Nucleic acid metabolism in the nucleus and cytoplasm of angiosperm meiocytes.
Over the course of flowering plant evolution many important stages in the life cycle have become compressed into the period between the premeiotic mitosis and gamete maturation. For this reason the changes in structure or chemistry that take place at this time are not easily ascribed to particular 'developmental stages'. During the meiotic process itself striking changes occur in levels of cytoplasmic and nuclear RNA, while DNA is synthesized in both the chromosomes and organelles. Evidence is presented indicating that a large proportion of the m- and rRNA is purged from the meiocyte cytoplasm by a combination of normal degradation and the activity of specific hydrolases. DNA synthesis in both the plastids and mitochondria precedes the re-differentiation of these organelles, while that in the chromosomes occurs in two major peaks, as has previously been reported (Hotta & Stern, 1971). High-resolution autoradiography did not indicate either peak of DNA synthesis to be localized within the synaptonemal complex, although final confirmation of this conclusion must await further investigation. Examination of these events in organisms where meiosis is removed from other stages of the life cycle suggests that the degradation of RNA and the chromosomal DNA synthesis are features which regularly accompany the meiotic process. The significance of these findings is discussed in terms of the hypothesis that there is at least one stage in the life cycle of all plants at which a purge of information-carrying molecules takes place.


LncRNA ANRIL negatively regulated chitooligosaccharide-induced radiosensitivity in colon cancer cells by sponging miR-181a-5p.
The radiosensitivity of colon cancer cells can be regulated by noncoding RNAs. In this study, the lncRNA antisense non-coding RNA in the INK4 locus (ANRIL) was selected to analyze its regulatory role in chitooligosaccharides (COS)-related radiosensitivity in colon cancer cells. The ANRIL expression in colon cancer cell lines was examined using real-time quantitative polymerase chain reaction (RT-qPCR), based on which we selected the cell line that presented the highest expression of ANRIL for radiosensitivity research. The cells were exposed to X-rays (0 Gy, 2 Gy, 4 Gy, and 6 Gy) and evaluated for changes in ANRIL and miR-181a-5p expression using RT-qPCR. Cell viability was evaluated using the CCK8 method, while apoptosis was detected with flow cytometry assays. Dual luciferase assays validated the binding between ANRIL and miR-181a-5p. The cell survival rates after differential COS treatments (0 mg/mL, 1.0 mg/mL, 2.0 mg/mL, 3.0 mg/mL, 4.0 mg/mL, and 5.0 mg/mL) were rated using CCK8 assay. The cells with the strongest dosage of COS (5.0 mg/mL) were selected to further investigate the role of ANRIL/miR-181a-5p in modulating the radiosensitivity observed with CCK-8 and flow cytometry assays. The ANRIL was highly expressed in colon cancer cells lines, especially in the SW480 cell line. Irradiation significantly decreased cell viability and ANRIL expression in a dose-dependent manner. The overexpression of ANRIL reduced the cell apoptosis rate after irradiation. MiR-181a-5p directly bound to ANRIL and was upregulated by irradiation in a dose-dependent manner. The suppression of miR-181a-5p decreased cell apoptosis. The COS treatment notably downregulated cell survival and promoted apoptosis in cells exposed to irradiation. The overexpression of ANRIL partially reversed COS-induced apoptosis and the inhibition rate; the upregulation of miR-181a-5p could counteract the impact of ANRIL regulation in cells. The ANRIL negatively regulated radiosensitivity induced by COS in colon cancer cells by sponging miR-181a-5p.


Integrated imaging approach with MEG and DTI to detect mild traumatic brain injury in military and civilian patients.
Traumatic brain injury (TBI) is a leading cause of sustained impairment in military and civilian populations. However, mild (and some moderate) TBI can be difficult to diagnose due to lack of obvious external injuries and because the injuries are often not visible on conventional acute MRI or CT. Injured brain tissues in TBI patients generate pathological low-frequency neuronal magnetic signal (delta waves 1-4 Hz) that can be measured and localized by magnetoencephalography (MEG). We hypothesize that abnormal MEG delta waves originate from gray matter neurons that experience de-afferentation due to axonal injury to the underlying white matter fiber tracts, which is manifested on diffusion tensor imaging (DTI) as reduced fractional anisotropy. The present study used a neuroimaging approach integrating findings of magnetoencephalography (MEG) and diffusion tensor imaging (DTI), evaluating their utility in diagnosing mild TBI in 10 subjects in whom conventional CT and MRI showed no visible lesions in 9. The results show: (1) the integrated approach with MEG and DTI is more sensitive than conventional CT and MRI in detecting subtle neuronal injury in mild TBI; (2) MEG slow waves in mild TBI patients originate from cortical gray matter areas that experience de-afferentation due to axonal injuries in the white matter fibers with reduced fractional anisotropy; (3) findings from the integrated imaging approach are consistent with post-concussive symptoms; (4) in some cases, abnormal MEG delta waves were observed in subjects without obvious DTI abnormality, indicating that MEG may be more sensitive than DTI in diagnosing mild TBI.


Generalizability of heterogeneous treatment effects based on causal forests applied to two randomized clinical trials of intensive glycemic control.
Purpose Machine learning is an attractive tool for identifying heterogeneous treatment effects (HTE) of interventions but generalizability of machine learning derived HTE remains unclear. We examined generalizability of HTE detected using causal forests in two similarly designed randomized trials in type II diabetes patients. Methods We evaluated published HTE of intensive versus standard glycemic control on all-cause mortality from the Action to Control Cardiovascular Risk in Diabetes study (ACCORD) in a second trial, the Veterans Affairs Diabetes Trial (VADT). We then applied causal forests to VADT, ACCORD, and pooled data from both studies and compared variable importance and subgroup effects across samples. Results HTE in ACCORD did not replicate in similar subgroups in VADT, but variable importance was correlated between VADT and ACCORD (Kendall's tau-b 0.75). Applying causal forests to pooled individual-level data yielded seven subgroups with similar HTE across both studies, ranging from risk difference of all-cause mortality of -3.9% (95% CI -7.0, -0.8) to 4.7% (95% CI 1.8, 7.5). Conclusions Machine learning detection of HTE subgroups from randomized trials may not generalize across study samples even when variable importance is correlated. Pooling individual-level data may overcome differences in study populations and/or differences in interventions that limit HTE generalizability.


A new generation of monoclonal and recombinant antibodies against cell-adherent prostate specific membrane antigen for diagnostic and therapeutic targeting of prostate cancer.
Prostate-specific membrane antigen (PSMA) is an excellent candidate for targeting prostate cancer by virtue of its restricted expression on prostatic epithelial cells and its upregulation on prostatic carcinoma cells. PSMA is expressed on the cell surface displaying a specific three-dimensional structure. Therefore, only antibodies with a high cell binding activity will have an important impact on antibody-based imaging and therapy. Monoclonal antibodies (mAbs) and single chain antibody fragments (scFvs) were prepared from spleen cells of mice that had been immunized either with purified PSMA or a cell lysate of prostate cancer LNCaP cells containing native PSMA. mAbs and scFvs were screened for reactivity with purified PSMA and binding to PSMA-expressing LNCaP cells. From mice immunized with purified PSMA, we obtained three mAbs (K7, K12, D20) and four scFvs (G0, G1, G2, G4), which were highly reactive with the isolated antigen, but showed weak or no reaction with viable LNCaP cells. From mice immunized with unpurified LNCaP lysate, we obtained three mAbs (3/E7, 3/F11, 3/A12), and one scFv (A5), which were reactive with purified PSMA, also showing a strong and specific binding to viable LNCaP cells and PSMA-transfected cells. Our results suggest that only the mAbs and scFvs, that were elicited with unpurified LNCaP lysate and not with purified PSMA will be useful agents for diagnostic imaging and therapeutic applications of prostate cancer.


Proteomic analysis of the cerebrospinal fluid of Parkinson's disease patients pre- and post-deep brain stimulation.
To investigate alterations in protein expression associated with deep brain stimulation (DBS) in an attempt to elucidate possible mechanisms of action . Cerebrospinal fluid (CSF), obtained from six Parkinson's disease (PD) patients (pre- and post-DBS) and from six normal healthy controls, was studied for differentially expressed proteins. 2-D DIGE, in combination with MALDI-TOF and TOF-TOF Mass Spectrometry (MS) or ESI-MS, was used to identify the changed proteins (3 PD patients and 3 controls). Selected proteins were further studied using western blotting (6 PD patients and 6 controls). Twenty-one proteins were identified after MS and protein database interrogation. Apart from apolipoprotein A-I (apoA-I), the expression levels of complement C4 (C4), IgA, tetranectin, and extracellular superoxide dismutase (EC-SOD), detected by western blotting, correlated well with the 2-D DIGE results. In the follow-up period, the expression levels of C4, apoA-I and IgA were stable whereas EC-SOD and tetranectin were significantly elevated. In addition, when DBS was ceased in one patient due to a suicide attempt, the levels of EC-SOD and tetranectin significantly decreased. Our preliminary results suggest that variations in the expression levels of EC-SOD and tetranectin in CSF is related to DBS.


New insights into heparin binding to vitronectin: studies with monoclonal antibodies.
Vitronectin is a plasma glycoprotein that binds to a variety of ligands. There is considerable debate regarding the dependency of these binding interactions upon the conformational status of vitronectin, the role of multimerization and how the binding of different ligands can change vitronectin's conformational state. We have developed a method of capturing vitronectin directly from fresh plasma using solid-phase monoclonal antibodies. Various biotin-labelled secondary monoclonal antibodies were used to quantify the bound vitronectin and to measure its degree of denaturation. Using these tools we demonstrated that one monoclonal antibody partially denatured vitronectin without direct multimerization. Treatment of vitronectin in plasma with soluble heparin produced a similar degree of denaturation. These results led to a proposed adaptation of the unfolding/refolding pathways for chemically denatured vitronectin originally presented by Zhuang and co-workers in 1996 [Zhuang, Blackburn and Peterson (1996) J. Biol. Chem. 271, 14323-14332 and Zhuang, Li, Williams, Wagner, Seiffert and Peterson (1996) J. Biol. Chem. 271, 14333-14343]. The adapted version allows for the production of a more stable partially unfolded intermediate, resulting from the binding of particular ligands. We also demonstrated that the avidity of heparin binding to vitronectin is governed by both the conformational state of the monomer and multimerization of the molecule.


