Use of synchrotron radiation for the measurement of fluorescence lifetimes with subpicosecond resolution.
A method for the measurement of fluorescence lifetimes with subpicosecond resolution is described. The temporal resolution is achieved by using the high harmonic content of the synchrotron radiation produced by an electron storage ring.


Biological control of dermatophytic fungus Trichophyton species by Bacillus subtilis.
A potent Bacillus subtilis-2 isolated from soil samples, showed 14 mm inhibition zone by spot inoculation method and 22 mm inhibition zone by well agar diffusion method against pathogenic test fungii. i.e. Trichophyton species. Among four media tested, the maximum growth and antibiotic production was found in trypticase soya broth (TSB) medium at 37 degrees C, pH 7 and 48 hrs of incubation. The MIC of antifungal substance was 35 microl/ml. The isolated Bacillus subtilis -2 has strong antifungal activity against dermatophytic fungi Trichophyton sp. and requires chemical characterization for its bioactive principle.


Computerized antimicrobial decision support for hospitalized patients with a bloodstream infection.
We developed a computerized antimicrobial decision support program founded on our local bacterial susceptibility data. In a retrospective analysis of patients with a bloodstream infection, we compared the actual antimicrobials prescribed to the antimicrobials recommended by the program. We found the computer-guided therapy to be clinically and statistically more effective than the therapy initiated by the physicians. We conclude that computerized decision support can improve the targeting of empiric antimicrobial therapy.


Child maltreatment and brain development.
A growing body of research has linked childhood experiences of maltreatment with a host of physical conditions that manifest in adulthood. In addition, newer neuroimaging techniques have documented structural changes that occur in the brains of individuals who suffer early maltreatment. This article briefly reviews the literature on these topics and outlines the connection between abuse in childhood and health problems in adulthood.


IL-23-producing CD68(+) macrophage-like cells predominate within an IL-17-polarized infiltrate in chronic periodontitis lesions.
To analyse antigen-presenting cells (APCs), such as dendritic cells (DCs), macrophages (Mo) or B cells depending on the regional site of chronic periodontitis (CP), and to investigate their relation to Th17 cells. Biopsies from oral mucosa as well as the coronal and bottom regions of CP were analysed by immunhistochemistry, immunofluorescence, flow cytometry and real-time PCR. A predominance of CD68(+) Mo-like cells and CD20(+) B cells and strong Th17 infiltration was observed in the bottom region of CP lesions, while CD1a(+) DCs were only detected in the coronal regions, where Th17 infiltration was low. Furthermore, CD68(+) Mo-like cells displayed CD163 expression as a typical Mo-marker, but expressed in parallel typical DCs markers, such as CD11c or CD209 and TLR4. Interestingly, Th17-inducing cytokine IL-23p19 was produced by CD68(+) Mo-like cells, but not CD20(+) B cells. Moreover, the stimulation of in vitro generated CD68(+) Mo-like cells by Porphyromonas gingivalis-derived (Pg) lipopolysaccharide resulted in the upregulation of their IL-23p19 mRNA expression, which was inhibited by the blockage of TLR4. In view of these data, a picture emerges that IL-17-producing cells in CP could be in part directed by CD68(+) Mo-like cells, which produce IL-23p19 upon TLR4 activation by Pg.


Leukocyte adhesion dynamics in shear flow.
Of the white blood cells that traverse the circulation, the polymorphonuclear leukocytes commonly called neutrophils, are the most numerous, numbering approximately 5000 per microliter of blood. In a sense, these cells are the emergency response unit within the body's circulatory highway since they are the first to be recruited at sites of tissue trauma, infection, or inflammation. The chief function of neutrophils is the capture, phagocytosis, and degradation of foreign invaders. To carry out this critical function, neutrophils sense infection via chemotactic receptors that trigger cellular activation upon ligation of as few as 10-100 bacterial or chemokine peptides. Despite this acute sensitivity to stimuli, neutrophils circulate in healthy individuals largely in a passive state, with a very low efficiency of capture and arrest on quiescent endothelium. Here, we define the efficiency as the fraction of all cells passing by a given length of vessel wall that is captured and achieves firm adhesion. Within seconds of chemotactic signaling, adhesion molecules expressed on the plasma membrane of neutrophils are activated and a rapid boost in the efficiency of stable adhesion is detected. This occurs for both homotypic neutrophil-neutrophil adhesion and neutrophil capture and adhesion on inflamed endothelium. With such large numbers of circulating neutrophils, and with their inherent capability to rapidly adhere to postcapillary venular endothelium, the body has evolved a diverse set of mechanisms to regulate the inflammatory cascade that begins with intracellular signaling and leads to cell arrest and extravasation at sites of tissue insult. In this article we will focus on the interplay between particle-fluid dynamics that involve the shear and normal forces that transport cells transversely and axially within the vessel, and the activation and interaction of adhesion molecules that involve receptor-ligand bond formation that enables the neutrophils to resist wall shear stress and adhere to specific sites of inflammation.


Reactive oxygen species-responsive silk sericin microcapsules used for antioxidative stress damage.
Silk sericin microcapsules loaded with horseradish peroxidase (HRP) are prepared through protein self-assembly in a green environment containing enzymes to protect liver cells from alcohol damage. Load content and release dynamics of HRP in sericin microcapsules are investigated. The role of HRP-loaded microcapsules in hydrogen peroxide (H2 O2 ) degradation is demonstrated using electrochemical method. Furthermore, the effect of the HRP-loaded microcapsules on cells and intracellular reactive oxygen species (ROS) level is evaluated using an alcohol damage model in vitro. Results show that HRP can be loaded effectively in the sericin microcapsules and can be released ROS-responsively from microcapsules. Cell survival rate increases after suffering from alcohol damage due to the presence of HRP-loaded microcapsule, and the active oxygen content in cells is maintained at a stable level even when it remained in an environment with high alcohol concentration. We believe that the internalized sericin microcapsules maintain HRP activity intracellularly, allow controlled HRP release within a host cell, and show excellent ability in antioxidative stress injury.


Conservation of the RD-BF-C2 organization in the pig MHC class-III region: mapping and cloning of the pig RD gene.
The RD gene, named after the arginine (R) and aspartic acid (D) repeat in the central part of its protein, was initially mapped in the mouse H-2S subregion between C4 and BF. It was later mapped in the same position in the human MHC and here we show it is also conserved in the pig MHC class III region, close to the complement BF gene. A pig RD genomic clone was isolated from a lambda-phage library. Hybridizations on genomic DNA separated with pulsed field gel electrophoresis identified common 220 kb NruI, 130 kb EagI and 200 kb MluI bands for RD, BF and C2. The RD gene has also a 17 kb Kp nI and 11 kb SacI fragment in common with BF but not with C2. The close linkage of the RD and BF genes was further established by hybridization of BF to a genomic lambda-phage clone also containing the RD gene. This genomic RD clone overlaps with a lambda-phage clone previously isolated and containing the complete BF gene and the 3' part of C2. The distance between RD and BF is about 6 kb. The junction between the two complement genes BF and C2 was sequenced and the BF 5' promoter region, overlapping the 3' noncoding region of C2, was compared with that of the human BF promoter. The overall homology was about 80% and all but one identified promoter elements were found in the same position in both genes. The results obtained demonstrate the RD-BF-C2 organization is strongly conserved between human, mouse and pig. No polymorphisms were detected in either the RD gene or in the BF promoter region using polymerase chain reaction and restriction fragment polymorphism analysis.


Buschke-Ollendorff syndrome in a three-generation family: influence of a novel LEMD3 mutation to tropoelastin expression.
Buschke-Ollendorff syndrome refers to the concomitant occurrence of connective tissue nevi, composed of elastic fibers in most cases, with osteopoikilosis. This autosomal dominant inherited disorder is caused by mutations in the gene LEMD3 on chromosome 12q14, which induces a rather heterogeneous clinical phenotype. Here, we report on the most proximal germline mutation found to date in the LEMD3 gene, p.Val94fs, in a three-generation Swiss family. Quantitative RNA analyses in affected and non-affected skin tissue from the proband demonstrate a comparable nonsense-mediated decay of mutant LEMD3 mRNA in both tissues; however, different levels of tropoelastin expression suggest that additional factors are involved in the development of the cutaneous lesions.


Categorical and continuous models of liability to externalizing disorders: a direct comparison in NESARC.
Patterns of genetic, environmental, and phenotypic relationships among antisocial behavior and substance use disorders indicate the presence of a common externalizing liability. However, whether this liability is relatively continuous and graded, or categorical and class-like, has not been well established. To compare the fit of categorical and continuous models of externalizing liability in a large, nationally representative sample. Categorical and continuous models of externalizing liability were compared using interview data from the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC). Face-to-face interviews conducted in the United States. Random sample of 43 093 noninstitutionalized adult civilians living in the United States. Lifetime and current (past 12 months) diagnoses of antisocial personality disorder, nicotine dependence, alcohol dependence, marijuana dependence, cocaine dependence, and other substance dependence. In the entire sample, as well as for males and females separately, using either lifetime or current diagnoses, the best-fitting model of externalizing liability was a continuous normal model. Moreover, there was a general trend toward latent trait models fitting better than latent class models, indicating that externalizing liability was continuous and graded, rather than categorical and class-like. Liability to externalizing spectrum disorders is graded and continuous normal in distribution. Research regarding etiology, assessment, and treatment of externalizing disorders should target externalizing liability over a range of severity. Current diagnoses represent extremes of this continuous liability distribution, indicating that conditions currently classified as subthreshold are likely to provide important information regarding liability to externalizing phenomena.


Impact of Grocery Store Proximity on Store Preference Among Atlanta SNAP-Ed Participants.
To assess the association between grocery store proximities and the individual's grocery store preferences among Supplemental Nutrition Assistance Program Education participants in Atlanta. University of Georgia Supplemental Nutrition Assistance Program Education participants (n = 615, response rate is 36%) in 3 counties provided their preferred grocery store chains. The association between store proximity (both network distance and driving time) and store preference was measured through logistic regression controlling for age, sex, and race. Descriptive statistics showed participants had widely varying proximities to grocery stores. Model results were significant for all smaller chains (Aldi, Big Bear, Wayfield, Food Depot, and Save-A-Lot), Kroger (P < 0.01), as well as for Walmart (time only, P = 0.002). Future studies might identify whether local groceries are more willing to partner on interventions or are more effective at reaching local residents. Surveys or techniques such as sketch mapping could also show whether individuals shop in neighborhoods close to work or friends and family.


