks to subtyping of HIV-1 (and HIV-2). There is now ample evidence that sexually transmitted diseases enhance the rate of transmission. Viral load in the plasma of the mother is highly predictive for perinatal transmission. Natural history studies have shown that true non-progressors are probably rare. Preliminary evidence indicates that the progression rate to AIDS and death does not differ by HIV-1 subtype. Some genetic factors are associated with the rate of disease progression and a few with susceptibility to HIV infection. Needle-exchange programmes as an intervention measure for injecting drug users were hotly debated and so were HIV (home) testing and counselling. Successes in prevention were reported from Thailand and Uganda, and also from small scale programmes.


Long Noncoding RNA SOCS2-AS Promotes Leukemogenesis in FLT3-ITD+ Acute Myeloid Leukemia Through miRNA-221.
LncRNAs play an important role in tumorigenesis and development in tumors, but the function of lncRNA SOCS2-AS in acute myeloid leukemia (AML) is unknown. In the present study, we used RT-PCR to detect the expression of SOCS2-AS in FLT3-ITD+, FLT3-ITD- AML patients and different AML cell lines. The colony formation and CCK-8 assay were performed to analyze the proliferation ability, and the flow cytometry was performed to analyze the capacity of apoptosis in Molm-13 and MV4-11 cells. The Western blot was applied to detect the expression of STAT5 and p-STAT5. The RNA pull-down and luciferase activity were used to investigate the interaction between SOCS2-AS and miR-221. The results indicate that SOCS2-AS shows overexpression in FLT3-ITD+ AML patients compared to FLT3-ITD- AML patients. Si-SOCS2-AS can inhibit the proliferation, boost the apoptosis and induce the cycle arrest in Molm-13 cells, and SOCS2-AS overexpression promotes proliferation and colony formation in MV4-11 cells. The miR-221 shows overexpression in FLT3-ITD+ AML patients compared to FLT3-ITD- AML patients. And the expression level of miR-221 and SOCS2-AS shows negative correlation in FLT3-ITD+ AML patients. Functionally, SOCS2-AS could be interacted with miR-221 in AML cells. After SOCS2-AS knockdown, the phosphorylation level of STAT5 was significantly decreased. Moreover, miR-221 inhibitor can rescue the viability in cells after si-SOCS2-AS transfection. And it is stated that SOCS2-AS regulates the STAT5 signal transduction pathway with sponging miR-221. In conclusion, this study confirms the molecular mechanism of SOCS2-AS in AML by targeting the miR-221/STAT5 signaling pathway. This indicates SOCS2-AS may serve as a potential therapeutic target for the treatment of AML.


The use of the multiplicity survey to identify migrants.
New approaches must be developed and tested to meet the needs for current data on population movement. One such approach is the survey with multiplicity (social networks). It allows respondents to report events (in this case, migration) that occurred to specified relatives living elsewhere. In order to test the value of the approach on a limited, exploratory basis, a two-stage research design was employed. A sample of persons resident in Cranston, Rhode Island who had changed address in the three years preceding the survey, was first identified and information was gathered in Round I on their mobility and on their relatives living in the State. In turn, these relatives (Round II) were asked about the mobility of their parents, siblings, and children. (The responses from Round II should have included the migrants in Round I.) Evaluation of the accuracy of Round II responses in comparison to those obtained in Round I was undertaken in terms of kin relationships, recency of move, and type of move. The results indicate that respondents had a low awareness of their relatives' movements, that moves of females were more likely to be reported than those of males, that recent moves were more frequently cited and that moves between communities were more likely to be reported than intracity mobility.


A study of a three-dimensional PLGA sponge containing natural polymers co-cultured with endothelial and mesenchymal stem cells as a tissue engineering scaffold.
The interaction between vascular endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) in a complex hemodynamic and mechanical environment plays an important role in the control of blood vessel growth and function. Despite the importance of VSMCs, substitutes are needed for vascular therapies. A potential VSMC substitute is human adult bone marrow derived mesenchymal stem cells (hMSCs). In this study, the effect of poly(lactic-co-glycolic acid) (PLGA) scaffolds containing three natural polymers (demineralized bone particles, silk, and small intestine submucosa) on the phenotype of MSCs and SMCs cultured with or without ECs was investigated. The study objective was to create a media equivalent for a tissue engineered blood vessel using PLGA, natural polymers, and MSCs co-cultured with ECs. The PLGA containing the natural polymers silk and SIS showed increased proliferation and cell adhesion. The presence of silk and DBP promoted a MSC phenotype change into a SMC-like phenotype at the mRNA level; however these differences at the protein level were not seen. Additionally, PLGA containing SIS did not induce SMC gene or protein upregulation. Finally, the effect of ECs in combination with the natural polymers was tested. When co-cultured with ECs, the mRNA of SMC specific markers in MSCs and SMCs were increased when compared to SMCs or MSCs alone. However, MSCs, when co-cultured with ECs on PLGA containing silk, exhibited significantly increased α-SMA and calponin expression when compared to PLGA only scaffolds. These results indicate that the natural polymer silk in combination with the co-culture of endothelial cells was most effective at increasing cell viability and inducing a SMC-like phenotype at the mRNA and protein level in MSCs.


[PET scan in neuro-intensive care and research].
The main objective of neurointensive care is to counteract the development of secondary brain ischemia. The management is focused on preventing, detecting and correcting secondary insults that are likely to produce ischemic brain damage. This requires intensive multimodality monitoring of the brain. There is no reliable technique available for continuous monitoring of cerebral blood flow and therefore intermittent methods have to be used in some cases for objective measurement of cerebral perfusion. PET allows measurements of both cerebral blood flow and metabolism, which is essential for the assessment of the energy metabolic state of the brain tissue. PET may be used to guide therapeutic intervention, to evaluate the effect of treatment, to validate new techniques for monitoring of the brain and to determine the efficacy of potential neuroprotective drugs. The possibilities that PET offers in neurointensive care and research are illustrated. The experiences from the application of PET in the evaluation of pharmacological treatment of increased intracranial pressure in head injured patients and the use of PET in combination with intracerebral microdialys, in an MCA-occlusion-reperfusion primate model and in patients with subarachnoid hemorrhage, to validate intracerebral microdialysis as an instrument for chemical monitoring of the brain during neurointensive care, are reported.


recA is required in the induction of pectin lyase and carotovoricin in Erwinia carotovora subsp. carotovora.
Pectin lyase (PNL) and the bacteriocin carotovoricin (CTV) were induced in Erwinia carotovora subsp. carotovora 71 by the DNA-damaging agents mitomycin C, nalidixic acid, and UV light. To determine whether the recA product was involved in the expression of these damage-inducible phenotypes, we cloned the E. carotovora subsp. carotovora recA+ gene, inactivated it by Tn5 insertion, and constructed an E. carotovora subsp. carotovora recA::Tn5 strain by gene replacement via homologous recombination. The RecA- strain was more sensitive to methyl methanesulfonate, nitroquinoline oxide, and UV light than its RecA+ parent. The recA mutation did not affect the production of pectate lyase, polygalacturonase, cellulase, and protease or the ability to cause soft rot of potato tubers. With this mutant, unlike with the RecA+ parent strain, PNL and CTV were not induced by mitomycin C or detected in potato tuber tissue. The RecA+ phenotype, including the inducibility of PNL and CTV, could, however, be restored in the mutant in trans by the recA+ gene from either E. carotovora subsp. carotovora or Escherichia coli. We conclude that, in E. carotovora subsp. carotovora, the recA product is required in the induction of PNL and CTV.