Independent living: methodology for measuring long-term outcomes.
A vexing issue in disability research today is the specification and measurement of long-term outcome measures relevant to the evaluation of independent living (IL) programs and services. The purpose of this article is to present 2 long-term outcome measures that are sensitive to both the philosophy of the IL movement and the policy making needs of funding sources. The method used in the study was to examine the specific outcomes of 111 spinal cord injured persons discharged from 10 medical rehabilitation centers. These outcomes were in turn ranked and weighted by an interagency panel consisting of providers, consumers, and funding agencies. The statistical results indicate a high level of agreement among panelists as to how each outcome should be ranked and weighted. The study concludes that the ranking and weighting methodology is a workable one that can take into account the value judgements implicit in the development of long-term outcome measures for IL and medical rehabilitation programs.


Protein mobilities and P-selectin storage in Weibel-Palade bodies.
Using fluorescence recovery after photobleaching (FRAP) we measured the mobilities of EGFP-tagged soluble secretory proteins in the endoplasmic reticulum (ER) and in individual Weibel-Palade bodies (WPBs) at early (immature) and late (mature) stages in their biogenesis. Membrane proteins (P-selectin, CD63, Rab27a) were also studied in individual WPBs. In the ER, soluble secretory proteins were mobile; however, following insertion into immature WPBs larger molecules (VWF, Proregion, tPA) and P-selectin became immobilised, whereas small proteins (ssEGFP, eotaxin-3) became less mobile. WPB maturation led to further decreases in mobility of small proteins and CD63. Acute alkalinisation of mature WPBs selectively increased the mobilities of small soluble proteins without affecting larger molecules and the membrane proteins. Disruption of the Proregion-VWF paracrystalline core by prolonged incubation with NH(4)Cl rendered P-selectin mobile while VWF remained immobile. FRAP of P-selectin mutants revealed that immobilisation most probably involves steric entrapment of the P-selectin extracellular domain by the Proregion-VWF paracrystal. Significantly, immobilisation contributed to the enrichment of P-selectin in WPBs; a mutation of P-selectin preventing immobilisation led to a failure of enrichment. Together these data shed new light on the transitions that occur for soluble and membrane proteins following their entry and storage into post-Golgi-regulated secretory organelles.


Double-minute chromatin bodies in HL-60 leukemia cells sensitive and resistant to differentiation inducing agents.
We studied the chromosome characteristics of HL-60 promyelocytic leukemia cells sensitive and resistant to differentiation inducing agents (DI). The karyotypic analysis of sensitive (HL-60 S) and resistant (HL-60 R) cells revealed the presence of identical chromosome abnormalities such as loss of chromosomes 5, 9, 10, 14, 16, 17 and X; and gain of chromosome 18. HL-60 S and HL-60 R cells also share five common markers. The difference between the two cell lines consisted essentially of the loss of an unidentifiable chromosome segment in the HL-60 R cell line. In addition, the two sublines showed marked differences in the content of double-minute chromatin bodies (DM), which were abundant in HL-60 S but rarely found in HL-60 R cells. Contrary to a previous report by others, there was no evidence of chromosome rearrangement of the DM as homogeneously stained regions (HSR) or abnormally banding regions (ABR) in the resistant HL-60 R cells. The presence of DM as an expression of gene amplification may be of relevance in the determination of susceptibility of HL-60 cells to DI.


Photochemically controlled synthesis of anisotropic Au nanostructures: platelet-like Au nanorods and six-star Au nanoparticles.
We report the shape-controlled synthesis of anisotropic Au nanostructures through TiO(2)-assisted photochemical reduction of HAuCl(4). By using this method, we have successfully synthesized the platelet-like Au nanorods and six-star Au nanoparticles. Importantly, the platelet Au nanorod exhibits the unique asymmetric five-twinned structure. The colloidal TiO(2) sols were used as both the photocatalyst to initiate the reaction and the stabilizing agent for the produced Au nanostructures. Significantly, in this photochemical process, the tunable irradiation intensity allows us to kinetically control the crystal evolution at various growth stages, leading to the shape difference of ultimate gold nanostructures. Our synthetic method shows a great potential as an alternative or supplement to the other wet chemical approaches for the shape-control of metallic nanostructures.


A new allele of engrailed, enNK14, causes supernumerary spermathecae in Drosophila melanogaster.
A spontaneous mutation, enNK14, was a new allele of engrailed (en) in Drosophila melanogaster. Females of enNK14 have three spermathecae, instead of two in wild type, under a wide range of developmental temperatures, while the males show no abnormal phenotype. Spermathecae of the mutant female can accept inseminated sperms, albeit with a delay of at least an hour until full acceptance compared with wild type. The time course of decrease in the number of stored sperms was thoroughly similar between the mutant and wild type. enNK14 females produced fewer progeny than wild type females despite storing a larger number of sperms. The delay of sperm entry and lower fecundity suggested some functional defects in secretory products of the spermathecae. In addition, some spermathecae in the mutant were accompanied by a mass of brown pigments in the adipose tissue surrounding the capsule. Six contiguous amino acids, Ser340-Ala345, were replaced by one Thr in enNK14. In another mutant, enspt, Ser325 was also shown to be substituted by a Cys. These amino acid changes were located within a serine-rich region, in which Ser325, Ser340 and Thr341 were suggested as targets of Protein Kinase C by an in silico analysis. The splicing pattern of en mRNA did not differ between enNK14 and wild type in embryo, larva, pupa or adult. Our results suggest that en plays an important role in determining the number of spermathecae as well as in sperm storage function in the Drosophila female.


Social Network Analysis of Factors Influencing Green Building Development in China.
Green buildings have been viewed as one of the most effective solutions to the negative environmental impacts of construction activities. For the sustainable development of the economy and the environment, many governments in the world have launched a variety of policies to encourage the development of green buildings. However, green targets achieved during the operational stage of green buildings are far below the expectations from the design stage. In addition, the development of green buildings is unevenly distributed in different cities. To help resolve these issues, this paper identifies 28 green building influencing factors from two perspectives, the life cycle and stakeholders. Then, a social network analysis is used to analyse their interactions and identify the critical factors. Our results show that government supervision, incremental cost, property management experience, and the awareness of environmental protection in green buildings are the critical influencing factors in promoting green building development. However, some factors related to contractors, designers and suppliers are not as important as perceived. Finally, some policy recommendations are proposed to promote green buildings in China.


Esmolol and landiolol, selective beta1-adrenoreceptor antagonists, provide neuroprotection against spinal cord ischemia and reperfusion in rats.
Paraplegia is a devastating and unpredictable complication occasionally resulting from surgery of the thoracic and thoracoabdominal aorta. Because ultrashort-acting selective beta(1)-adrenoreceptor antagonists provide neuroprotective effects after brain ischemia, we hypothesized that they would also ameliorate spinal cord injury after transient ischemia and reperfusion in rats. Male Sprague-Dawley rats were randomly assigned to one of the following 4 groups: saline (received IV infusion of 0.9% saline at a rate of 0.5 mL/h, n = 8), esmolol (esmolol 200 microg/kg/min, n = 8), landiolol (landiolol 50 microg/kg/min), or sham surgical (n = 6). Infusion of saline or drugs was initiated 30 minutes before spinal cord ischemia and continued for the subsequent 24-hour reperfusion. Spinal cord ischemia was induced by intraaortic balloon occlusion combined with proximal arterial hypotension for 10 minutes. The spinal cord was then reperfused for 24 hours. Ischemic injury was assessed in terms of the motor deficit index score of the hindlimb and the number of viable motor nerve cells in the anterior spinal cord at 24 hours after reperfusion. The motor deficit index scores were significantly lower in the esmolol and landiolol groups compared with the saline group (P < 0.05). Histopathologic evaluation of the spinal cord showed less damage in the esmolol and landiolol groups than in the saline group (P < 0.05). These data show that ultrashort-acting selective beta(1)-adrenoreceptor antagonists can reduce neurological injury in a rat model of spinal cord ischemia-reperfusion.


Stigmatization of Pregnant Individuals with Opioid Use Disorder.
Objective: The purpose of this study was to critically analyze the role of stigma in the care of pregnant and parenting individuals with opioid use disorder (OUD) through the theoretical lens of the Reproductive Justice (RJ) framework. Background: Overdose related maternal mortality, often involving opioids, is a national growing public health concern. OUD is a highly stigmatized condition that may negatively influence the well-being of pregnant/parenting individual's reproductive and human rights. Study Design: Secondary qualitative data analysis. Methods: A secondary data analysis was conducted using individual interviews (N = 23) from a larger study aimed at examining contextual factors surrounding pregnant/parenting individual's experiences with opioid use return to use and/or overdose. The RJ framework was used as a framework to examine the influence of OUD-related stigma and a person's right to bodily autonomy, their right to parent, and their right to parent the children they have in safe and sustainable environments. Results: The RJ framework supported the examination of factors that perpetuate stigma in this population. Individuals described stigmatizing experiences in the health care setting. Verbal and nonverbal interactions with health care providers and fear of child welfare involvement were counterproductive to recovery and potentially triggered OUD recurrence and/or overdose. Conclusions: Due to existing stigma, pregnant and parenting individuals with OUD often avoided health care and recovery support services; therefore, there should be the removal of barriers that prevent this population from accessing life-saving services. Future efforts should focus on health policy-related research to support structural changes within institutions.


Laryngeal foreign body mistreated as recurrent laryngitis and croup for one year.
Foreign body aspiration (FBA) is a common, dangerous problem among children of all ages. Laryngeal foreign bodies may present with less severe symptoms compared to lower respiratory tract foreign bodies, resulting in misdiagnosis, confusion and delay. We present the case of a 20-month-old girl with laryngeal foreign body unrecognized for one year. She was treated for recurrent laryngitis and infectious croup several times. A triangular-shaped, red plastic material with sharp edges was removed from the larynx by direct laryngoscopy. The clinical presentation and management are discussed.


Characterization of a β-1,3-glucanase active in the alkaline midgut of Spodoptera frugiperda larvae and its relation to β-glucan-binding proteins.
Spodoptera frugiperda β-1,3-glucanase (SLam) was purified from larval midgut. It has a molecular mass of 37.5 kDa, an alkaline optimum pH of 9.0, is active against β-1,3-glucan (laminarin), but cannot hydrolyze yeast β-1,3-1,6-glucan or other polysaccharides. The enzyme is an endoglucanase with low processivity (0.4), and is not inhibited by high concentrations of substrate. In contrast to other digestive β-1,3-glucanases from insects, SLam is unable to lyse Saccharomyces cerevisae cells. The cDNA encoding SLam was cloned and sequenced, showing that the protein belongs to glycosyl hydrolase family 16 as other insect glucanases and glucan-binding proteins. Multiple sequence alignment of β-1,3-glucanases and β-glucan-binding protein supports the assumption that the β-1,3-glucanase gene duplicated in the ancestor of mollusks and arthropods. One copy originated the derived β-1,3-glucanases by the loss of an extended N-terminal region and the β-glucan-binding proteins by the loss of the catalytic residues. SLam homology modeling suggests that E228 may affect the ionization of the catalytic residues, thus displacing the enzyme pH optimum. SLam antiserum reacts with a single protein in the insect midgut. Immunocytolocalization shows that the enzyme is present in secretory vesicles and glycocalyx from columnar cells.