The Straightened Splenic Vessels Method Improves Surgical Outcomes of Laparoscopic Distal Pancreatectomy.
In laparoscopic distal pancreatectomy (LDP), isolating the splenic artery and vein requires advanced techniques. This study aimed to assess the efficacy of a novel method termed the 'straightened splenic vessels' (SSV) method for isolating the splenic vessels in LDP. In SSV, to adjust the instrument axis, the splenic artery was straightened by grasping 2 points of its nerve sheath. Then, the layer between the splenic artery's nerve sheath and the pancreatic parenchyma was dissected. Next, the pancreas was mobilized from body to tail, and the splenic vein was straightened by 3-point retraction before isolation. To evaluate this method's efficacy, we investigated 51 patients who underwent LDP. In 39 patients who underwent LDP with splenectomy, the mean operating time was significantly shorter in the SSV group than in the conventional group (p = 0.004). In 12 patients who underwent LDP with preserving the splenic vessels, the mean intraoperative blood loss in the SSV group was 27.6 ml, which was significantly lower than that in the conventional group (p = 0.012). This method may be applied as a standard procedure with little blood loss and short operation time for LDP. Larger prospective studies are needed to further evaluate the feasibility.


p53 allelotypes and enhanced detection of allelic loss in ovarian cancer: lack of correlation with familial and clinical factors.
To develop a mapping strategy for improved detection of p53 allelic loss associated with ovarian carcinoma, we utilized multiple intragenic polymorphisms and single-strand conformation polymorphism (SSCP) analysis. p53 allelotype distributions were defined for 80 ovarian cancer patients from germ-line DNA. All polymorphic sites studied had polymorphism information content (PIC) rates of greater than 0.25. Tumor loss of heterozygosity (LOH) was determined from informative polymorphisms and migratory shifts on SSCP screening of exons 5-9. Of the four polymorphisms analyzed, the intron 1 (alu) was the most informative (PIC = 0.66). A novel allele of 110 base pairs was found in 4.4% of our ovarian cancer cohort at this site. The intron 3 (16-base pair repeat) and intron 6 (MSP1) polymorphisms were in relative equilibrium; thus, we chose to use only the intron 3 polymorphic site in the mapping strategy. Family cancer history did not influence the allelotype distribution frequencies. Overall, 56 of 61 tumors (91.8%) were informative for allelic loss, and LOH was observed in 66.1 %. A reduction to homozygosity at the p53 locus did not correlate with familial or clinical factors. These observations are consistent with the multiple mechanisms by which p53 dysfunction can occur.


Naproxen in the prevention of heterotopic ossification after total hip replacement.
The effectiveness of naproxen, indomethacin, and a placebo were compared for the prevention of heterotopic ossification after total hip arthroplasty. Eighty-four men at high risk of experiencing heterotopic ossification received randomly either naproxen 750 mg per day, indomethacin 75 mg per day, or a placebo for 6 consecutive weeks after surgery. Efficacy analysis showed that in 70% of patients treated with naproxen, in 34.8% of those treated with indomethacin, and in 15% of those treated with a placebo, no heterotopic ossification appeared on radiographs taken at 6 months. Naproxen was significantly more effective than the placebo or indomethacin in preventing the development of heterotopic ossification. Stratification into five categories, according to Brooker's classification at 6 months, showed that both drugs were equally superior to the placebo. Class III heterotopic ossification occurred only in patients who were given the placebo. The improvement in clinical criteria was comparable in the three groups, although at 6 months there was an improvement in abduction in patients treated with indomethacin compared with those treated with naproxen and the placebo. The overall tolerance was rated good by 87% of patients and 86% of physicians, with no difference between the groups. The results show that naproxen, given daily for 6 weeks, is an effective and safe medication for the prevention of heterotopic ossification after total hip replacement.


Mixed functional characteristics correlating with TCR-ligand koff -rate of MHC-tetramer reactive T cells within the naive T-cell repertoire.
The low frequency of antigen-specific naïve T cells has challenged numerous laboratories to develop various techniques to study the naïve T-cell repertoire. Here, we combine the generation of naïve repertoire-derived antigen-specific T-cell lines based on MHC-tetramer staining and magnetic-bead enrichment with in-depth functional assessment of the isolated T cells. Cytomegalovirus (CMV) specific T-cell lines were generated from seronegative individuals. Generated T-cell lines consisted of a variety of immunodominant CMV-epitope-specific oligoclonal T-cell populations restricted to various HLA-molecules (HLA-A1, A2, B7, B8, and B40), and the functional and structural avidity of the CMV-specific T cells was studied. Although all CMV-specific T cells were isolated based on their reactivity toward a specific peptide-MHC complex, we observed a large variation in the functional avidity of the MHC-tetramer positive T-cell populations, which correlated with the structural avidity measured by the recently developed Streptamer koff -rate assay. Our data demonstrate that MHC-tetramer staining is not always predictive for specific T-cell reactivity, and challenge the sole use of MHC-tetramers as an indication of the peripheral T-cell repertoire, independent of the analysis of functional activity or structural avidity parameters.


Osteoclast precursor differentiation by MCPIP via oxidative stress, endoplasmic reticulum stress, and autophagy.
Osteoclasts (OCs) are responsible for bone resorption in inflammatory joint diseases. Monocyte chemotactic protein-1 (MCP-1) has been shown to induce differentiation of monocytes to OC precursors, but nothing is known about the underlying mechanisms. Here, we elucidate how MCPIP, induced by MCP-1, mediates this differentiation. Knockdown of MCPIP abolished MCP-1-mediated expression of OC markers, tartrate-resistant acid phosphatase, and serine protease cathepsin K. Expression of MCPIP induced p47(PHOX) and its membrane translocation, reactive oxygen species formation, and induction of endoplasmic reticulum (ER) stress chaperones, up-regulation of autophagy marker, Beclin-1, and lipidation of LC3, and induction of OC markers. Inhibition of oxidative stress attenuated ER stress and autophagy, and suppressed expression of OC markers. Inhibition of ER stress by a specific inhibitor or by knockdown of IRE1 blocked autophagy and induction of OC markers. ER stress inducers, tunicamycin and thapsigargin, induced expression of OC markers. Autophagy inhibition by 3'-methyladenine, LY294002, wortmannin or by knockdown of Beclin-1 or Atg 7 inhibited MCPIP-induced expression of OC markers. These results strongly suggest that MCP-1-induced differentiation of OC precursor cells is mediated via MCPIP-induced oxidative stress that causes ER stress leading to autophagy, revealing a novel mechanistic insight into the role of MCP-1 in OCs differentiation.


Racial Differences of Psychosocial Characteristics, HIV Risk-Taking and HIV Prevention Uptake between Men Who Have Sex with Men Only and Men Who Have Sex with Men and Women: A Community-Based Study in Two US Cities.
Bisexual behavior preference plays an important role in shaping HIV risks among men who have sex with men (MSM), yet few studies have examined the racial differences in the patterns of sexual risk and HIV prevention uptake between men who have sex with men and women (MSMW) and men who have sex with men only (MSMO). We conducted a community-based study to identify and compare psychosocial characteristics, sexual risks, HIV testing frequency, and pre-exposure prophylaxis (PrEP) engagement between Black and White MSMW and MSMO in two US cities. Findings indicate that White MSMW were most likely to engage in HIV risk-taking behaviors (e.g., alcohol/drug use before sex, condomless/group sex), while Black MSMW were least likely to recently test for HIV or report awareness (aPR: 0.32, 95% CI: 0.17-0.61), willingness (aPR: 0.58, 95% CI: 0.30-0.98), and use (aPR: 0.59, 95% CI: 0.31-0.93) of PrEP. Findings from our study supplement salient information for designing culturally tailored HIV prevention interventions for specific MSM subgroups in the US.


Emergency department stabilization of pediatric patients with bacterial meningitis. Current advances.
The infant or child who presents to the Emergency Department with bacterial meningitis may have nonspecific vague symptoms with few signs of serious illness. However, the disease is often rapidly progressive and life-threatening, and may be associated with respiratory failure, circulatory failure, increased intracranial pressure, disseminated intravascular coagulation, or convulsions, any of which may lead to a fatal outcome. It is important for the triage technician in an Emergency Department to cautiously inspect each young patient who presents with illness, carefully considering whether the presenting syndrome of symptoms and signs might be consistent with early meningitis. If the young patient is triaged in a nonemergent category, then periodic assessments of the patients waiting to be seen may ensure that, when the infant or child with an obscure presentation develops evidence suggesting this diagnosis, the triage technician will promptly notify the appropriate definitive care providers who assume responsibility for immediate definitive evaluation and stabilization. Changes in delivery of lifesaving care to the life-threatened child are being impacted by current advances in the understanding of the biochemical basis of disease at the cellular and subcellular levels. Endotoxin release into the blood causes increased production of kinins, which results in vasodilatation and increased vascular permeability. Members of the leukotriene family may also enhance vascular permeability as well as produce augmented leukocyte aggregation to vascular endothelium, vasoconstriction, and bronchoconstriction. Endotoxin activates the complement cascade and induces platelets to form reversible aggregates that may be trapped in the pulmonary microcirculation; and endotoxemia-activated platelets release serotonin, which may be associated with pulmonary hypertension. Now that we have antibiotics that are effective against organisms whose degradation produces endotoxin, there is interest in lessening the host inflammatory response to endotoxin through use of dexamethasone as an anti-inflammatory agent. Clinical trials have revealed that patients who received dexamethasone became afebrile earlier and were less likely to acquire deafness after bacterial meningitis. Because administration of antibiotics is the current specific medical therapy for this life-threatening microbial invasion, it is reasonable to continue to strive to shorten the interval between recognition of disease and specific therapy. However, new studies suggest that consequences of the complex host inflammatory response (at the cellular and subcellular level) to microbial invasion and endotoxin release from bacterial degradation are increasingly important in determining survival or severity of morbidity. Therapeutic intervention with specific antibiotics and steroid anti-inflammatory agents for modulating host responses enhances outcome.