The protective effects of tea polyphenols and schisandrin B on nephrotoxicity of mercury.
Mercury (Hg) is an occupational and environmental contaminant that is a well-recognized health hazard. To approach the concrete mechanisms of mercury nephrotoxicity and find out a new way to prevent it, the rats were subcutaneously injected with different dosages of mercuric chloride (HgCl(2))--0, 2.2, 4.4, and 8.8 μmol/kg. The levels of Hg, blood urea nitrogen (BUN), urine protein, glutathione (GSH), malondialdehyde (MDA) and activities of N-acetyl-beta-D-glucosaminidase (NAG), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were investigated, and the levels of reactive oxygen species (ROS) and apoptosis and the pathological changes were also observed. In addition, the effects of 1 mmol/kg tea polyphenols (TP) and 0.04 mmol/kg schisandrin B (Sch B) were studied at 8.8 μmol/kg HgCl(2). It was observed that the levels of Hg, BUN, urine protein, GSH, and MDA and activities of NAG, ALP, and LDH increased significantly; the activities of SOD and GSH-Px decreased significantly; the levels of ROS and apoptosis increased obviously; and many pathological changes occurred dose-dependently in the HgCl(2) injection groups. Further investigation indicated that pretreatment with TP and Sch B significantly reversed the toxic effects of HgCl(2). These results suggested that TP and Sch B might antagonize the nephrotoxicity caused by HgCl(2) exposure.


Bioactive self-healing umbilical cord blood exosomes hydrogel for promoting chronic diabetic wound healing.
The deleterious effects of diabetes mellitus on wound healing have become a major public health concern worldwide. Given the complex microenvironment of diabetic wounds and the high prevalence of diabetes, the design and development of novel wound dressing materials with versatile capabilities is urgent. Extracellular vesicles (EVs) derived from human umbilical cord blood have demonstrated the potential to counter inflammation and accelerate wound healing. Herein, we explored the efficacy of incorporating human umbilical cord blood-derived exosomes (UCB-Exos) into an ABA-type amphiphilic hydrogel, which possesses the attributes of exosome (Exo) encapsulation, temperature-triggered reversible sol-gel conversion, and Exo-regulated release, for enhancing the stability and retention of Exos. We sought to examine the feasibility of this strategy in augmenting the therapeutic efficacy of UCB-Exos for the healing of diabetes-related wounds. The injectable hydrogel was conveniently applied directly onto the wound surface and the enclosed Exo significantly facilitated the healing process, resulting in faster wound closure, enhanced collagen deposition, accelerated re-epithelialization, and enhanced neo-vascularization within two weeks compared with the hydrogel-only treatment group. In summary, some hydrogels hold great promise for promoting wound healing in diabetics and represent a novel therapeutic option for diabetes-related ulcers.


Solvation Dynamics in Liquid Water. 1. Ultrafast Energy Fluxes.
Solvation dynamics in liquid water is addressed via nonequilibrium energy-transfer pathways activated after a neutral atomic solute acquires a unit charge, either positive or negative. It is shown that the well-known nonequilibrium frequency shift relaxation function can be expressed in a novel fashion in terms of energy fluxes, providing a clear-cut and quantitative account of the processes involved. Roughly half of the initial excess energy is transferred into hindered rotations of first hydration shell water molecules, i.e., librational motions, specifically those rotations around the lowest moment of inertia principal axis. After integration over all water solvent molecules, rotations account for roughly 80% of the energy transferred, while translations have a secondary role; transfer to intramolecular water stretch and bend vibrations is negligible. This picture is similar to that for relaxation of a single vibrationally or rotationally excited water molecule in neat liquid water, although solvation relaxation is more nonlocal. In addition, we find a remarkable independence of the main relaxation channels on the newly created charge's sign. Although the methodology is applied here to the simplest solute case, the approach is rather general, and it should be at least equally useful in more realistic and complex scenarios.


Cranial Nerve Injury After Carotid Endarterectomy: Incidence, Risk Factors, and Time Trends.
To review the incidence of post-carotid endarterectomy (CEA) cranial nerve injury (CNI), and to evaluate the risk factors associated with increased CNI risk. The study was a meta-analysis. Pooled rates with 95% confidence intervals (CIs) were calculated for CNIs after primary CEA. Odds ratios (ORs) were calculated for potential risk factors. A fixed-effects model or a random effects model (Mantel-Haenszel method) was used for non-heterogeneous and heterogeneous data, respectively. Meta-regression analysis was performed to examine the influence of publication year upon CNI rate. Twenty-six articles, published between 1970 and 2015, were included in the meta-analysis, corresponding to 20,860 CEAs. Meta-analysis revealed that the vagus nerve was the most frequently injured cranial nerve (pooled injury rate 3.99%, 95% CI 2.56-5.70), followed by the hypoglossal nerve (3.79%, 95% CI 2.73-4.99). Fewer than one seventh of these injuries are permanent (vagus nerve: 0.57% [95% CI 0.19-1.10]; hypoglossal nerve: 0.15% [95% CI 0.01-0.39]). A statistically significant influence of publication year on the vagus and hypoglossal nerve injury rate was found, with the injury rate having decreased from about 8% to 2% and 1%, respectively, over the last 35 years. Urgent procedures (OR 1.59, 95% CI 1.21-2.10; p = .001), as well as return to the operating room for a neurological event or bleeding (OR 2.21, 95% CI 1.35-3.61; p = .002) were associated with an increased risk of CNI, whereas no statistically significant association was found between CNIs and the type of anaesthesia, the use of a patch, redo operation, and the use of a shunt. The vagus nerve appears to be the most frequently injured cranial nerve after CEA, followed by the hypoglossal nerve, with only a small proportion of these injuries being permanent. The CNI rate has significantly decreased over the past 35 years to a point indicating that CNIs should not be considered a major influencing factor in the decision making process between CEA and stenting.


MeCP2 is the protector of epigenome integrity, membrane-less nuclear architecture, and stability of chromatin assembly.
Tweetable abstract MeCP2 is an epigenetic factor with global impact in epigenome integrity, membrane-less nuclear architecture, and chromatin stability. Our Editorial covers recent advances on these important topics.


Fracture characteristics of entrapped head impacts versus controlled head drops in infant porcine specimens.
In many forensic cases, the job of forensic pathologists and anthropologists is to determine whether pediatric death is due to an abusive act or an accidental fall. The goal of this study was to compare the cranial fracture patterns generated on the parietal bone of a developing, infant porcine (pig, Sus scrofa) model by a controlled energy head drop onto a plate versus previous data generated by blunt force impact at the same energy onto the head constrained to a plate. The results showed that blunt force impacts on a head constrained to a rigid plate produces more fracture, but the same general pattern, as that for a head dropped onto the plate with the same level of impact energy. The study suggests that head constraint may be an important factor to consider in the evaluation of death causation for blunt force impacts to the pediatric skull.


Anti-Ro/SS-A antibodies in the pathophysiology of congenital heart block in neonatal lupus syndrome, an experimental model. In vitro electrophysiologic and immunocytochemical studies.
To determine whether anti-Ro/SS-A antibodies selectively bind to neonatal cardiac cells and alter membrane repolarization. An in vitro electrophysiologic and immunocytochemical experimental model contrasting neonatal and rabbit cardiac tissue was employed. Sera and IgG-enriched fractions from anti-Ro/SS-A antibody-positive mothers of infants with neonatal lupus erythematosus and congenital heart block bind to neonatal, rather than adult, rabbit cardiac tissue and alter the transmembrane action potential (i.e., inhibit repolarization). The additional presence of anti-La/SS-B antibodies was not additive or synergistic for these immunocytochemical and electrophysiologic features. Sera containing other antibody specificities (i.e., anti-native DNA, cardiolipin, Sm, and nuclear RNP) failed to stain the neonatal cardiac tissue or produced alterations in membrane repolarization. Anti-Ro/SS-A antibodies may play a pathophysiologic role in the development of congenital heart block in neonatal lupus.