"Ground truth" for selection on CCR5-Delta32.
A much-celebrated story of positive selection in the human genome is the 32-bp deletion in the chemokine receptor CCR5, a variant that confers resistance to AIDS. This variant was postulated to be a relatively recent response to plague or smallpox. New research shows that the frequency of CCR5-Delta32 in Bronze Age samples is similar to that seen today, pushing the observed age of the allele back to at least 3000 and possibly 5000 years ago. Interestingly, the extent of heterozygosity, differentiation across populations and linkage disequilibrium in the CCR5 region is not dissimilar to other human genomic regions, challenging claims of recent positive selection.


Finger ridge count and finger print pattern in maturity onset diabetes mellitus.
Total finger ridge count, absolute finger ridge count and finger print pattern were studied in 150 maturity onset diabetes mellitus patients and compared to 120 controls. Significant findings were: in males, with both hands combined and separately (i) an increase in radial and ulnar loops and arches (ii) A decrease in whorls. (iii) In females, an increase in ulnar loops and a decrease in whorls in the left hand was observed.


Management of urologic cancers during the pandemic and potential impact of treatment deferrals on outcomes.
The COVID-19 pandemic-related constraints on healthcare access have raised concerns about adverse outcomes from delayed treatment, including the risk of cancer progression and other complications. Further, concerns were raised about a potentially significant backlog of patients in need of cancer care due to the pandemic-related delays in healthcare, further exacerbating any potential adverse outcomes. Delayed access to surgery is particularly relevant to urologic oncology since one-third of new cancers in men (20% overall) arise from the genitourinary (GU) tract and surgery is often the primary treatment. Herein, we summarize the prepandemic literature on deferred surgery for GU cancers and risk of disease progression. The aforementioned data on delayed surgery were gathered in the context of systemic delays present in certain healthcare systems, or occasionally, due to planned deferral in suboptimal surgical candidates. These data provide indirect, but sufficient insight to develop triage schemas for prioritization of uro-oncological cases. Herein, we outline the extent to which the pandemic-related triage guidelines had influenced urologic practice in various regions. To study the adverse outcomes in the pandemic-era, a survey of urologic oncologists was conducted regarding modifications in their initial management of urologic cancers and any delay-related adverse outcomes. While the adverse effects directly from COVID-19 related delays will become apparent in the coming years, the results showing short-term outcomes are quite instructive. Since cancer care was assigned a higher priority at most centers, this strategy may have avoided significant delays in care and limited the anticipated negative impact of pandemic-related constraints.


Association Between Job Stress and Organizational Commitment in Three Types of Chinese University Teachers: Mediating Effects of Job Burnout and Job Satisfaction.
Utilizing the Job Demands-Resources (JD-R) model as the theoretical framework, this study examines the relationship between job stress, job burnout, job satisfaction, and organizational commitment among 1,906 university teachers in China, and investigates teachers' differences across groups. The result of SEM indicates that job burnout and job satisfaction could play mediating roles between job stress and organizational commitment. The result of multi-group analysis shows that for national university teachers, the positive effect of job stress on job burnout is the highest among three types of university teachers, the negative effect of job burnout on organizational commitment is lower compared with provincial university teachers and the negative effect of job burnout on job satisfaction is lower compared with provincial university teachers. Only for provincial university teachers, the job stress can significantly positively predict organizational commitment, and the independent mediating effect of job burnout is significantly greater than job satisfaction. The practical advice to enhance Chinese university teachers' organizational commitment was provided in the end.


Loss of Endothelial Barrier Function in the Inflammatory Setting: Indication for a Cytokine-Mediated Post-Transcriptional Mechanism by Virtue of Upregulation of miRNAs miR-29a-3p, miR-29b-3p, and miR-155-5p.
Dysfunction of the endothelial barrier plays a central role in the pathogenesis of both acute and chronic inflammatory processes such as sepsis or atherosclerosis. Due to attenuation of endothelial cell contacts, there is an increased transfer of blood proteins and fluid into the surrounding tissue, which relates to edema formation and distribution disorders. However, the mechanisms underlying these responses are not fully understood. In this study, we used human endothelial cells to mimic the loss of barrier function in an inflammatory milieu. We found that a weakened endothelial barrier after cytokine stimulation was accompanied by a significantly changed transcriptome. Apparent was a depletion of mRNAs encoding cell adhesion molecules. Furthermore, we found that cytokine treatment of endothelial cells induced upregulation of miR-29a-3p, miR-29b-3p, and miR-155-5p. miRNAs are known to negatively affect stability and translational efficiency of target mRNAs. Remarkably, miR-29a-3p, miR-29b-3p, and miR-155-5p have already been described to target the mRNAs of central tight and adherent junction proteins including F11 receptor, claudin 1, β-catenin, p120-catenin, and eplin. This taken together points to the existence of a posttranscriptional mechanism for expression inhibition of central adhesion proteins, which is triggered by inflammatory cytokines and mediated by miR-29a-3p, miR-29b-3p, and miR-155-5p.


Has tamoxifen had its day?
Tamoxifen is probably the most important drug in the history of the management of breast cancer and its development is a tribute to cross talk between laboratory scientists and clinical investigators. Its use as adjuvant therapy has led to a decrease of 20-30% in age-adjusted cause-specific mortality in the developed world and it is approved in the USA for the chemoprevention of breast cancer in high-risk women. The recent ATAC and IBIS trials have challenged the supremacy of tamoxifen. The present paper is a personal view of the implications for the future use of this drug in competition with the oral aromatase inhibitors. In the opinion of the author tamoxifen will probably remain the mainstay for adjuvant therapy of postmenopausal women with hormone-responsive disease, but maturation of the ATAC data may allow a choice in selected cases. Anastrozole looks like a competitor for the future but we may have to wait another 10 years to find out.


Travelling between Two Worlds: Complement as a Gatekeeper for an Expanded Host Range of Lyme Disease Spirochetes.
Evading innate immunity is a prerequisite for pathogenic microorganisms in order to survive in their respective hosts. Concerning Lyme disease spirochetes belonging to the Borrelia (B.) burgdorferi sensu lato group, a broad range of diverse vertebrates serve as reservoir or even as incidental hosts, including humans. The capability to infect multiple hosts implies that spirochetes have developed sophisticated means to counter the destructive effects of complement of humans and various animals. While the means by which spirochetes overcome the hosts immune defense are far from being completely understood, there is a growing body of evidence suggesting that binding of the key regulator of the alternative pathway, Factor H, plays a pivotal role for immune evasion and that Factor H is an important determinant of host specificity. This review covers (i) the contribution of complement in host-specificity and transmissibility of Lyme disease spirochetes; (ii) the involvement of borrelial-derived determinants to host specificity; (iii) the interplay of human and animal Factor H with complement-acquiring surface proteins of diverse borrelial species; and (iv) the potential role of additional animal complement proteins in the immune evasion of spirochetes.


Optimizing Pelvic X-Ray indication in blunt trauma patients using clinical criteria.
to identify a subgroup of blunt trauma patients with very low chance of sustaining pelvic fractures based on clinical criteria. retrospective analysis of the trauma registry data, collected in a period of 24 months. We selected adult blunt trauma patients who had a PXR on admission. The frequency of pelvic fractures was calculated for the following groups: Normal neurological examination at admission (NNE), hemodynamical stability (HS), normal pelvic examination at admission (NPE), less than 60 years old (ID<60) and absence of distracting injuries (ADI). Logistic regression analysis was carried out in order to create a probability model of negative PXR. an abnormal PXR was identified in 101 (3.3%) out of the 3,055 patients who had undergone a PXR at admission. Out of these, 1,863 sustained a NNE, with 38 positive CXRs (2.0%) in this group. Considering only the 1,535 patients with NNE and HS, we found 28 positive PXRs (1.8%). Out of these, 1,506 have NPE, with 21 abnormal PXRs (1.4%). Of these, 1,202 were younger than 60 y, with 11 positive PXRs (0.9%). By adding all these criteria to the ADI, we found 2 abnormal PXRs in 502 (0.4%) cases. The probability model including all these variables had a 0,89 area under the ROC curve. by adding clinical criteria, it is possible to identify a group of trauma patients with very low chance of sustaining pelvic fractures. The necessity of PXR in these patients needs to be reassessed.


Validity of an Interactive Functional Reach Test.
Videogaming platforms such as the Microsoft (Redmond, WA) Kinect(®) are increasingly being used in rehabilitation to improve balance performance and mobility. These gaming platforms do not have built-in clinical measures that offer clinically meaningful data. We have now developed software that will enable the Kinect sensor to assess a patient's balance using an interactive functional reach test (I-FRT). The aim of the study was to test the concurrent validity of the I-FRT and to establish the feasibility of implementing the I-FRT in a clinical setting. The concurrent validity of the I-FRT was tested among 20 healthy adults (mean age, 25.8±3.4 years; 14 women). The Functional Reach Test (FRT) was measured simultaneously by both the Kinect sensor using the I-FRT software and the Optotrak Certus(®) 3D motion-capture system (Northern Digital Inc., Waterloo, ON, Canada). The feasibility of implementing the I-FRT in a clinical setting was assessed by performing the I-FRT in 10 participants with mild balance impairments recruited from the outpatient physical therapy clinic (mean age, 55.8±13.5 years; four women) and obtaining their feedback using a NASA Task Load Index (NASA-TLX) questionnaire. There was moderate to good agreement between FRT measures made by the two measurement systems. The greatest agreement between the two measurement system was found with the Kinect sensor placed at a distance of 2.5 m [intraclass correlation coefficient (2,k)=0.786; P<0.001] from the participant. Participants with mild balance impairments whose balance was assessed using the I-FRT software scored their experience favorably by assigning lower scores for the Frustration, Mental Demand, and Temporal Demand subscales on the NASA/TLX questionnaire. FRT measures made using the Kinect sensor I-FRT software provides a valid clinical measure that can be used with the gaming platforms.