Transgenic approaches to salivary gland research.
This review discusses the use of transgenic technology for the study of salivary gland function and development. Methods for gene transfer in the salivary gland of experimental mammals are described, including use of conventional and embryonic stem cell-derived transgenic mice and direct gene transfer techniques employing recombinant adenovirus vectors. Tissue-specific expression studies in transgenic mice have defined a number of salivary gland-specific promoter/enhancers that can be used to direct transcription into different salivary cell types. The consequences of expression of various oncogenes in the salivary glands of transgenic mice for the induction of neoplasia and hyperplasia are described. Recently, the use of adenoviral vectors has been demonstrated to be a powerful new approach to transfer genes into the salivary gland. This technology should have a major impact on the study of salivary gland gene expression, as well as provide a means to express therapeutic proteins in the salivary gland.


One-step preparation of highly monodisperse micron-size particles in organic solvents.
In this communication, we report the first simple and fast one-step method for synthesizing highly monodisperse micron-size PMMA particles in organic media through dispersion polymerization in the presence of PHSA (a polyhydroxyl-stearic-acid graft PMMA copolymer) as a stabilizer. There are two significant advantages of our method over earlier methods. First, by optimizing the composition of a solvent mixture of hexane and dodecane, we were able to increase the concentration of monomer up to 50-56% and obtain unusually large (up to 10 mum in diameter) PMMA particles. Second, by strictly controlling the nucleation time, we were able to make PMMA particles with a low polydispersity of around 1%, much lower than has ever before been achieved for such large particles. We also report an unusual apparent metastable state in the nucleation stage.


Mucogingival overgrowth in a geriatric patient.
Epulis fissuratum is a pathological condition caused by an ill-fitting denture. The mucogingival hyperplasia may be considered as a reactive condition of the oral mucosa to excessive mechanical pressure on the mucosa. Epulis fissuratum excision is a procedure usually done for prosthodontic reasons. The treatment of this benign entity is essential mainly for masticatory reasons. The use of diode laser for epulis removal without infiltrated anesthesia in a conscious geriatric patient is currently under investigation.


Olympic preparation in Brazilian judo athletes: description and perceived relevance of training practices.
The aim of this study was to describe the training routines used by judo athletes and their perception concerning the relevance, effort made, concentration needed, and pleasure obtained during the training sessions conducted 6 months before their Olympic participation and to compare with medal winners and other competitors in these aspects. Sixty-one Olympic Brazilian judo athletes (men = 39; women = 22), representing 66.3% of all Brazilian participants in this Olympic sport (from 1964 to 2008), including 10 medal winners (9 men and 1 woman) answered a questionnaire concerning their training routines. Mann-Whitney and Student's t-test for independent samples were used. Judo medalists and nonmedalists in the Olympic Games did not differ in: (a) the age when they started to practice and to compete in judo, (b) the age when they competed in the Olympic Games, (c) hours of training per week and per training session and the number of training sessions per day in their preparation for this event, (d) frequency and time spent for performing judo-specific and general exercises and their perceived relevance, effort, pleasure, and concentration for these activities performed during the preparation for the Olympic Games. The only differences found were the groundwork (ne-waza) randori practice, which was less frequently performed by medal winners, and perceived relevance attributed to this activity, which was considered less relevant by the medal winners compared with nonmedal winners. Thus, judo Olympic medal winners and nonmedalists did not differ in many training aspects in the final phase of their preparation to the Olympic Games.


Temperature-dependent water and ion transport properties of barley and sorghum roots : I. Relationship to leaf growth.
Root temperature strongly affects shoot growth, possibly via "nonhydraulic messengers" from root to shoot. In short-term studies with barley (Hordeum vulgare L.) and sorghum (Sorghum bicolor L.) seedlings, the optimum root temperatures for leaf expansion were 25 degrees and 35 degrees C, respectively. Hydraulic conductance (L(p)) of both intact plants and detached exuding roots of barley increased with increasing root temperature to a high value at 25 degrees C, remaining high with further warming. In sorghum, the L(p) of intact plants and of detached roots peaked at 35 degrees C. In both species, root temperature did not affect water potentials of the expanded leaf blade or the growing region despite marked changes in L(p). Extreme temperatures greatly decreased ion flux, particularly K(+) and NO(3) (-), to the xylem of detached roots of both species. Removing external K(+) did not alter short-term K(+) flux to the xylem in sorghum but strongly inhibited flux at high temperature in barley, indicating differences in the sites of temperature effects. Leaf growth responses to root temperature, although apparently "uncoupled" from water transport properties, were correlated with ion fluxes. Studies of putative root messengers must take into account the possible role of ions.


Exosomes isolation protocols: facts and artifacts for cardiac regeneration.
In recent years, exosomes have attracted increasing scientific interest and are no longer considered just as containers for cell waste, but as important mediators of intercellular communication. Among many biomedical research topics, a possible direct role of exosomes in the regenerative medicine field has been underlined in recent studies, including those regarding the so called "paracrine hypothesis". In this perspective, a therapeutic role and/or use of exosomes for tissue regeneration seems to be plausible. However, the majority of the cells isolated and cultured in vitro are exposed to an exogenous exosomes source because of the wide use of foetal bovine serum as cell culture supplement. Bovine serum has been gradually considered as a major biological stimulus, but with still unknown outcome. In this review, we present the state of the art about the role of exosomes in regenerative medicine, particularly for the cardiovascular system. We also analyse the most commonly used exosome isolation techniques that, since their discovery, have undergone continuous development to reach the highest degree of scalability for future clinical translation.


Cross-species hybridization with Fusarium verticillioides microarrays reveals new insights into Fusarium fujikuroi nitrogen regulation and the role of AreA and NMR.
In filamentous fungi, the GATA-type transcription factor AreA plays a major role in the transcriptional activation of genes needed to utilize poor nitrogen sources. In Fusarium fujikuroi, AreA also controls genes involved in the biosynthesis of gibberellins, a family of diterpenoid plant hormones. To identify more genes responding to nitrogen limitation or sufficiency in an AreA-dependent or -independent manner, we examined changes in gene expression of F. fujikuroi wild-type and DeltaareA strains by use of a Fusarium verticillioides microarray representing approximately 9,300 genes. Analysis of the array data revealed sets of genes significantly down- and upregulated in the areA mutant under both N starvation and N-sufficient conditions. Among the downregulated genes are those involved in nitrogen metabolism, e.g., those encoding glutamine synthetase and nitrogen permeases, but also those involved in secondary metabolism. Besides AreA-dependent genes, we found an even larger set of genes responding to N starvation and N-sufficient conditions in an AreA-independent manner. To study the impact of NMR on AreA activity, we examined the expression of several AreA target genes in the wild type and in areA and nmr deletion and overexpression mutants. We show that NMR interacts with AreA as expected but affects gene expression only in early growth stages. This is the first report on genome-wide expression studies examining the influence of AreA on nitrogen-responsive gene expression in a genome-wide manner in filamentous fungi.


Bilateral parotid tuberculosis.
Tuberculosis of parotid is a rare clinical entity, and cases of bilateral tubercular parotitis are even rarer. We present a case of bilateral primary parotid tuberculosis in a 49-year-old female. The patient received anti-tuberculosis treatment for six months, resulting in complete resolution of the disease. We also review the theories related to the pathogenesis of tubercular parotitis, and propose a novel hypothesis about greater involvement of parotid gland as compared to other salivary glands in primary tuberculosis.


A Two-Day Acceptance and Commitment Therapy (ACT) Workshop Increases Presence and Work Functioning in Healthcare Workers.
In this controlled naturalistic study performed in healthcare workers we examined the effect of a two-day acceptance commitment therapy (ACT) workshop on work presence and productivity, i.e. the influence the workshop had on treatment efficacy in a routine hospital care setting. To examine the influence of ACT on productivity and presence in healthcare workers. Study participants were all healthcare workers (nurses, medical doctors, physiotherapists, social workers, and art therapists) of four inpatient wards for depression. Half of the healthcare workers attended the workshop. Measures were evaluated 3 months after the intervention in the study participants and the patients treated by them in the same time period. A significantly higher treatment efficacy [as measured with HoNOS (Health of the nation outcome scales) change in the patients treated by the participants] has been observed in the healthcare workers who attended the ACT workshop when compared to the control group who did not attend the workshop. Moreover, the work presence of the participants of the ACT workshop was increased when compared with the time period before the intervention and with the presence of the control group. A cost analysis showed that ACT workshops lead to a significant return of investment for the employer as the costs for the workshop were ten times compensated by the increase of work presence in participants of the workshop. These findings provide support that ACT interventions motivate healthcare workers to work and increase their patients' treatment quality. To our knowledge this is the first study showing an ACT workshop in healthcare workers can influence HoNOS outcome in the treated patients.


Experimental and theoretical studies of 45Sc NMR interactions in solids.
Solid-state 45Sc NMR spectroscopy, ab initio calculations, and X-ray crystallography are applied to examine the relationships between 45Sc NMR interactions and molecular structure and symmetry. Solid-state 45Sc (I = 7/2) magic-angle spinning (MAS) and static NMR spectra of powdered samples of Sc(acac)3, Sc(TMHD)3, Sc(NO3)3.5H2O, Sc(OAc)3, ScCl3.6H2O, ScCl3.3THF, and ScCp3 have been acquired. These systems provide a variety of scandium coordination environments yielding an array of distinct 45Sc chemical shielding (CS) and electric field gradient (EFG) tensor parameters. Acquisition of spectra at two distinct magnetic fields allows for the first observations of scandium chemical shielding anisotropy (CSA). 45Sc quadrupolar coupling constants (CQ) range from 3.9 to 13.1 MHz and correlate directly with the symmetry of the scandium coordination environment. Single-crystal X-ray structures were determined for Sc(TMHD)3, ScCl3.6H2O, and Sc(NO3)3.5H2O to establish the hitherto unknown scandium coordination environments. A comprehensive series of ab initio calculations of EFG and CS tensor parameters are in excellent agreement with the observed parameters. Theoretically determined orientations of the NMR interaction tensors allow for correlations between NMR tensor characteristics and scandium environments. Solid-state 45Sc, 13C, and 19F NMR experiments are also applied to characterize the structures of the microcrystalline Lewis acid catalyst Sc(OTf)3 (for which the crystal structure is unknown) and a noncrystalline, microencapsulated, polystyrene-supported form of the compound.


