Initiation of leaf somatic embryogenesis involves high pectin esterification, auxin accumulation and DNA demethylation in Quercus alba.
Somatic embryogenesis is considered a convenient tool for investigating the regulating mechanisms of embryo formation; it is also a feasible system for in vitro regeneration procedures, with many advantages in woody species. Nevertheless, trees have shown recalcitrance to somatic embryogenesis, and its efficiency remains very low in many cases. Consequently, despite the clear potential of somatic embryogenesis in tree breeding programs, its application is limited since factors responsible for embryogenesis initiation have not yet been completely elucidated. In the present work, we investigated key cellular factors involved in the change of developmental program during leaf somatic embryogenesis initiation of white oak (Quercus alba), aiming to identify early markers of the process. The results revealed that pectin esterification, auxin accumulation and DNA demethylation were induced during embryogenesis initiation and differentially found in embryogenic cells, while they were not present in leaf cells before induction or in non-embryogenic cells after embryogenesis initiation. These three factors constitute early markers of leaf embryogenesis and represent processes that could be interconnected and involved in the regulation of cell reprogramming and embryogenesis initiation. These findings provide new insights into the mechanisms underlying plant cell reprogramming, totipotency and embryogenic competence acquisition, especially in tree species for which information is scarce, thus opening up the possibility of efficient manipulation of somatic embryogenesis induction.


Prosopagnosia: a defect in visual configural processing.
A patient with severe, lasting prosopagnosia could not get an immediate overview of a face sufficiently specific for successful identification. He also failed completely in tasks of visual closure despite adequate performances on numerous other tests of visual perception and memory. We conclude that prosopagnosia represents a loss of visual "configural processing"--a learned skill enabling immediate identification of individual members of a class without conscious visuospatial analysis or remembering. Prosopagnosia and agnosic alexia represent two distinct defects of configural processing: Alexics cannot identify items with distinctive features that are themselves identifiable. Prosopagnosics cannot identify objects whose critical distinguishing features have no independent identities.


Assessment of the association between periodontal disease and total cancer incidence and mortality: a meta-analysis.
Periodontal disease (PD) is a chronic inflammatory disease that leads to alveolar bone resorption and tooth loss. Many studies have reported the association between periodontal disease and various cancers including oral cancer, lung cancer, breast cancer and so on. However, there is still no specialized meta-analysis that assesses the association between periodontal disease and cancer incidence and mortality in-deepth. Thus, we conducted this meta-analysis. This meta-analysis was registered with PROSPERO: CRD42020183497. We searched five online databases for observational studies about the association between periodontal disease and breast, prostate, lung and bronchial, colorectal, and total cancers by July 2020. Then we evaluated quality of the included studies by the Newcastle-Ottawa scale. Risk ratios (HRs) and their 95% confidence intervals (CIs) were pooled to evaluate the strength of the association between periodontal disease and four cancers, total cancer incidence and mortality. In addition, we analyzed heterogeneity by subgroup analysis and sensitivity analysis. Finally, we inspected publication bias by Begg's and Egger's tests. None of the studies included in this meta-analysis were of poor quality. PD is not only related to breast cancer incidence (HR = 1.26,95%CI [1.11-1.43], I 2 = 75.8%, P = 0.000), but also connected with total cancer mortality (HR = 1.40,95%CI [1.24-1.58], I 2 = 0.0%, P = 0.718). Subgroup analyses showed that study population, study design, dental status, follow-up period, adjustment for smoking partially explained the heterogeneity between studies. The results of Begg's test and Egger's test were consistent and indicated that there is no publication bias in this study. In conclusion, this meta-analysis revealed a positive relationship between periodontal disease and breast cancer incidence and total cancer mortality. Further well-designed studies with specific inclusion and exclusion criteria are required to strengthen the conclusion of this meta-analysis. However, longer follow-up period, multi-center trials and even multinational studies are required to corroborate the results.


Acute feeding suppression and toxicity of raspberry ketone [4-(4-hydroxyphenyl)-2-butanone] in mice.
Raspberry ketone (RK; [4-(4-hydroxyphenyl)-2-butanone]) is used by the food and cosmetic industry as a flavoring agent. RK is also marketed as a dietary supplement for weight maintenance and appetite control. The purpose of the study was to characterize the acute feeding suppression with RK (64-640 mg/kg) by oral gavage in male and female C57BL/6J mice. Cumulative 24 h food intake was reduced at 200 mg/kg (24% feeding suppression) in males and reliably reduced at 640 mg/kg (49-77% feeding suppression). Feeding suppression was not associated with pica behavior over the range of doses or conditioned taste aversion. In a separate experiment, a single oral gavage of RK (640 mg/kg) resulted in approximate 43% mortality rate (6 out 14 male mice) within 2 days. Atrophy of white adipose tissue, splenic abnormalities, and thymus involution were noted after 2-4 days after oral gavage RK. Total white blood cell count, lymphocytes, monocytes, eosinophils were significantly lower, while mean red blood cells, hemoglobin, and hematocrit were significantly higher with RK treatment. Our findings indicated a dose-dependent feeding suppression with acute RK, but doses that reliable suppress food intake are associated with pathological changes.


Neuroglobin inhibits pancreatic cancer proliferation and metastasis by targeting the GNAI1/EGFR/AKT/ERK signaling axis.
Pancreatic cancer is an extremely aggressive malignancy with a very disappointing prognosis. Neuroglobin (NGB), a member of the globin family, has been demonstrated to have a significant role in a variety of tumor forms. The possible role of NGB as a tumor suppressor gene in pancreatic cancer was investigated in this work. Information from the public dataset TCGA combined with GTEx was used to analyze the finding that NGB was commonly downregulated in pancreatic cancer cell lines and tissues, correlating with patient age and prognosis. The expression of NGB in pancreatic cancer was investigated via RT-PCR, qRT-PCR, and Western blot experiments. In-vitro and in-vivo assays, NGB elicited cell cycle arrest in the S phase and apoptosis, hindered migration and invasion, reversed the EMT process, and suppressed cell proliferation and development. The mechanism of action of NGB was predicted via bioinformatics analysis and validated using Western blot and co-IP experiments revealed that NGB inhibited the EGFR/AKT/ERK pathway by binding to and reducing expression of GNAI1 and p-EGFR. In addition, pancreatic cancer cells overexpressing NGB showed increased drug sensitivity to gefitinib (EGFR-TKI). In conclusion, NGB inhibits pancreatic cancer progression by specifically targeting the GNAI1/EGFR/AKT/ERK signaling axis.


Prediction of amyloid PET positivity via machine learning algorithms trained with EDTA-based blood amyloid-β oligomerization data.
The tendency of amyloid-β to form oligomers in the blood as measured with Multimer Detection System-Oligomeric Amyloid-β (MDS-OAβ) is a valuable biomarker for Alzheimer's disease and has been verified with heparin-based plasma. The objective of this study was to evaluate the performance of ethylenediaminetetraacetic acid (EDTA)-based MDS-OAβ and to develop machine learning algorithms to predict amyloid positron emission tomography (PET) positivity. The performance of EDTA-based MDS-OAβ in predicting PET positivity was evaluated in 312 individuals with various machine learning models. The models with various combinations of features (i.e., MDS-OAβ level, age, apolipoprotein E4 alleles, and Mini-Mental Status Examination [MMSE] score) were tested 50 times on each dataset. The random forest model best-predicted amyloid PET positivity based on MDS-OAβ combined with other features with an accuracy of 77.14 ± 4.21% and an F1 of 85.44 ± 3.10%. The order of significance of predictive features was MDS-OAβ, MMSE, Age, and APOE. The Support Vector Machine using the MDS-OAβ value only showed an accuracy of 71.09 ± 3.27% and F-1 value of 80.18 ± 2.70%. The Random Forest model using EDTA-based MDS-OAβ combined with the MMSE and apolipoprotein E status can be used to prescreen for amyloid PET positivity.


Bladder Mass in Newborn: Case Report and Review of Literature.
Benign proliferative lesions of the bladder are exceptional in neonates. We describe a case of a 3-day-old neonate, presenting with bloody meconium and sonographic diagnosis of bladder mass. Cystoscopic biopsies were performed and a diagnosis of polypoid cystitis was made. The patient was treated conservatively and the lesion healed during follow-up. We review the literature of other cases of neonatal bladder masses.


The homeless mentally ill in Tucson: implications of early findings.
Homeless persons who received emergency psychiatric screening in a public general hospital exhibited patterns of gross geographic mobility. They did not uniformly avail themselves of community services, such as soup kitchens, and there were marked sex differences within the sample.


Clinical assessment of asthma progression in children and adults.
Asthma is a heterogeneous disorder with a variable course, characterized by episodes of cough, wheezing and shortness of breath, reversible airflow limitation, and bronchial hyperresponsiveness. It begins early in life in many subjects with intermittent symptoms occurring with viral respiratory tract infections. Over time, and in genetically susceptible children (those with an atopic predisposition), the disease becomes more persistent with symptoms occurring in the absence of respiratory tract infections. Children with persistent wheezing are eventually diagnosed with asthma, with those at greatest risk having developed allergic sensitization early in life. Among children with asthma, some will have lifelong asthma with active symptoms and progressive loss of lung function over time, whereas other children will undergo asthma remission in adolescence. Once in remission, the disease may remain quiescent, or it may relapse in midadult life. This review focuses on studies that have enhanced our understanding of the progression of asthma from infancy to adulthood. Studies evaluating progressive loss of lung function, the best-studied measure of asthma progression, are also reviewed, followed by a brief discussion of whether asthma progression can be modified by inhaled glucocorticoid therapy.


Passage of Gastrografin into the small mesenteric veins during an opaque enema.
An opaque enema was carried out with Gastrografin. The patient was a 2-year-old French boy. As the enema progressed we saw the small mesenteric veins opacify: no significant consequence followed. This is the first report of such an incident.