The effect of aleglitazar on the pharmacokinetics and pharmacodynamics of S- and R-warfarin in healthy male subjects.
: Aleglitazar acts through balanced activation of peroxisome proliferator-activated receptors α and γ; warfarin is a commonly prescribed anticoagulant. Given the extent of cardiovascular disease in patients with type 2 diabetes, cotreatment with aleglitazar and warfarin is likely in this population. This open-label, randomized, 2-period, crossover study in 12 healthy male subjects investigated the potential for drug-drug interactions between warfarin and aleglitazar (final data drawn from 11 white subjects). The primary objective was to investigate the effect of aleglitazar on the pharmacokinetic properties of S-warfarin and on the pharmacodynamics of the racemic mixture; the secondary objectives included the effect of aleglitazar on R-warfarin pharmacokinetics and of racemic warfarin on aleglitazar pharmacokinetics. Subjects were randomized to single-dose warfarin on day 1 or aleglitazar once daily (12 days) plus single-dose warfarin on day 6 followed by a 14-day washout period, then crossover. Coadministration of aleglitazar reduced S- and R-warfarin exposure (AUC0-∞) by 18% and 13%, respectively, but did not change its pharmacodynamic effects (prothrombin time and factor VII activity). After warfarin dosing, aleglitazar trough concentrations remained within the same range. These findings indicate that coadministration of aleglitazar and warfarin is unlikely to affect the efficacy or safety of either agent.


Phthalocyanines and Porphyrins/Polyaniline Composites (PANI/CuPctBu and PANI/TPPH2) as Sensing Materials for Ammonia Detection.
We combined a conducting polymer, polyaniline (PANI), with an organic semiconducting macrocyclic (MCs) material. The macrocycles are the phthalocyanines and porphyrins used to tune the electrical properties of the PANI, which benefits from their ability to enhance sensor response. For this, we proceeded by a simple ultrasonically assisted reaction involving the two components, i.e., the PANI matrix and the MCs, to achieve the synthesis of the composite nanostructure PANI/MCs. The composite nanostructure has been characterized and deposited on interdigitated electrodes (IDEs) to construct resistive sensor devices. The isolated nanostructured composites present good electrical properties dominated by PANI electronic conductivity, and the characterization reveals that both components are present in the nanostructure. The experimental results obtained under gas exposures show that the composite nanostructures can be used as a sensing material with enhanced sensing properties. The sensing performance under different conditions, such as ambient humidity, and the sensor's operating temperature are also investigated. Sensing behavior in deficient humidity levels and their response at different temperatures revealed unusual behaviors that help to understand the sensing mechanism. Gas sensors based on PANI/MCs demonstrate significant stability over time, but this stability is highly reduced after experiments in lower humidity conditions and at high temperatures.


Nonaccidental trauma presenting with respiratory distress and pleural effusion.
Nonaccidental trauma (NAT) is common and presents with varied symptoms. Pleural effusion as a complication of physical abuse has not been described in the past. We report the case of a 10-week-old infant who presented with multiple nonspecific complaints that included respiratory distress, refusal to feed, constipation, and lethargy. Sepsis was the working diagnosis on admission, but a massive pleural effusion and rib fractures seen on chest imaging ultimately led to the diagnosis of nonaccidental trauma. This interesting case highlights the importance of maintaining a high index of suspicion for abuse irrespective of atypical presenting signs.


[Goiter prevalence and urinary excretion of iodine in a sample of school age children in the city of Rome].
The aim of the study was the assessment of the urinary iodine excretion and the evaluation of thyroid volume compared with clinical examination in 1040 schoolchildren (6-14 years old), living in Rome. Mean urinary iodine excretion was 98.52 +/- 49.81 micrograms/l (median 92 micrograms/l). Thyroid enlargement, as assessed by palpation, was found to be grade 1A in 35.4% of the children, grade 1B in 9.6% and grade 2 in 0.2%. Thyroid volume, determined by ultrasound, increased with age, was significantly correlated with body surface area and was significantly higher in females, as compared to males, in the 11 and 12 years old group. Eleven children (1.9%) were negative at palpation (grade 0) but showed thyroid enlargement by ultrasound. The prevalence of goiter determined by ultrasound resulted to be 4.7%.


Contrast-enhanced MR imaging performed after successful lumbar disk surgery: prospective study.
A prospective study was undertaken to establish the normal spectrum and timing of gadolinium-enhanced magnetic resonance (MR) imaging findings in 15 patients who had resolution of symptoms after successful lumbar disk surgery. Enhancement of the facet joints (in 88% of disk levels) and paraspinal muscles (100%) decreased gradually after surgery. Enhancement of the decompressed nerve root tracked proximally toward the conus medullaris in 62% at 3 weeks and was absent in all by 6 months postoperatively. Areas of intermediate signal intensity with peripheral enhancement and mass effect were seen on T1-weighted images at the site of the original disk herniation in 38% at 3 weeks and 12% at 3 months, despite complete relief of leg pain. These results reveal that even in successfully treated (asymptomatic) patients, residual mass effect on the neural elements may frequently simulate a recurrent or residual disk fragment. There is an orderly progression of imaging changes during the first 6 months after lumbar surgery that limits the interpretation of MR examinations during that period.


Metastasizing Maxillary Ameloblastoma: Report of a Case with Molecular Characterization.
Ameloblastoma is a benign odontogenic tumour that may exhibit aggressive biological behaviour with local recurrence and metastasis following initial surgical resection. Surgery is the most acceptable modality of treatment, even if a biological approach is currently on study. We report a case of maxillary ameloblastoma with development of neck and brain metastases after repeated local recurrences. Molecular analysis was performed with the aim to better characterize this neoplasm and its peculiar behaviour. We investigated the status of tumour protein p53 (TP53), epidermal growth factor receptor (EGFR), B-Raf proto-oncogene (BRAF) and human epidermal growth factor receptor 2 (HER2) genes with immunohistochemical, fluorescent in situ hybridization and/or direct sequencing in order to clarify their possible role in the development of this neoplasm and the possibility of a targeted treatment. The histological appearance of the tumour was the same in the primary lesion, in the recurrence and in the metastases. EGFR positivity was present in the recurrence and the brain metastasis, while HER2 was negative in all samples tested. Fluorescent in situ hybridization analysis for EGFR showed disomy of neoplastic cells. Direct DNA sequencing of TP53 gene exons 5 - 9 was carried out in tumour samples from the infratemporal recurrence and brain metastasis, with no mutational alteration detected. Similarly, sequencing analysis of BRAF exon 15 (V600) and EGFR gene showed wild type results in all samples tested. Further studies are needed to identify molecular pathways that may provide an opportunity of alternative treatments and/or new potential predictive markers of local and distant spread of this rare tumour.


Motivation and readiness for therapeutic community treatment among adolescents and adult substance abusers.
A growing body of research has demonstrated the importance of motivation and readiness among drug abusers in seeking, complying with, and remaining in treatment. To date, however, there is little research on these factors among adolescent substance abusers. The present study reports findings from a large scale investigation of motivation and readiness differences across adolescent (range = 14-18 years, n > 1000) and adult (range = 19-26 + years, n > 1400) admissions to residential therapeutic communities (TCs). Data were collected with an instrument assessing circumstances, motivation, readiness, and suitability for TC treatment (i.e., CMRS). Results showed that: (1) there is a significant positive linear relationship between CMRS scores and age; (2) the CMRS scores were the largest and most consistent predictors of short term retention across all age groups. Although confined to TC samples, the present findings support clinical observations that adolescent drug abusers are less motivated to change or ready for treatment than adults; and they confirm the importance of motivational and readiness factors in the treatment process, regardless of age.


Increased prebeta-HDL levels, cholesterol efflux, and LCAT-mediated esterification in mice expressing the human cholesteryl ester transfer protein (CETP) and human apolipoprotein A-I (apoA-I) transgenes.
The effects of cholesteryl ester transfer protein (CETP) on the distribution of apolipoprotein (apo) A-I between high density lipoprotein (HDL) subspecies and its impact on efflux and esterification of cell-derived cholesterol was studied in transgenic mice expressing either the human apoA-I (HuAITg) or both the human apoA-I and CETP (HuAICETPTg) transgenes. The simultaneous expression of the human CETP and apoA-I transgenes induced a 2-fold increase in the proportion of human apoA-I in the prebeta-HDL fraction and 1.4- and 2.2-fold increases in the HDL3a and HDL3c fractions, respectively, at the expense of the larger HDL2b population. HuAICETPTg mouse plasma has a greater ability to cause efflux of cholesterol from 3H-labeled fibroblasts than plasma from HuAITg mice. Furthermore, the LCAT-mediated esterification of cell-derived cholesterol is increased 1.7-fold in mice expressing the human apoA-I and CETP transgenes compared to HuAITg mouse plasma. LCAT activity (measured with an exogenous substrate) was increased 1.4-fold and LCAT mRNA levels were increased 1.3-fold as a result of CETP expression. Taken together, these data indicate that in vivo, the expression of CETP resulted in an increase in the proportion of apoA-I in the prebeta-HDL fraction and a stimulation of the efflux and esterification of cell-derived cholesterol.


Transcriptomics and sequencing analysis of gene expression profiling for major depressive disorder.
Major depressive disorder (MDD) is a common psychiatric disorder characterized by constant sadness and a lack of interest in work and social interactions. Maintaining the transcriptome levels via the controlled regulation of mRNA processing and transport is essential to alleviating MDD. Various molecular phenotypes such as aberrant RNA splicing and stability are identified as critical determinants of MDD. This study aims to compare the mRNA expression profiles between major depressive disorder non-suicide (MDD), major depressive disorder suicide (MDD-S), and control groups using RNA-Seq. A transcriptomics and sequencing analysis of gene expression profiling was conducted in 9 patients with MDD, 10 patients with MDD-S, and 10 control patients. A comparison of the sample groups revealed that the PRKACB gene was upregulated in patients with MDD. At the same time, GRM3, DLGAP1, and GRIA2 were downregulated in these patients-these genes are majorly involved in the glutamatergic pathway. Five genes (GRIA1, CAMK2D, PPP3CA, MAPK10, and PPP2R2A) of the dopaminergic pathway were downregulated in patients with the MDD-S condition when compared with the MDD and control groups. Cholinergic synapses were altered in patients with MDD when compared to the control group due to the presence of dysregulated genes (KCNQ5, PLCB4, ADCY9, CAMK2D, PIK3CA, and GNG2). The results provide a new understanding of the etiology of depression in humans and identify probable depression-associated biomarkers.


