Quantification of in vivo metabolic kinetics of hyperpolarized pyruvate in rat kidneys using dynamic 13C MRSI.
With signal-to-noise ratio enhancements on the order of 10,000-fold, hyperpolarized MRSI of metabolically active substrates allows the study of both the injected substrate and downstream metabolic products in vivo. Although hyperpolarized [1-(13)C]pyruvate, in particular, has been used to demonstrate metabolic activities in various animal models, robust quantification and metabolic modeling remain important areas of investigation. Enzyme saturation effects are routinely seen with commonly used doses of hyperpolarized [1-(13)C]pyruvate; however, most metrics proposed to date, including metabolite ratios, time-to-peak of metabolic products and single exchange rate constants, fail to capture these saturation effects. In addition, the widely used small-flip-angle excitation approach does not correctly model the inflow of fresh downstream metabolites generated proximal to the target slice, which is often a significant factor in vivo. In this work, we developed an efficient quantification framework employing a spiral-based dynamic spectroscopic imaging approach. The approach overcomes the aforementioned limitations and demonstrates that the in vivo (13)C labeling of lactate and alanine after a bolus injection of [1-(13)C]pyruvate is well approximated by saturatable kinetics, which can be mathematically modeled using a Michaelis-Menten-like formulation, with the resulting estimated apparent maximal reaction velocity V(max) and apparent Michaelis constant K(M) being unbiased with respect to critical experimental parameters, including the substrate dose, bolus shape and duration. Although the proposed saturatable model has a similar mathematical formulation to the original Michaelis-Menten kinetics, it is conceptually different. In this study, we focus on the (13)C labeling of lactate and alanine and do not differentiate the labeling mechanism (net flux or isotopic exchange) or the respective contribution of various factors (organ perfusion rate, substrate transport kinetics, enzyme activities and the size of the unlabeled lactate and alanine pools) to the labeling process.


Toll-like receptor signalling pathways as key targets for mediating the anti-inflammatory and immunosuppressive effects of glucocorticoids.
Toll-like receptors (TLRs) play crucial roles in the induction of innate immune responses by recognising pathogen-associated molecular patterns. The engagement of TLRs by pathogens results in induction of co-stimulatory molecules that facilitate a specific immune response and also in the induction of pro-inflammatory proteins that will promote the elimination of pathogens from the body. TLRs employ many of the same signalling components as the type I interleukin (IL)-1 receptor (IL-1R). This is hardly surprising since the intracellular regions of TLRs and the IL-1R share a conserved Toll/IL-1R homology domain (TIR) that allows the receptors to recruit the intracellular TIR-containing adaptor protein Myd88. The latter then activates IL-1R-associated kinases that in turn recruit well-characterised downstream effectors culminating in activation of MAP kinases and transcription factors such as NFkappaB and AP-1. Since glucocorticoids are known to target the latter transcription factors and the MAP kinase cascades, this commentary highlights the likely crucial importance of Toll-like receptor signalling pathways as key targets for mediating the anti-inflammatory and immunosuppressive effects of steroids.


Isolated polycystic liver disease genes define effectors of polycystin-1 function.
Dominantly inherited isolated polycystic liver disease (PCLD) consists of liver cysts that are radiologically and pathologically identical to those seen in autosomal dominant polycystic kidney disease, but without clinically relevant kidney cysts. The causative genes are known for fewer than 40% of PCLD index cases. Here, we have used whole exome sequencing in a discovery cohort of 102 unrelated patients who were excluded for mutations in the 2 most common PCLD genes, PRKCSH and SEC63, to identify heterozygous loss-of-function mutations in 3 additional genes, ALG8, GANAB, and SEC61B. Similarly to PRKCSH and SEC63, these genes encode proteins that are integral to the protein biogenesis pathway in the endoplasmic reticulum. We inactivated these candidate genes in cell line models to show that loss of function of each results in defective maturation and trafficking of polycystin-1, the central determinant of cyst pathogenesis. Despite acting in a common pathway, each PCLD gene product demonstrated distinct effects on polycystin-1 biogenesis. We also found enrichment on a genome-wide basis of heterozygous mutations in the autosomal recessive polycystic kidney disease gene PKHD1, indicating that adult PKHD1 carriers can present with clinical PCLD. These findings define genetic and biochemical modulators of polycystin-1 function and provide a more complete definition of the spectrum of dominant human polycystic diseases.


Transient inhibition of BMP signaling by Noggin induces cardiomyocyte differentiation of mouse embryonic stem cells.
Embryonic stem (ES) cells are a promising source of cardiomyocytes, but clinical application of ES cells has been hindered by the lack of reliable selective differentiation methods. Differentiation into any lineage is partly dependent on the regulatory mechanisms of normal early development. Although several signals, including bone morphogenetic protein (BMP), Wnt and FGF, are involved in heart development, scarce evidence is available about the exact signals that mediate cardiomyocyte differentiation. While investigating the involvement of BMP signaling in early heart formation in the mouse, we found that the BMP antagonist Noggin is transiently but strongly expressed in the heart-forming region during gastrulation and acts at the level of induction of mesendoderm to establish conditions conducive to cardiogenesis. We applied this finding to develop an effective protocol for obtaining cardiomyocytes from mouse ES cells by inhibition of BMP signaling.


Benzene and Naphthalene Degrading Bacterial Communities in an Oil Sands Tailings Pond.
Oil sands process-affected water (OSPW), produced by surface-mining of oil sands in Canada, is alkaline and contains high concentrations of salts, metals, naphthenic acids, and polycyclic aromatic compounds (PAHs). Residual hydrocarbon biodegradation occurs naturally, but little is known about the hydrocarbon-degrading microbial communities present in OSPW. In this study, aerobic oxidation of benzene and naphthalene in the surface layer of an oil sands tailings pond were measured. The potential oxidation rates were 4.3 μmol L-1 OSPW d-1 for benzene and 21.4 μmol L-1 OSPW d-1 for naphthalene. To identify benzene and naphthalene-degrading microbial communities, metagenomics was combined with stable isotope probing (SIP), high-throughput sequencing of 16S rRNA gene amplicons, and isolation of microbial strains. SIP using 13C-benzene and 13C-naphthalene detected strains of the genera Methyloversatilis and Zavarzinia as the main benzene degraders, while strains belonging to the family Chromatiaceae and the genus Thauera were the main naphthalene degraders. Metagenomic analysis revealed a diversity of genes encoding oxygenases active against aromatic compounds. Although these genes apparently belonged to many phylogenetically diverse taxa, only a few of these taxa were predominant in the SIP experiments. This suggested that many members of the community are adapted to consuming other aromatic compounds, or are active only under specific conditions. 16S rRNA gene sequence datasets have been submitted to the Sequence Read Archive (SRA) under accession number SRP109130. The Gold Study and Project submission ID number in Joint Genome Institute IMG/M for the metagenome is Gs0047444 and Gp0055765.


[The molecular basis of infections associated to orthopedic implants].
Infections associated with the use of orthopedic implants are still the major complication associated with these devices and an unsolved challenge. The frequency of infection associated with orthopedic implant is relatively low, estimated to range between 0.5 to 5%. However, it has devastating consequences for the patient and health institutions. The majority of infections is secondary to gram-positive aerobic microorganisms belonging to the genus Staphylococcus. These bacteria establish chronic infections due to its ability of adhesion and biofilm formation. Biofilms are complex communities in a polysaccharide matrix. This structure retains nutrients and protects the bacteria against the immune response and antimicrobial agents. The study of molecular characteristics and biofilm formation regulation is vital for the understanding of judicious clinical management and the development of novel strategies for the prevention of infection.


Spray-congealed solid lipid microparticles as a new tool for the controlled release of bisphosphonates from a calcium phosphate bone cement.
The aim of this work was to develop an innovative drug delivery system potentially useful for the local delivery of Bisphosphonates to bone tissue. We propose the use of Solid Lipid Microparticles (MPs), up to now mainly used for oral and topical drug delivery, as carrier for bisphosphonates due to the favourable biocompatibility and lower toxicity of the lipids compared with many polymers. The delivery platform consisted of a biomimetic α-tricalcium phosphate-gelatin cement (CPC) enriched with alendronate loaded MPs (MPs-AL) produced by the spray congealing technology. Alendronate direct addition to cement composition is limited since Alendronate is able to sequester calcium from calcium phosphates, thus preventing the setting of the cements. At variance, this approach permitted to load a relatively high amount of the drug on the CPC and allowed the controlled release of the highly water soluble alendronate. A Design of Experiment (DoE) was employed for the screening of the effects of the formulation variables related to the presence of unloaded microparticle (MPs) on the cement most important mechanical properties. Then, MPs loaded with 10% w/w of alendronate were produced using five different carriers (Stearic Acid, Stearilic Alcohol, Cutina HR, Tristearin and Precirol ATO5). All MPs-AL exhibited a spherical shape, encapsulation efficiency higher than 90% and prevalent particle size ranging from 100 to 150μm. Solid state characterization (DSC, HSM and X-ray powder diffraction) demonstrated that encapsulation of alendronate into MPs did not alter its crystal structure. MPs-AL addition to the cement provoked a modest lengthening of the setting times and of the hardening reaction leading to the complete transformation of α-tricalcium phosphate into calcium-deficient hydroxyapatite, without significantly affect the cement mechanical properties. Moreover, the results of in vitro AL release study performed on cements enriched with MPs-AL showed that the system allows a controlled release of the drug over time.