[Detection of EGFR gene mutations in 100 non-small cell lung cancer clinical samples by a real-time polymerase chain reaction method using amplification refractory mutation system specific primers and Taqman fluorescence probes].
Epidermal growth factor receptor (EGFR) gene mutation is the most important predictor of the efficiency of EGFR-tyrosine kinase inhibitors in the treatment of non-small cell lung cancer (NSCLC). The detection of EGFR gene mutations can guide individual therapies for NSCLC. Numerous methods are used to detect EGFR gene mutation and each method has different features. This study aims to establish a real-time polymerase chain reaction (PCR) method for the detection of EGFR gene mutations using amplification refractory mutation system (ARMS) specific primers and Taqman fluorescence probes. ARMS specific primers for the two EGFR gene mutations (E746_A750 and L858R) and Taqman fluorescence probes for the detection of the target sequence were carefully designed by the Primer Premier 5.0 software. Then, using the recombinants containing E746_A750 and L858R mutations as the study objects, we further analyzed the sensitivity and lower limit of this method, and then determined the cutoff ΔCt value to evaluate specific or non-specific amplification. A total of 100 clinical samples were collected and used to detect the EGFR gene mutations using this method. The lower limit of this method for the detection of EGFR gene mutation was 10 copies if no interference of wild-type EGFR gene or background DNA existed. Regarding the method sensitivity, the detection resolution was as high as 1% and 0.1%-0.5% in the background of 500 and 5,000 copies/μL wild-type EGFR gene, respectively. Regarding the method specificity, non-specific amplifications were found when it was used to detect 21 L858R mutations in leukocyte DNA samples from healthy volunteers. However, the minimal ΔCt value was 14.48. Non-specific amplifications were not found when detecting 19 Del mutations. Among the 100 clinical samples, 39 mutations were detected (19 Del and 21 L858R were 21 and 18, respectively) using this method. The total mutation rate was 39.0%. The proposed ARMS-TaqMan real-time PCR method for the detection of 19 Del and 21 L858R mutations in EGFR gene was rapid, simple, sensitive, specific, and applicable in the clinical setting.


MOPP treatment of resistant Hodgkin's disease following ABVD failure.
Fourteen patients with Hodgkin's disease resistant to ABVD were treated with MOPP chemotherapy (nitrogen mustard, vincristine, procarbazine, prednisone). Complete remission was obtained in 6 patients (43%). Four of the 6 complete responders are disease free after 5, 20, 23, 35 months. The actuarial median survival after MOPP of all patients is 20 months. These data confirm that there is no "cross-resistance" among the drugs included in the two schedules.


Differential expression of type I collagen and cellular fibronectin isoforms in endothelial cell variants.
Endothelial cells in vivo and in vitro are inherently capable of manifesting more than one phenotype. Most endothelial cells in culture, even when maintained at confluent density for an extended period, exhibit only the typical polygonal morphology. Other cultures, albeit few, are characterized by rapid formation of variant "sprout" cells once the cobblestone monolayer is established. In vivo studies have revealed sprout cell formation to be one of the early events in neovascularization. Therefore, elucidation of the factors involved in this process is needed to gain an understanding of the sprouting phenomena. To this end, two morphologically distinct bovine endothelial cell clones derived from a fetal left ventricle were characterized. One culture of cells stably displayed the conventional polygonal phenotype, in contrast to the second culture in which an interconnecting network of sprout cells reproducibly developed beneath the cobblestone monolayer. Analysis of extracellular matrix proteins and RNA transcripts revealed that the sprouting clone showed induction of type I collagen and a shift in fibronectin RNA processing resulting in synthesis of the cellular isoforms. These collagenous and non-collagenous matrix molecules may serve in some manner to affect the processes required during angiogenesis, that is, cell division, cell migration and cell shape change.


[Heart and lung in X-ray images : Lost art (?)].
Echocardiography is the standard for the diagnosis of heart diseases. Nevertheless, thoracic X‑ray is a frequently used examination method, also for questions regarding the cardiac situation of patients. How relevant is the conventional radiological assessment of heart disease? Long-known knowledge about pathophysiology of heart disease and its radiological correlation are discussed. Knowledge of the normal anatomy of the heart is a basic prerequisite for the assessment of pathologies. For the question of heart disease, it is essential to also assess the pulmonary vascularization. The size of the entire heart and the individual cardiac cavities is assessed by direct and indirect signs, such as heart-lung ratio, cava triangle, vascular pedicle or the size of the aorta and of the main pulmonary artery. The most common cause of heart enlargement is valve disease, which shows various conventional radiological images. Conventional thoracic radiography still plays a central role in the diagnosis of cardiopulmonary diseases and is an important diagnostic tool for quickly obtaining an overview of the patient's cardiopulmonary situation. The interplay between physiology and X‑ray symptoms is complex, so close attention should be paid not only to the heart anatomy and configuration, but also primarily to the pulmonary blood flow.


Increased dopamine receptor binding in duodenal mucosa of duodenal ulcer patients.
High-affinity and saturable membrane-bound dopamine binding sites have been characterized in rat and human gastrointestinal tissues. Although their role in experimental ulcerogenesis has been suggested, dopamine receptor activity in peptic ulcer disease has not been investigated. Radioligand binding studies were performed with mucosal tissue homogenates obtained from the antrum and duodenum of six male healthy volunteers and six male duodenal ulcer patients. The binding assay was performed in triplicate with a crude membrane fraction using [3H]dopamine as a ligand at a final concentration of 1 nM at 22 degrees C in the dark. Nonspecific binding (which usually comprised about 30% of total binding) was determined in the presence of a 100-fold excess of unlabeled dopamine. A significant (P less than 0.05) increase of [3H]dopamine binding was found in duodenal mucosa of duodenal ulcer patients. [3H]Dopamine binding in stomach (antrum) of normal and duodenal ulcer patients did not differ significantly. These findings provide preliminary evidence for a role of dopamine receptors in duodenal ulcer and suggest that biochemical abnormalities of gut dopamine function may be operative in the pathogenesis of peptic ulcer disease.


Prolonged survival of a murine model of cerebral malaria by kynurenine pathway inhibition.
C57BL/6J mice infected with Plasmodium berghei ANKA develop neurological dysfunction and die within 7 days of infection. We show that treatment of infected mice with a kynurenine-3-hydroxylase inhibitor prevents them from developing neurological symptoms and extends their life span threefold until severe anemia develops.


Specialization of neural mechanisms underlying face recognition in human infants.
Newborn infants respond preferentially to simple face-like patterns, raising the possibility that the face-specific regions identified in the adult cortex are functioning from birth. We sought to evaluate this hypothesis by characterizing the specificity of infants' electrocortical responses to faces in two ways: (1) comparing responses to faces of humans with those to faces of nonhuman primates; and 2) comparing responses to upright and inverted faces. Adults' face-responsive N170 event-related potential (ERP) component showed specificity to upright human faces that was not observable at any point in the ERPs of infants. A putative "infant N170" did show sensitivity to the species of the face, but the orientation of the face did not influence processing until a later stage. These findings suggest a process of gradual specialization of cortical face processing systems during postnatal development.


An encoded N-terminal extension results in low levels of heterologous protein production in Escherichia coli.
The tdk gene (encoding deoxythymidine kinase) of the gamma-proteobacterium Xenorhabdus nematophila has two potential translation start sites. The promoter-distal start site was predicted to be functional based on amino acid sequence alignment with closely related Tdk proteins. However, to experimentally determine if either of the two possible start codons allows production of a functional Tdk, we expressed the "long-form" (using the promoter-proximal start codon) and "short-form" (using the promoter-distal start codon) X. nematophila tdk genes from the T7 promoter of the pET-28a(+) vector. We assessed Tdk production and activity using a functional assay in an Escherichia coli tdk mutant, which, since it lacks functional Tdk, is able to grow in 5-fluorodeoxyuridine (FUdR)-containing medium. Short-form Tdk complemented the E. coli tdk mutant strain, resulting in FUdR sensitivity of the strain. However, the E. coli tdk mutant expressing the long form of tdk remained FUdR resistant, indicating it did not have a functional deoxythymidine kinase enzyme. We report that long-form Tdk is at least 13-fold less abundant than short-form Tdk, the limited protein produced was as stable as short-form Tdk and the long-form transcript was 1.7-fold less abundant than short-form transcript. Additionally, we report that the long-form extension was sufficient to decrease heterologous production of a different X. nematophila protein, NilC. We conclude that the difference in the FUdR growth phenotype between the E. coli tdk mutant carrying the long-or short-form X. nematophila tdk is due to a difference in Tdk levels. The lower long-form protein level does not result from protein instability, but instead from reduced transcript levels possibly combined with reduced translation efficiency. Because the observed effect of the encoded N-terminal extension is not specific to Tdk production and can be overcome with induction of gene expression, these results may have particular relevance to researchers attempting to limit production of toxic proteins under non-inducing conditions.


Workplace Violence Against Emergency Nurses in Taiwan: A Cross-Sectional Study.
Workplace violence against emergency nurses is common worldwide. This study aimed to gain a more thorough overview of the point prevalence of workplace violence against emergency nurses and the preventive measures used in emergency departments in Taiwan. In this multicenter, descriptive, cross-sectional study, the questionnaire, Violence Against Nurses Working in Emergency Departments, was used to collect data from 407 nurses working at 5 emergency departments in Taiwan from May to October 2015. The results revealed that 378 emergency nurses (92.9%) experienced workplace violence over the last 2 years. The average visual analog scale score (1-10) of security effectiveness in preventing workplace violence was 5.0 (SD = 1.97). The average visual analog scale score of perceived safety level in terms of workplace violence was 4.38 (SD = 2.06). The average visual analog scale score of meeting nurses' needs was 5.72 (SD = 2.23). This questionnaire survey revealed that the current preventive measures for workplace violence against emergency nurses in Taiwan were not effective. The relevant measures should be improved, thereby reducing the prevalence and severity of workplace violence against emergency nurses.


Conditional knockdown of BCL2A1 reveals rate-limiting roles in BCR-dependent B-cell survival.
Bcl2 family proteins control mitochondrial apoptosis and its members exert critical cell type and differentiation stage-specific functions, acting as barriers against autoimmunity or transformation. Anti-apoptotic Bcl2a1/Bfl1/A1 is frequently deregulated in different types of blood cancers in humans but its physiological role is poorly understood as quadruplication of the Bcl2a1 gene locus in mice hampers conventional gene targeting strategies. Transgenic overexpression of A1, deletion of the A1-a paralogue or constitutive knockdown in the hematopoietic compartment of mice by RNAi suggested rate-limiting roles in lymphocyte development, granulopoiesis and mast cell activation. Here we report on the consequences of conditional knockdown of A1 protein expression using a reverse transactivator (rtTA)-driven approach that highlights a critical role for this Bcl2 family member in the maintenance of mature B-cell homeostasis. Furthermore, we define the A1/Bim (Bcl-2 interacting mediator of cell death) axis as a target of key kinases mediating B-cell receptor (BCR)-dependent survival signals, such as, spleen tyrosine kinase (Syk) and Brutons tyrosine kinase (Btk). As such, A1 represents a putative target for the treatment of B-cell-related pathologies depending on hyperactivation of BCR-emanating survival signals and loss of A1 expression accounts, in part, for the pro-apoptotic effects of Syk- or Btk inhibitors that rely on the 'BH3-only' protein Bim for cell killing.