Effect of Alkaline Peroxides on the Surface of Cobalt Chrome Alloy: An In Vitro Study.
Removable denture hygiene care is very important for the longevity of the rehabilitation treatment; however, it is necessary to analyze the effects that denture cleansers can cause on the surfaces of prostheses. Thus, this study evaluated the effect of alkaline peroxide-effervescent tablets on the surface of cobalt-chromium alloys (Co-Cr) used in removable partial dentures. Circular metallic specimens (12 × 3 mm) were fabricated and were immersed (n = 16) in: control, Polident 3 Minute (P3M), Steradent (S), Efferdent (E), Polident for Partials (PFP), and Corega Tabs (CT). The surface roughness (μm) (n = 10) was measured before and after periods of cleanser immersion corresponding to 0.5, 1, 2, 3, 4, and 5 years. Ion release was analyzed (n = 5) for Co, Cr, and molybdenum (Mo). Scanning electron microscopy (SEM) analysis and an Energy-dispersive X-ray spectroscopy (EDS) were conducted in one specimen. The surface roughness data were statistically analyzed (α = 0.05) with the Kruskal-Wallis test to compare the solutions, and the Friedman test compared the immersion durations. Ion release analysis was performed using 2-way ANOVA and Tukey's test. There was no significant surface roughness difference when comparing the solutions (p > 0.05) and the immersion durations (p = 0.137). Regarding ion release (μg/L), CT, E, and control produced a greater release of Co ions than S (p < 0.05). CT produced a greater release of Cr ions than control, S, and P3M (p < 0.05). Finally, E caused the greatest release of Mo ions (p < 0.05). SEM confirmed that the solutions did not damage the surfaces and EDS confirmed that there were no signs of oxidation. The various solutions tested did not have any deleterious effects on the Co-Cr alloy surface. Steradent, however, presented the smallest ionic release.


Curdlan, zymosan and a yeast-derived β-glucan reshape tumor-associated macrophages into producers of inflammatory chemo-attractants.
Anti-cancer T-cell responses are often halted due to the immune-suppressive micro-environment, in part related to tumor-associated macrophages. In the current study, we assessed indigestible β-glucans (oatβG, curdlan, grifolan, schizophyllan, lentinan, yeast whole glucan particles (yWGP), zymosan and two additional yeast-derived β-glucans a and b) for their physicochemical properties as well as their effects on the plasticity of human monocyte-derived macrophages that were polarized with IL-4 to immune-suppressive macrophages. Beta-glucans were LPS/LTA free, and tested for solubility, molecular masses, protein and monosaccharide contents. Curdlan, yeast-b and zymosan re-polarized M(IL-4) macrophages towards an M1-like phenotype, in particular showing enhanced gene expression of CCR7, ICAM1 and CD80, and secretion of TNF-α and IL-6. Notably, differential gene expression, pathway analysis as well as protein expressions demonstrated that M(IL-4) macrophages treated with curdlan, yeast-b or zymosan demonstrated enhanced production of chemo-attractants, such as CCL3, CCL4, and CXCL8, which contribute to recruitment of monocytes and neutrophils. The secretion of chemo-attractants was confirmed when using patient-derived melanoma-infiltrating immune cells. Taken together, the bacterial-derived curdlan as well as the yeast-derived β-glucans yeast-b and zymosan have the unique ability to preferentially skew macrophages towards a chemo-attractant-producing phenotype that may aid in anti-cancer immune responses.


Eukaryotic translation initiation factor 4E (eIF4E) binding site and the middle one-third of eIF4GI constitute the core domain for cap-dependent translation, and the C-terminal one-third functions as a modulatory region.
The mammalian eukaryotic initiation factor 4GI (eIF4GI) may be divided into three roughly equal regions; an amino-terminal one-third (amino acids [aa] 1 to 634), which contains the poly(A) binding protein (PABP) and eIF4E binding sites; a middle third (aa 635 to 1039), which binds eIF4A and eIF3; and a carboxy-terminal third (aa 1040 to 1560), which harbors a second eIF4A binding site and a docking sequence for the Ser/Thr kinase Mnk1. Previous reports demonstrated that the middle one-third of eIF4GI is sufficient for cap-independent translation. To delineate the eIF4GI core sequence required for cap-dependent translation, various truncated versions of eIF4GI were examined in an in vitro ribosome binding assay with beta-globin mRNA. A sequence of 540 aa encompassing aa 550 to 1090, which contains the eIF4E binding site and the middle region of eIF4GI, is the minimal sequence required for cap-dependent translation. In agreement with this, a point mutation in eIF4GI which abolished eIF4A binding in the middle region completely inhibited ribosomal binding. However, the eIF4GI C-terminal third region, which does not have a counterpart in yeast, modulates the activity of the core sequence. When the eIF4A binding site in the C-terminal region of eIF4GI was mutated, ribosome binding was decreased three- to fourfold. These data indicate that the interaction of eIF4A with the middle region of eIF4GI is necessary for translation, whereas the interaction of eIF4A with the C-terminal region plays a modulatory role.


Focal cerebral ischemia activates neurovascular restorative dynamics in mouse brain.
Cerebral ischemia triggers regeneration of neural stem/progenitor cells (NSCs/NPCs), which are associated with neovascularization and white matter repair in the brain. This study analyzed the dynamics of neurogenesis, neovascularization, and white matter injury/repair after middle cerebral artery occlusion (MCAO) and elucidated their temporal association. Mice were subjected to MCAO for 60 minutes and sacrificed up to 28 days after reperfusion. Neurogenesis and angiogenesis, as measured by double staining of 5-bromo-2-deoxyuridine (BrdU) with DCX or tomato lectin, respectively, were substantially activated soon after ischemia and persisted for 4 weeks. Despite the moderate recovery of functional vessels in infarct margin from 7 days post-ischemia, a significant decrease in vascular density remained over time. Clusters of immature neurons localized proximal to angiogenic blood vessels beginning 14 days after ischemia, suggesting interplay between neurogenesis and revascularization. Progenitors of oligodendrocytes (NG2+) constitutively presented in the normal brain and proliferated soon after ischemia. However, axon damage and the loss of white matter integrity after ischemic stroke were almost irreversible, as revealed by sustained decreases of myelin basic protein (MBP) and neurofilament-200 expression.


Ultra-high contrast frontend for high peak power fs-lasers at 1030 nm.
We present the results from a new frontend within a double-chirped pulse amplification architecture (DCPA) utilizing crossed-polarized wave generation (XPW) for generating ultra-high contrast, 150 μJ-level, femtosecond seed pulses at 1030 nm. These pulses are used in the high energy class diode-pumped laser system Polaris at the Helmholtz Institute in Jena. Within this frontend, laser pulses from a 75 MHz oscillator-pulse train are extracted at a repetition rate of 1 Hz, temporally stretched, amplified and then recompressed reaching a pulse energy of 2 mJ, a bandwidth of 12 nm and 112 fs pulse duration at a center wavelength of 1030 nm. These pulses are temporally filtered via XPW in a holographic-cut BaF₂ crystal, resulting in 150 μJ pulse energy with an efficiency of 13 %. Due to this non-linear filtering, the relative intensity of the amplified spontaneous emission preceding the main pulse is suppressed to 2×10⁻¹³. This is, to the best of our knowledge, the lowest value achieved in a high peak power laser system operating at 1030 nm center wavelength.


Hemispheric asymmetry in stress processing in rat prefrontal cortex and the role of mesocortical dopamine.
The prefrontal cortex (PFC) is known to play an important role not only in the regulation of emotion, but in the integration of affective states with appropriate modulation of autonomic and neuroendocrine stress regulatory systems. The present review highlights findings in the rat which helps to elucidate the complex nature of prefrontal involvement in emotion and stress regulation. The medial PFC is particularly important in this regard and while dorsomedial regions appear to play a suppressive role in such regulation, the ventromedial (particularly infralimbic) region appears to activate behavioral, neuroendocrine and sympathetic autonomic systems in response to stressful situations. This may be especially true of spontaneous stress-related behavior or physiological responses to relatively acute stressors. The role of the medial PFC is somewhat more complex in conditions involving learned adjustments to stressful situations, such as the extinction of conditioned fear responses, but it is clear that the medial PFC is important in incorporating stressful experience for future adaptive behavior. It is also suggested that mesocortical dopamine plays an important adaptive role in this region by preventing excessive behavioral and physiological stress reactivity. The rat brain shows substantial hemispheric specialization in many respects, and while the right PFC is normally dominant in the activation of stress-related systems, the left may play a role in countering this activation through processes of interhemispheric inhibition. This proposed basic template for the lateralization of stress regulatory systems is suggested to be associated with efficient stress and emotional self-regulation, and also to be shaped by both early postnatal experience and gender differences.


Randomization analysis of dental data characterized by skew and variance heterogeneity.
Many distributions of dental variables exhibit positive skew and variance heterogeneity. Although the use of parametric tests on data with these disturbances usually does not pose a problem with respect to alpha error level, transformation of data sometimes increases power. Occasionally one or both of these expectations do not hold and it becomes problematic as to whether data should be transformed and which analyses should be considered valid. Simulated dental trials were conducted on independent samples (n = 25 per group) drawn from exponential distributions with the same or different population means, t-tests on raw, log, and square root transformed data; the Mann-Whitney-Wilcoxon ranking test; and a randomization test, all exhibited satisfactory levels of alpha error but the randomization test exhibited the greatest power followed closely by the t-test on raw scores. The present results demonstrate that the log transformation does not increase power for all forms of positively skewed data. Randomization tests, not being subject to normality and variance homogeneity assumptions, may yield greater confidence in validity in these cases. Some implications for analytical strategy are discussed.


Litomosoides silvai (Nematoda: Onchocercidae) parasitizing Akodon montensis (Rodentia: Cricetidae) in the southern region of Brazil.
In the present study, Litomosoides silvai parasitizing Akodon montensis in the southern region of Brazil is reported for the first time. New morphological information is provided for some structures of this nematode species, such as a flattened cephalic extremity, presence of two dorsal cephalic papillae, female tail with a constriction at its tip, "s" shaped vagina, spicules characteristic of the carinii species group and microfilaria tail constricted at the tip. This nematode was found parasitizing the thoracic cavity with a prevalence of 10% (2/20), mean intensity of 4 (6/2), mean abundance of 0.4 (8/20) and range of infection of 2-6 specimens per host, in southern Brazil. This occurrence of L. silvai in A. montensis is a new geographical record for southern Brazil, in the Upper Paraná Atlantic Forest ecoregion of the northwestern region of Rio Grande do Sul, which is part of the Atlantic Forest biome.