Preclinical safety profile of RC88-ADC:a novel mesothelin-targeted antibody conjugated with Monomethyl auristatin E.
Mesothelin (MSLN) is an attractive therapeutic target for antibody drug conjugates (ADC) because of significant differences in expression pattern between diseased and normal tissues. RC88-ADC is a novel ADC, targeting MSLN, and inhibits tumor growth significantly in mice xenograft models. We performed an 11-week repeated dose toxicity study of RC88-ADC via intravenous injection in Cynomolgus Monkeys with an 8-Week recovery period according to International Conference on Harmonization (ICH) S9 and S6(R1). RC88-ADC was administered to groups of 5 male and 5 female monkeys at dose levels of 2.5, 5, and 10 mg/kg/2 weeks, meanwhile vehicle, naked antibody, small molecule groups were set up as the control. 4 animals died in 10 mg/kg group of RC88-ADC. The clinical symptoms mainly included ocular toxicity, weight loss and food intake decrease in the middle and high dose groups of RC88-ADC. RC88-ADC caused dose-related reversible myelosuppression, manifested as hematologic toxicity, which was consistent with the small molecule toxicity profile of its coupling. The highest non-severely toxic dose of RC88-ADC was 5 mg/kg in monkeys after repeated dosing. Nonetheless, the integrated analysis showed that RC88-ADC demonstrated an acceptable safety profile and provided an improved treatment window. These results pave the way for further investigation of RC88-ADC in humans.


The impact of COVID-19 on the elderly dependent population in Spain with special reference to the residential care sector.
The objective of this study is to analyze the residential care crisis in Spain in the context of the COVID-19 pandemic and its impact on high mortality and abandonment of the user population. The direct, indirect and structural causes are analyzed. Specifically, precarious employment in residences over the past decade was analyzed as one of the main explanatory causes of the structural crisis of nursing homes. The theoretical focus of analysis is the comprehensive and person-centered care (CPCC) model based on the autonomy of people and the centrality of their rights. The methodology combines a quantitative analysis of employment and a qualitative analysis of documents and debates. The study concludes by proposing a comprehensive reform of long-term care that includes both a change in residential care in the form of small cohabitation units and reinforcement of care in the home and the community as a growing preference for the elderly population. An optimal combination of residential and home care is the basic proposal of this work.


An operator at -280 base pairs that is required for repression of araBAD operon promoter: addition of DNA helical turns between the operator and promoter cyclically hinders repression.
A site has been found that is required for repression of the Escherichia coli araBAD operon. This site was detected by the in vivo properties of deletion mutants. In vitro protection studies with DNase I and dimethylsulfate showed that araC protein can specifically bind in this area to nucleotides lying at position -265 to -294 with respect to the araBAD operon promoter (PBAD) transcription start point. The previously known sites of protein binding in the ara operon lie between +20 and -160. Since the properties of deletion strains show that all the sites required for araBAD induction lie between +20 and -110, the new site at -280 exerts its repressive action over an unusually large distance along the DNA. Insertions of -16, -8, 0, 5, 11, 15, 24, and 31 base pairs of DNA between the new site and PBAD were constructed. Repression was impaired in those cases in which half-integral turns of the DNA helix were introduced, but repression was nearly normal for the insertions of 0, +11, and +31 base pairs.


Exploiting drug-resistant enzymes as tools to identify thienopyrimidinone inhibitors of human immunodeficiency virus reverse transcriptase-associated ribonuclease H.
The thienopyrimidinone 5,6-dimethyl-2-(4-nitrophenyl)thieno[2,3-d]pyrimidin-4(3H)-one (DNTP) occupies the interface between the p66 ribonuclease H (RNase H) domain and p51 thumb of human immunodeficiency virus reverse transcriptase (HIV RT), thereby inducing a conformational change incompatible with catalysis. Here, we combined biochemical characterization of 39 DNTP derivatives with antiviral testing of selected compounds. In addition to wild-type HIV-1 RT, derivatives were evaluated with rationally designed, p66/p51 heterodimers exhibiting high-level DNTP sensitivity or resistance. This strategy identified 3',4'-dihydroxyphenyl (catechol) substituted thienopyrimidinones with submicromolar in vitro activity against both wild type HIV-1 RT and drug-resistant variants. Thermal shift analysis indicates that, in contrast to active site RNase H inhibitors, these thienopyrimidinones destabilize the enzyme, in some instances reducing the Tm by 5 °C. Importantly, catechol-containing thienopyrimidinones also inhibit HIV-1 replication in cells. Our data strengthen the case for allosteric inhibition of HIV RNase H activity, providing a platform for designing improved antagonists for use in combination antiviral therapy.


Effects of Narrative Messages on Key COVID-19 Protective Responses: Findings From a Randomized Online Experiment.
We investigated the effectiveness of narrative vs non-narrative messages in changing COVID-19-related perceptions and intentions. The study employed a between-subjects two-group (narratives vs non-narratives) experimental design and was administered online. 1804 U.S. adults recruited via Amazon MTurk in September 2020 were randomly assigned to one of two experimental conditions and read either three narrative or three non-narrative messages about social distancing, vaccination, and unproven treatments. Perceptions and intentions were assessed before and after message exposure (7-point scales). Using multivariable regression, we assessed main effects of the experimental condition (controlling for baseline measures) and interactions between the condition and pre-exposure perceptions/intentions in predicting post-exposure outcomes. Compared to non-narratives, narratives led to (1) less positive perceptions about the benefits of unproven treatments (Mnarrative = 3.60, Mnon-narrative = 3.77, P = .007); and (2) less willingness to receive an unproven drug (Mnarrative = 3.46, Mnon-narrative = 3.77, P < .001); this effect was stronger among individuals with higher baseline willingness to receive unproven drugs (baseline willingness = 2.09: b = -.06, P = .461; baseline willingness = 3.90: b = -.30, P < .001; baseline willingness = 5.71: b = -.55, P < .001). Narratives also led to more positive perceptions of vaccine safety/effectiveness, but only among individuals with lower baseline vaccine perceptions (baseline perceptions = 4.51: b = .10, P = .008; baseline perceptions = 5.89: b = .04, P = .167; baseline perceptions = 7: b = -.01, P = .688). Narratives are a promising communication strategy, particularly for topics where views are not entrenched and among individuals who are more resistant to recommendations.


When the voluntary mind meets the irresistible event: stimulus-response correspondence effects on task selection during voluntary task switching.
In the present study, we investigated how task selection is biased by inherent stimulus characteristics in the voluntary task-switching paradigm. We used digits as the task stimuli, since they may automatically induce spatially horizontal representations of numbers. Specifically, we examined whether an irrelevant spatial representation of a number coincides with its associated response codes and whether such a stimulus-response (S-R) correspondence effect biases task selection for a digit. Participants were randomly assigned to one of two different action code layout conditions: Two numerical tasks were arranged as inner/outer in the horizontal layout condition or as upper/down in the vertical layout condition. Participants in the horizontal layout condition were more likely to choose a task when the task's action code and the digit's spatial representation corresponded, as compared with when they did not. On the other hand, no selection bias was observed in the vertical layout condition, since there was no overlapping spatial representation between the stimulus and response. The present study extends previous findings by considering the influence of the stimulus-driven effect on task selection with regard to the S-R correspondence effect.


Peripheral neuropathy as the presenting feature of tyrosinaemia type I and effectively treated with an inhibitor of 4-hydroxyphenylpyruvate dioxygenase.
A 21 month old girl presented with a short history of frequent falls and a right sided foot drop. She went on to suffer recurrent episodes of distal weakness in her arms and legs with hyporeflexia. Electrophysiological studies were consistent with inflammatory demyelinating polyradiculoneuropathy (IDP) and treatment with corticosteroids appeared to lead to an improvement. However, the development of hypertension, evidence of tubulopathy, and hepatomegaly led to re-evaluation. A diagnosis of type I tyrosinaemia was made, based on increased urinary excretion of succinylacetone and decreased activity of fumarylacetoacetase in her cultured skin fibroblasts. A low tyrosine diet did not prevent life-threatening exacerbations of neuropathy but intravenous haemarginate appeared to aid her recovery from one exacerbation. An immediate improvement in strength was seen after starting treatment with 2-(2-nitro-4-trifluoro-methyl-benzoyl)-1,3-cyclohexanedione (NTBC), an inhibitor of 4-hydroxy-phenylpyruvate dioxygenase. A liver transplant was performed but the patient died of immediate postoperative complications. Tyrosinaemia needs to be considered in a child with recurrent peripheral neuropathy because (i) the signs of liver disease and renal tubular dysfunction may be subtle; (ii) acute exacerbations may be life threatening; (iii) specific forms of treatment are available.


Loss of function of cinnamyl alcohol dehydrogenase 1 leads to unconventional lignin and a temperature-sensitive growth defect in Medicago truncatula.
There is considerable debate over the capacity of the cell wall polymer lignin to incorporate unnatural monomer units. We have identified Tnt1 retrotransposon insertion mutants of barrel medic (Medicago truncatula) that show reduced lignin autofluorescence under UV microscopy and red coloration in interfascicular fibers. The phenotype is caused by insertion of retrotransposons into a gene annotated as encoding cinnamyl alcohol dehydrogenase, here designated M. truncatula CAD1. NMR analysis indicated that the lignin is derived almost exclusively from coniferaldehyde and sinapaldehyde and is therefore strikingly different from classical lignins, which are derived mainly from coniferyl and sinapyl alcohols. Despite such a major alteration in lignin structure, the plants appear normal under standard conditions in the greenhouse or growth chamber. However, the plants are dwarfed when grown at 30 °C. Glycome profiling revealed an increased extractability of some xylan and pectin epitopes from the cell walls of the cad1-1 mutant but decreased extractability of others, suggesting that aldehyde-dominant lignin significantly alters cell wall structure.


SYVN1, an ERAD E3 Ubiquitin Ligase, Is Involved in GABAAα1 Degradation Associated with Methamphetamine-Induced Conditioned Place Preference.
Abuse of methamphetamine (METH), a powerful addictive amphetamine-type stimulants (ATS), is becoming a global public health problem. The gamma-aminobutyric acid (GABA)ergic system plays a critical role in METH use disorders. By using rat METH conditioned place preference (CPP) model, we previously demonstrated that METH-associated rewarding memory formation was associated with the reduction of GABAAα1 expression in the dorsal straitum (Dstr), however, the underlying mechanism was unclear. In the present study, we found that METH-induced CPP formation was accompanied by a significant increase in the expression of Synovial apoptosis inhibitor 1 (SYVN1), an endoplasmic reticulum (ER)-associated degradation (ERAD) E3 ubiquitin ligase, in the Dstr. The siRNA knockdown of SYVN1 significantly increased GABAAα1 protein levels in both primary cultured neurons and rodent Dstr. Inhibition of proteasomal activity by MG132 and Lactacystin significantly increased GABAAα1 protein levels. We further found that SYVN1 knockdown increased GABAAα1 in the intra-ER, but not in the extra-ER. Accordingly, endoplasmic reticulum stress (ERS)-associated Glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP) increased. Thus, this study revealed that SYVN1, as the ERAD E3 ubiquitin ligase, was associated with Dstr GABAAα1 degradation induced by METH conditioned pairing.