Plasma metabolites to profile pathways in noncommunicable disease multimorbidity.
Multimorbidity, the simultaneous presence of multiple chronic conditions, is an increasing global health problem and research into its determinants is of high priority. We used baseline untargeted plasma metabolomics profiling covering >1,000 metabolites as a comprehensive readout of human physiology to characterize pathways associated with and across 27 incident noncommunicable diseases (NCDs) assessed using electronic health record hospitalization and cancer registry data from over 11,000 participants (219,415 person years). We identified 420 metabolites shared between at least 2 NCDs, representing 65.5% of all 640 significant metabolite-disease associations. We integrated baseline data on over 50 diverse clinical risk factors and characteristics to identify actionable shared pathways represented by those metabolites. Our study highlights liver and kidney function, lipid and glucose metabolism, low-grade inflammation, surrogates of gut microbial diversity and specific health-related behaviors as antecedents of common NCD multimorbidity with potential for early prevention. We integrated results into an open-access webserver ( https://omicscience.org/apps/mwasdisease/ ) to facilitate future research and meta-analyses.


Under the weather?-The direct effects of climate warming on a threatened desert lizard are mediated by their activity phase and burrow system.
For ectotherms such as lizards, the importance of behavioural thermoregulation in avoiding thermal extremes is well-established and is increasingly acknowledged in modern studies of climate warming and its impacts. Less appreciated and understood are the buffering roles of retreat sites and activity phase, in part because of logistical challenges of studying below-ground activity. Burrowing and nocturnal activity are key behavioural adaptations that have enabled a diverse range of reptiles to survive extreme environmental temperatures within hot desert regions. Yet, the direct impact of recent global warming on activity potential has been hypothesised to have caused extinctions in desert lizards, including the Australian arid zone skink Liopholis kintorei. We test the relevance of this hypothesis through a detailed characterisation of the above- and below-ground thermal and hydric microclimates available to, and used by, L. kintorei. We integrate operative temperatures with observed body temperatures to construct daily activity budgets, including the inference of subterranean behaviour. We then assess the likelihood that contemporary and future local extinctions in this species, and those of similar burrowing habits, could be explained by the direct effects of warming on its activity budget and exposure to thermal extremes. We found that L. kintorei spent only 4% of its time active on the surface, primarily at dusk, and that overall potential surface activity will be increased, not restricted, with climate warming. The burrow system provides an exceptional buffer to current and future maximum extremes of temperature (≈40°C reduction from potential surface temperatures), and desiccation (burrows near 100% humidity). Therefore, any climate warming impacts on this species are likely to be indirect. Our findings reflect the general buffering capacity of underground microclimates, therefore, our conclusions for L. kintorei are more generally applicable to nocturnal and crepuscular ectotherms, and highlight the need to consider the buffering properties of retreat sites and activity phase when forecasting climate change impacts.


Dengue/dengue haemorrhagic fever: history and current status.
Dengue fever (DF) is an old disease; the first record of a clinically compatible disease being recorded in a Chinese medical encyclopaedia in 992. As the global shipping industry expanded in the 18th and 19th centuries, port cities grew and became more urbanized, creating ideal conditions for the principal mosquito vector, Aedes aegypti. Both the mosquitoes and the viruses were thus spread to new geographic areas causing major epidemics. Because dispersal was by sailing ship, however, there were long intervals (10-40 years) between epidemics. In the aftermath of World War II, rapid urbanization in Southeast Asia led to increased transmission and hyperendemicity. The first major epidemics of the severe and fatal form of disease, dengue haemorrhagic fever (DHF), occurred in Southeast Asia as a direct result of this changing ecology. In the last 25 years of the 20th century, a dramatic global geographic expansion of epidemic DF/DHF occurred, facilitated by unplanned urbanization in tropical developing countries, modern transportation, lack of effective mosquito control and globalization. As we go into the 21st century, epidemic DF/DHF is one of the most important infectious diseases affecting tropical urban areas. Each year there are an estimated 50-100 million dengue infections, 500000 cases of DHF that must be hospitalized and 20000-25 000 deaths, mainly in children. Epidemic DF/DHF has an economic impact on the community of the same order of magnitude as malaria and other important infectious diseases. There are currently no vaccines nor antiviral drugs available for dengue viruses; the only effective way to prevent epidemic DF/DHF is to control the mosquito vector, Aedes aegypti.


Left Recumbent Position Decreases Heart Rate without Alterations in Cardiac Autonomic Nervous System Activity in Healthy Young Adults.
Some studies have reported that recumbent position may have advantages in patients with heart disease and in pregnancy. However, it remains controversial whether recumbent position affects autonomic nervous system activity and hemodynamics in healthy adults. The aim of this study was to evaluate alterations in heart rate variability (HRV) and hemodynamics in the supine, left recumbent and right recumbent positions in healthy young adults. A total of 80 participants aged 22.8 ± 3.1 years were enrolled in this observational study. Fifty-eight volunteers (29 men and 29 women) maintained the supine position followed by the left and right recumbent positions, while electrocardiographic data were recorded for spectral analysis of HRV to assess cardiac vagal nerve and sympathetic nerve activities. The heart rate (HR) was significantly lower in the left recumbent position than in the other positions. There were no statistically significant differences in HRV among the three positions. Considering the possibility that the echographic procedure affects autonomic nervous system (ANS) activity, the other 22 participants (11 men and 11 women) underwent an echographic evaluation of hemodynamics in the heart and inferior vena cava (IVC) across the three positions. Although a low HR was also observed, there were no statistically significant differences in the IVC or the heart blood volume between the supine and the left recumbent positions. A postural change to the left recumbent position does not affect the cardiac blood circulation or ANS activity, though it does decrease HR in healthy young adults. This finding indicates that the lower HR in the left recumbent position is not attributable to the ANS activity.


Altered functional connectivity during visual working memory state in patients with mild cognitive impairment.
Patients with mild cognitive impairment (MCI) suffer from severe memory function impairment, especially working memory [1]. Based on Electroencephalogram (EEG), this study used power spectral density and large-scale network analysis to reveal the frequency changes of brain areas and the diverse network patterns during the visual WM coding stages between MCI and normal controls (NC). The results showed, compared to NC, the left and right prefrontal lobes and central regions has significant synchronization in the θ frequency band, while the left temporal lobe has significant desynchronization during the working memory coding state among MCI. Brain network analysis revealed the diverse network patterns during the WM coding stage between two group. Compared with the NC, the brain of MCI patients reduced the top-down visual network connection flow derived from frontal lobe to parietal lobe, and recruited a more up-down mechanism with a much stronger information flow from frontal lobe to occipital lobe during the WM coding state. This result provides a new perspective for the neural mechanism of WM deficits in MCI patients.Clinical Relevance-Abnormal EEG rhythm and connectivity of brain regions may be important indicators of WM disorders in patients with MCI.


Community quorum sensing signalling and quenching: microbial granular biofilm assembly.
Recent reports exploring the role of gradients of quorum sensing (QS) signals in functional activated sludge have raised the question of whether shared systems of signalling synthesis and degradation, or quorum quenching (QQ), across the community inform of the means by which QS biology regulate floccular and granular biofilm assembly. In this study, we aimed to explore the species origin and interactive role of QS and QQ activities in such highly diverse microbial biofilm communities. Here, such aims were addressed systematically by a comprehensive multi-pronged RNA-sequencing, microbiological and analytical chemistry experimental approach, using two related but independently evolved floccular and granular sludge communities. Our data revealed a distinct difference between the QS and QQ potentials of the two communities, with different species largely displaying either QS or QQ functions. The floccular sludge community showed a high rate of QQ activity, and this rate was dependent on the acyl chain length demonstrating specificity of degradation. When the floccular biomass was transformed into the granular sludge, the QQ activity of the community was reduced by 30%. N-acyl homoserine lactones with four to eight carbons on the acyl chain accumulated at the granular stage, and their concentrations were at least threefold higher than those of the floccular stage. These findings corroborated meta-community analysis where a major shift in the dominant species from potential signal quenchers to producers was observed during the transition from flocs to granules, indicating the role of species composition and associated signalling activities in coordinating community behaviours. This study suggests that QQ has an important function in regulating community level QS signalling, and provides a mechanistic insight into the role of QS biology in complex community assembly.