Cathepsin K inhibition induces Raptor destabilization and mitochondrial dysfunction via Syk/SHP2/Src/OTUB1 axis-mediated signaling.
The Raptor signaling pathway is a critical point of intervention in the invasion and progression of cancer. The non-receptor tyrosine kinase Src-mediated phosphorylation of OTUB1-Y26 plays a critical role in Raptor stabilization, whereas cathepsin K inhibitor (odanacatib; ODN) and knockdown (siRNA) induce Raptor destabilization. However, the mechanisms involved in cathepsin K inhibition-induced OTUB1-Y26 phosphorylation in Raptor stabilization have not been yet elucidated. This study showed that cathepsin K inhibition activates SHP2, a tyrosine phosphatase, that dephosphorylates OTUB1 and destabilizes Raptor, whereas SHP2 deletion and pharmacological inhibition increase OTUB1-Y26 phosphorylation and Raptor expression. SHP2 deletion also led to the inhibition of ODN-induced mitochondrial ROS, fusion, and dysfunction. Furthermore, cathepsin K inhibition phosphorylated spleen tyrosine kinase (Syk) at Y525 and Y526, resulting in the SHP2-mediated dephosphorylation of OTUB1-Y26. Collectively, our findings identified Syk not only as an upstream tyrosine kinase required for SHP2 activation but also showed a critical mechanism that regulates ODN-induced Raptor downregulation and mitochondrial dysfunction. In conclusion, Syk/SHP2/Src/OTUB1 axis-mediated signaling can act as a therapeutic target in cancer management.


Kinesiophobia in patients with angina pectoris of coronary artery disease: A cross-sectional survey.
In the field of chronic pain research, kinesiophobia is defined as avoidance behavior due to fear of pain, but this perspective seems to be neglected in the field of coronary artery disease (CAD). To investigate the status quo of angina pectoris (AP) and kinesiophobia and factors associated with kinesiophobia in patients with AP of CAD. This was a cross-sectional study. Participants were recruited by convenience sampling, and patients with AP of CAD in four wards of the Cardiology Department of a hospital were enrolled in this study. Participants completed questionnaires and scales face-to-face with researchers. Most patients with AP of CAD suffered from at least moderate pain episodes (60.2%) of at least 5 minutes per episode (53.8%), but less than 1/3 of patients reached 5 instances of pain per week (29.1%). The total score of kinesiophobia in patients was 40.80±6.65, and the vast majority of patients had moderate to high levels of kinesiophobia (75.7%). The results of multiple linear stepwise regression analysis showed that personal monthly income, New York Heart Function Assessment (NYHA) classification, pain intensity, and pain resilience were independent factors associated with kinesiophobia, and these factors explained 30.2% of the variation in total scores of kinesiophobia. The symptoms of AP were prominent in terms of pain intensity and duration of pain. The level of kinesiophobia was moderate, and this was affected by multiple factors. Health care providers and researchers seldom pay attention to the kinesiophobia of patients with AP of CAD. This study refocused on the effect of "fear of pain" in kinesiophobia in patients with CAD. It opens up new horizons for the application of fear-avoidance models in CAD patients and helps to raise awareness of kinesiophobia in AP patients with CAD and provides guidance for reducing the level of kinesiophobia in the future.


X-linked Leigh's syndrome.
Two male half siblings developed rapid progression of neurologic symptoms at 11/2 and 21/2 years of age. Neither boy had a metabolic acidosis. Characteristic features of subacute necrotizing encephalomyelopathy, the neuropathologic basis of Leigh's syndrome, were demonstrated at autopsy. X-linkage of the disorder was considered because the boys had different fathers. An X-linked form of Leigh's syndrome was supported by a review of the literature, which showed an unexplained male/female ratio in Leigh's syndrome of 1.83/1, and a significant excess of male-male siblings. An X-linked form of Leigh's syndrome would explain the excess of males, and may account for some of the clinical and biochemical heterogeneity.


Improved production of butyl butyrate with lipase from Thermomyces lanuginosus immobilized on styrene-divinylbenzene beads.
Two immobilized preparations from Thermomyces lanuginosus lipase (TLL) were compared in the synthesis of butyl butyrate. The commercial Lipozyme TL-IM, and TLL immobilized on styrene-divinylbenzene beads (MCI-TLL) were tested in the esterification reaction using n-hexane as solvent. The variables temperature (30-60°C), substrate molar ratio (1:1 to 5:1), added water (0-1%), and biocatalyst content (3-40%) were evaluated in terms of initial reaction rate for each biocatalyst. SDS-PAGE analysis revealed that MCI-TLL had an immobilized enzymatic load twice as high as Lipozyme TL-IM, but with an activity 3-fold higher. MCI-TLL presented high initial reaction rates up to 1.0 M butyric acid, while Lipozyme TL-IM showed a decrease in its activity above 0.5 M. Moreover, MCI-TLL allowed a productivity of 14.5 mmol g(-1) h(-1), while Lipozyme TL-IM 3.2 mmol g(-1) h(-1), both by mass of biocatalyst.


O2 microsensors for minimally invasive tissue monitoring.
Tissue oxygenation is a key factor ensuring normal tissue functions and viability. Continuous real-time monitoring of the partial pressure of oxygen, pO(2), in tissues gives insight into the dynamic fluctuations of O(2) supplies to tissues by blood circulation. Small oxygen sensors enable investigations of the spatial variation of pO(2) in tissues at different locations in relation to local microvessels. In this paper, pO(2) measurement using microelectrodes and biocompatible sensorsv is discussed and recent progress of their application in human skin is reviewed. Emphasis is given to working principles of a number of existing oxygen sensors and their potential application in vivo and in tissue engineering. Results on spatial and temporal variations of the pO(2) in human skin introduced by localized ischaemia-reperfusion are presented when the surface of the skin is covered by an oxygen-free paraffin oil layer and the range of the tissue pO(2) is deduced to be between 0 and 60 mmHg. In the study, pO(2) increases from 8.0 +/- 3.2 mmHg (n = 6) at the surface of the skin to 35.2 +/- 8.0 mmHg (n = 9) at a depth just above the subpapillary plexus. Temporal decay in pO(2) following tissue compression and rise in pO(2) following pressure release can be described using mono-exponential functions. The time constant for the exponential decay, tau = 8.44 +/- 1.53 s (n = 7) is consistently greater than that for the exponential rises, tau' = 4.75 +/- 0.82 s (n = 6). The difference in pO2 change with the time following tissue compression and pressure release reveals different dynamic mechanisms involved in the two transient phases. The elevated steady state pO(2) following reperfusion, which is approximately 20% higher than the pre-occlusion value, indicates localized reactive hyperaemia. Possible applications of O(2) microsensors in diseases, e.g. tumours, pressure ulcers, are also discussed.


Respiratory modulation of baroreceptor and chemoreceptor reflexes affecting heart rate through the sympathetic nervous system.
1. Brief stimuli were delivered to the carotid body chemoreceptors or the carotid sinus baroreceptors at different phases of the respiratory cycle in anaesthetized dogs. Chemoreceptor stimulation was achieved by injecting small volumes (0.2-0.5 ml.) of warmed saline equilibrated with CO(2) near to the carotid bodies on both sides. Baroreceptor stimulation was achieved by injecting larger volumes (2-5 ml.) of saline equilibrated with air into the region of the carotid bifurcation on both sides, after first clamping the common carotid arteries.2. When the vagus nerves were intact, but sympathetic nervous effects on heart rate were blocked by administration of propranolol, there was a prompt and pronounced bradycardia evoked when either baroreceptor or chemoreceptor stimuli were given in expiration, but little or no change in heart rate when they were given in inspiration.3. When the vagus nerves were cut, but sympathetic nervous function was intact, respiratory modulation of both baroreceptor and chemoreceptor reflex effects on heart rate could still be demonstrated. The bradycardia evoked by either stimulus was more marked for stimuli given in expiration than for stimuli given in inspiration. A complementary response pattern for brief decreases in baroreceptor stimulation (carotid occlusions) was demonstrated: the tachycardia evoked by occlusions timed during inspiration was greater than that evoked by occlusions timed during expiration. All the reflex effects were mediated by the sympathetic system because they were abolished by administration of propranolol.4. Typically, the sympathetic reflex effects were slight in comparison with the vagal reflexes evoked by either chemoreceptor or baroreceptor stimuli.


High-D-dimer plasma levels predict poor outcome in esophageal variceal bleeding.
Variceal bleeding carries a high-mortality rate in patients with liver cirrhosis. Since coagulation and fibrinolysis are abnormal in these patients we evaluated whether or not abnormalities of these haemostasis systems were independently related to mortality. Global coagulation, coagulation activation and fibrinolysis measurements were performed in 43 cirrhotics bleeding from esophageal varices at baseline and during follow-up and in 43 non-bleeding cirrhotic patients at baseline only. Baseline measurements of coagulation activation and fibrinolysis were more impaired in bleeders. In bleeders, prothrombin time, tissue type plasminogen activator antigen and D-dimer plasma levels were persistently more abnormal in patients who died. High-D-dimer, infection, Child-Pugh C class and MELD score >or=17 were the significant predictors of death at univariate analysis. Two different multivariate analyses to assess the independent prognostic value of these variables, one including the Child-Pugh class, the other including MELD, were performed. Independent predictors of death were high-D-dimer and infection, but not Child-Pugh class, in the former; MELD and infection, but not D-dimer, in the latter. Beside infection, high-D-dimer is a stronger predictor of death as compared to Child-Pugh C class, but not to a MELD score >or=17.


Arginine methylation of the c-Jun coactivator RACO-1 is required for c-Jun/AP-1 activation.
c-Jun, the major component of the AP-1 transcription factor complex, has important functions in cellular proliferation and oncogenic transformation. The RING domain-containing protein RACO-1 functions as a c-Jun coactivator that molecularly links growth factor signalling to AP-1 transactivation. Here we demonstrate that RACO-1 is present as a nuclear dimer and that c-Jun specifically interacts with dimeric RACO-1. Moreover, RACO-1 is identified as a substrate of the arginine methyltransferase PRMT1, which methylates RACO-1 on two arginine residues. Arginine methylation of RACO-1 promotes a conformational change that stabilises RACO-1 by facilitating K63-linked ubiquitin chain formation, and enables RACO-1 dimerisation and c-Jun interaction. Abrogation of PRMT1 function impairs AP-1 activity and results in decreased expression of a large percentage of c-Jun target genes. These results demonstrate that arginine methylation of RACO-1 is required for efficient transcriptional activation by c-Jun/AP-1 and thus identify PRMT1 as an important regulator of c-Jun/AP-1 function.


Occupational and Environmental Carcinogenesis.
Occupational carcinogens have been shown to cause a considerable disease burden at a national and global level [...].