Changing indications for penetrating keratoplasty: a newly developed country's experience.
To study the changing indications for penetrating keratoplasty in Singapore and compare local indications with developed countries with established corneal graft programs. A descriptive study of all penetrating keratoplasty cases performed in the Singapore National Eye Center over a 5 year period, from 1 January 1991 to 31 December 1995 using records of the Singapore Eye Bank registry. A total of 327 penetrating keratoplasties were performed. Bullous keratopathy was the indication in more than a quarter of all cases (26.3%). Aphakic bullous keratopathy (11.6%) and pseudophakic bullous keratopathy (11.3%) accounted for the majority of the bullous keratopathy cases. The other leading indications were regrafts (11.9%), corneal dystrophies (10.4%), traumatic scarring (10.1%) and keratoconus (9.8%). A higher proportion of aphakic and pseudophakic bullous keratopathy as well as regrafts was noted in this series compared to a previous report on corneal transplantation in the 1980s. A rising indication rate of pseudophakic bullous keratopathy, in conjunction with a declining indication rate of herpetic keratitis was observed over the 5 year study period. The indications for penetrating keratoplasty in Singapore appear to follow the trend seen in developed Western countries over the past few decades.


Vacuum Insulated Probe Heated Electrospray Ionization Source Enhances Microflow Rate Chromatography Signals in the Bruker timsTOF Mass Spectrometer.
By far the largest contribution to ion detectability in liquid chromatography-driven mass spectrometry-based proteomics is the efficient generation of peptide molecular ions by the electrospray source. To maximize the transfer of peptides from the liquid to gaseous phase and allow molecular ions to enter the mass spectrometer at microspray flow rates, an efficient electrospray process is required. Here we describe the superior performance of newly design vacuum insulated probe heated electrospray ionization (VIP-HESI) source coupled to a Bruker timsTOF PRO mass spectrometer operated in microspray mode. VIP-HESI significantly improves chromatography signals in comparison to electrospray ionization (ESI) and nanospray ionization using the captivespray (CS) source and provides increased protein detection with higher quantitative precision, enhancing reproducibility of sample injection amounts. Protein quantitation of human K562 lymphoblast samples displayed excellent chromatographic retention time reproducibility (<10% coefficient of variation (CV)) with no signal degradation over extended periods of time, and a mouse plasma proteome analysis identified 12% more plasma protein groups allowing large-scale analysis to proceed with confidence (1,267 proteins at 0.4% CV). We show that the Slice-PASEF VIP-HESI mode is sensitive in identifying low amounts of peptide without losing quantitative precision. We demonstrate that VIP-HESI coupled with microflow rate chromatography achieves a higher depth of coverage and run-to-run reproducibility for a broad range of proteomic applications. Data and spectral libraries are available via ProteomeXchange (PXD040497).


In vitro antimicrobial activities of lactoferrin, its concomitant use with cefpodoxime proxetil and clinical effect of cefpodoxime proxetil.
As one of the biodefense mechanisms, lactoferrin (LFN) in the secreta of female genital organ may be an interesting biological material in view of its antimicrobial activity. In the present study, we investigated antimicrobial activities of LFN and its combination with cefpodoxime proxetil (CPDX-PR), we also as evaluated clinical effect of CPDX-PR. The following results were obtained. 1. Antimicrobial activities of LFN were tested against 15 strains of 10 species of bacteria, and potent activities against Staphylococcus aureus 209P, Escherichia coli, Klebsiella pneumoniae and Proteus spp. were found. 2. In a concomitant use of LFN with CPDX-PR (a checkerboard method), synergistic actions were observed against S. aureus 209P, E. coli STf, K. pneumoniae 602 and Pseudomonas aeruginosa 1046, and additive actions against E. coli NIHJ and Providencia rettgeri 1603. In 3 strains, the MICs of CPDX-PR in the presence of LFN were reduced to < 1/64. 3. In the evaluation of clinical effect of CPDX-PR, efficacy rates were 53/57 (92.9%) in a patient group with infections. The incidence of adverse reaction was 0/57.


Accuracy of rapid point-of-care serological tests for leprosy diagnosis: a systematic review and meta-analysis.
Leprosy is a chronic infectious disease, still endemic in many countries that may lead to neurological, ophthalmic, and motor sequelae if not treated early. Access to timely diagnosis and multidrug therapy (MDT) remains a crucial element in the World Health Organization's strategy to eliminate the disease as a public health problem. This systematic review aims to evaluate the accuracy of rapid point-of-care (POC) tests for diagnosis of leprosy. Searches were carried out in electronic databases (PubMed, EMBASE, CRD, Cochrane Library and LILACS) in April 2021 for patients with suspicion or confirmatory diagnostic of leprosy, classified in multibacillary (MB) or paucibacillary (PB) cases, performing rapid POC serological tests compared to clinical evaluation, smear microscopy and immunohistochemistry analysis. Methodological quality was assessed using the Quality Assessment of Diagnostic Accuracy Studies tool (QUADAS-2). A meta-analysis was undertaken to generate pooled estimates of diagnostic parameters, presenting sensitivity, specificity and diagnostic odds ratio (DOR) values. The review protocol was registered at PROSPERO, CRD # 42014009658. From 893 potentially relevant references, 12 articles were included reporting 16 diagnostic tests accuracy studies with 5395 individuals enrolled. Meta-analysis of NDO-LID and PGL-I tests data in MB patients showed sensitivity and specificity [95% confidence interval (CI)] of 0.83 (0.71-0.91), 0.91 (0.72-0.97); and 0.92 (0.86-0.96), 0.93 (0.78-0.98); respectively, with high heterogeneity among the studies. Our results can inform policymakers regarding the possibility of implementing accurate, rapid POC tests for leprosy in public health services, especially within primary health care.


Colocalization of inhibitory mediators, NO, VIP and galanin, in canine enteric nerves.
The colocalization of three putative inhibitory mediators of enteric nerves, vasoactive intestinal peptide (VIP), galanin (GAL) and nitric oxide synthase (nNOS), was examined in the myenteric plexus of canine antrum, intestine and colon. Many ileal and colonic neurons contained nNOS-immunoreactive (nNOS-IR) activity with some also containing VIP-IR; only a few neurons also contained GAL-IR. Ileal and colonic VIP-IR nerves often appeared to be interneurons innervating nNOS nerves. Many antral neurons contained VIP-IR with nearly all also containing GAL-IR. A few also contained nNOS-IR. The predominance of nNOS-IR neurons relative to VIP-IR and GAL-IR neurons in the ileal and colonic, but not the antral, myenteric plexus is consistent with NO being the primary inhibitory mediator in the intestine but not in the antrum.


Urinary fibrin (-ogen) degradation products in Russell's viper (Daboia russelii siamensis) bite victims.
Serum and urine concentrations of fibrin (-ogen) degradation products (FDP) were estimated in 20 proven Russell's viper bite (RVB) cases with severe defribination. All patients had similar degrees of high serum FDP levels. However, the ten who developed into acute renal failure (ARF) had significantly (p < 0.001) higher urinary FDP levels than those who did not. The urinary FDP levels of ARF cases increased correspondingly with high serum FDP levels but not in cases without ARF. Serial comparison of serum and urinary FDP levels in RVB cases with severe defibrination may be of value in predicting the likelihood of developing ARF. The present study favored disseminated intravascular coagulation as the main cause of ARF in Myanmar RVB cases.