Closed reductions and percutaneus 'k' wire fixation for adolescent intercondylar fractures of the distal humerus.
Intercondylar (T-Condylar) fractures of the humerus are very rare in children. The usual mechanism of the injury which is involved is a fall on a flexed elbow. The treatment options vary, depending upon the degree of the displacement and the amount of comminution and; they may range from a simple application of traction to open reduction and internal fixation with plates and screws. Open reduction is associated with significant post-operative stiffness of the elbow and subsequent less than satisfactory functional results. In this publication, we are presenting our experience of treating seven adolescent T-Condylar fractures of the distal humerus with closed reductions and internal fixations with the percutaneus Kirchner ('K') wire. Seven patients (M:F-6:1) with an average age of 14(range 12-16) years were treated by closed reduction and internal fixation with a 2.0 mm 'K' wire fixation under an image intensifier. Following the surgery, all the elbows were immobilised in plaster of Paris (POP) splints for three weeks. At three weeks after the surgery, the pop splints were removed and gentle elbow exercises were started. Five weeks after the surgery, the 'K' wires were removed, based on the radiological evidence of a bridging callus formation. Six patients (85%) out of seven showed satisfactory functional results on the Mayo Elbow Performance Score (MEPS), with a good range of motion at the elbow joint. Three (42%) patients had mild elbow stiffness, which resolved on subsequent treatment, while one (14%) had a mild pin tract infection, who was managed conservatively. There was one patient (14%) who was lost to follow-up, who ultimately had a painful and a stiff elbow. The remaining six patients were able to return to the premorbid sate. We recommend closed reduction and percutaneous 'K' wire fixation for adolescent T- Condylar fractures of the humerus as an easy and inexpensive procedure with satisfactory functional results.


Non-Surgical Endodontic Therapy as Treatment of Choice for a Misdiagnosed Recurring Extraoral Sinus Tract.
The purpose of this article is to present the treatment of an odontogenic cutaneous sinus tract with exuberant extraoral granulation tissue and its successful endodontic treatment and follow up with Cone-Beam Computed Tomography (CBCT). In this case, a 31-year-old woman was referred for management of a reddish nodule on her chin. Previous surgical and antibiotic intervention by the dermatologist had not resolved the problem. Profound clinical and radiological examination (including CBCT) revealed apical periodontitis of tooth 32 to be the cause of the recurring cutaneous sinus tract. Conservative non-surgical root canal treatment was performed. With the aid of a topical corticosteroid and supplemental antibiotic therapy, healing of the apical periodontitis and resolution of the granulation tissue was evident after 1 year both clinically and radiographically. This case report emphasises the need for more awareness by dermatologists and other medical practitioners for the differential diagnosis of extraoral sinus tracts. Correct diagnosis of the dental cause can prevent unnecessary and multiple antibiotic and surgical interventions. Antibiotic therapy should never be administered without addressing the underlying dental cause. Conservative non-surgical endodontic treatment is the treatment of choice for an extraoral sinus tract of endodontic origin.


Anti-inflammatory activity of pergolide, a dopamine receptor agonist.
Pergolide, a dopamine agonist effective in the treatment of Parkinson's disease, has been shown to have anti-inflammatory activity in the carrageenan paw edema assay in rats at p.o. doses greater than or equal to 0.3 mg/kg. Studies were done to investigate the mechanism of action and to determine the pharmacologic significance of this finding. Because pergolide elevates circulating glucocorticoids, the effect of pergolide on carrageenan-induced paw swelling was assessed in adrenalectomized rats. Pergolide retained its anti-inflammatory activity in adrenalectomized carrageenan-injected rats, thus eliminating corticosterone induction as a possible mechanism of action. Pergolide treatment also did not decrease thromboxane B2, prostaglandin E2 or leukotriene B4 production, ruling out direct effects on arachnoid acid inflammatory mediators. Interactions with the autonomic nervous system were suggested, in that an alpha adrenergic agonist (clonidine) mimicked the activity of pergolide in the carrageenan assay, and an alpha adrenergic antagonist (phenoxybenzamine) blocked the anti-inflammatory activity of pergolide in this assay. Dopamine receptor antagonists (haloperidol or sulpiride) partially inhibited the effect of pergolide in the carrageenan model. However, the peripherally restricted dopamine antagonist, domperidone, was ineffective, suggesting that a central dopamine receptor was involved in the effect. Experiments in chronic inflammation models such as lipoidal-amine induced arthritis in rats and picryl chloride-induced delayed type hypersensitivity in mice also revealed an anti-inflammatory effect of pergolide. Activity in the carrageenan system and the lipoidalamine model demonstrated that the anti-inflammatory effects of pergolide were separable from potential immunosuppressive effects. Multiple dose studies indicated that tolerance might develop to the anti-inflammatory effect of pergolide.(ABSTRACT TRUNCATED AT 250 WORDS)


[Experimental allergy to penicillin].
The inhalation method for penicillin sensitization with a long-term excitation of the respiratory tract mucosa with formalin was studied in experiments with rabbits. It was shown that inhalation of formalin significantly increased the frequency of the animal sensitization to penicillin. It is suggested that inflammation due to the formalin effect promoted penicillin binding by tissue proteins with formation of full value antigens from penicillin which resulted in the high frequency of the animal sensitization. The data may be used in studies on the allergenic properties of drugs.


A comparative stereological study of the term placenta in the donkey, pony and Thoroughbred.
The aim of the study was to compare horse and donkey placentae using stereological techniques. Term placentae were collected at spontaneous foaling from seven Thoroughbred mares, seven pony mares, and six jenny donkeys. Maternal and foal weights were recorded and the mass, volume, and gross area of each allantochorion was also recorded. Ten random biopsies were recovered and processed for light microscopy from which the surface density of the microcotyledons (S(v)) and the total microscopic area of fetomaternal contact were calculated stereologically. Gestation length was longer in the donkeys than the other two groups (median values: 371 vs. 327 and 341 days, P < 0.05). There were significant correlations between foal birthweight and gross area (rho = 0.89; n = 20; P < 0.05), mass (rho = 0.84; n = 20; P < 0.05) and volume (rho = 0.89; n = 20; P < 0.05) of the allantochorion. S(v) was higher in the donkey placenta than the other groups (median values: 0.05 vs. 0.03 and 0.04 microm(-1), P < 0.05) although placental efficiency was lower in the donkeys (median values: 0.87 vs. 1.33 and 1.32 kg/m2, P < 0.01). The results of the study confirmed that, although strong morphological similarities exist between the allantochorion of the horse and donkey, that of the donkey develops more complex microcotyledons, as judged stereologically, and exhibits a lower placental efficiency. These differences may be related to maternal genotype and/or the longer gestation length shown by the donkey compared to the horse, but a negative correlation (rho = -0.92, P < 0.01) was also found between age and placental efficiency in donkeys.


Comorbidities.
The term comorbidities or mixed pathologies is used when brain tissue, a surgical sample, or postmortem brain displays a mixture of protein alterations or other pathologies. Most of the alterations when seen in sufficient extent are considered causative, are related to a certain clinical phenotype, i.e., when hyperphosphorylated τ (HPτ) is observed in occipital cortex concomitant with β-amyloid (Aβ), the diagnosis is Alzheimer disease (AD). When HPτ is observed in hippocampal structures in a subject with extensive and widespread α-synuclein pathology, a Lewy body disease (LBD), the HPτ pathology is considered as a concomitant alteration. There are numerous reports indicating that when "concomitant" pathologies are seen in a subject with certain neurodegenerative diseases, the clinical phenotype might be altered. In addition there are those cases where many alterations are seen in a sparse extent, but jointly they lead to a clinical syndrome. Thus today it is not sufficient to confirm a certain pathology to be seen, i.e., AD- or LBD-related; in addition the concomitant aging-related alterations have to be looked for.


Assignment of Listeria grayi and Listeria murrayi to a single species, Listeria grayi, with a revised description of Listeria grayi.
The genomic relatedness between Listeria grayi and Listeria murrayi was reevaluated by using DNA-DNA hybridization, multilocus enzyme electrophoresis, and rRNA restriction fragment length polymorphism techniques. The high levels of similarity observed between the strains of these two species confirmed the data published since 1973 and indicated that they should be considered members of a single species. On grounds of priority, the species should be named L. grayi.