Evaluating the coupling effects of climate aridity and vegetation restoration on soil erosion over the Loess Plateau in China.
In this study, the coupling effects of climate aridity and vegetation restoration on runoff and sediment yield over the Loess Plateau were examined and characterized. To take into consideration the complexity of drought, as well as the varied strengths and weaknesses of different drought measures, two drought indices are selected to identify and evaluate drought variability. The Normalized Difference Vegetation Index (NDVI) data were obtained to monitor and express spatiotemporal variations in vegetation cover. The results show that most regions of the Loess Plateau experienced increasingly severe droughts over the past 40years, and these regions comprise the major source of the Yellow River sediment. Climatic drying initially occurred in the 1990s, and became statistically significant in 2000s. The increasingly severe droughts could negatively impact surface and groundwater supplies as well as soil water storage, but may also minimize surface runoff yield, which is one of the major causes of soil erosion on the Loess Plateau. Vegetation cover on the Loess Plateau was significantly improved after the implementation of "Grain for Green" project, which were helpful for controlling severe soil erosion. With the impacts of the construction of check dams, terraces and large reservoirs, runoff and sediment yield over the Loess Plateau initially exhibited downward trends between 1970 and 1990. After 1990, with the effects of the climate warming and drying, a second sharp reduction in runoff and sediment yield occurred. The coupling effects of climate aridity and vegetation restoration have led to a third significant decrease in runoff and sediment yield over the Loess Plateau after 2000.


Clinical development of dabrafenib in BRAF mutant melanoma and other malignancies.
The development of selective BRAF inhibitors in patients with metastatic BRAF V600 mutant melanoma has proven to be an effective therapeutic strategy. While vemurafenib was the first approved BRAF inhibitor for this indication, another selective BRAF inhibitor, dabrafenib, has demonstrated efficacy in patients with BRAF mutant melanoma, including those with active brain metastases and other malignancies. This review covers the current role of BRAF inhibitors in patients with metastatic melanoma and the clinical development of dabrafenib. The pharmacological, safety and efficacy data are discussed from the Phases I, II and III studies of dabrafenib. In addition, the results of the Phase II study of dabrafenib in melanoma patients with active brain metastases (BREAK-MB) and the Phase I/II study of dabrafenib/trametinib are examined. While dabrafenib has demonstrated comparable efficacy to vemurafenib in BRAF V600E mutant melanoma patients, the BREAK-MB and dabrafenib/trametinib studies have taken BRAF inhibitor strategies further with evidence of disease activity in patients with metastatic melanoma brain metastases and potential abrogation of BRAF inhibitor resistance.


Microbial degradation of 4-monobrominated diphenyl ether with anaerobic sludge.
Polybrominated diphenyl ethers (PBDEs) are widely used flame retardant additives for many plastic and electronic products. Owing to their ubiquitous distribution in the environment, multiple toxicity to humans, and increasing accumulation in the environment, the fate of PBDEs is of serious concern for public safety. In this study, the degradation of 4-monobrominated diphenyl ether (BDE-3) in anaerobic sludge and the effect of carbon source addition were investigated. BDE-3 can be degraded by two different anaerobic sludge samples. The by-products, diphenyl ether (DE) and bromide ions, were monitored, indicating the reaction of debromination within these anaerobic samples. Co-metabolism with glucose facilitated BDE-3 biodegradation in terms of kinetics and efficiency in the Jhongsing sludge. Through the pattern of amplified 16S rRNA gene fragments in denatured gradient gel electrophoresis (DGGE), the composition of the microbial community was analyzed. Most of the predominant microbes were novel species. The fragments enriched in BDE-3-degrading anaerobic sludge samples are presumably Clostridium sp. This enrichment coincides with the H(2) gas generation and the facilitation of debromination during the degradation process. Findings of this study provide better understanding of the biodegradation of brominated DEs and can facilitate the prediction of the fate of PBDEs in the environment.


Selective radiosensitization and cytotoxicity of human melanoma cells using halogenated deoxycytidines and tetrahydrouridine.
The halogenated pyrimidines 5-chloro-2'-deoxycytidine (CldCyd) and 5-bromo-2'-deoxycytidine (BrdCyd) can act as radiosensitizers and cytotoxic agents. It was hypothesized that tumor cells and normal cells might use different metabolic pathways to incorporate these halogenated deoxycytidines into DNA. This difference could potentially be exploited to produce selective radiosensitization and cytotoxicity of human tumor cells compared to normal human fibroblasts. This hypothesis was tested using two human melanoma cell lines and two normal fibroblast cell lines. Either CldCyd or BrdCyd alone caused both cytotoxicity and radiosensitization of tumor and normal cells. The addition of the cytidine deaminase inhibitor tetrahydrouridine (H4U) significantly protected the normal cells but had relatively little effect on the tumor cells. These data indicate that it may be possible to exploit differences between the pyrimidine metabolism of normal cells and melanoma cells to improve the therapeutic index of halogenated pyrimidines both as radiosensitizers and as cytotoxic agents.


Objective and subjective changes in voice quality after radiotherapy for early (T1 or T2, N0) laryngeal cancer: a pilot prospective cohort study.
Limited data exist on long-term voice quality after treatment for early laryngeal cancer. We report on the feasibility of establishing a dedicated service to assess changes after radiotherapy. From June 1999 to December 2005, all patients referred for radical radiotherapy for early laryngeal cancers were invited to join a prospective cohort study. Subjective and objective voice quality tests were performed before and after treatment and at 3, 6, 12, and 24 months after treatment. Twenty-seven patients were screened for the study. Sufficient data were available for 15 (56%) patients. Both subjective and objective assessments improved significantly. This pilot study shows that it is possible to collect prospective voice quality data. There are both subjective and objective improvements in voice that persist to at least 2 years after radiotherapy. We propose the use of the methods described here to study voice outcomes in a lager cohort of patients treated with both surgery and radiotherapy.


Stochastic theory of size exclusion chromatography by the characteristic function approach.
A general stochastic theory of size exclusion chromatography (SEC) able to account for size dependence on both pore ingress and egress processes, moving zone dispersion and pore size distribution, was developed. The relationship between stochastic-chromatographic and batch equilibrium conditions are discussed and the fundamental role of the 'ergodic' hypothesis in establishing a link between them is emphasized. SEC models are solved by means of the characteristic function method and chromatographic parameters like plate height, peak skewness and excess are derived. The peak shapes are obtained by numerical inversion of the characteristic function under the most general conditions of the exploited models. Separate size effects on pore ingress and pore egress processes are investigated and their effects on both retention selectivity and efficiency are clearly shown. The peak splitting phenomenon and peak tailing due to incomplete sample sorption near to the exclusion limit is discussed. An SEC model for columns with two types of pores is discussed and several effects on retention selectivity and efficiency coming from pore size differences and their relative abundance are singled out. The relevance of moving zone dispersion on separation is investigated. The present approach proves to be general and able to account for more complex SEC conditions such as continuous pore size distributions and mixed retention mechanism.


Correction to: Progression of frailty and prevalence of osteoporosis in a community cohort of older women-a 10-year longitudinal study.
The Open Access license, which previously read.


Endoscopic mucosal resection with an over-the-counter hyaluronate preparation.
Hyaluronic acid (HA) provides a long-lasting and distinct mucosal elevation for EMR, but expense and inconvenience have limited its adoption. To evaluate the safety and efficacy of an over-the-counter 0.15% HA preparation for EMR. Retrospective study. Veterans Administration Hospital and university hospital. 30 patients with a total of 32 colonic lesions and 1 duodenal lesion. EMR by using HA. En bloc resection rate and complications. EMR was successful in all cases. En bloc resection was achieved in 26 of the 28 lesions up to 25 mm in diameter. Two lesions, both with fibrosis from prior attempted resection, had trace residual tissue necessitating cauterization with argon plasma. Five lesions measuring 30 mm to 60 mm all required piecemeal resection. There was one complication, a postpolypectomy bleed. Small number of patients and retrospective design. EMR may be performed safely and effectively by using an inexpensive, over-the-counter 0.15% HA preparation. Further studies are needed to verify the results of this study and to compare the safety and efficacy of this HA preparation with saline solution.


Reactive Oxygen-Forming Nox5 Links Vascular Smooth Muscle Cell Phenotypic Switching and Extracellular Vesicle-Mediated Vascular Calcification.
Vascular calcification, the formation of calcium phosphate crystals in the vessel wall, is mediated by vascular smooth muscle cells (VSMCs). However, the underlying molecular mechanisms remain elusive, precluding mechanism-based therapies. Phenotypic switching denotes a loss of contractile proteins and an increase in migration and proliferation, whereby VSMCs are termed synthetic. We examined how VSMC phenotypic switching influences vascular calcification and the possible role of the uniquely calcium-dependent reactive oxygen species (ROS)-forming Nox5 (NADPH oxidase 5). In vitro cultures of synthetic VSMCs showed decreased expression of contractile markers CNN-1 (calponin 1), α-SMA (α-smooth muscle actin), and SM22-α (smooth muscle protein 22α) and an increase in synthetic marker S100A4 (S100 calcium binding protein A4) compared with contractile VSMCs. This was associated with increased calcification of synthetic cells in response to high extracellular Ca2+. Phenotypic switching was accompanied by increased levels of ROS and Ca2+-dependent Nox5 in synthetic VSMCs. Nox5 itself regulated VSMC phenotype as siRNA knockdown of Nox5 increased contractile marker expression and decreased calcification, while overexpression of Nox5 decreased contractile marker expression. ROS production in synthetic VSMCs was cytosolic Ca2+-dependent, in line with it being mediated by Nox5. Treatment of VSMCs with Ca2+ loaded extracellular vesicles (EVs) lead to an increase in cytosolic Ca2+. Inhibiting EV endocytosis with dynasore blocked the increase in cytosolic Ca2+ and VSMC calcification. Increased ROS production resulted in increased EV release and decreased phagocytosis by VSMCs. We show here that contractile VSMCs are resistant to calcification and identify Nox5 as a key regulator of VSMC phenotypic switching. Additionally, we describe a new mechanism of Ca2+ uptake via EVs and show that Ca2+ induces ROS production in VSMCs via Nox5. ROS production is required for release of EVs, which promote calcification. Identifying molecular pathways that control Nox5 and VSMC-derived EVs provides potential targets to modulate vascular remodeling and calcification in the context of mineral imbalance. Graphic Abstract: A graphic abstract is available for this article.