Beyond survival: reclaiming self after leaving an abusive male partner.
Research on the process of leaving an abusive male partner has focused on surviving abuse and the crisis of leaving. Little is known about the experience of women who have left abusive male partners and not gone back. In this feminist grounded theory study of women leaving abusive partners, the researchers discovered the basic social-psychological process of reclaiming self in which women voyaged through 4 stages: counteracting abuse, breaking free, not going back, and moving on. The focus of this paper is the last stage, moving on, during which women move beyond framing their lives as survivors of an abusive relationship through the processes of figuring it out, putting it in its rightful place, launching new relationships, and taking on a new image. The findings extend our knowledge of the leaving process by delineating the ways in which the abuse experience and the survival process are displaced as the centre of the woman's intra-psychic, interpersonal, and social existence. Questions are raised about how nurses and other health professionals can avoid revictimizing women who have moved on.


Exploring the Impact of Quality of Life on Survival: A Case Study in Total Knee Replacement Surgery.
Background. There is growing evidence that quality of life (QoL) has a strong association with mortality. However, incorporation of QoL is uncommon in standard survival modeling. Methods. Using data extracted from a registry of patients undergoing total knee replacement (TKR), the impact of incorporating QoL in survival modeling was explored using 4 parametric survival models. QoL was incorporated and tested in 2 forms, which are baseline and change in QoL due to intervention. Life expectancy and quality-adjusted life years (QALYs) were calculated and comparisons made to a reference model (no QoL) to translate the findings in the context of modeled economic evaluations. Results. A total of 2858 TKR cases (2309 patients) who had TKR between 2006 and 2015 were included in this analysis. Increases in baseline and change in QoL were associated with a reduction in mortality. Compared to the reference model, differences of up to 0.32 life years and 0.53 QALYs were observed, and these translated into a 9.5% change in incremental effectiveness. These differences were much larger as the strength of the association between QoL and mortality increased. Conclusions. This work has demonstrated that the inclusion of QoL measures (at baseline and change from baseline) when extrapolating survival does matter. It can influence health outcomes such as life expectancy and QALYs, which are relevant in cost-effectiveness analysis. This is important because neglecting the correlation between QoL and mortality can lead to imprecise extrapolations and thus risk misleading results affecting subsequent decisions made by policy makers.


SIRT1 activation by resveratrol ameliorates cisplatin-induced renal injury through deacetylation of p53.
Nephrotoxicity is one of the important dose-limiting factors during cisplatin treatment. There is a growing body of evidence that activation of p53 has a critical role in cisplatin-induced renal apoptotic injury. The nicotinamide adenine dinucleotide-dependent protein deacetylase SIRT1 decreases apoptosis through deacetylating of p53, and resveratrol is known as an activator of SIRT1. To study the role of SIRT1 in cisplatin-induced renal injury through interaction with p53, mouse proximal tubular cells (MPT) were treated with cisplatin and examined the expression level of SIRT1, acetylation of p53, PUMA-α, Bax, the cytosolic/mitochondrial cytochrome c ratio, and active caspase-3. The expression of SIRT1 was decreased by cisplatin. Resveratrol, a SIRT1 activator, ameliorated cisplatin-induced acetylation of p53, apoptosis, and cytotoxicity in MPT cells. In addition, resveratrol remarkably blocked cisplatin-induced decrease of Bcl-xL in MPT cells. Further specific SIRT1 inhibition with EX 527 or small interference RNA specific to SIRT1 reversed the effect of resveratrol on cisplatin-induced toxicity. Inhibition of p53 by pifithrin-α reversed the effect of EX527 in protein expression of PUMA-α, Bcl-xL, and caspase-3 and cytotoxicity in MPT cells. SIRT1 protein expression after cisplatin treatment was significantly decreased in the kidney. SIRT1 activation by resveratrol decreased cisplatin-induced apoptosis while improving the glomerular filtration rate. Taken together, our findings suggest that the modulation of p53 by SIRT1 could be a possible target to attenuate cisplatin-induced kidney injury.


Immunoquantitation of thromboxane synthase in human tissues.
Although the formation of TxB2 has been reported for different cells such as monocytes, lymphocytes, granulocytes or fibroblasts and tissues like lung and liver, the corresponding enzyme has not yet been measured. We have established an enzyme-linked immunosorbent assay (ELISA) for measurement of human Tx-synthase with a monospecific polyclonal and a purified monoclonal antibody. ELISA data were verified by either western blots or immunohistochemistry. Next to platelets, blood monocytes were found to have the highest content of Tx-synthase, whereas lung fibroblasts and promyelocytic cells possess only very low levels. In granulocytes and lymphocytes neither Tx-synthase nor its activity could be demonstrated. Among different tissues, lung and liver showed the highest content of Tx-synthase. These data correlate TxB2-synthase activity in a given human tissue with its enzyme content of Tx-synthase for the first time. In view of the frequent involvement of TxA2 in pathological conditions, the technique developed may prove valuable for the characterization of disease states, particularly inflammatory events.


Promoting sexual health services to young men: findings from focus group discussions.
Recently, increasing interest has been shown in men's reproductive health, sexual behaviour and use of contraception. As the majority of sexual health service clients are female, however, little research has been done on the characteristics and needs of male clients. Using data from focus group discussions, this paper considers whether young men need sexual health services, whether current services are appropriate and accessible, and what promotion strategies might increase service uptake. Nine focus group discussions with 75 men aged 13-21 years at different locations in England. Young men's decision-making around sexual health may involve seeking advice from a close friend, but is less likely in some male social groups. Use of services by young men is most likely to obtain free condoms, or to remedy a crisis situation. While a young man is becoming familiar with obtaining condoms from a service, the need for a quick, straightforward service seems important. The stereotypical view within the groups was that sexual health services are women-oriented. However, suggestions are given to make services more youth- and male-friendly. Promotion should aim to increase awareness and advance a positive image of a sexual health service. Effective promotion campaigns (designed with the input of local young men), combined with appropriate and accessible services, should help to increase service use among young men.


Bevacizumab in stage II-III colon cancer: 5-year update of the National Surgical Adjuvant Breast and Bowel Project C-08 trial.
The National Surgical Adjuvant Breast and Bowel Project trial C-08 was designed to investigate the safety and efficacy of adding bevacizumab to fluorouracil, leucovorin, and oxaliplatin (FOLFOX6) for the adjuvant treatment of patients with stage 2-3 colon cancer. Our report summarizes the primary and secondary end points of disease-free and overall survival, respectively, with 5 years median follow-up time. Patients received modified FOLFOX6 once every 2 weeks for a 6-month period (control group) or modified FOLFOX6 for 6 months plus bevacizumab (5 mg/kg) once every 2 weeks for a 12-month period (experimental group). The primary end point of the study was disease-free survival (DFS) and overall survival (OS) was a secondary end point. Of 2,673 analyzed patients, demographic factors were well-balanced by treatment. With a median follow-up of 5 years, the addition of bevacizumab to mFOLFOX6 did not result in an overall significant increase in DFS (hazard ratio [HR], 0.93; 95% CI, 0.81 to 1.08; P = .35). Exploratory analyses found that the effect of bevacizumab on DFS was different before and after a 1.25-year landmark (time-by-treatment interaction P value <.0001). The secondary end point of OS was no different between the two study arms for all patients (HR, 0.95; 95% CI, 0.79 to 1.13; P = .56) and for those with stage 3 disease (HR, 1.0; 95% CI, 0.83 to 1.21; P = .99). Bevacizumab for 1 year with modified FOLFOX6 does not significantly prolong DFS or OS in stage 2-3 colon cancer. We observed no evidence of a detrimental effect of exposure to bevacizumab. A transient effect on disease-free survival was observed during bevacizumab exposure in the study's experimental arm.


Parkin is not regulated by the unfolded protein response in human neuroblastoma cells.
Mutations in the parkin gene cause the majority of cases of familial-linked Parkinson's disease, and mounting evidence suggests that parkin may play a role in idiopathic disease. Previous reports suggest that parkin may respond to and relieve, via E3-ligase activity, cellular stress at the endoplasmic reticulum caused by the accumulation of unfolded proteins. However, parkin's relationship to the mammalian unfolded protein response is unclear. Here, we comprehensively evaluate endogenous parkin in SH-SY5Y neuroblastomas at the promoter, RNA, and protein levels in response to unfolded protein stress induced by tunicamycin. While we find strong up-regulation of genes linked to the unfolded protein stress pathway, we detect no significant changes in parkin. These data suggest a lack of association between parkin and the unfolded protein response in SH-SY5Y cells.