Recent advances in nanotechnology-based COVID-19 vaccines and therapeutic antibodies.
COVID-19 has caused a global pandemic and millions of deaths. It is imperative to develop effective countermeasures against the causative viral agent, SARS-CoV-2 and its many variants. Vaccines and therapeutic antibodies are the most effective approaches for preventing and treating COVID-19, respectively. SARS-CoV-2 enters host cells through the activities of the virus-surface spike (S) protein. Accordingly, the S protein is a prime target for vaccines and therapeutic antibodies. Dealing with particles with dimensions on the scale of nanometers, nanotechnology has emerged as a critical tool for rapidly designing and developing safe, effective, and urgently needed vaccines and therapeutics to control the COVID-19 pandemic. For example, nanotechnology was key to the fast-track approval of two mRNA vaccines for their wide use in human populations. In this review article, we first explore the roles of nanotechnology in battling COVID-19, including protein nanoparticles (for presentation of protein vaccines), lipid nanoparticles (for formulation with mRNAs), and nanobodies (as unique therapeutic antibodies). We then summarize the currently available COVID-19 vaccines and therapeutics based on nanotechnology.


Prevalence of, and factors associated with, gastroesophageal reflux disease: a population-based study in Shanghai, China.
The prevalence of gastroesophageal reflux disease (GERD) in China is lower than that in the Western countries, but appears to be increasing. The aim of this pilot study was to evaluate the prevalence of GERD in Shanghai, China, and to explore which population characteristics where associated with GERD. A sample of 1200 adult inhabitants of Shanghai, selected using randomized, stratified, multi-stage sampling, completed Mandarin translations of the Reflux Disease Questionnaire and GERD Impact Scale. Potential risk factors for GERD were examined by nested case-control analysis, using Cochran-Armitage trend testing and logistic regression analysis. The response rate was 86.2%; 919 responses were suitable for analysis. The prevalence of GERD, defined as heartburn and/or regurgitation of any frequency during the previous week, was 6.2% in Shanghai. Obesity and urban dwelling were associated with GERD (odds ratio 3.4, 95% confidence interval 1.3-9.3; and odds ratio 3.6, 95% confidence interval 1.2-10.4, respectively). The prevalence of GERD in Shanghai agreed with previous Chinese studies. GERD in Shanghai was associated with obesity and residency in an urban environment.


Enhanced stimulation of human bone marrow macrophage colony formation in vitro by recombinant human macrophage colony-stimulating factor in agarose medium and at low oxygen tension.
Recombinant (r) and natural human (h) macrophage colony-stimulating factor (M-CSF, CSF-1) have been considered poor stimulators of macrophage progenitor cells present in human marrow, although they are potent stimulators of these cells in mouse marrow. We compared the growth characteristics of rhM-CSF-responsive human macrophage progenitor cells placed in semisolid agarose or agar culture medium and incubated for 14 days at ambient (approximately 20%) or lowered (5%) O2 tension. By itself, rhM-CSF was found to be a good stimulator of macrophage colony formation by human bone marrow cells cultured in agarose but not in agar; this growth was enhanced by incubation at 5% O2. Maximal numbers (up to 115/10(5) nonadherent low density cells plated) of macrophage colonies (50 to greater than 500 cells per colony) were stimulated by 500 to 1,000 units rhM-CSF/mL, with 1/2 maximal numbers stimulated by 250 to 500 units/mL. With agarose as the support medium, rhM-CSF was two- to fourfold more active on mouse than on human macrophage colony formation, in contrast to previous reports of 10- to 100-fold greater activity when agar was used as the support medium. Using nonadherent low density T lymphocyte-depleted human bone marrow cells growing in agarose at 5% O2, greater than additive effects on colony formation were observed when 31 to 500 units rhM-CSF were used in combination with either 10 ng rh interleukin-1 alpha (IL-1 alpha), 20, or 200 units rh granulocyte-macrophage (GM)-CSF or rhG-CSF. The agarose assay system should be useful for evaluating factors regulating the proliferation of human macrophage progenitor cells in vitro and during clinical trials with rhM-CSF.


An analysis of two island groups as potential sites for trials of transgenic mosquitoes for malaria control.
Considerable technological advances have been made towards the generation of genetically modified mosquitoes for vector control. In contrast, less progress has been made towards field evaluations of transformed mosquitoes which are critical for evaluating the success of, and hazards associated with, genetic modification. Oceanic islands have been highlighted as potentially the best locations for such trials. However, population genetic studies are necessary to verify isolation. Here, we used a panel of genetic markers to assess for evidence of genetic isolation of two oceanic island populations of the African malaria vector, Anopheles gambiae s.s. We found no evidence of isolation between the Bijagós archipelago and mainland Guinea-Bissau, despite separation by distances beyond the known dispersal capabilities of this taxon. Conversely, the Comoros Islands appear to be genetically isolated from the East African mainland, and thus represent a location worthy of further investigation for field trials. Based on assessments of gene flow within and between the Comoros islands, the island of Grande Comore was found to be genetically isolated from adjacent islands and also exhibited local population structure, indicating that it may be the most suitable site for trials with existing genetic modification technologies.


Unmet care demands as perceived by stroke patients: deficits in health care?
To describe unmet care demands as perceived by stroke patients and to identify sociodemographic and health characteristics associated with these unmet demands to investigate the appropriateness of health care. Sample of patients who participated in a multicentre study (23 hospitals) on quality of care in The Netherlands. Non-institutionalised patients who had been admitted to hospital because of stroke. Patients were interviewed six months (n = 382) and five years (n = 224) after stroke. Six months after stroke data were collected on: (a) sociodemographic characteristics in terms of age, sex, living arrangement, educational level, and regional level of urbanisation; (b) health characteristics in terms of cognitive function, disability, emotional distress, and general health perception; (c) utilisation of professional care; and (d) unmet care demands as perceived by patients. Data on utilisation of care and unmet demands were also collected five years after stroke. Data were collected from June 1991 until December 1996. The percentage of unmet care demands was highest at six months after stroke (n = 120, 31%). Multiple logistic regression analyses showed that disabled patients were more likely to be unmet demanders for therapy, (I)ADL care and aids (range odds ratio (OR) = 3.5 to 7.9) than to be no demanders, whereas emotionally distressed patients were more likely to be unmet demanders for psychosocial support (OR = 3.8). When comparing unmet demanders with care users only for (instrumental) activities of daily living (I)ADL care differences were found: men (OR = 3.8), disabled patients (OR = 3.0), and emotionally distressed patients (OR = 6.5) were more likely to be users. Patients who perceived an unmet care demand do appear genuinely to have an unmet care need as supported by assessment of their health status: (a) types of unmet care demands correspond with types of health problems and (b) unmet demanders were in general unhealthier than no demanders and more comparable with care users for health characteristics. To improve an equitable distribution of healthcare services, guidelines for indicating and allocating health care have to be developed and should be based on scientific evidence and consensus meetings including professionals' and patients' perspectives.


Lifestyle changes during adolescence and risk of breast cancer: an ecologic study of the effect of World War II in Norway.
There are biologic reasons to believe that the period between the larche and the first full-term pregnancy is a particularly sensitive period in a woman's life regarding the development of breast cancer. In this ecologic study, data provided by the Norwegian Cancer Registry were analyzed to compare risk of breast cancer among women who experienced this sensitive period before, during, or after World War II. An ordinary age-cohort model and a model where the cohort was described by exposure by calendar period and sensitivity to this exposure at different ages, were fitted to the data. The incidence of breast cancer was lower than expected among women who experienced puberty during the war. The estimated configuration of the exposure variable showed an increase in exposure up to the start of WWII to twice the level in 1916, dropped by 13 percent during the war, and increased again after the war. The level in 1975 was approximately 2.7 times higher than the level in 1916. The results indicate that one or more lifestyle factors that changed among adolescent women during the war, influenced their risk of breast cancer. Dietary intake of energy, fat, meat, milk, fish, fresh vegetables, and potatoes, in addition to physical activity level and height, are important factors to consider in relation to breast cancer risk.