Ultrastructure of the corpus cardiacum and corpus allatum of the house cricket Acheta domesticus.
The ultrastructure of the corpus cardiacum (CC) and corpus allatum (CA) of the house cricket, Acheta domesticus, is described. Axon profiles within the CC contain neurosecretory granules 160-350 nm in diameter which are indistinguishable from those found in type I neurosecretory cells of the pars intercerebralis and in the nervus corporis cardiaci I. The CC itself contains two cell types: intrinsic neurosecretory cells and glial cells. Intrinsic NSC cytoplasm contains Golgi bodies and electron dense neurosecretory granules 160-350 nm in diameter. Synaptoid configurations with 20-50 nm diameter electron lucent vesicles were observed within axon profiles of the CC. The structure of the CA is relatively uniform with one cell type predominating. Typical CA cells possess large nucleoli, active Golgi complexes, numerous mitochondria, and occassional microtubules. Groups of dark staining cells scattered throughout the CA of some animals were interpreted as evidence of cellular death.


Ontogeny of the chick embryo spleen--a cytological study.
The differentiation of the chick embryo spleen was investigated from E4 till after hatching, using a number of cytological markers. The reticular fiber network, disclosed by silver staining, was a loose array of short fibers until E15. It developed into a complex pattern closely related to the specialized segments of the vascular tree between E15 and E18. While the very first cells belonging to the hemopoietic lineage could be identified as early as E4, steady state hemopoiesis set in by E10. Of the two cell lineages maturing in the organ, erythroid cells disappeared by E15, and granulocytes emigrated within 3 to 4 days around hatching. The process of lymphocyte homing, detected by specific markers, began just before hatching and was completed within the first week of life. Thus differentiation of the stromal anatomy appears as the prerequisite for the transition of embryonic (= hemopoietic) to adult (= secondary lymphoid) functions.


Craniofacial and dental characteristics of three Japanese individuals with genetically diagnosed SATB2-associated syndrome.
Craniofacial defects are one of the most frequent phenotypes in syndromic diseases. More than 30% of syndromic diseases are associated with craniofacial defects, which are important for the precise diagnosis of systemic diseases. Special AT-rich sequence-binding protein 2 (SATB2)-associated syndrome (SAS) is a rare syndromic disease associated with a wide variety of phenotypes, including intellectual disability and craniofacial defects. Among them, dental anomalies are the most frequently observed phenotype and thus becomes an important diagnostic criterion for SAS. In this report, we demonstrate three Japanese cases of genetically diagnosed SAS with detailed craniofacial phenotypes. The cases showed multiple dental problems, which have been previously reported to be linked to SAS, including abnormal crown morphologies and pulp stones. One case showed a characteristic enamel pearl at the root furcation. These phenotypes add new insights for differentiating SAS from other disorders.


PHI-1 induced enhancement of myosin phosphorylation in chicken smooth muscle.
Herein, we provide evidence that in chicken smooth muscle, G-protein stimulation by a Rho-kinase pathway leads to an increase in myosin light chain phosphorylation. Additionally, G-protein stimulation did not increase MYPT1 phosphorylation at Thr695 or Thr850, and CPI-17, was not expressed in chicken smooth muscle. However, PHI-1 was present in chicken smooth muscle tissues. Both agonist and GTP(gamma)S stimulation result in an increase in PHI-1 phosphorylation, which is inhibited by inhibitors to both Rho-kinase (Y-27632) and (PKC) GF109203x. These data suggest that PHI-1 may act as a CPI-17 analog in chicken smooth muscle and inhibit myosin phosphatase activity during G-protein stimulation to produce Ca2+ sensitization.


The middle fossa approach and extended middle fossa approach: technique and operative nuances.
The middle fossa approach and extended middle fossa approach, also known as the anterior transpetrosal approach, are cranial base techniques for addressing small vestibular schwannomas, medial temporal bone lesions, midbasilar trunk aneurysms, and selected petroclival lesions. To provide an outline of a number of technical nuances that are important to correct application of these approaches, maximizing exposure, and limiting potential morbidity. Via a temporal craniotomy, the petrous apex is removed in variable degrees, depending on the exposure requirements of the lesion. The technique is described in detail with appropriate nuances of the technique provided. The described nuances of technique in the performance of the approaches have resulted in successful application of these techniques in a significant number of cases. Significant familiarity and practice with these surgical approach techniques are critical to applying them safely to clinical problems. A number of technical details can assist the surgeon in achieving optimal exposure and limited morbidity.


Cortical rhythms reactivity in AD, LBD and normal subjects: a quantitative MEG study.
The present study evaluated the reactivity of cortical rhythms in 15 Alzheimer's disease (AD) patients, 7 Lewy body dementia (LBD) patients and 9 control subjects using a 165 SQUID whole-head MEG system. The absolute power values of the rhythms recorded over different areas over the brain (frontal, parietal, temporal, occipital) were analysed in the 3-47Hz frequency range. The cortical reactivity of the alpha (9-14Hz) and pre-alpha rhythms (7-9Hz) during open and closed eyes conditions differentiated the control group from the patient groups and moderate AD from severe AD and LBD groups, respectively. The cortical reactivity of the slow-band (3-7Hz) obtained by comparing a simple mental task and the rest discriminated the severe AD group from the other groups. In addition, spectral coherence analysis in the alpha band showed that the loss of coherence in AD and LBD patients mainly involved long connections. These results suggest that investigations on rhythms reactivity and spectral coherence might help on the study of the dementias with different etiology.


Low molecular weight xanthan gum for treating osteoarthritis.
Osteoarthritis (OA) is one of the most common chronic diseases and characterized by degradation of articular cartilage. We have previously reported xanthan gum (XG) injection preparation with high molecular weights (Mw) in ranging from 3×106Da to 5×106Da (HM-XG) could enhance the viscosity of synovial fluid, protect joint cartilage in rabbit, and the therapeutical effect has no significance difference with an existing clinical medication (sodium hyaluronate, SH) at the same injection frequency (once weekly for 5 weeks). Herein, we prepared a XG injection preparation with a low Mw (LM-XG) in ranging from 1×106Da to 1.5×106Da, and evaluated the therapeutical effect for OA therapy at once every 2 weeks for 5 weeks with an SH at once weekly for 5 weeks as reference. The model of OA was induced using anterior cruciate ligament transection (ACLT) in a rabbit in vivo and also using sodium nitroprusside (SNP) in cell culture in vitro. The results showed that LW-XG could also protect cartilage from damage, decrease the concentration of nitric oxide (NO) in synovial fluid and reverse the amplification of the knee joint width similar to HM-XG as our previously reported. At the cellular level, LW-XG promotes proliferation while decreases apoptosis of chondrocytes. Mechanistically at the molecular level, these effects are elicited via down-regulation of the protein levels of caspase-3 and bax and up-regulation of the protein levels of bcl-2 in cartilage in both in vivo and in vitro. These results showed that LW-XG maybe become an excellent candidate long-acting drug for treating OA.


A Leigh syndrome caused by compound heterozygous mutations on NDUFAF5 induce early infant death: A case report.
The NADH:ubiquinone oxidoreductase complex assembly factor gene (NDUFAF5) has been linked to the occurrence of Leigh syndrome, but few causative mutations have been identified. Here we report a rare case of Leigh syndrome in an infant who died in the early postnatal period. We performed whole-exome sequencing (WES) and mutation analysis of NDUFAF5 to obtain genetic data on the patient and describe the clinical and genetic characteristics. The proband was a 2-month-old male infant who suffered from recurrent vomiting and persistent seizure and died at 2 months of age after early medical support and treatment. His parents reported the unexplained death of the infant's older brother at 1 year of age. WES of the patient's DNA revealed c.357C>G and c.611C>T compound heterozygous mutations in NDUFAF5; analysis with the MutationTaster application indicated that both were pathogenic (p = 0.99). Significant structural changes in the transport domain of the protein were predicted using SWISS-MODEL. We estimated the stability of the mutant protein using a mutation cutoff scanning matrix and found reductions in Gibbs free energy (-0.623 kcal/mol for p.D119E and -0.813 kcal/mol for p.A204V), indicating that the mutations led to an unstable protein structure. We speculated that the patient died as a result of impaired mitochondrial function caused by the NDUFAF5 mutations, and made a diagnosis of Leigh syndrome. Our results demonstrate that molecular genetic screening is useful for the diagnosis of mitochondrial diseases, especially in children with a positive family history. Leigh syndrome should be considered in the diagnosis of patients presenting with severe recurrent vomiting and feeding difficulties with persistent seizure. Our findings expand the mutation spectrum of the NDUFAF5 gene and contribute to the genotype-phenotype map of mitochondrial respiratory chain complex I deficiency.


Substrate as a source of thermodynamic nonideality in enzyme kinetic studies: invertase-catalyzed hydrolysis of sucrose.
Expressions for the effects of thermodynamic nonideality arising from the use of high concentrations of small substrate in enzyme kinetic studies are derived. Their application to experimental results for the hydrolysis of sucrose by yeast invertase (pH 4.9, 37 degrees C) signifies that the progressive decrease in initial velocity at high sucrose concentration is consistent with the occurrence of isomeric expansion during the transition of an enzyme-substrate complex to its activated state. Ultracentrifuge studies on the yeast enzyme preparation are then used to establish the physical acceptability of the volume change required to account for the kinetic effects in these terms: the postulated expansion of 1.3 liter/mol would represent a mere 0.16% increase in hydrated volume (or a corresponding increase in extent of asymmetry). Finally, although originally interpreted to signify an effect of sucrose on water concentration, published results for the invertase-sucrose system [J. M. Nelson and M. P. Schubert (1928) J. Amer. Chem. Soc. 50, 2188-2193] also find a rational explanation in terms of the present analysis based on effects of thermodynamic nonideality in enzyme kinetic studies.


Free tissue transfers at Hutt Hospital.
The Wellington Regional Plastic, Maxillofacial & Burns Unit based at Hutt Hospital provides comprehensive reconstructive services to central New Zealand with a population of 1.1 million. Free tissue transfer procedures in the Unit were audited to determine the indications and rate of usage in our population, our success and complication rates, and how these compare with published series. Prospectively collected data on all free tissue transfer procedures between January 2006 and September 2010 were analysed. Two hundred and seven free flaps including 17 flap types being performed on 186 consecutive patients including 199 primary and 8 salvage flaps. Eighty-three percent were elective and 17% were acute cases. The majority of the flaps were used for head and neck (48%) and breast (31.5%) reconstruction. Ulnar forearm flap was the most commonly used fasciocutaneous flap. 18.8% of patients had major complications requiring return to theatre. Microsurgical revision was performed in nine (4.3%) flaps of which six were successfully salvaged. Overall, 13 flaps (6.3%) failed completely, giving an overall success rate of 93.7%. Haematoma requiring formal drainage occurred in 12 (5.8%) cases. The wide variety of flaps used reflects the very broad range of defects requiring free flap reconstruction. We show a free flap success rate of 93.7% in our medium-sized regional unit. Our microsurgical revision rate of 4.3% is lower than the revision rate of 10% in reported series with high overall success rates. More consistent early detection of failing flaps is likely to further improve our overall success rate.