Multifaceted behavior of PEST sequence enriched nuclear proteins in cancer biology and role in gene therapy.
The amino acid sequence enriched with proline (P), glutamic acid (E), serine (S), and threonine (T) (PEST) is a signal-transducing agent providing unique features to its substrate nuclear proteins (PEST-NPs). The PEST motif is responsible for particular posttranslational modifications (PTMs). These PTMs impart distinct properties to PEST-NPs that are responsible for their activation/inhibition, intracellular localization, and stability/degradation. PEST-NPs participate in cancer metabolism, immunity, and protein transcription as oncogenes or as tumor suppressors. Gene-based therapeutics are getting the attention of researchers because of their cell specificity. PEST-NPs are good targets to explore as cancer therapeutics. Insights into PTMs of PEST-NPs demonstrate that these proteins not only interact with each other but also recruit other proteins to/from their active site to promote/inhibit tumors. Thus, the role of PEST-NPs in cancer biology is multivariate. It is hard to obtain therapeutic objectives with single gene therapy. An especially designed combination gene therapy might be a promising strategy in cancer treatment. This review highlights the multifaceted behavior of PEST-NPs in cancer biology. We have summarized a number of studies to address the influence of structure and PEST-mediated PTMs on activation, localization, stability, and protein-protein interactions of PEST-NPs. We also recommend researchers to adopt a pragmatic approach in gene-based cancer therapy.


The Hand Eczema Trial (HET): Design of a randomised clinical trial of the effect of classification and individual counselling versus no intervention among health-care workers with hand eczema.
Hand eczema is the most frequently recognized occupational disease in Denmark with an incidence of approximately 0.32 per 1000 person-years. Consequences of hand eczema include chronic severe eczema, prolonged sick leave, unemployment, and impaired quality of life. New preventive strategies are needed to reduce occupational hand eczema. We describe the design of a randomised clinical trial to investigate the effects of classification of hand eczema plus individual counselling versus no intervention. The trial includes health-care workers with hand eczema identified from a self-administered questionnaire delivered to 3181 health-care workers in three Danish hospitals. The questionnaire identifies the prevalence of hand eczema, knowledge of skin-protection, and exposures that can lead to hand eczema. At entry, all participants are assessed regarding: disease severity (Hand Eczema Severity Index); self-evaluated disease severity; number of eruptions; quality of life; skin protective behaviour, and knowledge of skin protection. The patients are centrally randomised to intervention versus no intervention 1:1 stratified for hospital, profession, and severity score. The experimental group undergoes patch and prick testing; classification of the hand eczema; demonstration of hand washing and appliance of emollients; individual counselling, and a skin-care programme. The control group receives no intervention. All participants are reassessed after six months. The primary outcome is observer-blinded assessment of disease severity and the secondary outcomes are unblinded assessments of disease severity; number of eruptions; knowledge of skin protection; skin-protective behaviour, and quality of life. The trial is registered in ClinicalTrials.Gov, NCT01012453.


[Inhaled nitric oxide (NO) for vasoreactivity testing in pulmonary hypertension].
Thirty patients (24F, 6M, aged 40.5 +/- 15.4 years) with chronic precapillary pulmonary hypertension (primary pulmonary hypertension--PPH)--20 pts, associated with connective tissue disease--6 pts, due to thromboembolism--2 pts and Eisenmenger syndrome--2 pts) were submitted to 5 min inhalation of 10 and 20 ppm of nitric oxide (NO) during right heart catheterization. For the whole group this test resulted in minimal changes in directly assessed hemodynamics. However, individual reactions differed markedly between patients. Assuming three levels of criteria of preserved reactivity (liberal, standard, demanding) 11/30 (37%) patients had fall in PAP and PVR > or = 20% from baseline, 4/30 (13%) decreased PAP and PVR by > or = 20% and 30% respectively and 2 (7%) patients normalized PAP and decreased PVR > or = 50% from baseline, respectively. Interestingly, even liberal criteria were fulfilled positive test was observed exclusively in patients with primary PH. In one patient with connective tissue disease and systemic hypertension acute pulmonary congestion appeared during NO testing, with concomitant increase in systemic arterial pressure and pulmonary wedge pressure. No other complications were noted and no rebound PH after discontinuation of NO was observed in any of the patients. 1. Acute inhaled nitric oxide test appears to be safe in patients with PH and no coexisting left heart dysfunction. 2. It allows for stratification of patients according to the degree of preserved pulmonary vascular reactivity. However, criteria which would permit to select optimal treatment based on the results of NO test require further studies.


Multi-Institutional Prospective Study of Reirradiation with Proton Beam Radiotherapy for Locoregionally Recurrent Non-Small Cell Lung Cancer.
The management of recurrent NSCLC in the setting of prior radiation therapy is challenging. Proton radiotherapy (PRT) is ideally suited to minimize toxicity to previously irradiated organs. We report the safety/feasibility of PRT for NSCLC reirradiation in a prospective multi-institutional study. Between October 2010 and December 2015, 57 patients with recurrent NSCLC in or near their prior radiation field were treated at three proton centers. Patients were classified by tumor volume, location, and clinical characteristics. Toxicities were scored using the National Cancer Institute Common Terminology Criteria for Adverse Events, version 4.0. Survival outcomes were estimated by using Kaplan-Meier analysis. Fifty-two patients (93%) completed the reirradiation course. Their median age was 65 years (41-86). Patients with high tumor volume (clinical target volume-to-internal target volume ratio ≥250 cm3) were closed to enrollment owing to infeasibility in August 2012. Concurrent systemic therapy was delivered to 67% of patients. Fourteen patients (25%) had evidence of local (n = 9) or regional (n = 5) recurrence. Distant metastases after reirradiation developed in six patients (11%). The 1-year rates of overall and progression-free survival were 59% and 58%, respectively. In total, grade 3 or higher acute and/or late toxicity developed in 24 patients (42%), acute toxicity developed in 22 (39%), and late toxicity developed in seven (12%). Six grade 5 toxicities were observed. Increased overlap with the central airway region, mean esophagus and heart doses, and concurrent chemotherapy were associated with significantly higher rates of grade 3 or higher toxicity. Decreased overall survival was seen with increased mean esophagus dose (p = 0.007). In this prospective study, PRT for recurrent NSCLC is feasible but can be associated with significant toxicity. Providers should remain cautious in reirradiating NSCLC, paying close consideration to tumor volume, location, and relevant dosimetric parameters. Further research is needed for optimal patient selection to improve overall outcomes.


Lop-sided Raman-Nath diffraction in PT-antisymmetric atomic lattices.
Two-dimensional (2D) optical lattices of driven cold atoms can provide a useful platform to construct 2D electromagnetically induced grating (EIG) with parity-time (PT) antisymmetry. This atomic grating is achieved by the spatial modulations of the atomic density and frequency detunings in the four-level double-Λ atomic system. Gain-assisted PT antisymmetry allows us to realize lop-sided Raman-Nath diffraction with high diffraction efficiency at the exception point. It is shown that the nontrivial phenomenon originates from non-Hermitian degeneracy of PT antisymmetry. Our scheme may provide the possibility for active all-optical control and conversion of the spatial beam in optics.


Renal Neoplasia in Tuberous Sclerosis: A Study of 41 Patients.
To study the clinical features and identify unique renal neoplasia subtypes and their prognostic implications in individuals with tuberous sclerosis complex (TSC). The Mayo Clinic nephrectomy registry included 37 patients with TSC diagnosed between 1970 and 2018. Four additional patients were identified from the pathology consultation and autopsy files. All available renal tumors were further characterized using immunohistochemistry and fluorescence in situ hybridization. Clinicopathologic features and follow-up were obtained from the medical record. The American Association for Cancer Research Project GENIE registry was accessed using cBioPortal for molecular profiling of angiomyolipoma (AML). A total of 276 renal tumors from 41 patients were analyzed. Renal tumors were classified into 9 distinct morphological subtypes, with AML predominating (238 [86%]). Interestingly, all these tumors acted in a benign fashion except one renal cell carcinoma with clear cells and fibromyomatous stroma and one epithelioid AML that metastasized. Molecular profiling studies revealed that epithelioid AMLs were enriched for alterations of TP53, RB1, and ATRX. Eight patients died of direct complications of TSC, including 3 of end-stage renal disease. To date, none have died of a renal epithelial neoplasm. The identification of unique renal neoplasia subtypes may provide important clues to establish a diagnosis of TSC, and in the somatic setting, this finding has important implications for accurate prognostication. These tumors tend to be indolent, and only 2 of 276 tumors in our study exhibited metastatic behavior. Our results support multidisciplinary management with a focus on preservation of renal function.


An exploratory study of reiki experiences in women who have cancer.
To explore the perceptions and experiences of reiki for women who have cancer and identify outcome measures for an intervention study. A cross-sectional qualitative study of 10 women who had received reiki after cancer treatment was conducted. Interviews were audiotaped, transcribed and coded using framework analysis. Key themes identified were: limited understanding of reiki prior to receiving any reiki; release of emotional strain during reiki-feelings of a release of energy, a clearing of the mind from cancer, inner peace/relaxation, hope, a sense of being cared for; experience of physical sensations during reiki, such as pain relief and tingling; physical, emotional and cognitive improvements after reiki, such as improved sleep, a sense of calm and peace, reduced depression and improved self-confidence. Findings suggest that reiki could be a beneficial tool in the self-management of quality of life issues for women who have cancer.


Assessment of Risk Factors for Drug Resistance of Dual Anti Platelet Therapy After PCI.
Since aspirin and clopidogrel are the widely and conventionally used drugs to treat acute myocardial infarction after percutaneous coronary intervention (PCI), it is important to explore potential risk factors of their resistance. The platelet aggregation rate with arachidonic acid (AA, PAg-AA%) and adenosine diphosphate (ADP, PAg-ADP%) of 219 PCI patients were measured after standard treatment for 24 h. The disease history and laboratory data (before PCI) were obtained. We found 101 (46.12%) patients to be aspirin-resistant, and PAg-ADP% was the most prominent risk factor of aspirin resistance. Clopidogrel resistance was present in 157 of 219 patients. Patients in the clopidogrel-resistant group carried more CYP2C19*3 or *2, which was associated with higher clopidogrel resistance in this group (69.11%, 47/68) than in the control group (64.29%, 36/56). Platelet count (109/L) and hemoglobin (g/L) were the prominent risk factors of clopidogrel resistance. Among the 219 patients, 98 showed dual antiplatelet drug resistance, for which platelet count (109/L) and monocyte count (g/L) were the risk factors. Aspirin resistance was found to usually accompany clopidogrel resistance.