Understanding the Relationship between Depression and Chronic Diseases Such as Diabetes and Hypertension: A Grounded Theory Study.
There is a bi-directional relationship between depression and chronic illnesses such as diabetes and hypertension. This comorbidity is associated with higher mortality risk and diminishes the efficacy of interventions. The specific mechanisms of this mutual influence are still not fully understood, and most intervention protocols address these conditions separately. This study aims to improve our understanding of this relationship. We interviewed 18 patients and 24 health care professionals, focusing on understanding the different ways in which depression and chronic illness could influence each other. Our results show a common cyclical pattern and specific situations where the reported bi-directional relationship does not occur. We discuss the importance of opening a space for the patient's grief process after the chronic illness diagnosis, managing the demands and stress of the patient's treatment, and how to adjust the treatment to the different needs and possibilities of each person.


Effect of methoxamine on spontaneous motility of the isolated rat uterus.
In the present study the effect of methoxamine and its modification by several adrenergic antagonists is described in the isolated oestrogenized rat uterus. Methoxamine produced a dose-related increase in uterine activity, prazosin produced a rightward displacement of the dose-response curve of methoxamine reaching the same maximal effect. Yohimbine abolished the contractile response to methoxamine. propranolol or reserpine failed to modify the effect of methoxamine. Pretreatment with reserpine abolished the inhibitory effect of yohimbine. These results suggest the presence of excitatory alpha-adrenoceptors in the rat uterus.


Family Complexity among Children in the United States.
Researchers largely have relied on a measure of family structure to describe children's living arrangements, but this approach captures only the child's relationship to the parent(s), ignoring the presence and composition of siblings. We develop a measure of family complexity that merges family structure and sibling composition to distinguish between simple two-biological-parent families, families with complex-sibling (half or stepsiblings) arrangements, and complex-parent (stepparent, single-parent) families. Using the Survey of Income and Program Participation (SIPP), we provide a descriptive profile of changes in children's living arrangements over a 13-year span (1996-2009). SIPP sample sizes are sufficiently large to permit an evaluation of changes in the distribution of children in various (married, cohabiting, and single-parent) simple and complex families according to race/ethnicity and parental education. The article concludes by showing that we have reached a plateau in family complexity and that complexity is concentrated among the most disadvantaged families.


The NfeD protein family and its conserved gene neighbours throughout prokaryotes: functional implications for stomatin-like proteins.
NfeD-like proteins are widely distributed throughout prokaryotes and are frequently associated with genes encoding stomatin-like proteins (slipins). Here, we reveal that the NfeD family is ancient and comprises three major groups: NfeD1a, NfeD1b and truncated NfeD1b. Members of each group are associated with one of four conserved gene partners, three of which have eukaryotic homologues that are membrane raft associated, namely stomatin, paraslipin (previously SLP-2) and flotillin. The first NfeD group (NfeD1b), comprises proteins of approximately 460-aa long that have three functional domains: an N-terminal protease, a middle membrane-spanning region and a soluble C-terminal region rich in beta-strands. The nfeD1b gene is adjacent to eoslipin in prokaryotic genomes except in Firmicutes and Deinococci, where yqfA replaces eoslipin. Proteins in the second major group (NfeD1a) are homologous to the C-terminus of NfeD1b which forms a beta-barrel-like domain, and their genes are associated with paraslipin. Using OrthoMCL clustering, we show that nfeD1b genes have become truncated on many independent occasions giving rise to the third major group. These short NfeD homologues frequently remain associated with their ancestral gene neighbour, resembling NfeD1a in structure, yet are much more related to full-length NfeD1b; we term these "truncated NfeD1b". These conserved associations suggest that NfeD proteins are dependent on gene partners for their function and that the site of interaction may lie within the C-terminal portion that is common to all NfeD homologues. Although NfeD homologues are confined to prokaryotes, this conserved association could represent an excellent system to study slipin and flotillin proteins.


[Visual art, creativity and dementia].
Visual art is an expression of neurological function and how it organizes and interprets perception. The art is predominantly in the right hemisphere, in contrast, the left side, have inhibitory effects on artistic expression. In normal subjects, inhibitory and excitatory mechanisms could interact in a complex harmony, reflecting a paradoxical functional facilitation. Brain diseases such as dementia could change this harmony and then, alter the artistic abilities. Evaluate the art expression in the degenerative diseases. Artistic abilities of 3 painters with degenerative diseases were assessment. Patient 1: A 83 - year old right handed female, diagnosis: Alzheimer's disease. Artistic description: low productivity, simplified versions of earlier and alteration of the visuospatial organization. Patient 2: A 78-year-old right handed female, diagnosis: Primary Progressive Aphasia (PPA); Artistic description: oversimplified drawings which maintaining overall spatial organization, without impair artistic skills. Patient 3: A 68 year-old right handed woman, diagnosis: Fronto-Temporal Dementia (FTD). Artistic description: Increased artistic activity, originality, freedom, utilization of intense colours with perseverative and repetitive copying of similar paintings of her own work. Visual art in Alzheimer's disease is a consequence of visuospatial and constructive disabilities. In contrast, the conservation of this cognitive functions and left asymmetrical involved, in FTD and PPA respectively, suggest artistic preservation, independently of the language injury. The disproportionate functional prevalence of the right over the left could lead to a release of novelty - seeking in art and can contribute to emergent creativity. These observations suggest an organization for art in the brain and proposed bases for further investigations in dementias.


Intraoperative Flap Complications in LASIK Surgery Performed by Ophthalmology Residents.
To report the rate of flap-related complications in LASIK surgery performed by in-training ophthalmology residents and to analyze the risk factors for these complications. We analyzed 273 flap dissections in 145 patients from March 2013 to February 2014. We included all LASIK surgeries performed by 32 ophthalmology residents using a Moria M2 microkeratome. All the flap-related complications were noted. Comparison between both groups with and without complications was performed with an independent Student's t-test and relative risks were calculated. There were 19 flap-related complications out of the 273 flap dissections (6.95%). The most common complication was incomplete flap dissection (n = 10; 3.66%), followed by free-cap (n = 5; 1.83%), and flap-buttonhole (n = 2; 0.73%). There was no significant difference between the complicated and uncomplicated cases in terms of the right versus the left eye, pachymetry results, white-to-white diameter, and spherical equivalent. But this difference was significant for mean keratometry (P = 0.008), K-min (P = 0.01), and K-max (P = 0.03) between these groups. Final visual acuity after rescheduling laser treatment was similar in both groups. Relative risks for flap-related complications were 2.03 for the first LASIK surgery (CI 95% 0.64 to 6.48; P = 0.22) and 1.26 (CI 95% 0.43 to 3.69; P = 0.66) for the surgeon's flap-related complications. Female gender presented an odds ratio of 2.48 (CI 95% 0.68 to 9.00; P = 0.16) for complications. Flap-related complications are common intraoperative event during LASIK surgery performed by in-training ophthalmologists. Keratometries and surgeon's first procedure represent a higher probability for flap related complications than some other biometric parameters of patient's eye.


Targeted Manganese doped silica nano GSH-cleaner for treatment of Liver Cancer by destroying the intracellular redox homeostasis.
Background: Glutathione (GSH), the primary antioxidant in cells, could fight against oxidative stress. Tumor cells display a higher GSH level than normal cells for coping with the hyperoxidative state, which meets the requirements of enhanced metabolism and vicious proliferation. Therefore, the consumption of GSH will lead to cell redox imbalance and impede life activities. Herein, targeted sorafenib (SFB) loaded manganese doped silica nanoparticle (FaPEG-MnMSN@SFB) was constructed, which could destroy the intracellular redox homeostasis by consuming GSH. Methods: In this study, MnMSN was prepared by an optimized one-pot Stober's method for loading SFB, and FaPEG chain was modified on the surface of MnMSN to achieve long circulation and targeted delivery. The anticancer efficacy and mechanism of the designed FaPEG-MnMSN@SFB were assessed both in vitro and in vivo.Results: FaPEG-MnMSN@SFB exhibited efficient antitumor activity by dual depleting intracellular GSH (the degradation of MnMSN would consume intracellular GSH and the SFB would inhibit the effect of Xc- transport system to inhibit GSH synthesis). Moreover, disruption of redox balance would lead to apoptosis and reactive oxygen species (ROS)-dependent ferroptosis of tumor cells. Conclusion: Such a GSH-starvation therapeutic strategy would cause multi-path programmed cell death and could be a promising strategy for cancer therapy.


Chronic intestinal pseudoobstruction associated with autoantibodies against proliferating cell nuclear antigen.
Autoantibodies against proliferating cell nuclear antigen (PCNA) have been detected almost exclusively in sera from patients with systemic lupus erythematosus (SLE). Because of the extremely low prevalence of anti-PCNA, however, it has been difficult to establish its disease specificity and clinical associations. We present 2 cases in which patients with anti-PCNA developed severe gastrointestinal dysfunction that led to chronic intestinal pseudoobstruction. Neither patient had manifestations suggestive of SLE. Although sclerodermatous skin changes were minimal or absent in these patients, physical and radiographic findings indicated that systemic sclerosis was a likely cause of the gastrointestinal dysfunction. The similarity of the clinical features in these patients implies that the unusual detection of this unique autoantibody may not be a coincidence.