Cloning and expression of sheep DNA methyltransferase 1 and its development-specific isoform.
Unlike the mouse embryo, where loss of DNA methylation in the embryonic nucleus leaves cleavage stage embryos globally hypomethylated, sheep preimplantation embryos retain high levels of methylation until the blastocyst stage. We have cloned and sequenced sheep Dnmt1 and found it to be highly conserved with both the human and mouse homologues. Furthermore, we observed that the transcript normally expressed in adult somatic tissues is highly abundant in sheep oocytes. Throughout sheep preimplantation development the protein is retained in the cytoplasm whereas Dnmt1 transcript production declines after the embryonic genome activation at the 8-16 cell stage. Attempts to clone oocyte-specific 5' regions of Dnmt1, known to be present in the mouse and human gene, were unsuccessful. However, a novel ovine Dnmt1 exon, theoretically encoding 13 amino acids, was found to be expressed in sheep oocytes, preimplantation embryos and early fetal lineages, but not in the adult tissue. RNAi-mediated knockdown of this novel transcript resulted in embryonic developmental arrest at the late morula stage, suggesting an essential role for this isoform in sheep blastocyst formation.


Data of enzymatic activities of the electron transport chain and ATP synthase complexes in mouse hepatoma cells following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).
2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is the most widely studied ligand of the aryl hydrocarbon receptor (AHR). The AHR-dependent TCDD-induced mitochondrial hyperpolarization (Tappenden et al., 2011) [1] and reduced oxygen consumption rate of intact mouse hepatoma cells (Huang et al., in press) [2] in the previous studies suggest that these alterations can be related to enzymatic activities of the electron transport chain (ETC) and ATP synthase in oxidative phosphorylation (OXPHOS) system. Here, we evaluated the activity of each complex in the OXPHOS system using in vitro enzymatic assays. The calculated enzymatic activity of each complex was normalized against the activity of citrate synthase. To combine each value from an independent experiment, normalized enzyme activities from cells exposed to TCDD were converted to fold changes via comparison to the activity relative to time-matched vehicle control. The averaged fold change for each treatment suggests more replicates are needed in order to clearly evaluate a difference between treatments.


Changes over time in survival of children after AIDS diagnosis in New York City.
We determined whether survival of children following AIDS diagnosis is improving over time through 1991. AIDS surveillance data from New York City Department of Health on 914 pediatric AIDS patients, diagnosed between 1979 and 1991 and presumed due to maternal-infant HIV transmission, were analyzed. Survival following AIDS diagnosis, established from hospital records and death certificates, was compared by calendar year of initial diagnosis using Kaplan-Meier lifetable analysis. Cox Proportional Hazards regression models were used to compare survival for patients diagnosed earlier or later in the decade, controlling for age at diagnosis, presenting opportunistic illness, and gender of the child. Patients diagnosed with AIDS from October 1987 to September 1989 survived longer, median survival 17 months after diagnosis, than patients diagnosed before September 1987, median survival 10 months (relative risk [RR] = 0.76; 95% confidence intervals [CI] = 0.62, 0.93). Patients diagnosed from October 1989 to December 1991 also survived a median of 17 months. Secular improvements in survival after AIDS diagnosis remained after controlling for age at diagnosis, presenting diagnosis, and gender, even if deaths within three months of diagnosis were excluded. These data suggest that for recent years, survival following AIDS diagnosis in those contracting the infection through maternal-infant transmission has been prolonged. Possible explanations for these findings include both methodological issues (changes in diagnostic criteria, incomplete ascertainment of deaths) and substantive issues (developments in therapeutic interventions and management of pediatric AIDS).


Amiloride evokes significant natriuresis and weight loss in kidney transplant recipients with and without albuminuria.
Albuminuria in kidney transplant recipients (KTRs) is associated with hypertension and aberrant glomerular filtration of serine proteases that may proteolytically activate the epithelial Na+ channel (ENaC). The present nonrandomized, pharmacodynamic intervention study aimed to investigate if inhibition of ENaC increases Na+ excretion and reduces extracellular volume in KTRs dependent on the presence of albuminuria. KTRs with and without albuminuria (albumin-to-creatinine ratio > 300 mg/g, n = 7, and <30 mg/g, n = 7, respectively) were included and ingested a diet with fixed Na+ content (150 mmol/day) for 5 days. On the last day, amiloride at 10 mg was administered twice. Body weight, 24-h urine electrolyte excretion, body water content, and ambulatory blood pressure as well as plasma renin, angiotensin II, and aldosterone concentrations were determined before and after amiloride. Amiloride led to a significant decrease in body weight, increase in 24-h urinary Na+ excretion, and decrease in 24-h urinary K+ excretion in both groups. Urine output increased in the nonalbuminuric group only. There was no change in plasma renin, aldosterone, and angiotensin II concentrations after amiloride, whereas a significant decrease in nocturnal systolic blood pressure and increase in 24-h urine aldosterone excretion was observed in albuminuric KTRs only. There was a significant correlation between 24-h urinary albumin excretion and amiloride-induced 24-h urinary Na+ excretion. In conclusion, ENaC activity contributes to Na+ and water retention in KTRs with and without albuminuria. ENaC is a relevant pharmacological target in KTRs; however, larger and long-term studies are needed to evaluate whether the magnitude of this effect depends on the presence of albuminuria.NEW & NOTEWORTHY Amiloride has a significant natriuretic effect in kidney transplant recipients (KTRs) that relates to urinary albumin excretion. The epithelial Na+ channel may be a relevant direct pharmacological target to counter Na+ retention and hypertension in KTRs. Epithelial Na+ channel blockers should be further investigated as a mean to mitigate Na+ and water retention and to potentially obtain optimal blood pressure control in KTRs.


Acute hemodynamic changes during far infrared treatment of the arteriovenous fistula in hemodialysis patients.
The arteriovenous fistula (AVF) is the preferred vascular access for hemodialysis (HD) treatment and preservation of a stable vascular access is crucial. Long term Far Infrared Radiation (FIR) has been found to increase access flow together with an enhanced maturation and patency of the AVF. The acute effects of FIR on access flow have been sparsely described and the results are contradictory, perhaps due to differences in measurement conditions and other factors of importance for access flow. Twenty patients in HD with an AVF were included. Each patient was randomized to receive either FIR (FIR group) or no FIR (control group). The acute changes in access flow were investigated in both groups on the second dialysis day of the week and during the first 1.5 h of the dialysis session. Concomitant changes in hemodynamic parameters of importance for access flow were also explored. There was no significant change in access flow in the FIR group compared with the control group (median (Interquartile Range)) (-10 (-413.8; 21.3) ml/min vs -17.5 (-83.8; 76.3) ml/min, p = 0.58). There was no significant difference in any of the hemodynamic parameters between the FIR and the control group; cardiac output (-0.7 (-1.2; -0.2) l/min vs -0.4 (-0.9; 0.1) l/min, p = 0.58), cardiac index (-0.3 (-0.5; -0.1)) l/min/m2 vs -0.3 (-0.4; 0) l/min/m2, p = 0.68), mean arterial pressure (5.5 (-1.8; 8.4) mmHg vs 1.5 (-3; 6.3) mmHg, p = 0.35) and total peripheral resistance (2 (1.8; 3.4) mmHg × min/l vs 1 (-0.3; 3.1) mmHg × min/l, p = 0.12). In this trial, with a highly standardized set-up, one session of FIR did not result in any acute changes in access flow. This was not due to differences in the hemodynamic parameters between the groups.


Connectome Reorganization Associated With Surgical Outcome in Temporal Lobe Epilepsy.
To identify the distinct pattern of anatomical network reorganization in surgically refractory mesial temporal lobe epilepsy (MTLE) patients using a longitudinal design. We collected longitudinal diffusion-weighted images of 19 MTLE patients before and after anterior temporal lobectomy. Patients were classified as seizure-free (SF) or nonseizure-free (NSF) at least 1 year after surgery. We constructed whole-brain anatomical networks derived from white matter tractography and evaluated network connectivity measures by graph theoretical analysis. The reorganization trajectories of network measures in SF and NSF patients were investigated by two-way mixed analysis of variance, with factors "group" (SF vs NSF) and "treatment" (presurgery vs postsurgery). Widespread brain structures showed opposite reorganization trajectories in FS and NSF groups (interaction effect). Most of them showed group difference before surgery and then converge after surgery, suggesting that surgery remodeled these structures into a similar status. Conversly, contralateral amygdala-planum-temporale and thalamic-parietal tracts showed higher connectivity strength in NSF than in SF patients after surgery, indicating maladaptive neuroplastic responses to surgery in NSF patients. Our findings suggest that surgical outcomes are associated not only with the preoperative pattern of anatomical connectivity, but also with connectome reconfiguration following surgery. The reorganization of contralateral temporal lobe and corticothalamic tracts may be particularly important for seizure control in MTLE.


Weight loss in conservative treatment of obesity in women is associated with physical activity and circadian phenotype: a longitudinal observational study.
The study investigates the association between circadian phenotype (CP), its stability (interdaily stability - IS) and physical activity (PA) in a weight loss (WL) programme. Seventy-five women in WL conservative treatment (BMI ≥ 25 kg/m2) were measured (for about 3 months in between 2016 and 2018) by actigraphy. We observed a difference in time of acrophase (p = 0.049), but no difference in IS (p = 0.533) between women who lost and did not lose weight. There was a difference in PA (mesor) between groups of women who lost weight compared to those who gained weight (p = 0.007). There was a relationship between IS and PA parametres mesor: p0.001; and the most active 10 h of a day (M10): p < 0.001 - the more stable were women in their rhythm, the more PA they have. Besides confirming a relationship between PA and WL, we also found a relation between WL and CP based on acrophase. Although no direct relationship was found for the indicators of rhythm stability (IS), they can be considered very important variables because of their close connection to PA - a main factor that contributes to the success of the WL programme. According to the results of the study, screening of the CP and its stability may be beneficial in the creation of an individualized WL plan.