Isolation and structure elucidation of phenazine derivative from Streptomyces sp. strain UICC B-92 isolated from Neesia altissima (Malvaceae).
Endophytic actinomycetes have been known as a promising source for new antibiotics discovery against susceptible and resistant forms of pathogenic microorganisms. This study was aimed at determining antibacterial compound from Streptomyces sp. strain B-92 isolated from a medicinal plant Neesia altissima. Streptomyces sp. strain UICC B-92 was endophytic actinomycetes of N. altissima that obtained from Universitas Indonesia Culture Collection (UICC). Isolation and determination of bioactive compound were carried out using thin layer chromatography (TLC), nuclear magnetic resonance spectroscopy (NMR), and liquid chromatography mass spectrometry (LC-MS) analyses. An in vitro antibacterial assay of pure bioactive compound from the endophytic actinomycetes strain was performed against Bacillus cereus strain ATCC 10876, Escherichia coli strain ATCC 25922, Salmonella typhimurium strain ATCC 25241, Shigella flexneri strain ATCC 12022 and Staphylococcus aureus strain ATCC 25923. The bioactive compound was identified as 4-((3S,4R,5S)-3,4,5-trihydroxy-6-(hydroxymethyl) tetrahydro-2H-pyran-2-yloxy) phenazine-1-carboxylic acid. In vitro antimicrobial assay showed that bioactive compound of Streptomyces sp. strain UICC B-92 exhibited antagonistic activities against two Gram-positive bacteria, viz, B. cereus strain ATCC 10876 and S. aureus strain ATCC 25923. The findings of this research showed that, bioactive compound of Streptomyces sp. strain UICC B-92 is suggested a new compound based on glycoside structure and its position.


Correlation of Objective Pupillometry to Midline Shift in Acute Stroke Patients.
Pupillary dysfunction is recognized as a sign of acute neurological deterioration due to worsening mass effect in patients with hemispheric strokes. Recent neuroimaging studies suggest that horizontal displacement of brain structures may be more important than vertical displacement in explaining these pupillary findings. Pupillometers allow objective and standardized evaluation of the pupillary light reflex. We hypothesized that pupillary data (Neurological Pupil index [NPi] and constriction velocity [CV]) obtained with a hand-held pupilometer, correlate with horizontal intracranial midline shift in patients with ischemic and hemorrhagic strokes. The ENDPANIC registry is a prospective database of pupillometer readings in neurological patients. There were 134 patients in the database with an acute ischemic stroke or intracerebral hemorrhage who had at least 2 neurologic imaging studies (CT or MRI) and pupillometer assessments performed within 6 hours of the imaging. Horizontal shift of the septum pellucidum (SPS) was measured in 293 images. We computed the correlation between SPS and the following pupillary variables: size, NPi, CV (left, right, and left-right difference), followed by a regression model to control for confounders. There were 94 patients (70.1%) with an ischemic stroke and 40 patients (29.9%) had an intracerebral hemorrhage. After controlling for age, race, and gender, there was a significant correlation between the SPS and NPi (left [P < .001], right [P < .001]), CV (left [P < .005], right [P < .001]) pupillary asymmetry (absolute difference between right and left; P < .05), but not between SPS and pupillary size (left or right). There was a significant correlation between the NPi and CV for the right pupil when there was a right-to-left SPS (P < .001 and P < .05, respectively), but none between the NPi and CV for the left pupil and left-to-right SPS. In patients with ischemic and hemorrhagic strokes, there is a significant correlation between SPS and the NPi, CV and pupillary asymmetry, but not with pupillary size.


Determinants of HIV/AIDS risk behaviors in expectant fathers in Haiti.
Haiti has an HIV/AIDS epidemic of the highest magnitude outside of sub-Saharan Africa. Factors such as relationship power imbalances, traditional gender role acceptance, and patriarchal belief systems that devalue women's sexuality have increased Haitian women's vulnerability to HIV infection. Because of these influences and since the HIV epidemic is largely heterosexually transmitted, it is important to understand the role that men's beliefs and behaviors play in the continuing risk of young men and women in Haiti. The purpose of this study was to gather information from male community members through semi-structured interviews in order to describe the prevalence of HIV/AIDS risk behaviors (e.g., condom use, number of sexual partners) among expectant fathers in Haiti and identify predictive psychosocial variables of HIV/AIDS risk behaviors. Results from this study showed that men who were not married (OR = 0.22, p = 0.05) and men who had medium (OR = 22.50, p < 0.001) and high sexual communication (OR = 36.51, p < 0.001) were more likely to use condoms. This study also showed that high stigma associated with HIV (OR = 16.07, p < 0.05), low HIV knowledge (OR = 0.10, p < 0.01), and high decision making power (OR = 62.52, p < 0.001) were predictors of multiple sex partners for the expectant fathers in the sample. HIV prevention programs should be designed to increase knowledge about HIV transmission, treatment, prevention and personal risk of contraction as well as correct misconceptions about individuals with HIV or AIDS and promote sex communication among partners.


Decreased type 2 corticotropin-releasing hormone receptor mRNA expression in the ventromedial hypothalamus during repeated immobilization stress.
Chronic or repeated stress results in reduction of food intake and body weight in rats. Stress-induced anorexia has been attributed to increased corticotropin-releasing hormone (CRH) function in the central nervous system. To explore possible roles of other neuropeptides and peripheral hormones involved in food intake and energy utilization during continuing stress, we examined the impact of repeated immobilization stress on expression of mRNAs coding for CRH, neuropeptide Y (NPY), galanin and pro-opiomelanocortin (POMC) mRNAs in such hypothalamic nuclei as the paraventricular nucleus (PVN), arcuate nucleus (ARC) and dorsomedial hypothalamus (DMH), as well as plasma insulin and leptin concentrations. Changes in type 2 CRH receptor (CRHR-2) mRNA in the ventromedial hypothalamus (VMH), a possible target of anorectic CRH effect, were also examined. Rats were immobilized for 2 h daily for 6 days and sacrificed 24 h after the last immobilization. Immobilized rats had lower food intake and body weight and higher levels of PVN CRH mRNA than controls. Repeated immobiliza tion also lowered plasma insulin and leptin concentrations and VMH CRHR-2 mRNA levels. These results provide additional evidence linking VMH CRHR-2 mRNA levels to plasma leptin concentration. ARC NPY and DMH galanin mRNAs increased following repeated immobilization, while ARC POMC mRNA decreased. DMH NPY mRNA and ARC galanin mRNA were unaltered by immobilization. Since NPY and galanin are considered orexigenic, while the POMC-melanocortin-4 receptor system is apparently anorexigenic, the changes in neuropeptide mRNAs and VMH CRHR-2 mRNA may play counterregulatory roles against anorectic CRH effects.


A subtilisin-like protein from soybean contains an embedded, cryptic signal that activates defense-related genes.
Among the arsenal of plant-derived compounds activated upon attack by herbivores and pathogens are small peptides that initiate and amplify defense responses. However, only a handful of plant signaling peptides have been reported. Here, we have isolated a 12-aa peptide from soybean (Glycine max) leaves that causes a pH increase of soybean suspension-cultured cell media within 10 min at low nanomolar concentrations, a response that is typical of other endogenous peptide elicitors and pathogen-derived elicitors. The amino acid sequence was determined and was found to be derived from a member of the subtilisin-like protease (subtilase) family. The sequence of the peptide was located within a region of the protein that is unique to subtilases in legume plants and not found within any other plant subtilases thus far identified. We have named this peptide signal Glycine max Subtilase Peptide (GmSubPep). The gene (Glyma18g48580) was expressed in all actively growing tissues of the soybean plant. Although transcription of Glyma18g48580 was not induced by wounding, methyl jasmonate, methyl salicylate, or ethephon, synthetic GmSubPep peptide, when supplied to soybean cultures, induced the expression of known defense-related genes, such as Cyp93A1, Chib-1b, PDR12, and achs. GmSubPep is a unique plant defense peptide signal, cryptically embedded within a plant protein with an independent metabolic role, providing insights into plant defense mechanisms.


Functional genomics of osteoarthritis: on the way to evaluate disease hypotheses.
Functional genomics is a challenging new way to address complex diseases such as osteoarthritis on a molecular level. This complements previous research and will open up new areas of so far unrecognized molecular networks. In this respect, articular cartilage is a good target for functional genomics as it contains only one cell type to which all expression signals can be attributed to. Despite considerable limitations at present, such as a low sensitivity and insensitivity to alternative splicing, posttranscriptional regulation, and posttranslational modification, cDNA-array technology provides a powerful tool to obtain an overview on gene expression patterns hardly achievable with other techniques. This has been shown to be true for known genes as well as for the identification of new genes of interest. Therefore, gene expression analysis will help to identify single genes depending on the disease and experimental conditions investigated. However, the expression pattern of the plethora of expressed genes will paint a picture (network) of disease context, maybe even more pushing forward our understanding of complex diseases such as osteoarthritis.