Global health surveillance.
Awareness of the importance of global health surveillance increased in the latter part of the 20th century with the global emergence of human immunodeficiency virus and novel strains of influenza. In the first decade of the 21st century, several events further highlighted global shared interests in and vulnerability to infectious diseases. Bioterrorist use of anthrax spores in 2001 raised awareness of the value of public health surveillance for national security. The epidemic of severe acute respiratory syndrome (SARS) in 2003, re-emergence of a panzootic of avian influenza A H5N1 in 2005, and the sudden emergence of pandemic H1N1 in North America in 2009 all highlighted the importance of shared global responsibility for surveillance and disease control. In particular, in 2003, SARS precipitated changes in awareness of the world's collective economic vulnerability to epidemic shocks.


Chlorantraniliprole resistance in the diamondback moth (Lepidoptera: Plutellidae).
The wide application of chlorantraniliprole, which selectively targets insect ryanodine receptors (RyR), for control of the diamondback moth, Plutella xylostella (L.), has led to increasingly prominent development of resistance to this insecticide. Although much work has been carried out on the structure and function of RyR, the molecular mechanisms of resistance to chlorantraniliprole in diamondback moth still needs further investigation. P. xylostella strains with medium and high resistance to chlorantraniliprole were obtained by laboratory selection and field collection. The biological activity of chlorantraniliprole against the third-instar larvae of susceptible and resistant strains was tested, and resistance development and biological fitness were investigated. The realized heritability (h2) of resistance showed the diamondback moth has a high risk of resistance to chlorantraniliprole. RyR transcript levels were lower in resistant strains than in susceptible strains, indicating that decreased expression of PxRyR may be associated with chlorantraniliprole resistance in P. xylostella. A 4,400 bp fragment of the RyR cDNA, which encodes most of the functional domains of RyR, was cloned and characterized from four strains (S, F18, BY, and ZC). A 14 amino acid (Q4546-S4559) deletion was found in three resistant strains (F18, BY, and ZC). A point mutation resulting in a glycine to glutamate substitution, as reported in a previously published article, was also found in the carboxyl-terminal region of two resistant strains (BY and ZC). These results indicated that decreased transcriptional level of RyR mRNA and combined with the site mutation might be related to chlorantraniliprole resistance in P. xylostella.


[Evaluation of long-term results of vidian neurectomy in the treatment of perennial rhinitis].
Vidian neurectomies by transnasal and transseptal approaches were performed on 21 patients with perennial rhinitis and with a follow-up of 5 years. The results showed that sneezing and watery rhinorrhea relapsed in 85.7% of the operated series and 66.7 per cent in one year after operation. The causes of recurrence were discussed and analysed. We consider that symptoms of sneezing and watery rhinorrhea are induced by a series of factors so that the efficacy of single vidian neurectomy needs further evaluation.


Specific sites within the ligand-binding domain and ion channel linkers modulate NMDA receptor gating.
Gating in the NMDA receptor is initiated in the extracellular ligand-binding domain (LBD) and is ultimately propagated via three linkers-S1-M1, M3-S2, and S2-M4-to the ion channel. M3-S2 directly couples LBD movements into channel gating, but the functional and structural contributions of S1-M1 and S2-M4 to the overall gating process are unknown. A scan of substituted cysteines in and around the NMDA receptor S1-M1 and S2-M4 with a bulky cysteine-reactive reagent identified numerous positions that showed potentiation of glutamate-activated as well as leak currents. As indexed by MK801 (dizocilpine hydrogen maleate), an open channel blocker, this potentiation was attributable to an increase in open probability, an interpretation confirmed for a subset of positions with single-channel recordings. The magnitude of this gating effect, acting through S1-M1 or S2-M4, was dependent on the intrinsic gating properties of the NMDA receptors, being more effective in the inherently low open probability GluN2C- than the higher open probability GluN2A-subunit-containing receptors. For the majority of these potentiation positions, we propose that alteration of gating arises from steric destabilization of contact interfaces where close apposition of the contacting partners is necessary for efficient channel closure. Our results therefore indicate that the NMDA receptor S1-M1 and S2-M4 linkers are dynamic during gating and can modulate the overall energetics of this process. Furthermore, the results conceptualize a mechanistic, as well as a possible structural, framework for pharmacologically targeting the linkers through noncompetitive and subunit-specific modes of action.


Alcohol-related morbidity and mortality.
Alcohol use is related to a wide variety of negative health outcomes including morbidity, mortality, and disability. Research on alcohol-related morbidity and mortality takes into account the varying effects of overall alcohol consumption and drinking patterns. The results from this epidemiological research indicate that alcohol use increases the risk for many chronic health consequences (e.g., diseases) and acute consequences (e.g., traffic crashes), but a certain pattern of regular light-to-moderate drinking may have beneficial effects on coronary heart disease. Several issues are relevant to the methodology of studies of alcohol-related morbidity and mortality, including the measurement of both alcohol consumption and the outcomes studied as well as study design. Broad summary measures that reflect alcohol's possible effects on morbidity, mortality, and disability may be more useful than measures of any one outcome alone.


Acoustic destruction of a microcapsule having a hard plastic shell.
Acoustic destruction of a microcapsule having a hard plastic shell is discussed. In an ultrasonic drug delivery system, microcapsules having thin elastic shells release drugs that are contained therein when the shell is destroyed. In this paper, two subjects related to capsule destruction are discussed: the driving pulse duration for capsule destruction and the frequency dependence of capsule destruction. Optical observation of microcapsule destruction is performed with a high-speed video camera. In the case of capsule destruction by a pulse wave, the internal gas of the microcapsule cannot be ejected completely, and a portion of the internal gas remains inside the broken shell. It is found that capsule destruction by pulse waves depends on both the amplitude of the driving pressure and the pulse duration. The frequency dependence of microcapsule destruction also is investigated. In the case of capsule destruction by a low-amplitude acoustic wave, the destruction rate under the resonance condition is higher than under nonresonance conditions. By controlling the driving frequency, selective capsule destruction can be achieved.


Automated scoring of nematode nictation on a textured background.
Entomopathogenic nematodes, including Steinernema spp., play an increasingly important role as biological alternatives to chemical pesticides. The infective juveniles of these worms use nictation-a behavior in which animals stand on their tails-as a host-seeking strategy. The developmentally-equivalent dauer larvae of the free-living nematode Caenorhabditis elegans also nictate, but as a means of phoresy or "hitching a ride" to a new food source. Advanced genetic and experimental tools have been developed for C. elegans, but time-consuming manual scoring of nictation slows efforts to understand this behavior, and the textured substrates required for nictation can frustrate traditional machine vision segmentation algorithms. Here we present a Mask R-CNN-based tracker capable of segmenting C. elegans dauers and S. carpocapsae infective juveniles on a textured background suitable for nictation, and a machine learning pipeline that scores nictation behavior. We use our system to show that the nictation propensity of C. elegans from high-density liquid cultures largely mirrors their development into dauers, and to quantify nictation in S. carpocapsae infective juveniles in the presence of a potential host. This system is an improvement upon existing intensity-based tracking algorithms and human scoring which can facilitate large-scale studies of nictation and potentially other nematode behaviors.