Glycogen synthase kinase 3-β inhibition induces lymphangiogenesis through β-catenin-dependent and mTOR-independent pathways.
Lymphatic vessels play an important role in health and in disease. In this study, we evaluated the effects of GSK3-β inhibition on lung lymphatic endothelial cells in vitro. Pharmacological inhibition and silencing of GSK3-β resulted in increased lymphangiogenesis of lung lymphatic endothelial cells. To investigate mechanisms of GSK3-β-mediated lymphangiogenesis, we interrogated the mammalian/mechanistic target of rapamycin pathway and found that inhibition of GSK3-β resulted in PTEN activation and subsequent decreased activation of AKT, leading to decreased p-P70S6kinase levels, indicating inhibition of the mTOR pathway. In addition, consistent with a negative role of GSK3-β in β-catenin stability through protein phosphorylation, we found that GSK3-β inhibition resulted in an increase in β-catenin levels. Simultaneous silencing of β-catenin and inhibition of GSK3-β demonstrated that β-catenin is required for GSK3-β-induced lymphangiogenesis.


Regulation of isotype immunoglobulin and interleukin production by adjuvants in vitro.
An in vitro system has been developed in which antibodies to fluorescein isothiocyanate (FITC)-labelled human gamma globulin (HGG) or dextran sulfate (DXS), are produced in the presence or absence of different adjuvants. The antibody response of in vitro cultures was measured by assaying the total Ig-secreting cells and FITC-specific plaque-forming cells (PFC). The presence of low levels of antigen and various cytokines were necessary for the production of isotypes other than IgM. Our results indicate that the regulation of isotype switching in vitro is dependent upon the non-specific stimuli-adjuvants, which probably activate different types of cells for cytokine production. The pre-activation of spleen cells, in our system, by antigen, seems to play a decisive role in the subclass of IgG produced. The adjuvant may still be able to influence the precommitted cell isotype to switch to another subclass but only in a "down-stream" direction i.e., IgG3----IgG1----IgG2b----IgG2a.


Physiological effect of prone positioning in mechanically ventilated SARS-CoV-2- infected patients with severe ARDS: An observational study.
Mechanical ventilation in prone position was associated with a reduction in mortality and increase in arterial oxygenation in acute respiratory distress syndrome (ARDS) patients. However, physiological effects of prone position in COVID ARDS patients are unknown. In this prospective observational study, data of n = 47 consecutive real time RT- PCR confirmed SARS-CoV-2-infected patients with severe ARDS were included. Respiratory mechanics and oxygenation data of recruited patients were collected before and after prone position. Median (Interquartile range, IQR) age of the recruited patients was 60 (50-67) years and median (IQR) PaO2/FiO2 ratio of 61.2 (54-80) mm Hg with application of median (IQR) positive end expiratory pressure (PEEP) of 12 (10-14) cm H2O before initiation of prone position. Out of those patients, 36 (77%) were prone responders at 16 hours after prone session, evident by increase of PaO2 by at least 20 mm Hg or by 20% as compared to baseline, and 73% patients were sustained responders (after returning to supine position). Plateau airway pressure (p < 0.0001), peak airway pressure (p < 0.0001), and driving pressure (p < 0.0001) were significantly reduced in prone position, and static compliance (p = 0.001), PaO2/FiO2 ratio (p < 0.0001), PaO2 (p = 0.0002), and SpO2 (p = 0.0004) were increased at 4 hours and 16 hours since prone position and also after returning to supine position. In SARS-CoV-2-infected patients, mechanical ventilation in prone position is associated with improvement in lung compliance and oxygenation in almost three-fourth of the patients and persisted in supine position in more than 70% of the patients.


Daptomycin experience in patients with human immunodeficiency virus and resistant gram-positive infections.
This study was conducted to evaluate the clinical experience with daptomycin in the treatment of resistant gram-positive infections (GPIs) in patients with HIV infection. Using a retrospective, multicenter, and observational registry study, investigators assessed outcomes following daptomycin therapy in 78 patients (62 efficacy evaluable) infected with HIV and with resistant GPIs. Overall, success rates by infection type were bacteremia 91% (20 of 22), endocarditis 91% (10 of 11), and bone/joint 100% (9 of 9). Success by pathogen was 93% (39 of 42), 93% (14 of 15), and 100% (5 of 5) for methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, and methicillin-resistant coagulase-negative staphylococci, respectively. Daptomycin appeared to be well tolerated, with 9% having an adverse event possibly related to daptomycin and 4% discontinuing daptomycin. In HIV-infected patients, daptomycin appears to be a useful agent for treating resistant GPIs.


NeuN: a useful neuronal marker for diagnostic histopathology.
The monoclonal antibody A60 specifically recognizes the DNA-binding, neuron-specific protein NeuN, which is present in most neuronal cell types of vertebrates. In this study we demonstrate the potential use of NeuN as a diagnostic neuronal marker using a wide range of formalin-fixed, paraffin-embedded human surgical and autopsy specimens from the central and peripheral nervous system. After microwave antigen retrieval, almost all neuronal populations revealed strong immunoreactivity for NeuN in nuclei, perikarya, and some proximal neuronal processes, whereas more distal axon cylinders and dendritic ramifications were not stained. The stain greatly enhanced the gray matter architecture. NeuN immunoreactivity was not detected in Purkinje cells, most neurons of the internal nuclear layer of the retina, and in sympathetic chain ganglia. We examined nine gangliogliomas and 14 dysembryoplastic neuroepithelial tumors, one ganglioneuroma, and one dysplastic cerebellar gangliocytoma. The neuronal component of all of these lesions showed marked immunoreactivity for NeuN. In addition, NeuN immunoreactivity was focally seen in one of seven medulloblastomas with prominent neuronal differentiation. There was no staining of non-neuronal structures. The results indicate that NeuN immunoreactivity is a sensitive and specific neuronal marker in formalin-fixed, paraffin-embedded tissues, and may be useful in diagnostic histopathology.


Role of TH-17 cells in rheumatic and other autoimmune diseases.
In humans multiple pathways can induce TH-17 cell differentiation, whereas in mice this process is mostly modulated by IL-6 and TGF-β. IL-17 produced by TH-17 cells has been associated with a number of inflammatory autoimmune diseases including psoriasis, systemic lupus erythematosus, inflammatory bowel disease, multiple sclerosis, and rheumatoid arthritis. In this review, we have primarily focused on the role of TH-17 cells/IL-17 in the pathogenesis of rheumatoid arthritis and experimental arthritis. The potential role of TH-17 cells in rheumatoid arthritis progression has been demonstrated by correlating the percent TH-17 cells or levels of IL-17 with rheumatoid arthritis disease activity score and C-reactive protein levels. Further, previous studies suggest that IL-17 mediated vascularization may lay the foundation for rheumatoid arthritis joint neutrophil and monocyte recruitment as well as cartilage and bone destruction. The profound role of IL-17 in the pathogenesis of experimental arthritis may be due to its synergistic effect with TNF-α and IL-1β. Although the initial clinical trial employing anti-IL-17 antibody has been promising for rheumatoid arthritis, future studies in humans will shed more light on how anti-IL-17 therapy affects rheumatoid arthritis and other autoimmune disease pathogenesis.


Modulation of mammalian circadian rhythms by tumor necrosis factor-α.
Systemic low doses of the endotoxin lipopolysaccharide (LPS, 100 µg/kg) administered during the early night induce phase-delays of locomotor activity rhythms in mice. Our aim was to evaluate the role of tumor necrosis factor (Tnf)-alpha and its receptor 1/p55 (Tnfr1) in the modulation of LPS-induced circadian effects on the suprachiasmatic nucleus (SCN). We observed that Tnfr1-defective mice (Tnfr1 KO), although exhibiting similar circadian behavior and light response to that of control mice, did not show LPS-induced phase-delays of locomotor activity rhythms, nor LPS-induced cFos and Per2 expression in the SCN and Per1 expression in the paraventricular hypothalamic nucleus (PVN) as compared to wild-type (WT) mice. We also analyzed Tnfr1 expression in the SCN of WT mice, peaking during the early night, when LPS has a circadian effect. Peripheral inoculation of LPS induced an increase in cytokine/chemokine levels (Tnf, Il-6 and Ccl2) in the SCN and in the PVN. In conclusion, in this study, we show that LPS-induced circadian responses are mediated by Tnf. Our results also suggest that this cytokine stimulates the SCN after LPS peripheral inoculation; and the time-related effect of LPS (i.e. phase shifts elicited only at early night) might depend on the increased levels of Tnfr1 expression. We also confirmed that LPS modulates clock gene expression in the SCN and PVN in WT but not in Tnfr1 KO mice. We demonstrate a fundamental role for Tnf and its receptor in circadian modulation by immune stimuli at the level of the SCN biological clock.


[The action of helium-neon laser radiation on the growth of Mycobacterium tuberculosis].
The growth properties of M. tuberculosis subjected to the action of helium-neon laser radiation was studied. Laser radiation was shown to change the quantitative and qualitative composition of mycobacterial population. Disturbances in the viability of mycobacteria appear as a consequence of changes in the morphological structure of mycobacterial cells. The maximum effect of helium-neon laser radiation was achieved after the irradiation of M. tuberculosis culture on days 2-3 after inoculation. These results made it possible to suggest that the effect of helium-neon laser radiation was most pronounced in cells at the stage of mitosis (the logarithmic stage of growth) with the highest degree of metabolism.


Are Titania Photocatalysts and Titanium Implants Safe? Review on the Toxicity of Titanium Compounds.
Titanium and its compounds are broadly used in both industrial and domestic products, including jet engines, missiles, prostheses, implants, pigments, cosmetics, food, and photocatalysts for environmental purification and solar energy conversion. Although titanium/titania-containing materials are usually safe for human, animals and environment, increasing concerns on their negative impacts have been postulated. Accordingly, this review covers current knowledge on the toxicity of titania and titanium, in which the behaviour, bioavailability, mechanisms of action, and environmental impacts have been discussed in detail, considering both light and dark conditions. Consequently, the following conclusions have been drawn: (i) titania photocatalysts rarely cause health and environmental problems; (ii) despite the lack of proof, the possible carcinogenicity of titania powders to humans is considered by some authorities; (iii) titanium alloys, commonly applied as implant materials, possess a relatively low health risk; (iv) titania microparticles are less toxic than nanoparticles, independent of the means of exposure; (v) excessive accumulation of titanium in the environment cannot be ignored; (vi) titanium/titania-containing products should be clearly marked with health warning labels, especially for pregnant women and young children; (vi) a key knowledge gap is the lack of comprehensive data about the environmental content and the influence of titania/titanium on biodiversity and the ecological functioning of terrestrial and aquatic ecosystems.