Sub-chronic restraint stress exposure in adult rats: An insight into possible inhibitory mechanism on testicular function in relation to germ cell dynamics.
Psychological stress is now widely recognized as one of the major risk factors for male fertility. Its impact on the dynamics of testicular germ cells, however, has yet to be fully investigated. Therefore, we used the rat restraint stress (RS) model as a psychological stressor to assess the impact of psychological stress on testicular germ cell dynamics. Adult male SD rats were exposed to sub-chronic RS for 1.5 and 3 h per day for 30 days. The quality of cauda epididymis spermatozoa was adversely affected by RS exposure, and the frequency of spermatozoa with tail abnormalities was higher than that of spermatozoa with head abnormalities. RS exposure adversely affected testicular daily sperm production by disturbing the meiotic and post meiotic germ cell kinetics in the testis. The histomorphology of the testis was altered by loosening and vacuolization in the seminiferous epithelium, germ cell exfoliation and the presence of giant cells. Seminiferous tubules of stage I-VI and VII-VIII were severely affected in rats exposed to RS for 3 h. By interfering with steroidogenic enzymes, RS exposure disrupts testosterone biosynthesis. The testicular oxidative balance was also disturbed by RS exposure, which disrupted the levels/activities of lipid peroxidation, Nrf-2, superoxide dismutase and catalase. There was also an increase in caspase-3 activity and a decrease in the Bax-Bcl2 ratio. In conclusion, our findings suggest that psychological stressors like RS impair testicular functions in rats by disrupting germ cell dynamics, downregulating testicular androgenesis and increasing oxidative stress and apoptosis.


Anthracycline derivatives inhibit cardiac CYP2J2.
Anthracycline chemotherapeutics are highly effective, but their clinical usefulness is hampered by adverse side effects such as cardiotoxicity. Cytochrome P450 2J2 (CYP2J2) is a cytochrome P450 epoxygenase in human cardiomyocytes that converts arachidonic acid (AA) to cardioprotective epoxyeicosatrienoic acid (EET) regioisomers. Herein, we performed biochemical studies to understand the interaction of anthracycline derivatives (daunorubicin, doxorubicin, epirubicin, idarubicin, 5-iminodaunorubicin, zorubicin, valrubicin, and aclarubicin) with CYP2J2. We utilized fluorescence polarization (FP) to assess whether anthracyclines bind to CYP2J2. We found that aclarubicin bound the strongest to CYP2J2 despite it having large bulky groups. We determined that ebastine competitively inhibits anthracycline binding, suggesting that ebastine and anthracyclines may share the same binding site. Molecular dynamics and ensemble docking revealed electrostatic interactions between the anthracyclines and CYP2J2, contributing to binding stability. In particular, the glycosamine groups in anthracyclines are stabilized by binding to glutamate and aspartate residues in CYP2J2 forming salt bridge interactions. Furthermore, we used iterative ensemble docking schemes to gauge anthracycline influence on EET regioisomer production and anthracycline inhibition on AA metabolism. This was followed by experimental validation of CYP2J2-mediated metabolism of anthracycline derivatives using liquid chromatography tandem mass spectrometry fragmentation analysis and inhibition of CYP2J2-mediated AA metabolism by these derivatives. Taken together, we use both experimental and theoretical methodologies to unveil the interactions of anthracycline derivatives with CYP2J2. These studies will help identify alternative mechanisms of how anthracycline cardiotoxicity may be mediated through the inhibition of cardiac P450, which will aid in the design of new anthracycline derivatives with lower toxicity.


Functional and structural alterations in the pain-related circuit in major depressive disorder induced by electroconvulsive therapy.
Approximately two-thirds of major depressive disorder (MDD) patients have pain, which exacerbates the severity of depression. Electroconvulsive therapy (ECT) is an efficacious treatment that can alleviate depressive symptoms; however, treatment for pain and the underlying neural substrate is elusive. We enrolled 34 patients with MDD and 33 matched healthy controls to complete clinical assessments and neuroimaging scans. MDD patients underwent second assessments and scans after ECT. We defined a pain-related network with a published meta-analysis and calculated topological patterns to reveal topologic alterations induced by ECT. Using the amplitude of low-frequency fluctuations (ALFFs), we probed local function aberrations of pain-related circuits in MDD patients. Subsequently, we applied gray matter volume (GMV) to reveal structural alterations of ECT relieving pain. The relationships between functional and structural aberrations and pain were determined. ECT significantly alleviated pain. The neural mechanism based on pain-related circuits indicated that ECT weakened the circuit function (ALFF: left amygdala and right supplementary motor area), while augmenting the structure (GMV: bilateral amygdala/insula/hippocampus and anterior cingulate cortex). The topologic patterns became less efficient after ECT. Correlation analysis between the change in pain and GMV had negative results in bilateral amygdala/insula/hippocampus. Similarity, there was a positive correlation between a change in ALFF in the left amygdala and improved clinical symptoms. ECT improved pain by decreasing brain local function and global network patterns, while increasing structure in pain-related circuits. Functional and structural alterations were associated with improvement in pain.


S. cerevisiae induces apoptosis in human metastatic breast cancer cells by altering intracellular Ca2+ and the ratio of Bax and Bcl-2.
We have recently demonstrated that non-metastatic human breast cancer cell lines undergo apoptosis following phagocytosis of S. cerevisiae. In this study, we investigated the apoptotic effect of heat-killed yeast against human metastatic breast cancer (MBC) cells, MDA-MB-231 in vitro, and the underlying mechanistic bases of this effect. Results show that monolayer MDA-MB-231 cells phagocytized yeast (50% at 16 h) and underwent apoptosis (32% compared with 7.6% of untreated cells, representing a 4.2-fold increase). The increase in apoptosis was associated with an elevation of [Ca2+]I. Addition of 2-aminoethoxydiphenyl borate (2APB), a pharmacological inhibitor of Ca2+ release from the endoplasmic reticulum, effectively diminished yeast-induced apoptosis. Furthermore, yeast caused a substantial decrease in expression of Bcl-2 and an increase in Bax resulting in alteration in the Bax:Bcl-2 ratio. However, yeast had no effect on NO levels. In conclusion, yeast induces apoptosis of human MBC cells in vitro by a mechanism involving intracellular Ca2+ and Bax:Bcl-2.


Development of visual predictive checks accounting for multimodal parameter distributions in mixture models.
The assumption of interindividual variability being unimodally distributed in nonlinear mixed effects models does not hold when the population under study displays multimodal parameter distributions. Mixture models allow the identification of parameters characteristic to a subpopulation by describing these multimodalities. Visual predictive check (VPC) is a standard simulation based diagnostic tool, but not yet adapted to account for multimodal parameter distributions. Mixture model analysis provides the probability for an individual to belong to a subpopulation (IPmix) and the most likely subpopulation for an individual to belong to (MIXEST). Using simulated data examples, two implementation strategies were followed to split the data into subpopulations for the development of mixture model specific VPCs. The first strategy splits the observed and simulated data according to the MIXEST assignment. A shortcoming of the MIXEST-based allocation strategy was a biased allocation towards the dominating subpopulation. This shortcoming was avoided by splitting observed and simulated data according to the IPmix assignment. For illustration purpose, the approaches were also applied to an irinotecan mixture model demonstrating 36% lower clearance of irinotecan metabolite (SN-38) in individuals with UGT1A1 homo/heterozygote versus wild-type genotype. VPCs with segregated subpopulations were helpful in identifying model misspecifications which were not evident with standard VPCs. The new tool provides an enhanced power of evaluation of mixture models.


Making my mark.
Neil Brimblecombe explains how his first research project was carried out by accident, and how he has come to learn why stubbornness and resilience are such important characteristics for the nurse researcher to possess.


The National Security Implications of Cyberbiosecurity.
The cyber- and biological sciences are converging rapidly, creating benefits, new and advantageous applications, and increasing risks to all nations. The parts of the public and private sectors that should be responsible for cyberbiosecurity are not yet sufficiently organized or supported financially. This article addresses the need to ensure that national security policy: (1) assesses cyberbiological risk and incorporates deterrent and enforcement measures; (2) sets forth clear consequences for those individuals and countries that conduct cyberbiological attacks or otherwise compromise cyberbiosecurity, without imperiling the legitimate sharing of scientific data and information; (3) establishes voluntary cyberbiosecurity standards in partnership with the private sector; (4) identifies cyberbiosecurity threats, vulnerabilities, consequences, and solutions; and (5) results from the combined efforts of all branches of government and the private sector.


Peptide display in functional genomics.
The completion of the human genome project has opened novel scientific avenues in functional genomics, structural genomics and proteomics. These areas have a common goal: the identification of all the proteins acting and cross-talking in a single cell at a defined moment of its lifecycle. The expansion of these areas in bioscience has been facilitated by the rapid development of high throughput screening (HTS) methods which has, in turn, attracted the business community to make investments in this novel business segment of biotechnology. By using these HTS methods, the hope is that novel targets will be validated much more rapidly speeding up the development of novel drugs. Numerous techniques and tools have emerged over the past decade for the identification of small target-specific molecular ligands that exploit a common feature: the exploration of molecular diversity using combinatorial methods. While chemists developed new methods for rapidly and efficiently synthesising and screening large collections of small molecules, biologists used recombinant DNA techniques for selecting displayed repertoires. To this end, the discovery of new low molecular weight peptides is becoming increasingly important, not only as molecular tools for the understanding of protein-protein interactions but also for the generation of lead compounds.


Effects of an aqueous extract of Aloe buettneri, Justicia insularis, Hibiscus macranthus, Dicliptera verticillata on some physiological and biochemical parameters of reproduction in immature female rats.
Different doses of aqueous extracts from the leaves of Aloe buettneri, Justicia insularis, Hibiscus macranthus and Dicliptera verticillata, locally used to regulate the menstrual cycle and to treat dysmenorrhea or infertility in women, were given daily to 22 day old rats for 5, 10, 15, 20 and 25 days by gastric intubation. At the end of each experimental period, the weights of ovaries and uteri, levels of uterine and ovarian proteins, ovarian cholesterol and serum oestradiol were evaluated. The results showed a decrease in growth rate of animals treated with 94 mg/kg per day at the end of the experimental period. The ovarian and uterine weights were high in all treated groups especially within the pubertal period (36-41 days old) when compared to the respective controls. During the same period, the parameters analysed showed significant differences in treated rats when compared to the controls. This was seen with ovarian and uterine protein levels, as well as with serum oestradiol whose level was especially high in the groups given 49 or 94 mg/kg per day of the plant extracts (52 and 42%, respectively when compared with the respective controls). A concomitant decrease in ovarian cholesterol was observed in the same treated groups. These results suggested a possible presence of oestrogenic compounds in the plant extracts.