Racial and Ethnic Disparities in Home Health Referral Among Adult Medicare Patients.
Home health care (HHC) services following hospital discharge provide essential continuity of care to mitigate risks of posthospitalization adverse outcomes and readmissions, yet patients from racial and ethnic minority groups are less likely to receive HHC visits. To examine how the association of nurse assessments of patients' readiness for discharge with referral to HHC services at the time of hospital discharge differs by race and ethnic minority group. Secondary data analysis from a multisite study of the implementation of discharge readiness assessments in 31 US hospitals (READI Randomized Clinical Trial: 09/15/2014-03/31/2017), using linear and logistic models adjusted for patient demographic/clinical characteristics and hospital fixed effects. All Medicare patients in the study's intervention arm (n=14,684). Patient's race/ethnicity and discharge disposition code for referral to HHC (vs. home) from electronic health records. Patient's Readiness for Hospital Discharge Scale (RHDS) score (0-10 scale) assessed by the discharging nurse on the day of discharge. Adjusted RHDS scores were similar for non-Hispanic White (8.21; 95% CI: 8.18-8.24), non-Hispanic Black (8.20; 95% CI: 8.12-8.28), Hispanic (7.92; 95% CI: 7.81-8.02), and other race/ethnicity patients (8.09; 95% CI: 8.01-8.17). Non-Hispanic Black patients with low RHDS scores (6 or less) were less likely than non-Hispanic White patients to be discharged with an HHC referral (Black: 26.8%, 95% CI: 23.3-30.3; White: 32.6%, 95% CI: 31.1-34.1). Despite similar RHDS scores, Black patients were less likely to be discharged with HHC. A better understanding of root causes is needed to address systemic structural injustice in health care settings.


Radiographic management of juvenile angiofibromas.
Juvenile angiofibromas are benign, vascular, locally aggressive neoplasms that are preferably treated by surgical resection, or irradiation if surgery is not possible. Adequate surgery in the past has been limited by incomplete knowledge of the anatomy of the tumor and technical difficulties related to the vascularity. To better define the tumor, 12 patients with juvenile angiofibroma have been studied by axial and coronal high resolution computed tomography (CT). The extent of the neoplasm was better demonstrated by CT than by other techniques. Based on the CT findings, we propose an anatomic classification that is helpful in determining treatment methods. Nine patients were considered operable, and eight of these underwent preoperative embolization with Silastic spheres and Gelfoam. The preoperative embolization significantly reduced operative difficulty and the necessity for blood transfusions. Based on these cases, we believe the current radiographic management of juvenile angiofibromas should consist of plain films, CT, angiography and, in surgical cases, preoperative embolization.


[Mycobacterium tuberculosis and atypical mycobacterioses].
This brief review describes the current status of the tuberculosis endemics in Belgium as well as the position of this country in the management of the disease: there is no come back of Mycobacterium tuberculosis but a stagnation which further postpones the time of eradication. Some very general considerations about multiresistance are then presented, owing to the raising possibility of being confronted to this problem in the next future, together with more and more numerous cases of so called atypical mycobacteriosis. Simple and general guidelines from the recent literature about tuberculosis, multiresistance and non tuberculous bacteria are briefly recalled.


Prognostic value of pulmonary hemodynamic parameters in cardiac transplant candidates.
Pulmonary hypertension is a contradiction for heart transplantation (HTx). The aim of the study was to examine prognostic significance of pulmonary hemodynamic variables in patients with severe chronic heart failure (HF) considered for HTx. Patients with HF were qualified to HTx in Poland. We measured pulmonary artery systolic pressure (PASP), pulmonary capillary wedge pressure (PCWP), transpulmonary gradient (TPG), cardiac output (CO), pulmonary vascular resistance (PVR) and systemic vascular resistance (SVR). We performed biochemical evaluation, 6-min walking test, VO2max. Death or emergency HTx were assumed as the endpoints in the follow-up. Death or any kind of HTx were considered an end of observation. Survival analysis was conducted using Kaplan-Meier curves (long rank test with strait defined by terciles of analyzed hemodynamic parameters). Six hundred and fifty-eight patients were qualified to HTx between 2003 and 2007. The mean follow-up: 601 days. 87.8% male. Mean age was under 50, III and IV NYHA class. Mean PASP was 44.3 ± 16.9 mm Hg, TPG 10.0 ± 6.6 mm Hg and PVR 2.9 mm Hg, PCWP20.9 ± 9.3 mm Hg. PASP and PCWP had influence on survival or emergency HTx. There was a significant difference in survival between patients with PCWP > 25 mm Hg and PCWP < 25 mm Hg. The worst prognosis was with PASP higher than 50 mm Hg. One-, two-, and three-year survival was 75%, 58% and 48% compared to patients with PASP < 35 mm Hg (80%, 70%, and 68%, respectively). In patients qualified to HTx, pathological values of pulmonary hemodynamic parameters have a significant influence on survival. The worst prognosis have patients with PASP > 50 mm Hg, and PCWP > 25 mm Hg. Pulmonary hemodynamic parameters are important during allocation process to HTx.


Functional and metabolic disorders in celiac disease: new implications for nutritional treatment.
Celiac disease (CD) is a chronic disease causing the inflammation of the proximal small intestine, in genetically predisposed individuals. This is triggered by the consumption of the gluten protein and the side effects of the disease are mitigated by a lifelong gluten-free diet (GFD) treatment. The predominant consequence of CD is malnutrition due to malabsorption (with diarrhea, weight loss, nutritional deficiencies, and altered blood parameters), especially in patients who do not show strict adherence to GFD treatment. Recent evidence shows that, despite a lifelong GFD, some functional disorders persist, such as compromised gallbladder function and motility, exocrine pancreatic insufficiency, increased gut permeability, small-intestinal bowel overgrowth, nonalcoholic fatty liver disease (NAFLD), lactose intolerance, and milk allergy. These abnormalities may predispose to the occurrence of overweight and obesity even in CD patients. This review focuses on the principal functional and metabolic disorders in both treated and untreated CD, ranging from alterations of the gastrointestinal system to impaired glucose and lipid metabolism and insulin secretion with the aim of providing new implications beyond a GFD, for an ad hoc nutrition treatment in these patients.


The nature of genetic variation in sex and reproduction-related genes among sibling species of the Drosophila melanogaster complex.
Much is known about the biology of Drosophila melanogaster. As a model organism, a comprehensive understanding of its development, physiology and reproduction has been acquired. As a result, a broad variety of transferable genetic tools and information has allowed sibling species of the D. melanogaster complex to emerge as an important speciation model system. By comparing D. melanogaster with its close relative, Drosophila simulans, as well as its other sibling species, we are beginning to understand the nature of genetic changes during the early stages of speciation. In general, we find that genes and traits involved in sex and reproduction are more variable. A large assortment of genes and traits that are involved in various aspects of mating and fertility reveal diagnostic differences between these sibling species. Sex and reproduction-related (SRR) genes are, on average, more diverged than genes with no apparent reproductive function. Furthermore, SRR genes appear more permissive at opting in novel function. These results follow a general trend observed in other taxa and demonstrate the preferential involvement of SRR genes in reproductive isolation and species formation.


Ecological traits influence the phylogenetic structure of bird species co-occurrences worldwide.
The extent to which species' ecological and phylogenetic relatedness shape their co-occurrence patterns at large spatial scales remains poorly understood. By quantifying phylogenetic assemblage structure within geographic ranges of >8000 bird species, we show that global co-occurrence patterns are linked - after accounting for regional effects - to key ecological traits reflecting diet, mobility, body size and climatic preference. We found that co-occurrences of carnivorous, migratory and cold-climate species are phylogenetically clustered, whereas nectarivores, herbivores, frugivores and invertebrate eaters tend to be more phylogenetically overdispersed. Preference for open or forested habitats appeared to be independent from the level of phylogenetic clustering. Our results advocate for an extension of the tropical niche conservatism hypothesis to incorporate ecological and life-history traits beyond the climatic niche. They further offer a novel species-oriented perspective on how biogeographic and evolutionary legacies interact with ecological traits to shape global patterns of species coexistence in birds.