Autozygosity-driven genetic diagnosis in consanguineous families from Italy and the Greater Middle East.
Autozygosity-driven exome analysis has been shown effective for identification of genes underlying recessive diseases especially in countries of the so-called Greater Middle East (GME), where high consanguinity unravels the phenotypic effects of recessive alleles and large family sizes facilitate homozygosity mapping. In Italy, as in most European countries, consanguinity is estimated low. Nonetheless, consanguineous Italian families are not uncommon in publications of genetic findings and are often key to new associations of genes with rare diseases. We collected 52 patients from 47 consanguineous families with suspected recessive diseases, 29 originated in GME countries and 18 of Italian descent. We performed autozygosity-driven exome analysis by detecting long runs of homozygosity (ROHs > 1.5 Mb) and by prioritizing candidate clinical variants within. We identified a pathogenic synonymous variant that had been previously missed in NARS2 and we increased an initial high diagnostic rate (47%) to 55% by matchmaking our candidate genes and including in the analysis shorter ROHs that may also happen to be autozygous. GME and Italian families contributed to diagnostic yield comparably. We found no significant difference either in the extension of the autozygous genome, or in the distribution of candidate clinical variants between GME and Italian families, while we showed that the average autozygous genome was larger and the mean number of candidate clinical variants was significantly higher (p = 0.003) in mutation-positive than in mutation-negative individuals, suggesting that these features influence the likelihood that the disease is autozygosity-related. We highlight the utility of autozygosity-driven genomic analysis also in countries and/or communities, where consanguinity is not widespread cultural tradition.


A natural plasmid uniquely encodes two biosynthetic pathways creating a potent anti-MRSA antibiotic.
Understanding how complex antibiotics are synthesised by their producer bacteria is essential for creation of new families of bioactive compounds. Thiomarinols, produced by marine bacteria belonging to the genus Pseudoalteromonas, are hybrids of two independently active species: the pseudomonic acid mixture, mupirocin, which is used clinically against MRSA, and the pyrrothine core of holomycin. High throughput DNA sequencing of the complete genome of the producer bacterium revealed a novel 97 kb plasmid, pTML1, consisting almost entirely of two distinct gene clusters. Targeted gene knockouts confirmed the role of these clusters in biosynthesis of the two separate components, pseudomonic acid and the pyrrothine, and identified a putative amide synthetase that joins them together. Feeding mupirocin to a mutant unable to make the endogenous pseudomonic acid created a novel hybrid with the pyrrothine via "mutasynthesis" that allows inhibition of mupirocin-resistant isoleucyl-tRNA synthetase, the mupirocin target. A mutant defective in pyrrothine biosynthesis was also able to incorporate alternative amine substrates. Plasmid pTML1 provides a paradigm for combining independent antibiotic biosynthetic pathways or using mutasynthesis to develop a new family of hybrid derivatives that may extend the effective use of mupirocin against MRSA.


Accuracy of ultrasound in the diagnosis of acute appendicitis compared with the surgeon's clinical impression.
To compare the accuracy of the surgeon's clinical diagnosis of acute appendicitis with that of an ultrasonographic examination of the abdomen. Prospective trial. US Navel Hospital, San Diego, Calif. One hundred ten patients admitted to the hospital with suspected appendicitis from May 1990 to June 1992. Symptoms and signs for each patient were recorded, along with the surgeon's clinical impression of immediate surgery or observation. The patient then underwent an ultrasound examination performed by a staff radiologist. On the basis of the ultrasound findings the patient was placed into one of three categories: appendicitis, normal examination results, or other conditions. Patients with an ultrasound-based diagnosis of appendicitis proceeded to the operation, regardless of the surgeon's clinical impression. Those with other conditions diagnosed with ultrasonography were treated as was appropriate for the condition. The ultrasound-derived diagnosis of appendicitis had a sensitivity of 85.5%, a specificity of 84.4%, a positive predictive value of 88.3%, a negative predictive value of 80.1%, and an overall accuracy of 85.0%. The surgeon's clinical impression at the time of admission had a sensitivity of 62.9%, a specificity of 82.2%, a positive predictive value of 82.9%, a negative predictive value of 61.7%, and an overall accuracy of 71.2%. The overall accuracy of ultrasonography in the diagnosis of appendicitis was statistically superior to that of the surgeon's clinical impression (P < .0001). However, 24% of the patients with normal ultrasound findings were ultimately found to have appendicitis at operation, emphasizing the point that ultrasonography cannot be relied on to the exclusion of the surgeon's careful and repeated evaluation.


Evaluation of methods for generating parametric (R-[11C]PK11195 binding images.
Activated microglia can be visualised using (R)-[(11)C]PK11195 (1-[2-chlorophenyl]-N-methyl-N-[1-methyl-propyl]-3-isoquinoline carboxamide) and positron emission tomography (PET). In previous studies, various methods have been used to quantify (R)-[(11)C]PK11195 binding. The purpose of this study was to determine which parametric method would be best suited for quantifying (R)-[(11)C]PK11195 binding at the voxel level. Dynamic (R)-[(11)C]PK11195 scans with arterial blood sampling were performed in 20 healthy and 9 Alzheimer's disease subjects. Parametric images of both volume of distribution (V(d)) and binding potential (BP) were obtained using Logan graphical analysis with plasma input. In addition, BP images were generated using two versions of the basis function implementation of the simplified reference tissue model, two versions of Ichise linearisations, and Logan graphical analysis with reference tissue input. Results of the parametric methods were compared with results of full compartmental analysis using nonlinear regression. Simulations were performed to assess accuracy and precision of each method. It was concluded that Logan graphical analysis with arterial input function is an accurate method for generating parametric images of V(d). Basis function methods, one of the Ichise linearisations and Logan graphical analysis with reference tissue input provided reasonably accurate and precise estimates of BP. In pathological conditions with reduced flow rates or large variations in blood volume, the basis function method is preferred because it produces less bias and is more precise.


[Chronobiological analysis of behavior of the intact and pinealectomized rats in the open field test].
It was shown that adaptation to experimental conditions does not result in complete extinction of rat exploratory activity in the open-field test, as it was thought earlier. Behavior of rats is rhythmic. It can be described by a set of infradian oscillations with the periods about 30.4, 18.2, 13.5, 9.3, 7.3, 5.7, and 3.5 days. Pinealectomy does not abolish the infradian rhythmicity of behavioral parameters but modifies it. These findings allow the pineal gland to be considered as an organizer of biorhythms of animal behavior within a wide range of periods including the infradian as well as circadian and seasonal oscillations.


Identification and Function of Apicoplast Glutaredoxins in Neospora caninum.
Glutaredoxins (GRXs), important components of the intracellular thiol redox system, are involved in multiple cellular processes. In a previous study, we identified five GRXs in the apicomplexan parasite, Neospora caninum. In the present study, we confirmed that the GRXs S14 and C5 are located in the apicoplast, which suggests unique functions for these proteins. Although single-gene deficiency did not affect the growth of parasites, a double knockout (Δgrx S14Δgrx C5) significantly reduced their reproductive capacity. However, there were no significant changes in redox indices (GSH/GSSG ratio, reactive oxygen species and hydroxyl radical levels) in double-knockout parasites, indicating that grx S14 and grx C5 are not essential for maintaining the redox balance in parasite cells. Key amino acid mutations confirmed that the Cys203 of grx S14 and Cys253/256 of grx C5 are important for parasite growth. Based on comparative proteomics, 79 proteins were significantly downregulated in double-knockout parasites, including proteins mainly involved in the electron transport chain, the tricarboxylic acid cycle and protein translation. Collectively, GRX S14 and GRX C5 coordinate the growth of parasites. However, considering their special localization, the unique functions of GRX S14 and GRX C5 need to be further studied.