The relationship between LAPTM4B polymorphisms and cancer risk in Chinese Han population: a meta-analysis.
LAPTM4B is a newly cloned gene that shows an active role in many solid tumors progression in substantial researches, mainly through the autophage function. Accumulated studies have been conducted to determine the association of LAPTM4B polymorphism with cancer risk. While the results are inconsistent, we conducted the meta-analysis to determine the strength of the relationship. Results showed that allele*2 carriers exhibited a significantly increased risk of cancer development with comparison to allele*1 homozygote (for *1/2, OR = 1.55, 95% CI 1.367-1.758; for *2/2, OR = 2.093, 95%CI 1.666-2.629; for *1/2 + *2/2, OR = 1.806, 95%CI 1.527-2.137). We also observed a significant association between *2/2 homozygote and cancer risk with comparison to allele*1 containing genotypes (OR = 1.714, 95%CI 1.408-2.088). Allele*2 is a risk factor for cancer risk (OR = 1.487, 95%CI 1.339-1.651). Stratified analysis by tumor type exhibits the significant association of this genetic variants with various cancers. In conclusion, LAPTM4B polymorphism is associated with cancer risk and allele*2 is a risk factor.


Pachytene analysis in a 17;21 reciprocal translocation carrier: role of the acrocentric chromosomes in male sterility.
Pachytene analysis was undertaken in an infertile male, heterozygous for a 17;21 reciprocal translocation. The quadrivalent was identified by its configuration and chromomere pattern. A non-random association was found between the quadrivalent and the sex vesicle in 77% of the pachytene nuclei analysed. In 13.1% of the cells the contact with the sex vesicle was established by the terminal chromomere of the two chromosomes 21; in 63.9% of the cells, the entire region of the breakpoints was completely hidden by the sex vesicle. In some nuclei asynapsis was found in the region of the breakpoints. The nature of the contact between the quadrivalent and the sex vesicle is discussed in this paper. It is proposed that the acrocentric chromosome favours the contact between the quadrivalent and the sex vesicle, and increases the risk of sterility in male carriers of Robertsonian translocations and of reciprocal translocations involving one acrocentric chromosome.


Transcription of the Drosophila CKI gene dacapo is regulated by a modular array of cis-regulatory sequences.
Organogenesis requires coordination between developmental programs and cell cycle progression. The Cip/Kip families of cyclin-dependent kinase inhibitor (CKI) are main effectors linking these two programs. In many instances, expression of Cip/Kip CKIs are increased abruptly in cells entering their last mitotic cycle, suggesting that CKI expression is developmentally regulated. Expression of Dacapo (Dap), a Drosophila Cip/Kip CKI, is transiently up-regulated immediately before cells enter G1 arrest. Here we report that dap transcription is controlled by modular arrays of tissues specific cis-regulatory elements. Furthermore, we identified pan-neural Prospero as a regulator of dap transcription in the developing nervous system, providing an example how tissue-specific developmental programs can be linked to cell cycle progression.


Messenger RNA activities of four acute phase proteins during inflammation.
Poly(A)+ RNA isolated from the livers of normal rats and of rats suffering from an acute inflammation was translated in a cell-free translation system from rabbit reticulocytes. The translation products were immunoprecipitated with specific antisera against alpha 1-acid glycoprotein, alpha 2-macroglobulin, transferrin, alpha 1-proteinase inhibitor and albumin. 15 to 21 h after intramuscular injection of turpentine 73-, 66-, 2.8-, and 2-fold increases in translatable mRNAs for alpha 1-acid glycoprotein, alpha 2-macroglobulin, transferrin and alpha 1-proteinase inhibitor, respectively, were observed. For albumin a decrease in translatable mRNA to about 30% of controls was measured.


Serological study of ovine toxoplasmosis in Colombia: prevalence of haemagglutinating antibodies to toxoplasma in sheep.
A total of 1655 ovine sera from the six principal sheep rearing areas of Colombia were examined for toxoplasma antibodies using the indirect haemagglutination test. It was shown that 58 per cent of these samples had titres of 1/16 or greater. Seropositive animals were distributed throughout all the areas studied. The possible factors affecting the distribution of seropositive sheep in one of the areas are discussed.


Cells positive for the O4 surface antigen isolated by cell sorting are able to differentiate into astrocytes or oligodendrocytes.
Cells expressing the surface antigen O4 were isolated as pure populations from cultures of murine brain or cerebellum using fluorescence activated cell sorting. When these O4-positive cells were further cultured in the presence of fetal calf serum (FCS) many cells expressed both O4 and the astrocyte marker glial fibrillary acidic protein (GFAP) after 4 days of culture. Cells not exposed to FCS expressed O4, but never GFAP. GFAP-positive cells in the presence of fetal calf serum very rarely expressed the myelin associated glycoprotein (MAG) or O1, both of which are expressed on more differentiated oligodendrocytes, and never expressed O10 that is characteristic of even more mature oligodendrocytes. These results show that glial cells expressing O4, but not MAG, O1, O10 or GFAP are bipotential precursor cells able to differentiate into astrocytes or oligodendrocytes depending on the culture conditions and suggest that bipotentiality of glial precursor cells is retained up to a later developmental stage than that of the O2A progenitor cell.


Basics of compounding: considerations for implementing United States pharmacopeia chapter 797 pharmaceutical compounding-sterile preparations, part 16: suggested standard operating procedures.
The standard operating procedures suggested in this article are presented to compounding pharmacies to ensure the quality of the environment in which a CSP is prepared. Since United States Pharmacopeia Chapter 797 provides minimum standards, each facility should aim for best practice gold standard. The standard operating procedures should be tailored to meet the expectations and design of each facility. Compounding personnel are expected to know and understand each standard operating procedure to allow for complete execution of the procedures.


Long- and short-term mortality rates in patients who primarily survive cardiac arrest compared with a normal population.
This is a follow-up study of 180 survivors after cardiac arrest outside intensive care and coronary care units. The follow-up extended over 0.5 to 8.5 years (averaged 4.3 years) after the primary cardiac arrest. Of the patients, 72 (40%) were discharged from hospital, 13 with anoxic brain damage. Thirty-ourred in 43% after 2 years, and in 50% after 3 years. After that time, the mortality was similar to that of a comparable normal population. At the end of the study, 34 patients were still alive, including eight with neurological sequelae. Their present cardiac function was satisfactory in the majority (59%) of the patients falling into group I or II of the American Heart Association classification. Their social situation, however, left much to be desired, as only 50% had fully or partly regained their previous level of activity.


MicroRNA-338-3p helps regulate ovarian function by affecting granulosa cell function and early follicular development.
Follicular development in mammalian ovaries is a complex and dynamic process, and the interactions and regulatory-feedback loop between the follicular microenvironment, granulosa cells (GCs), and oocytes can affect follicular development and normal ovary functions. Abnormalities in any part of the process may cause abnormal follicular development, resulting in infertility. Hence, exploring the pathogenesis of abnormal follicular development is extremely important for diagnosing and treating infertile women. RNA sequencing was performed with ovarian cortical tissues established in vitro. In situ-hybridization assays were performed to study microRNA-338-3p (miR-338-3p) expressed in GCs and oocytes. In vitro culture models were established with GCs and neonatal mouse ovaries to study the biological effects of miR-338-3p. We also performed in vivo experiments by injecting adeno-associated virus vectors that drive miR-338-3p overexpression into the mouse ovarian bursae. Sequencing analysis showed that miR-338-3p was expressed at significantly higher levels in ovarian cortical tissues derived from patients with ovarian insufficiency than in cortical tissues derived from patients with normal ovarian function; miR-338-3p was also significantly highly expressed in the GCs of patients with diminished ovarian reserve (P < 0.05). In situ-hybridization assays revealed that miR-338-3p was expressed in the cytoplasm of GCs and oocytes. Using in vitro culture models of granulosa cells, we found that miR-338-3p overexpression significantly suppressed the proliferation and oestradiol-production capacity of GCs (P < 0.05). In vitro culture models of neonatal mouse ovaries indicated that miR-338-3p overexpression suppressed the early follicular development in mouse ovaries. Further analysis revealed that miR-338-3p might be involved in transforming growth factor β-dependent regulation of granulosa cell proliferation and, thus, early follicular development. Injecting miR-338-3p-overexpression vectors into the mouse ovarian bursae showed that miR-338-3p down-regulated the oocyte mitochondrial membrane potential in mice and disrupted mouse oestrous cycles. miR-338-3p can affect early follicular development and normal ovary functions by interfering with the proliferation and oestradiol production of GCs. We systematically elucidated the regulatory effect of miR-338-3p on follicular development and the underlying mechanism, which can inspire new studies on the diagnosis and treatment of diseases associated with follicular development abnormalities.


Prolyl 4-hydroxylase subunit alpha 3 facilitates human colon cancer growth and metastasis through the TGF-β/Smad signaling pathway.
Prolyl 4-hydroxylase subunit alpha 3 (P4HA3) has been known to be associated with a variety of human cancers. However, the role of P4HA3 on colon cancer growth and metastasis is unclear. In this study, we investigated the effect of P4HA3 on the growth and metastasis of colon cancer and its possible molecular mechanism. First of all, we demonstrated that P4HA3 expression was greatly higher in cells and tissues of colon cancer than that in non-tumor tissues and cells, and the prognosis of patients who had higher P4HA3 was distinctively poorer than patients who had lower level of P4HA3. Second, it was shown that P4HA3 knockdown strongly inhibited the migration, proliferation and invasion ability of colon cancer cells. However, P4HA3 over-expression accelerated the abilities. Meanwhile, P4HA3 could promote subcutaneous tumorigenesis in nude mice in vivo. In addition, P4HA3 knockdown significantly decreased mesenchymal markers Vimentin, N-cadherin and Snail expression and increased epithelial marker E-cadherin expression. And conversely, over-expression of P4HA3 produced the opposite effects. In the current study, there was further evidence that down-regulating P4HA3 significantly reduced both TGF-β and its following molecules including p-Smad2 as well as p-Smad3. However, overexpression of P4HA3 showed the opposite effect. In conclusion, this study shows that P4HA3 promotes the human colon cancer growth and metastasis by affecting TGF-β/Smad signaling pathway. P4HA3 may become a new target for early diagnosis, treatment and prognosis assessment of colon cancer.


Cavity-enabled enhancement of ultrafast intramolecular vibrational redistribution over pseudorotation.
Vibrational strong coupling (VSC) between molecular vibrations and microcavity photons yields a few polaritons (light-matter modes) and many dark modes (with negligible photonic character). Although VSC is reported to alter thermally activated chemical reactions, its mechanisms remain opaque. To elucidate this problem, we followed ultrafast dynamics of a simple unimolecular vibrational energy exchange in iron pentacarbonyl [Fe(CO)5] under VSC, which showed two competing channels: pseudorotation and intramolecular vibrational-energy redistribution (IVR). We found that under polariton excitation, energy exchange was overall accelerated, with IVR becoming faster and pseudorotation being slowed down. However, dark-mode excitation revealed unchanged dynamics compared with those outside of the cavity, with pseudorotation dominating. Thus, despite controversies around thermally activated VSC modified chemistry, our work shows that VSC can indeed alter chemistry through a nonequilibrium preparation of polaritons.