California's zero-emission vehicle adoption brings air quality benefits yet equity gaps persist.
Zero-emission vehicle (ZEV) adoption is a key climate mitigation tool, but its environmental justice implications remain unclear. Here, we quantify ZEV adoption at the census tract level in California from 2015 to 2020 and project it to 2035 when all new passenger vehicles sold are expected to be ZEVs. We then apply an integrated traffic model together with a dispersion model to simulate air quality changes near roads in the Greater Los Angeles. We found that per capita ZEV ownership in non-disadvantaged communities (non-DACs) as defined by the state of California is 3.8 times of that in DACs. Racial and ethnic minorities owned fewer ZEVs regardless of DAC designation. While DAC residents receive 40% more pollutant reduction than non-DACs due to intercommunity ZEV trips in 2020, they remain disproportionately exposed to higher levels of traffic-related air pollution. With more ZEVs in 2035, the exposure disparity narrows. However, to further reduce disparities, the focus must include trucks, emphasizing the need for targeted ZEV policies that address persistent pollution burdens among DAC and racial and ethnic minority residents.


Action potentials are required for the lateral transmission of glycinergic transient inhibition in the amphibian retina.
Transient lateral inhibition (TLI), the suppression of responses of a ganglion cell to light stimuli in the receptive field center by changes in illumination in the receptive field surround, was studied in light-adapted mud puppy and tiger salamander retinas using both eyecup and retinal slice preparations. In the eyecup, TLI was measured in on-off ganglion cells as the ability of rotating, concentric windmill patterns of 500-1200 micron inner diameter to suppress the response to a small spot stimulus in the receptive field center. Both the suppression of the spot response and the hyperpolarization produced in ganglion cells by rotation of the windmill were blocked in the presence of 2 microM strychnine or 500 nM tetrodotoxin (TTX), but not by 150 microM picrotoxin. In the slice preparation in which GABA-mediated currents were blocked with picrotoxin, IPSCs elicited by diffuse illumination were blocked by strychnine and strongly reduced by TTX. The TTX-resistant component was probably attributable to illumination of the receptive field center. TTX had a much greater effect in reducing the glycinergic inhibition elicited by laterally displaced stimulation versus nearby focal electrical stimulation. Strychnine enhanced light-evoked excitatory currents in ganglion cells, but this was not mimicked by TTX. The results suggest that local glycinergic transient inhibition does not require action potentials and is mediated by synapses onto both ganglion cell dendrites and bipolar cell terminals. In contrast, the lateral spread of this inhibition (at least over distances >250 micron) requires action potentials and is mainly onto ganglion cell dendrites.


The influence of oxygen tension on the structure and function of isolated liver sinusoidal endothelial cells.
Liver sinusoidal endothelial cells (LSECs) are specialized scavenger cells, with crucial roles in maintaining hepatic and systemic homeostasis. Under normal physiological conditions, the oxygen tension encountered in the hepatic sinusoids is in general considerably lower than the oxygen tension in the air; therefore, cultivation of freshly isolated LSECs under more physiologic conditions with regard to oxygen would expect to improve cell survival, structure and function. In this study LSECs were isolated from rats and cultured under either 5% (normoxic) or 20% (hyperoxic) oxygen tensions, and several morpho-functional features were compared. Cultivation of LSECs under normoxia, as opposed to hyperoxia improved the survival of LSECs and scavenger receptor-mediated endocytic activity, reduced the production of the pro-inflammatory mediator, interleukin-6 and increased the production of the anti-inflammatory cytokine, interleukin-10. On the other hand, fenestration, a characteristic feature of LSECs disappeared gradually at the same rate regardless of the oxygen tension. Expression of the cell-adhesion molecule, ICAM-1 at the cell surface was slightly more elevated in cells maintained at hyperoxia. Under normoxia, endogenous generation of hydrogen peroxide was drastically reduced whereas the production of nitric oxide was unaltered. Culture decline in high oxygen-treated cultures was abrogated by administration of catalase, indicating that the toxic effects observed in high oxygen environments is largely caused by endogenous production of hydrogen peroxide. Viability, structure and many of the essential functional characteristics of isolated LSECs are clearly better preserved when the cultures are maintained under more physiologic oxygen levels. Endogenous production of hydrogen peroxide is to a large extent responsible for the toxic effects observed in high oxygen environments.


A Combined Spectroscopic and In Silico Approach to Evaluate the Interaction of Human Frataxin with Mitochondrial Superoxide Dismutase.
Frataxin (FXN) is a highly conserved mitochondrial protein whose deficiency causes Friedreich's ataxia, a neurodegenerative disease. The precise physiological function of FXN is still unclear; however, there is experimental evidence that the protein is involved in biosynthetic iron-sulfur cluster machinery, redox imbalance, and iron homeostasis. FXN is synthesized in the cytosol and imported into the mitochondria, where it is proteolytically cleaved to the mature form. Its involvement in the redox imbalance suggests that FXN could interact with mitochondrial superoxide dismutase (SOD2), a key enzyme in antioxidant cellular defense. In this work, we use site-directed spin labelling coupled to electron paramagnetic resonance spectroscopy (SDSL-EPR) and fluorescence quenching experiments to investigate the interaction between human FXN and SOD2 in vitro. Spectroscopic data are combined with rigid body protein-protein docking to assess the potential structure of the FXN-SOD2 complex, which leaves the metal binding region of FXN accessible to the solvent. We provide evidence that human FXN interacts with human SOD2 in vitro and that the complex is in fast exchange. This interaction could be relevant during the assembly of iron-sulfur (FeS) clusters and/or their incorporation in proteins when FeS clusters are potentially susceptible to attacks by reactive oxygen species.


PFN1 and integrin-β1/mTOR axis involvement in cornea differentiation of fibroblast limbal stem cells.
Ex vivo limbal stem cell transplantation is the main therapeutic approach to address a complete and functional re-epithelialization in corneal blindness, the second most common eye disorder. Although important key points were defined, the molecular mechanisms involved in the epithelial phenotype determination are unclear. Our previous studies have demonstrated the pluripotency and immune-modulatory of fibroblast limbal stem cells (f-LSCs), isolated from the corneal limbus. We defined a proteomic profile especially enriched in wound healing and cytoskeleton-remodelling proteins, including Profilin-1 (PFN1). In this study we postulate that pfn-1 knock down promotes epithelial lineage by inhibiting the integrin-β1(CD29)/mTOR pathway and subsequent NANOG down-expression. We showed that it is possible modulate pfn1 expression levels by treating f-LSCs with Resveratrol (RSV), a natural compound: pfn1 decline is accompanied with up-regulation of the specific differentiation epithelial genes pax6 (paired-box 6), sox17 (sex determining region Y-box 17) and ΔNp63-α (p63 splice variant), consistent with drop-down of the principle stem gene levels. These results contribute to understand the molecular biology of corneal epithelium development and suggest that pfn1 is a potential molecular target for the treatment of corneal blindness based on epithelial cell dysfunction.


Betanin, a beetroot component, induces nuclear factor erythroid-2-related factor 2-mediated expression of detoxifying/antioxidant enzymes in human liver cell lines.
Our recent study has shown that beetroot juice protects against N-nitrosodimethylamine (NDEA)-induced liver injury and increases the activity of phase II enzymes, suggesting the activation of the nuclear factor erythroid-2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway. The aim of the present study was to further explore the mechanism of the activity of beetroot by evaluating the cytoprotective effects of its major component. The influence of betanin (BET) on the activation of Nrf2 and the expression of GSTA, GSTP, GSTM, GSTT, NQO1 and HO-1 was assessed in two hepatic cell lines: non-tumour THLE-2 and hepatoma-derived HepG2 cell lines. The level of the tumour suppressor p53 in both cell lines and the methylation of GSTP in HepG2 cells were also evaluated. Treatment of both cell lines with 2, 10 and 20 μm of BET resulted in the translocation of Nrf2 from the cytosol to the nucleus. The mRNA and nuclear protein levels of Nrf2 and the binding of Nrf2 to ARE sequences were increased only in the THLE-2 cells and were accompanied by the phosphorylation of serine/threonine kinase (AKT), c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK). BET also significantly increased the mRNA and protein levels of GSTP, GSTT, GSTM and NQO1 in these cells. Conversely, besides the translocation of Nrf2 from the cytosol to the nucleus, BET did not modulate any of the other parameters measured in the HepG2 cells. BET did not change the methylation of GSTP1 in these cells either. These results indicate that BET through the activation of Nrf2 and subsequent induction of the expression of genes controlled by this factor may exert its hepatoprotective and anticarcinogenic effects. Moreover, the activation of mitogen-activated protein kinases may be responsible for the activation of Nrf2 in the THLE-2 cells.