Lanosterol Synthase Prevents EMT During Lens Epithelial Fibrosis Via Regulating SREBP1.
Epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs) is a predominant pathological process underlying fibrotic cataracts. Here we investigated the role and mechanism of lanosterol synthase (LSS), a key rate-limiting enzyme in sterol biosynthesis, in EMT of LECs. Human lens epithelial explants, primary rabbit LECs, and whole rat lenses were treated with TGFβ2. RNA-sequencing was conducted to explore genetic changes during fibrosis of human lens epithelial explants. Loss- and gain-of-function studies were performed in primary LECs to investigate roles and mechanisms of LSS, lanosterol and sterol regulatory element binding transcription protein 1 (SREBP1) in EMT. Rat lenses were applied to evaluate the potential effect of lanosterol on lens fibrosis. Expression of LSS, SREBP1, EMT-related regulators, and markers were analyzed by Western blot, qRT-PCR, or immunofluorescent staining. LSS and steroid biosynthesis were downregulated in TGFβ2-induced lens fibrosis. LSS inhibition directly triggered EMT by inducing Smad2/3 phosphorylation and nucleus translocation, an overexpression of LSS protected LECs from EMT by inhibiting Smad2/3 activation. Moreover, LSS inhibition decreased the expression of SREBP1, which regulated EMT via intervening TGFβ2/Smad2/3 transduction. Furthermore, lanosterol protected LECs from EMT caused by both TGFβ2 treatment and LSS inhibition via suppressing Smad2/3 activation and maintained lens transparency by preventing fibrotic plaques formation. We first identified that LSS protected LECs from EMT and played an antifibrotic role to maintain lens transparency. Additionally, lanosterol and sterol biosynthesis regulation might be promising strategies for preventing and treating fibrotic cataracts.


The formation of free radicals by cardiac myocytes under oxidative stress and the effects of electron-donating drugs.
The interaction of myoglobin with H2O2 leads via a two-electron oxidation process to the formation of ferryl myoglobin. Metmyoglobin is more readily activated than oxymyoglobin to the ferryl states, which are capable of inducing peroxidative damage to membranes. E.p.r. and optical spectroscopic studies show that the thiol-containing compounds N-(2-mercaptopropionyl) glycine and N-acetylcysteine and the trihydroxamate desferrioxamine attenuate these processes by reducing the ferryl myoglobin species to metmyoglobin, with the formation of thiyl radicals and the desferrioxamine nitroxide radical respectively. Biochemical investigations of the potential for myoglobin in ruptured myocytes to be involved in radical generation, when under oxidative stress, and of the nature of the resulting species, were also undertaken. E.p.r. spectroscopic studies revealed the formation of a radical species which is capable of inducing membrane lipid peroxidation. The interaction of the thiol compounds and desferrioxamine with components of myocardial tissue under these conditions results in the generation of thiol-derived radical species and the desferrioxamine nitroxide radical respectively. These data, along with those obtained using optical spectrocopy, support the assignment of the identity of the radical species generated from the myocytes as the ferryl myoglobin radical.


Attitudes towards human papillomavirus vaccination among African parents in a city in the north of England: a qualitative study.
Human papillomavirus (HPV) is sexually transmitted and has been conclusively linked to cervical cancer and genital warts. Cervical cancer is attributed to approximately 1100 deaths annually in UK, and is the second most common female cancer globally. It has been suggested that black African women are more predisposed to HPV infection and cervical cancer. A vaccine has been developed to reduce HPV infection, and in the UK, has been offered to 12-13 year old adolescent girls through schools as part of their childhood immunization programme since 2008. Upon programme initiation, it was noted that vaccine uptake was lower in schools where girls from ethnic minority groups were proportionately higher. The study's objectives were to explore factors influencing UK based African parents' acceptance or decline of the HPV vaccine, whether fathers and mothers share similar views pertaining to vaccination and any interfamily tensions resulting from differing views. A qualitative study was conducted with five African couples residing in north England. Face to face semi-structured interviews were carried out. Participants were parents to at least one daughter aged between 8 and 14 years. Recruitment was done through purposive sampling using snowballing. HPV and cervical cancer awareness was generally low, with awareness lower in fathers. HPV vaccination was generally unacceptable among the participants, with fear of promiscuity, infertility and concerns that it's still a new vaccine with yet unknown side effects cited as reasons for vaccine decline. There was HPV risk denial as religion and good cultural upbringing seemed to result in low risk perceptions, with HPV and cervical cancer generally perceived as a white person's disease. Religious values and cultural norms influenced vaccine decision-making, with fathers acting as the ultimate decision makers. Current information about why the vaccine is necessary was generally misunderstood. Tailored information addressing religious and cultural concerns may improve vaccine acceptability in African parents.


Drug resistance occurred in a newly characterized preclinical model of lung cancer brain metastasis.
Cancer metastasis and drug resistance have traditionally been studied separately, though these two lethal pathological phenomena almost always occur concurrently. Brain metastasis occurs in a large proportion of lung cancer patients (~ 30%). Once diagnosed, patients have a poor prognosis surviving typically less than 1 year due to lack of treatment efficacy. Human metastatic lung cancer cells (PC-9-Br) were injected into the left cardiac ventricle of female athymic nude mice. Brain lesions were allowed to grow for 21 days, animals were then randomized into treatment groups and treated until presentation of neurological symptoms or when moribund. Prior to tissue collection mice were injected with Oregon Green and 14C-Aminoisobutyric acid followed by an indocyanine green vascular washout. Tracer accumulation was determined by quantitative fluorescent microscopy and quantitative autoradiography. Survival was tracked and tumor burden was monitored via bioluminescent imaging. Extent of mutation differences and acquired resistance was measured in-vitro through half-maximal inhibitory assays and qRT-PCR analysis. A PC-9 brain seeking line (PC-9-Br) was established. Mice inoculated with PC-9-Br resulted in a decreased survival time compared with mice inoculated with parental PC-9. Non-targeted chemotherapy with cisplatin and etoposide (51.5 days) significantly prolonged survival of PC-9-Br brain metastases in mice compared to vehicle control (42 days) or cisplatin and pemetrexed (45 days). Further in-vivo imaging showed greater tumor vasculature in mice treated with cisplatin and etoposide compared to non-tumor regions, which was not observed in mice treated with vehicle or cisplatin and pemetrexed. More importantly, PC-9-Br showed significant resistance to gefitinib by in-vitro MTT assays (IC50 > 2.5 μM at 48 h and 0.1 μM at 72 h) compared with parental PC-9 (IC50: 0.75 μM at 48 h and 0.027 μM at 72 h). Further studies on the molecular mechanisms of gefitinib resistance revealed that EGFR and phospho-EGFR were significantly decreased in PC-9-Br compared with PC-9. Expression of E-cadherin and vimentin did not show EMT in PC-9-Br compared with parental PC-9, and PC-9-Br had neither a T790M mutation nor amplifications of MET and HER2 compared with parental PC-9. Our study demonstrated that brain metastases of lung cancer cells may independently prompt drug resistance without drug treatment.


Cerebral white matter lesions are not associated with apoE genotype but with age and female sex in Alzheimer's disease.
Cerebral white matter lesions, such as leukoaraiosis, may be a result of damage from cerebral ischaemia, and may also be associated with the degenerative process in Alzheimer's disease. The apolipoprotein epsilon4 (apoepsilon4) genotype is a genetic risk factor for both Alzheimer's disease and ischaemic brain damage through acceleration of atherosclerosis. The aim was to determine whether apoepsilon4 may be related to the formation of cerebral white matter lesions in Alzheimer's disease. The association of apoE genotype, sex, age, and the presence of several vascular risk factors, with the presence of white matter lesions in 55 patients clinically diagnosed with Alzheimer's disease was investigated. The cerebral white matter lesions were identified by T2 weighted MRI and classified on a 4 grade scale from no lesion to diffuse lesion. The odds ratio (OR) of the factors mentioned above to the presence of white matter lesions was determined and tested by Fisher's exact test. The association of the lesion grades with these factors was analysed by non-parametric tests. The apoE 4 genotype was strongly associated with Alzheimer's disease (p=0.0001), but not associated with the presence or the degree of cerebral white matter lesions in Alzheimer's disease (OR=1.09, p>0.99). Aging (>70 years old) was a significant risk factor for white matter lesions (OR=7.2, p=0.0006) and age was significantly correlated with the lesion (p=0.0075). The OR of female sex to the lesion grades was 2.89 (p=0.084) and the lesion grade of female sex was significantly higher than that of the male sex (p=0.047). Other vascular risk factors were not significantly associated with the presence of white matter lesions. These findings suggest that there is a sex difference in white matter pathology in Alzheimer's disease.