Adult-Onset Still's Disease: A Case Report.
Adult-onset Still's disease (AOSD) is a disorder that is occasionally seen. Autoinflammation is a generally accepted pathogenic mechanism leading to systemic signs and symptoms. We report the case of a young female presenting with high-grade fever, rash, and arthralgias. After a thorough assessment, the diagnosis of adult-onset Still's disease was made based on presenting symptoms and elevated serum ferritin, c-reactive protein (CRP), and absence of serologic markers and confirmed based on Yamaguchi criteria. She was treated with corticosteroids and achieved complete clinical remission.


Transition-Tempered Metadynamics Is a Promising Tool for Studying the Permeation of Drug-like Molecules through Membranes.
Metadynamics is an important enhanced sampling technique in molecular dynamics simulation to efficiently explore potential energy surfaces. The recently developed transition-tempered metadynamics (TTMetaD) has been proven to converge asymptotically without sacrificing exploration of the collective variable space in the early stages of simulations, unlike other convergent metadynamics (MetaD) methods. We have applied TTMetaD to study the permeation of drug-like molecules through a lipid bilayer to further investigate the usefulness of this method as applied to problems of relevance to medicinal chemistry. First, ethanol permeation through a lipid bilayer was studied to compare TTMetaD with nontempered metadynamics and well-tempered metadynamics. The bias energies computed from various metadynamics simulations were compared to the potential of mean force calculated from umbrella sampling. Though all of the MetaD simulations agree with one another asymptotically, TTMetaD is able to predict the most accurate and reliable estimate of the potential of mean force for permeation in the early stages of the simulations and is robust to the choice of required additional parameters. We also show that using multiple randomly initialized replicas allows convergence analysis and also provides an efficient means to converge the simulations in shorter wall times and, more unexpectedly, in shorter CPU times; splitting the CPU time between multiple replicas appears to lead to less overall error. After validating the method, we studied the permeation of a more complicated drug-like molecule, trimethoprim. Three sets of TTMetaD simulations with different choices of collective variables were carried out, and all converged within feasible simulation time. The minimum free energy paths showed that TTMetaD was able to predict almost identical permeation mechanisms in each case despite significantly different definitions of collective variables.


Hypoplastic Left Ventricle: Definition, Morphology, and Classification of the Cardiac Phenotypes.
A prior manuscript provided information about hypoplastic left heart syndrome (HLHS) and related malformations, including definitions, morphology, and classification, based on the 2021 International Paediatric and Congenital Cardiac Code (IPCCC) and the Eleventh Revision of the International Classification of Diseases (ICD-11). This manuscript provides information about the related cardiac phenotypes associated with "hypoplastic left ventricle" that might be suitable for biventricular repair, including definitions, morphology, and classification of the cardiac phenotypes of the following four congenital cardiac malformations that can all include a hypoplastic left ventricle: (1) hypoplastic left heart complex (HLHC), (2) critical aortic stenosis with left ventricular hypoplasia, (3) aortic atresia + ventricular septal defect, and (4) hypoplastic left ventricle with severely unbalanced atrioventricular septal defect.


Selenium and the thyroid: how the relationship was established.
Several hypotheses concerning consequences of selenium deficiency on iodine metabolism can be proposed on the basis of experimental studies in rats and from epidemiological and experimental studies in humans. By decreasing intracellular GSH peroxidase activity, selenium deficiency may increase hydrogen peroxide (H2O2) supply and lead over several weeks to the thyroid atrophy observed in myxoedematous cretins. By improving thyroid hormone synthesis and by decreasing peripheral thyroxin (T4) deiodination, selenium deficiency could protect fetal brain T4 supply and thus prevent neurologic cretinism. Selenium deficiency may protect against iodine deficiency by decreasing T4 metabolism--and thus iodide leakage and--perhaps also by increasing H2O2 supply and thyroid hormone synthesis and thus thyroid efficiency.


[How are the Blaschko lines arranged on the scalp?].
Linear inherited or nevoid skin lesions mostly follow a characteristic pattern which was delineated by Blaschko in 1901. The system of lines published by this author covered the entire human body, with the exception of the scalp because of lack of pertinent case reports regarding this area. We here report an observation which enables us to fill the blank area in the atlas of the lines of Blaschko. On the scalp of a girl affected with the oral-facial-digital syndrome, we observed an alopecia distributed in several spiral streaks. The pattern resembled that of the normal vertex, but the direction of the hairless streaks did not correspond completely to the direction of hair growth. As the oral-facial-digital syndrome is inherited as an X-linked dominant trait, the linear alopecia probably reflects functional X-chromosome mosaicism. Arguments are presented in favor of the assumption that the hairless streaks of this child follow the lines of Blaschko.


A quantitative polymerase chain reaction assay for rapid detection of 9 pathogens directly from stools of travelers with diarrhea.
Every year, 80 million tourists traveling to tropical and subtropical areas contract traveler's diarrhea (TD). Forty percent to 80% of cases are caused by bacteria, yet clinical diagnostic tests are available to identify only a few of the strains that cause TD. We aimed to develop a quantitative polymerase chain reaction (qPCR) assay to identify all major pathogens in stool samples. We developed a low-cost, high-throughput, multiplex qPCR assay for simultaneous detection of 9 bacterial pathogens in stool samples: Salmonella, Yersinia, Campylobacter, and Vibrio cholerae, as well as Shigella or enteroinvasive Escherichia coli, enterohemorrhagic E coli, enterotoxigenic E coli (ETEC), enteroaggregative E coli (EAEC), and enteropathogenic E coli (EPEC). The assay was validated using positive (n = 245) and negative (n = 243) control strains, as well as preselected positive and negative stool samples. In addition, stool samples were collected from 96 returning travelers with TD. The findings were compared with those from routine diagnostic tests. The assay detected the bacterial strains with 100% sensitivity and specificity, compared with results from the reference tests. Of all stool samples collected from travelers with TD, EPEC was found in 47%, EAEC in 46%, ETEC in 22%, enterohemorrhagic E coli in 7%, Campylobacter in 6%, Shigella or enteroinvasive E coli in 2%, and Salmonella in 2%. Multiple pathogens were found in 37% of all samples. We developed a low-cost, high-throughput qPCR assay for use in routine diagnostic analysis and research. It detects the pathogenic bacteria most commonly associated with TD in stool samples with 100% sensitivity and specificity, compared with reference methods. The assay requires 4 hours, whereas current detection methods require 1 to 7 days. At least 1 TD pathogen was identified in stool samples from 76% of returning travelers, whereas conventional methods found a pathogen in only 17%. The most commonly detected bacteria were EPEC, EAEC, and ETEC.


Ultrafast ultrasonic imaging coupled to rheometry: principle and illustration.
We describe a technique coupling standard rheology and ultrasonic imaging with promising applications to characterization of soft materials under shear. Plane wave imaging using an ultrafast scanner allows to follow the local dynamics of fluids sheared between two concentric cylinders with frame rates as high as 10 000 images per second, while simultaneously monitoring the shear rate, shear stress, and viscosity as a function of time. The capacities of this "rheo-ultrasound" instrument are illustrated on two examples: (i) the classical case of the Taylor-Couette instability in a simple viscous fluid and (ii) the unstable shear-banded flow of a non-Newtonian wormlike micellar solution.


A comparison of adolescent and adult defense styles.
Despite the great need for assessment of the adaptive and defensive style of adolescents, there are few instruments available to assess these characteristics. We assessed Vaillant's defense hierarchial model in adolescents, using Bond's self-report questionnaire (DSQ) for adults adopted for adolescents. In all cases, the factor structure in adolescents differed from adults, which may represent a manifestation of development.


Isolation and identification of a novel bacterium, Pseudomonas sp. ZyL-01, involved in the biodegradation of CL-20.
Hexanitrohexaazaisowurtzitane (CL-20) is a compound with a polycyclic cage and an N-nitro group that has been shown to play an unfavorable role in environmental fate, biosafety, and physical health. The aim of this study was to isolate the microbial community and to identify a single microbial strain that can degrade CL-20 with desirable efficiency. Metagenomic sequencing methods were performed to investigate the dynamic changes in the composition of the community diversity. The most varied genus among the microbial community was Pseudomonas, which increased from 1.46% to 44.63% during the period of incubation (MC0-MC4). Furthermore, the new strain was isolated and identified from the activated sludge by bacterial morphological and 16s rRNA sequencing analyses. The CL-20 concentrations decreased by 75.21 μg/mL and 74.02 μg/mL in 48 h by MC4 and Pseudomonas sp. ZyL-01, respectively. Moreover, ZyL-01 could decompose 98% CL-20 of the real effluent in 14 day's incubation with the glucose as carbon source. Finally, a draft genome sequence was obtained to predict possible degrading enzymes involved in the biodegradation of CL-20. Specifically, 330 genes that are involved in energy production and conversion were annotated by Gene Ontology functional enrichment analysis, and some of these candidates may encode enzymes that are responsible for CL-20 degradation. In summary, our studies indicate that microbes might be a valuable biological resource for the treatment of environmental contamination caused by CL-20 and that Pseudomonas sp. ZyL-01 might be a promising candidate for eradicating CL-20 to achieve a more biosafe environment and improve public health.


Primary brain amyloidoma, both a neoplastic and a neurodegenerative disease: a case report.
Scattered extracellular deposits of amyloid within the brain parenchyma can be found in a heterogeneous group of diseases. Its condensed accumulation in the white matter without evidence for systemic amyloidosis is known as primary brain amyloidoma (PBA). Although originally considered as a tumor-like lesion by its space-occupying effect, this condition displays also common hallmarks of a neurodegenerative disorder. A 50-year-old woman presented with a mild cognitive decline and seizures with a right temporal, irregular and contrast-enhancing mass on magnetic resonance imaging. Suspecting a high-grade glioma, the firm tumor was subtotally resected. Neuropathological examination showed no glioma, but distinct features of a neurodegenerative disorder. The lesion was composed of amyloid AL λ aggregating within the brain parenchyma as well as the adjacent vessels, partially obstructing the vascular lumina. Immunostaining confirmed a distinct perivascular inflammatory reaction. After removal of the PBA, mnestic impairments improved considerably, the clinical course and MRI-results are stable in the 8-year follow-up. Based on our histopathological findings, we propose to regard the clinicopathological entity of PBA as an overlap between a neoplastic and neurodegenerative disorder. Since the lesions are locally restricted, they might be amenable to surgery with the prospect of a definite cure.