Pbx and Prdm1a transcription factors differentially regulate subsets of the fast skeletal muscle program in zebrafish.
The basic helix-loop-helix factor Myod initiates skeletal muscle differentiation by directly and sequentially activating sets of muscle differentiation genes, including those encoding muscle contractile proteins. We hypothesize that Pbx homeodomain proteins direct Myod to a subset of its transcriptional targets, in particular fast-twitch muscle differentiation genes, thereby regulating the competence of muscle precursor cells to differentiate. We have previously shown that Pbx proteins bind with Myod on the promoter of the zebrafish fast muscle gene mylpfa and that Pbx proteins are required for Myod to activate mylpfa expression and the fast-twitch muscle-specific differentiation program in zebrafish embryos. Here we have investigated the interactions of Pbx with another muscle fiber-type regulator, Prdm1a, a SET-domain DNA-binding factor that directly represses mylpfa expression and fast muscle differentiation. The prdm1a mutant phenotype, early and increased fast muscle differentiation, is the opposite of the Pbx-null phenotype, delayed and reduced fast muscle differentiation. To determine whether Pbx and Prdm1a have opposing activities on a common set of genes, we used RNA-seq analysis to globally assess gene expression in zebrafish embryos with single- and double-losses-of-function for Pbx and Prdm1a. We find that the levels of expression of certain fast muscle genes are increased or approximately wild type in pbx2/4-MO;prdm1a-/- embryos, suggesting that Pbx activity normally counters the repressive action of Prdm1a for a subset of the fast muscle program. However, other fast muscle genes require Pbx but are not regulated by Prdm1a. Thus, our findings reveal that subsets of the fast muscle program are differentially regulated by Pbx and Prdm1a. Our findings provide an example of how Pbx homeodomain proteins act in a balance with other transcription factors to regulate subsets of a cellular differentiation program.


Granulopoiesis in severe congenital neutropenia.
The pathogenesis of the granulopoietic failure in three children with severe congenital neutropenia was studied. Mature neutrophils were absent from both peripheral blood and bone marrow. Assay of bone marrow granulocyte colony-forming cells (CFU-C) in a methylcellulose tissue culture system using colony-stimulating activity (CSA) from peripheral blood leukocytes demonstrated normal or increased concentrations of CFU-C compared to those from marrows of 60 age-matched controls. Colonies were of normal size and by light microscopy appeared to contain granulocytes in all stages of maturation including the mature polymorphonuclear neutrophil. CFU-C from peripheral blood of two patients were normal. Production and activity of CSA from the patients' peripheral blood leukocytes and urinary CSA excretion were normal. No serum inhibitors against CFU-C or CSA could be demonstrated using both control and autologous marrow. The defect did not appear to be due to a lack of granulocytic stem cells, a reduction of humoral stimulators of granulopoiesis, nor the presence of an inhibitor as measured by these techniques.


Population bottlenecks and nonequilibrium models in population genetics. II. Number of alleles in a small population that was formed by a recent bottleneck.
A model is presented in which a large population in mutation/drift equilibrium undergoes a severe restriction in size and subsequently remains at the small size. The rate of loss of genetic variability has been studied. Allelic loss occurs more rapidly than loss of genic heterozygosity. Rare alleles are lost especially rapidly. The result is a transient deficiency in the total number of alleles observed in samples taken from the reduced population when compared with the number expected in a sample from a steady-state population having the same observed heterozygosity. Alternatively, the population can be considered to possess excess gene diversity if the number of alleles is used as the statistical estimator of mutation rate. The deficit in allele number arises principally from a lack of those alleles that are expected to appear only once or twice in the sample. The magnitude of the allelic deficiency is less, however, than the excess that an earlier study predicted to follow a rapid population expansion. This suggests that populations that have undergone a single bottleneck event, followed by rapid population growth, should have an apparent excess number of alleles, given the observed level of genic heterozygosity and provided that the bottleneck has not occurred very recently. Conversely, such populations will be deficient for observed heterozygosity if allele number is used as the sufficient statistic for the estimation of 4Nev. Populations that have undergone very recent restrictions in size should show the opposite tendencies.


Parathyroid Hormone in Pregnancy: Vitamin D and Other Determinants.
We aimed to assess the parathyroid hormone (PTH) concentration in pregnant women at the beginning of pregnancy (1st trimester) and within days before delivery (3rd trimester) and evaluate its determinants. From September 2014 through December 2015 in a cross-sectional study, 204 women in the 1st trimester of pregnancy and 203 women in the 3rd trimester of pregnancy were recruited. Blood samples were collected to measure PTH and circulating 25-hydroxy-vitamin D (25(OH)D) concentrations. Lifestyle and demographic data were collected using a questionnaire. Serum 25(OH)D and PTH were inversely correlated in both early and late pregnancy. Our analyses suggest that in the 3rd trimester of pregnancy, a 25(OH)D level of 18.9 ng/mL (47.3 nmol/L) could serve as an inflection point for the maximal suppression of PTH. Statistically significant determinants of PTH concentrations in multiple regression were 25(OH)D concentrations, season, multiparity and education of the partner (all p < 0.05) in early pregnancy. In late pregnancy, 25(OH)D concentrations and country of origin were statistically significant determinants of PTH concentrations (all p < 0.05). These factors and their effect on PTH appear to be vastly determined by 25(OH)D; however, they might also affect PTH through other mechanisms besides 25(OH)D.


Effects of forskolin on insulin release and cyclic AMP content in rat pancreatic islets.
The effects of forskolin, a diterpene stimulator of adenylate cyclase, on cyclic AMP content and insulin release, have been studied in rat pancreatic islets. In the presence of 2.8 mM glucose, 0.3 to 30 microM forskolin gave concentration-dependent increases in cyclic AMP content to as much as seven-fold without causing any significant increase in insulin release. With 5.6 mM glucose, cyclic AMP contents were elevated by forskolin to the same extent as with 2.8 mM glucose but, under these conditions, insulin release was stimulated. 3 microM forskolin was a maximal concentration with respect to insulin release, but not for the elevation of cyclic AMP levels. Concentration-dependent increases in both insulin release and cyclic AMP content were detected over the range of 0.3 to 3 microM forskolin. The effect of forskolin to stimulate insulin release was rapid and was reversible within 15 minutes of removal of the drug. When tested with 16.7 mM glucose, forskolin potentiated both phases of stimulated release, with the greater effect upon the first phase.


GBDT_KgluSite: An improved computational prediction model for lysine glutarylation sites based on feature fusion and GBDT classifier.
Lysine glutarylation (Kglu) is one of the most important Post-translational modifications (PTMs), which plays significant roles in various cellular functions, including metabolism, mitochondrial processes, and translation. Therefore, accurate identification of the Kglu site is important for elucidating protein molecular function. Due to the time-consuming and expensive limitations of traditional biological experiments, computational-based Kglu site prediction research is gaining more and more attention. In this paper, we proposed GBDT_KgluSite, a novel Kglu site prediction model based on GBDT and appropriate feature combinations, which achieved satisfactory performance. Specifically, seven features including sequence-based features, physicochemical property-based features, structural-based features, and evolutionary-derived features were used to characterize proteins. NearMiss-3 and Elastic Net were applied to address data imbalance and feature redundancy issues, respectively. The experimental results show that GBDT_KgluSite has good robustness and generalization ability, with accuracy and AUC values of 93.73%, and 98.14% on five-fold cross-validation as well as 90.11%, and 96.75% on the independent test dataset, respectively. GBDT_KgluSite is an effective computational method for identifying Kglu sites in protein sequences. It has good stability and generalization ability and could be useful for the identification of new Kglu sites in the future. The relevant code and dataset are available at https://github.com/flyinsky6/GBDT_KgluSite .


A monoclonal antibody based capture ELISA for botulinum neurotoxin serotype B: toxin detection in food.
Botulism is a serious foodborne neuroparalytic disease, caused by botulinum neurotoxin (BoNT), produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A-H) have been described. The majority of human cases of botulism are caused by serotypes A and B followed by E and F. We report here a group of serotype B specific monoclonal antibodies (mAbs) capable of binding toxin under physiological conditions. Thus, they serve as capture antibodies for a sandwich (capture) ELISA. The antibodies were generated using recombinant peptide fragments corresponding to the receptor-binding domain of the toxin heavy chain as immunogen. Their binding properties suggest that they bind a complex epitope with dissociation constants (KD's) for individual antibodies ranging from 10 to 48 × 10-11 M. Assay performance for all possible combinations of capture-detector antibody pairs was evaluated and the antibody pair resulting in the lowest level of detection (L.O.D.), ~20 pg/mL was determined. Toxin was detected in spiked dairy samples with good recoveries at concentrations as low as 0.5 pg/mL and in ground beef samples at levels as low as 2 ng/g. Thus, the sandwich ELISA described here uses mAb for both the capture and detector antibodies (binding different epitopes on the toxin molecule) and readily detects toxin in those food samples tested.