Epithelial-mesenchymal transitions in bronchopulmonary dysplasia of newborn rats.
Bronchopulmonary dysplasia (BPD) is a major threat to the health of premature infants yet its pathogenesis is not fully understood. Epithelial-mesenchymal transition (EMT) of lung epithelial cells may lead to BPD. To investigate the potential occurrence of EMT in a newborn rat model of BPD. Newborn rats were exposed to a hyperoxic environment within 12 hr of birth. Lung tissue and isolated alveolar epithelial type II cells (AT2 cells) were collected on Days 1, 3, 7, 14, and 21 after hyperoxic exposure. Pathological changes in lung tissue, alveolar development, ultrastructural changes in AT2 cells, co-expression of surfactant associated surfactant protein C (SPC), and α-smooth muscle actin (α-SMA) were investigated. The relative expression of SPC, α-SMA, E-cadherin, and N-cadherin were investigated in lung tissue and isolated AT2 cells. In lung tissue, alveolar development was attenuated from Day 7 onwards in the BPD model group; co-expression of SPC and α-SMA and ultrastructural changes typical of EMT were observed in AT2 cells from rats in the BPD group. SPC and α-SMA expression levels were higher in tissue samples from the BPD group than in control samples. Beginning on Day 7, evidence of a switch from E-cadherin to N-cadherin expression was observed in BPD lung tissue sample and in isolated AT2 cells. EMT of AT2 cells occurred in the hyperoxia-induced newborn rat BPD model and resulted in attenuated alveolar development as a portion of the myofibroblasts accumulated in the lung originated from AT2 cells via EMT.


In patients with combined clavicle and multiple rib fractures, does fracture fixation of the clavicle improve clinical outcomes? A multicenter prospective cohort study of 232 patients.
Clavicle and rib fractures are often sustained concomitantly. The combination of injuries may result in decreased stability of the chest wall, making these patients prone to (respiratory) complications and prolonged hospitalization. This study aimed to assess whether adding chest wall stability by performing clavicle fixation improves clinical outcomes in patients with concurrent clavicle and rib fractures. A prospective multicenter study was performed including all adult patients admitted between January 2018 and March 2021 with concurrent ipsilateral clavicle and rib fractures. Patients treated operatively versus nonoperatively for their clavicle fracture were matched using propensity score matching. The primary outcome was hospital length of stay (HLOS). Secondary outcomes were intensive care unit length of stay, duration of mechanical ventilation, pain, complications, and quality of life at 6 weeks and 12 months of follow-up. In total, 232 patients with concomitant ipsilateral clavicle and rib fractures were included. Fifty-two patients (22%) underwent operative treatment of which 39 could be adequately matched to 39 nonoperatively treated patients. No association was observed between clavicle plate fixation and HLOS (mean difference, 2.3 days; 95% confidence interval, -2.1 to 6.8; p = 0.301) or any secondary endpoint. Eight of the 180 nonoperatively treated patients (4%) had a symptomatic nonunion, for which 5 underwent secondary clavicle fixation. We found no evidence that, in patients with combined clavicle and multiple rib fractures, plate fixation of the clavicle reduces HLOS, pain, or (pulmonary) complications, nor that it improves quality of life. Therapeutic/Care Management; Level III.


Glial and glycoconjugate boundaries during postnatal development of the central nervous system.
The localization of glycosylated molecules and glia has been studied during early postnatal development in the mouse central nervous system (CNS) using autoradiographic detection of radiolabeled fucose incorporation, and in sections processed either for histochemistry or immunocytochemistry following binding of labeled lectins or an antibody to glial fibrillary acidic protein. Radiolabeled sugar incorporation, lectin binding of glycoconjugates, and glial labeling all reveal borders between nuclei within the diencephalon, midbrain, and brainstem through the first postnatal week. Glycoconjugate and glial boundaries exist throughout the CNS during pattern formation events, and they also are seen in relation to fine aspects of developing functional organization within individual structures (e.g. segmentation associated with the representation of mystacial vibrissae within the brainstem trigeminal complex). The observation that each of the probes employed in this study fails to label boundary organization during later postnatal times suggests that the distribution and chemistry of the glial/glycoconjugate network are dynamic, and they change in accordance with distinct maturational states of the nervous system.


Prognosis of spontaneously ruptured hepatocellular carcinoma: a propensity score matching study.
Spontaneous rupture is a fatal complication of hepatocellular carcinoma (HCC). This study compared the prognosis of spontaneously ruptured HCC (srHCC) with that of non-ruptured HCC (nrHCC). A total of 185 srHCC patients and 1085 nrHCC patients treated by hepatectomy between February 2005 and December 2017 at Zhongshan Hospital were retrospectively reviewed and enrolled. The overall survival (OS) and time to recurrence (TTR) were evaluated. A 1:2 propensity score matching (PSM) analysis was performed using the nearest neighbor matching with a caliper of 0.2. Before PSM, patients with srHCC who underwent hepatectomy (n = 185) had a poorer prognosis than those with nrHCC (n = 1085; 5-year OS, 39.1% vs 59.2%, P < 0.001; 5-year TTR, 83.8% vs 54.9%, P < 0.001). After PSM, patients with srHCC (n = 156) had higher 5-year TTR (83.2% vs 69.0%, P < 0.001) but comparable 5-year OS with those with nrHCC (n = 312, 44.0% vs 46.0%, P = 0.600). Univariate and multivariate analyses demonstrated spontaneous rupture as an independent risk factor for TTR (hazard ratio [HR], 1.681; 95% confidence interval [CI] 1.326-2.132; P < 0.001), but not for OS (HR: 1.074; 95% CI 0.823-1.401; P = 0.600). Further analysis revealed that srHCC was not appropriate to be assigned as T4 stage in American Joint Committee on Cancer classification. Spontaneous rupture of HCC is not a risk factor for survival. If resected eventually, srHCC may achieve comparable survival with nrHCC.


Taenia crassiceps: temperature, polycations, polyanions, and cysticercal endocytosis.
The effect of various temperatures, poly-L-lysine, and poly-L-glutamic acid on endocytosis of smooth micropinocytotic vesicles (pinosomes) in the tegument of the cysticercus of Taenia crassiceps has been investigated stereologically. The temperature regimes used were 0, 5, 10, 20, 30, and 40 C. Maximum volume, surface density, and number per unit volume were found at 40 C, and minimum surface-to-volume ratio and numbers at 10 C. At 10 C, mean pinosome volume and mean surface area per pinosome were maximal, but volume and surface density did not differ significantly from 40 C. It is proposed that this anomalous finding for 10 C incubations was due to this being a critical temperature at which a slower rate of pinosome formation was compensated for by the formation of larger individual pinosomes. Poly-L-lysine was shown to be a stimulant of pinosome formation, leading to a significant increase in numbers per unit volume. However, volume and surface density, surface-to-volume ratio, mean volume, and mean surface area per pinosome were not significantly different in poly-L-lysine-incubated samples, when compared to controls (fresh from the mouse) or incubations in medium only or samples returned to medium after poly-L-lysine incubation, the only exception being surface to volume ratio and mean volume of pinosomes in the 75-min incubation. These anomalous results were explained by a marked reduction in the form ellipse values, which indicated the production of more elliptical-shaped pinosomes under poly-L-lysine stimulation. Incubation in poly-L-glutamine acid did not have any significant effect at any incubation time.


On-Going Frontal Alpha Rhythms Are Dominant in Passive State and Desynchronize in Active State in Adult Gray Mouse Lemurs.
The gray mouse lemur (Microcebus murinus) is considered a useful primate model for translational research. In the framework of IMI PharmaCog project (Grant Agreement n°115009, www.pharmacog.org), we tested the hypothesis that spectral electroencephalographic (EEG) markers of motor and locomotor activity in gray mouse lemurs reflect typical movement-related desynchronization of alpha rhythms (about 8-12 Hz) in humans. To this aim, EEG (bipolar electrodes in frontal cortex) and electromyographic (EMG; bipolar electrodes sutured in neck muscles) data were recorded in 13 male adult (about 3 years) lemurs. Artifact-free EEG segments during active state (gross movements, exploratory movements or locomotor activity) and awake passive state (no sleep) were selected on the basis of instrumental measures of animal behavior, and were used as an input for EEG power density analysis. Results showed a clear peak of EEG power density at alpha range (7-9 Hz) during passive state. During active state, there was a reduction in alpha power density (8-12 Hz) and an increase of power density at slow frequencies (1-4 Hz). Relative EMG activity was related to EEG power density at 2-4 Hz (positive correlation) and at 8-12 Hz (negative correlation). These results suggest for the first time that the primate gray mouse lemurs and humans may share basic neurophysiologic mechanisms of synchronization of frontal alpha rhythms in awake passive state and their desynchronization during motor and locomotor activity. These EEG markers may be an ideal experimental model for translational basic (motor science) and applied (pharmacological and non-pharmacological interventions) research in Neurophysiology.


Immunological evidence for an ACTH-like antigen in the nervous ganglion of a protochordate Ciona intestinalis (Ascidiacea).
Nervous fibers and neurons containing an ACTH-like antigen could be demonstrated in the nervous ganglion of a Protochordate Ciona intestinalis, using indirect immunofluorescence and anti (17-39)-ACTH and anti (25-39)-ACTH antisera.


Investigation of the HIV-1 matrix interactome during virus replication.
Like all viruses, human immunodeficiency virus type 1 (HIV-1) requires host cellular factors for productive replication. Identification of these factors may lead to the development of novel cell-based inhibitors. A Strep-tag was inserted into the C-terminus of the matrix (MA) region of the HIV-1 gag gene. The resultant virus was replication competent and used to infect Jurkat T-cells. MA complexes were affinity purified with Strep-Tactin agarose. Protein quantification was performed using sequential window acquisition of all theoretical fragment ion spectra (SWATH) MS, data were log2 -transformed, and Student t-tests with Bonferroni correction used to determine statistical significance. Several candidate proteins were validated by immunoblot and investigated for their role in virus infection by siRNA knockdown assays. A total of 17 proteins were found to be statistically different between the infected versus uninfected and untagged control samples. X-ray repair cross-complementing protein 6 (Ku70), X-ray repair cross-complementing protein 5 (Ku80), and Y-box binding protein 1 (YB-1) were confirmed to interact with MA by immunoblot. Knockdown of two candidates, EZRIN and Y-box binding protein 1, enhanced HIV infection in vitro. The Strep-tag allowed for the capture of viral protein complexes in the context of virus replication. Several previously described factors were identified and at least two candidate proteins were found to play a role in HIV-1 infection. These data further increase our understanding of HIV host -cell interactions.