PostMod: sequence based prediction of kinase-specific phosphorylation sites with indirect relationship.
Post-translational modifications (PTMs) have a key role in regulating cell functions. Consequently, identification of PTM sites has a significant impact on understanding protein function and revealing cellular signal transductions. Especially, phosphorylation is a ubiquitous process with a large portion of proteins undergoing this modification. Experimental methods to identify phosphorylation sites are labor-intensive and of high-cost. With the exponentially growing protein sequence data, development of computational approaches to predict phosphorylation sites is highly desirable. Here, we present a simple and effective method to recognize phosphorylation sites by combining sequence patterns and evolutionary information and by applying a novel noise-reducing algorithm. We suggested that considering long-range region surrounding a phosphorylation site is important for recognizing phosphorylation peptides. Also, from compared results to AutoMotif in 36 different kinase families, new method outperforms AutoMotif. The mean accuracy, precision, and recall of our method are 0.93, 0.67, and 0.40, respectively, whereas those of AutoMotif with a polynomial kernel are 0.91, 0.47, and 0.17, respectively. Also our method shows better or comparable performance in four main kinase groups, CDK, CK2, PKA, and PKC compared to six existing predictors. Our method is remarkable in that it is powerful and intuitive approach without need of a sophisticated training algorithm. Moreover, our method is generally applicable to other types of PTMs.


Occurrence and Characteristics of Mcrs among Gram-Negative Bacteria Causing Bloodstream Infections of Infant Inpatients between 2006 and 2019 in China.
The aim of this study was to determine the occurrence of mobilized colistin resistance (mcr) genes in Gram-negative bacteria causing bloodstream infections of child inpatients in China. Bacteria were collected between 2006 and 2019 in a maternal and child health hospital, and mcr genes were screened by PCR. Five of 252 isolates were mcr-positive, including one mcr-1-positive colistin-resistant Escherichia coli isolate, two mcr-9-positive colistin-susceptible Salmonella enterica isolates, and two mcr-9-positive colistin-susceptible Enterobacter hormaechei isolates. These were obtained from two neonate and three infant patients admitted between 2009 and 2018. The E. coli isolate was obtained from a neonate aged 20 min, suggestive of a possible mother-to-neonate transmission. The five mcr-positive isolates were multidrug resistant, and two S. enterica and one E. hormaechei isolate showed a hypervirulent phenotype compared to a hypervirulent Klebsiella pneumoniae type strain in a Galleria mellonella infection model. The mcr-1 gene was carried by an IncX4-type pA1-like epidemic plasmid, and the mcr-9 gene was detected on IncHI2/2A-type novel plasmids co-carrying multiple resistance genes. The four IncHI2/2A-type plasmids shared a backbone and a high similarity (≥77% coverage and ≥ 90% nucleotide identity), suggesting that they were derived from a common ancestor with cross-species transmission and have circulated locally over a long period. The conjugation assay showed that the mcr-1-encoding plasmid and one mcr-9-encoding plasmid were self-transmissible to E. coli with high conjugation frequencies. Our findings demonstrate that mcr genes have disseminated in the community and/or hospitals, mediated by epidemic/endemic plasmids over a long period. The study shows that continuous monitoring of mcr genes is imperative for understanding and tackling their dissemination. IMPORTANCE Antimicrobial resistance, especially the spread of carbapenemase-producing Enterobacteriaceae (CPE), represents one of the largest challenges to One Health coverage of environmental, animal, and human sectors. Colistin is one of the last-line antibiotics for clinical treatment of CPE. However, the emergence of the mobilized colistin resistance (mcr) gene largely threatens the usage of colistin in the clinical setting. In this study, we investigated the existence of mcr genes in 252 Gram-negative bacteria collected between 2006 and 2019 which caused bloodstream infections of child inpatients in China. We found a high prevalence of mcr carriage among children inpatients in the absence of professional exposure, and mcr might have widely disseminated in the community via different routes. This study emphasizes the importance of rational use of colistin in the One Health frame, and highlights both the urgent need for understanding the prevalence and dissemination of mcr genes in different populations and the importance of effective measures to control their spread.


Pathophysiology, recognition and management of pneumoretroperitoneum.
The medical records of 15 patients found to have pneumoretroperitoneum in a 10-year period were reviewed. The cause of retroperitoneal emphysema was infection in six patients, trauma in five and iatrogenic in four. Emphysema was confined to the retroperitoneum in 11 patients, and extended to the mediastinum in four and to the soft tissue of the neck in three. Failure to diagnose pneumoretroperitoneum resulted in delayed intervention in two patients. One patient with pneumoretroperitoneum and pneumoperitoneum secondary to a lung lesion underwent unnecessary laparotomy. Outcome was favourable in these three patients. The presence of air in the retroperitoneum is not dangerous but its early recognition and detection of the source are important as septic conditions may be involved.


Should trustees be members of medical staff committees? Its' debatable.
Trustee membership on medical staff committees is not an appropriate way of ensuring liasion between the medical staff and the governing board. All assurances concerning quality of care should come to the governing board from the organized medical staff, to whom such accountability is and should be delegated.


Hot flushes: are there effective alternatives to estrogen?
Hot flushes are the most common indication for the prescription of hormone replacement therapy (HRT) since it is effective in over 80% of cases. In 1995, 37% of American women took HRT, principally for this purpose. However, over the last five years, publications such as those from the Women's Health Initiative (WHI) have caused concern among women since they perceive that the risks outweigh the benefits. Following this publication, half of the women taking HRT in the UK, USA and New Zealand discontinued HRT. With the discontinuation of estrogen many women re-developed hot flushes; however only a small number (18%) of women report restarting hormone therapy. The majority of these (76%) for the recurrence of severe hot flushes or night sweats. Alternatives are available, but limited knowledge on aetiology and mechanisms of hot flushing represents a major obstacle for the development of new, targeted, non-hormonal treatments, and no current alternatives are as effective as estrogen.


Accessing young adolescents and their families for research.
To describe the strategies used to obtain and retain a sample of young adolescents and their families. The sample was 847 families with young adolescents aged 11 to 14 years who resided in a large urban Midwestern community of the U.S. The first design strategy was becoming credible within the community. For 5 years before conducting the intervention research in the community, surveys were conducted of the needs of middle-school students as they entered adolescence, and programs were delivered to meet the assessed needs. The second design strategy was instituting developmentally appropriate recruitment and retention strategies based upon direct consultation with groups of young adolescents. The consultants recommended referring to young adolescents as young adults and holding the training sessions in sites not associated with schools. The third design strategy was setting up participation methods that were nonburdensome to schools and families. The successful recruitment of over 800 families and the retention of 80% of them for survey participation and 77% of them for the intervention protocol can be attributed to three major strategies: (a) community involvement; (b) adherence to principles of adolescent development; and (c) ease of participation for school personnel and families.


Natural killer cell proliferation and renal disease: a functional and phenotypic study.
We describe a 57-year-old woman who presented with a constitutional syndrome, glomerulonephritis, and lymphocytosis. The phenotypic study, using flow cytometry, showed an expansion of natural killer (NK) cells (CD2+, CD3-, CD16+, CD56+, and CD7+). We performed a functional study of peripheral blood mononuclear cells (PBMCs) and of purified CD16+ cells (NK cells) and CD3+ cells (normal T cells). The expanded NK cell population, CD16+, did not proliferate with phytohemagglutinin (PHA) or anti-CD3 but showed a dose-dependent proliferation with recombinant interleukin-2 (rIL-2) and also proliferated with phorbol dibutyrate. This population showed very strong NK and lymphokine-activated killer cell (LAK) activities. The patient's symptoms resolved spontaneously without treatment. Three years later, however, there is still abnormal renal function, and the expansion of NK cells persists, although with no indication of malignancy. We review the features of the different large granular lymphocyte proliferations and their seldom described relationship with renal disease.


Assessment of antioxidants in foods and biological samples: a short critique.
Antioxidants can be defined on a biological basis by their ability to inhibit the oxidation of susceptible biomolecules such as proteins, lipids and DNA. There is observational evidence on an inverse association between dietary antioxidant intake and the development of chronic conditions, although randomized trials have not confirmed these results. The antioxidant potential of a biological material can be evaluated by methods categorized into three groups. The first group includes the determination of the individual antioxidants present in the investigated sample. The second focuses on 'total antioxidant capacity', which is considered a convenient tool for obtaining an overall estimate of the antioxidant potential of the sample. The third group involves the quantification of in vivo markers of oxidative stress, such as the products of protein, lipid and DNA oxidation. Due to differences between the assays, however, current research suggests the application of a combination of such methods for a more informative assessment of the oxidation status of the organism.


Structural basis for the functional roles of critical residues in human cytochrome p450 aromatase.
Cytochrome P450 aromatase (CYP19A1) is the only enzyme known to catalyze the biosynthesis of estrogens from androgens. The crystal structure of human placental aromatase (pArom) has paved the way toward understanding the structure-function relationships of this remarkable enzyme. Using an amino terminus-truncated recombinant human aromatase (rArom) construct, we investigate the roles of key amino acids in the active site, at the intermolecular interface, inside the access channel, and at the lipid-protein boundary for their roles in enzyme function and higher-order organization. Replacing the active site residue D309 with an N yields an inactive enzyme, consistent with its proposed involvement in aromatization. Mutation of R192 at the lipid interface, pivotal to the proton relay network in the access channel, results in the loss of enzyme activity. In addition to the distal catalytic residues, we show that mutation of K440 and Y361 of the heme-proximal region critically interferes with substrate binding, enzyme activity, and heme stability. The D-E loop deletion mutant Del7 that disrupts the intermolecular interaction significantly reduces enzyme activity. However, the less drastic Del4 and point mutants E181A and E181K do not. Furthermore, native gel electrophoresis, size-exclusion chromatography, and analytical ultracentrifugation are used to show that mutations in the intermolecular interface alter the quaternary organization of the enzyme in solution. As a validation for interpretation of the mutational results in the context of the innate molecule, we determine the crystal structure of rArom to show that the active site, tertiary, and quaternary structures are identical to those of pArom.


Vocal cues to male androgen levels in giant pandas.
Little is known about the potential of non-human mammal vocalizations to signal information on the hormonal status of the caller. In the current study, we used endocrine data and acoustic analyses to determine whether male giant panda bleats provide reliable information about the caller's current androgen levels. Our results revealed significant relationships between acoustic features of male giant panda bleats and the caller's faecal androgen metabolite concentrations. To our knowledge, this constitutes the first demonstration that the acoustic structure of a non-human mammal call has the potential to yield information about the caller's current androgen levels. We go on to discuss the anatomical basis for our findings and the potential functional relevance of signalling information on male androgen levels in giant panda sexual communication.