Low molecular weight organic compounds of chestnut wood (Castanea sativa L.) and corresponding aged brandies.
Oak and chestnut species have been largely used for the aging of brandies, but nowadays chestnut is rarely used. There have been no previous studies regarding the cooperage utilization of chestnut wood. This study provides, for the first time, specific information about the characterization of the northern Portuguese Castanea sativa wood and examines the influence of this wood and its heat treatment on the chemical composition of two-year-aged brandies, by the quantitative determination (HPLC) of low molecular weight phenolic compounds. The predominance of gallic acid among the analyzed extractable compounds both in chestnut wood and in the corresponding aged brandies was remarkable. The heat treatment has a very significant influence on the majority of extractable compounds analyzed. Thus, it could be responsible for the related sensorial properties of aged brandies and greatly affect their general balance.


How Does Urban Green Space Impact Residents' Mental Health: A Literature Review of Mediators.
In recent years, the interest in the relationship between urban green space and residents' mental health has gradually risen. A number of researchers have investigated the causal relationship and possible mediators between the two, although few have summarized these mediators. For this reason, we searched for relevant studies and filtered them by criteria and quality score, and analyzed the mediators and paths of the impact of urban green space on residents' mental health. The mediators can be divided into environmental factors, outdoor activity, and social cohesion. From the perspective of heterogeneity, both individual characteristics (e.g., age and gender) and group characteristics (e.g., level of urban development and urban density) of residents are considered to be the cause of various mediating effects. Types of urban green space tend to affect residents' mental health through different paths. Furthermore, this review discusses the details of each part under the influence paths. Finally, the policy implications for urban green space planning from three mediator levels are put forward based on an analysis of the situation in different countries.


Transplastomic Plants Homoplasmic for Foreign Transgenes.
Scientists pay more and more attention to the research on plastid engineering for its following advantages foreign genes can be integrated site-specifically into the plastid genome (plastome) there are no position effects as experienced with random insertion of transgenes in nuclear transformation pollen-mediated dispersion of transgenes can be avoided because chloroplasts are maternally transmitted gene silencing does not occur in plastids and therefore transgene expression is stable in progeny of transplastomic plants and the high ploidy level of the plastome in leaf cells makes high levels of transgene expression feasible. At the same time, however, the highly polyploid plastome makes it difficult to get transplastomic plants homoplasmic for foreign transgenes. In this work, chloroplast transforming vector pTRCH205, which carries two psbA5'-nifH-psbA3'and Prrn-aadA-TpsbA cassettes flanked by plastid DNA sequence to target their insertion between psbA and trnK operons, was constructed. Plastid transformation of Nicotiana tabacum was carried out by the biolistic delivery of transforming plasmid pTRCH205 DNA into leaf cells. Integration of nifH and aadA by two homologous recombination events via the flanking ptDNA sequences, and selective amplification of the transplastomes on MS medium with high concentration of spectinomycin, yielded resistant cell lines. All the independent transplastomic lines were subjected to three additional rounds of regeneration and were subcultured for 6--10 times through stem sections on MS medium containing 500 mg/L spectinomycin, to obtain homoplasmic tissues. The results of PCR assay and Southern blot hybridization, probed with 0.9 kb BglII/SnaBI homologous fragment, indicated that foreign genes had been integrated into the plastomes of transgenic plants, which finally became homoplasmic for foreign transgenes.


Neurite outgrowth-regulating properties of GABA and the effect of serum on mouse spinal cord neurons in culture.
Time-lapse photography was used to examine the effects of gamma-aminobutyric acid (GABA) on the outgrowth and motility of neurites in cultures from mouse spinal cord. GABA at concentrations of 100, 10 and 1 microM caused significant inhibition of neurite outgrowth and the motility of growth cones was significantly reduced by treatment with 100 and 10 microM GABA. This effect was mimicked by the GABA(B) receptor agonist baclofen, whereas the GABA(A) receptor agonist muscimol had no effect. The effect of GABA on outgrowth and motility seems to be dependent on the type of serum employed. The results reported here were obtained only when heat-inactivated serum was used and not when non heat-inactivated serum was added to the culture medium. They suggest that GABA has a role in the regulation of process outgrowth within the embryonic mouse spinal cord.


Kinetic analysis of two types of Na+ channels in rat dorsal root ganglia.
1. The gating properties of two types of Na+ channels were studied in neurones isolated from rat dorsal root ganglia using the whole cell variation of the patch electrode voltage-clamp technique. 2. Two types of Na+ currents (INa) were identified on the basis of their sensitivity to tetrodotoxin (TTX). One type was insensitive to TTX (up to 0.1 mM), while the other type was blocked by 1 nM of TTX. Whereas they were both insensitive to 50 microM Cd2+, a high concentration (2 mM) of Co2+ selectively inhibited the TTX-insensitive type. 3. The activation thresholds were about -60 and -40 mV for the TTX-sensitive and the TTX-insensitive INa, respectively. Activation of the TTX-sensitive INa developed with a sigmoidal time course which was described by m3 kinetics, whereas the activation of the TTX-insensitive INa was described by a single exponential function. A deactivation process, as measured by the tail current upon repolarization, followed an exponential decay in either type of INa. 4. The rate constant of activation indicated that under comparable membrane potential conditions, the TTX-insensitive channels open 4-5 times slower than the TTX-sensitive ones upon depolarization. Likewise, the rate constant of inactivation indicated that the TTX-insensitive channels inactivate 3-7 times more slowly than the TTX-sensitive ones upon repolarization. 5. The steady-state activation curve for the TTX-insensitive INa was shifted about 20 mV in the positive direction from that for the TTX-sensitive INa. 6. The steady-state inactivation curve for the TTX-insensitive INa as obtained with a 0.5 s prepulse was shifted about 26 mV in the positive direction from that for the TTX-sensitive INa, indicating a greater availability for the TTX-insensitive INa in depolarized membrane. However, on increasing the duration of prepulse, the inactivation curve for the TTX-insensitive INa, but not for the TTX-sensitive INa, shifted in the negative direction due to an extremely slow inactivation process in the TTX-insensitive INa. Consequently, an overlap between the activation and inactivation curves which causes a steady influx of Na+ (window current) became progressively reduce. 7. The time course of INa decay was best described by a single exponential process in either the TTX-sensitive or TTX-insensitive INa, whereas the development of inactivation and the recovery from inactivation, which were measured by a conventional double-pulse protocol, followed a second order process in either channel type.(ABSTRACT TRUNCATED AT 400 WORDS)


[Dilemmas in definition and classification of adenocarcinoma of esophagogastric junction: from history to current status].
In recent years, adenocarcinoma of esophagogastric junction (AEG) has received increased attention from the academic community. However, the esophagogastric junction (EGJ) straddles two anatomical regions: the thoracic cavity and the abdominal cavity. The histological features of the EGJ are different from those of the esophagus and stomach. There are general disagreements among the related disciplines regarding the definition and classification of AEG. By summarizing the views of different disciplines, including endoscopy, radiography, and pathology, a more comprehensive definition of the EGJ was formulated in the Japanese Classification of Gastric Carcinoma (the 15th edition), and the principle of endoscopic diagnostic priority was established. In recent years, with the development of physiological and anatomical studies, the EGJ has gradually expanded conceptually into a complex functional anatomical region covering the distal esophagus to the proximal stomach. The venous and lymphatic vessels in the EGJ are characterized by bidirectional flow, which is an important anatomical basis for the invasion and metastasis patterns of tumors in this region. The clinical practice of EGJ cancer has been promoted by the creation of Nishi and Siewert classification systems. With the support of a series of clinical studies for its scientificity and effectiveness, the Siewert classification is widely accepted by the international community, and successively introduced into major international practice guidelines. In general, the staging and management of Siewert Ⅰ and Ⅱ AEG are recommended as esophageal cancer, while Siewert Ⅲ AEG is recommended for gastric cancer. However, in the Japanese guidelines for the treatment of esophageal and gastric cancers, the Nishi classification is still used to define and classify EGJ cancer. Recent year, a Chinese consensus on the surgical treatment of AEG was formulated by multidisciplinary experts. The main controversies were summarized in the consensus, and proposals that incorporate the domestic situation were also presented. At present, only by returning to the basic anatomical and physiological perspectives, strengthening multidisciplinary communication and cooperation, and with the help of emerging bioinformatics, digital, and material technology, can it be possible to get out of the dilemma faced by traditional AEG classification and staging system.


Spinal cord oligodendrocytes develop from a limited number of migratory highly proliferative precursors.
Oligodendrocytes are responsible for myelin formation in spinal cord white matter. In the mature spinal cord, the majority of white matter is localized peripherally. During early development, however, the first oligodendrocyte precursors arise in the ventral ventricular zone of the developing cord. Thus, prior to myelination, both migration and proliferation of oligodendrocyte precursors must occur. When and where these events occur is currently unclear. In the chick spinal cord, oligodendrocyte precursors express antigens recognized by the monoclonal antibody O4. Here we show that all chick spinal cord oligodendrocytes are derived from O4+ cells and all O4+ cells appear to give rise to oligodendrocytes. Analysis of the number and distribution of oligodendrocyte precursors in chick spinal cord at different stages of development suggests that relatively few cells migrate from the ventricular source which then proliferate extensively in white matter. This migration is guided by general dispersive cues. Clonal analysis of oligodendrocyte development in cultures derived from different regions of the rodent spinal cord indicated that the cells that initially populate dorsal and peripheral spinal cord retained similar clonal properties to those in ventral spinal cord, suggesting the migrating cells were immature, highly proliferative precursors. Consistent with these results, BrdU incorporation studies indicate that glial proliferation is extensive and persistent in postnatal rat spinal cord white matter. Together, these studies suggest that spinal cord white matter is initially populated by very immature precursors that then undergo extensive local proliferation prior to myelination.


Diagnosis of brainstem abscess in the cerebritis stage by magnetic resonance imaging--case report.
A 52-year-old male presented with a brainstem abscess manifesting as high fever, diplopia, and left hemiparesis. Magnetic resonance (MR) imaging with gadolinium diethylenetriamine pentaacetic acid showed the lesion as a ring-like enhanced mass consisting of a necrotic center with surrounding edema, whereas postcontrast computed tomography revealed no such confirmatory findings. He was treated with antibiotics as the lesion had been detected in the acute cerebritis stage. Serial MR images showed that the lesion decreased remarkably in size. MR imaging can detect brain abscess in the earliest inflammatory stage.