Selective hydroxylation of highly branched fatty acids and their derivatives by CYP102A1 from Bacillus megaterium.
Highly branched fatty acids, the main components of the preen-gland waxes of the domestic goose and the Muscovy duck, and their derivatives are promising chiral precursors for the synthesis of macrolide antibiotics. The key step in the utilisation of these compounds is their regioselective hydroxylation, which cannot be achieved in a classical chemical approach. Three P450 monooxygenases, CYP102A1, CYP102A2 and CYP102A3, demonstrating high turnover numbers in the hydroxylation of iso and anteiso fatty acids (>400 min(-1)), were tested for their activity towards these substrates. CYP102A1 from Bacillus megaterium and its A74G F87V L188Q triple mutant hydroxylate a variety of these substrates with high activity and regioselectivity. In all cases, the triple mutant showed much higher activities than the wild-type enzyme. The binding constants, determined for wild-type CYP102A1 and the triple mutant with tetramethylnonanol as substrate, were >200 microM and approximately 23 microM, respectively. Data derived from binding analysis support the differences in activity found for the wild-type CYP102A1 and the triple mutant. Surprisingly, CYP102A2 and CYP102A3 from Bacillus subtilis did not show any activity. Substrate binding spectra, recorded to investigate substrate accessibility to the enzyme's active sites, revealed that the substrates either could not access the active site of the Bacillus subtilis monooxygenases, or did not come into proximity with the heme.


Early diagnosis of trisomy 21, trisomy 18 and trisomy 13 using nuchal translucency thickness and ductus venosus blood flow waveform in West China.
Abnormal nuchal translucency (NT) thickness and ductus venosus (DV) blood flow have been associated with trisomy 21, 18 and 13. However, the association of abnormal DV with these aneuploidies varies among clinical studies. The present study examined the possibility of using NT combined with DV for the early diagnosis of the three aforementioned aneuploidies in the Western Chinese population, focusing on three aspects: Biological pathway analysis, theoretical statistical analysis and clinical data analysis from 1,962 first‑trimester pregnant women from Western China. The pathway and statistical analyses performed suggested the reliability of integrating NT and DV in the prediction of the three aneuploidies. The clinical data analysis suggested that integrating NT and DV, compared with NT alone, resulted in increased predictive power (34.09 vs. 22.45%), better rejection probability (0.21 vs. 0.44%), increased specificity (96.71 vs. 94.07%) and increased sensitivity (89.47 vs. 80.49%). The present results suggested the reliability of integrating NT and DV for the early diagnosis of trisomy 21, 18 and 13 for the Western Chinese population. The present results provided novel statistical analyses to the field of prenatal diagnosis in the Western Chinese population.


Effects of preeclampsia on maternal and cord blood clotting activity.
This study was undertaken to evaluate the effect of preeclampsia on maternal and cord blood clotting parameters. Pregnant controls and preeclamptics plus their offspring had plasma analyzed for prothrombin time, partial thromboplastin time, fibrinogen, and clotting activity for factors II, V, VII, VIII, IX, X, XII, plus factor II antigen. Maternal data was consistent with that previously reported by numerous authors. The cord blood data showed that neonates of the preeclamptic mother had elevated fibrinogen and decreased II, V and VII activity, plus decreased II antigen. The data indicates that abnormal clotting parameters seen in the neonates from preeclampic mothers may result from impaired liver function and not as previously thought, from a deficiency in neonatal vitamin K.


Introducing consumer directed care in residential care settings for older people in Australia: views of a citizens' jury.
Objectives Health services worldwide are increasingly adopting consumer directed care approaches. Traditionally, consumer directed care models have been implemented in home care services and there is little guidance as to how to implement them in residential care. This study used a citizens' jury to elicit views of members of the public regarding consumer directed care in residential care. Methods A citizens' jury involving 12 members of the public was held over two days in July 2016, exploring the question: For people with dementia living in residential care facilities, how do we enable increased personal decision making to ensure that care is based on their needs and preferences? Jury members were recruited through a market research company and selected to be broadly representative of the general public. Results The jury believed that person-centred care should be the foundation of care for all older people. They recommended that each person's funding be split between core services (to ensure basic health, nutrition and hygiene needs are met) and discretionary services. Systems needed to be put into place to enable the transition to consumer directed care including care coordinators to assist in eliciting resident preferences, supports for proxy decision makers, and accreditation processes and risk management strategies to ensure that residents with significant cognitive impairment are not taken advantage of by goods and service providers. Transparency should be increased (perhaps using technologies) so that both the resident and nominated family members can be sure that the person is receiving what they have paid for. Conclusions The views of the jury (as representatives of the public) were that people in residential care should have more say regarding the way in which their care is provided and that a model of consumer directed care should be introduced. Policy makers should consider implementation of consumer directed care models that are economically viable and are associated with high levels of satisfaction among users.


Achieving superelasticity in additively manufactured NiTi in compression without post-process heat treatment.
Shape memory alloys (SMAs), such as Nitinol (i.e., NiTi), are of great importance in biomedical and engineering applications due to their unique superelasticity and shape memory properties. In recent years, additive manufacturing (AM) processes have been used to produce complex NiTi components, which provide the ability to tailor microstructure and thus the critical properties of the alloys, such as the superelastic behavior and transformation temperatures (TTs), by selection of processing parameters. In biomedical applications, superelasticity in implants play a critical role since it gives the implants bone-like behavior. In this study, a methodology of improving superelasticity in Ni-rich NiTi components without the need for any kind of post-process heat treatments will be revealed. It will be shown that superelasticity with 5.62% strain recovery and 98% recovery ratio can be observed in Ni-rich NiTi after the sample is processed with 250 W laser power, 1250 mm/s scanning speed, and 80 µm hatch spacing without, any post-process heat treatments. This superelasticity in as-fabricated Ni-rich SLM NiTi was not previously possible in the absence of post-process heat treatments. The findings of this study promise the fast, reliable and inexpensive fabrication of complex shaped superelastic NiTi components for many envisioned applications such as patient-specific biomedical implants.


Predatory cues drive colony size reduction in marine diatoms.
Colony formation is a common feature among nonmotile marine phytoplankton. Several theories exist around the potential benefits of larger colonies.Here, we test the hypothesis that predation is one of the drivers behind colony formation and chain length plasticity. We exposed cultures of Thalassiosira rotula, Chaetoceros curvisetus, and Chaetoceros affinis to copepodamides, a chemical alarm cue released by copepods and perceived as an indicator of predation threat by their prey. This was coupled with a grazing experiment, which compared copepod grazing rates on different chain lengths.Our results show that T. rotula and C. curvisetus decreased their chain lengths by 79% and 49%, respectively, in response to copepodamides. Single cells and short chains were grazed at lower rates compared with long chains, and the copepodamide-driven size shift led to 30% and 12% lower grazing in T. rotula and C. curvisetus, respectively. In contrast, C. affinis showed a slight increased chain length in response to copepodamides although nonsignificant.We found that 2 of 3 studied species reduce their chain length in response to the presence of copepod grazers. Altered size structure has implications for the route of carbon in the marine food webs and carbon export to deeper strata.


Relative expression of genes of terpene metabolism in different tissues of Artemisia annua L.
Recently, Artemisia annua L. (annual or sweet wormwood) has received increasing attention due to the fact that the plant produces the sesquiterpenoid endoperoxide artemisinin, which today is widely used for treatment of malaria. The plant produces relatively small amounts of artemisinin and a worldwide shortage of the drug has led to intense research in order to increase the yield of artemisinin. In order to improve our understanding of terpene metabolism in the plant and to evaluate the competition for precursors, which may influence the yield of artemisinin, we have used qPCR to estimate the expression of 14 genes of terpene metabolism in different tissues. The four genes of the artemisinin biosynthetic pathway (amorpha-4,11-diene synthase, amorphadiene-12-hydroxylase, artemisinic aldehyde ∆11(13) reductase and aldehyde dehydrogenase 1) showed remarkably higher expression (between ~40- to ~500-fold) in flower buds and young leaves compared to other tissues (old leaves, stems, roots, hairy root cultures). Further, dihydroartemisinic aldehyde reductase showed a very high expression only in hairy root cultures. Germacrene A and caryophyllene synthase were mostly expressed in young leaves and flower buds while epi-cedrol synthase was highly expressed in old leaves. 3-Hydroxy-3-methyl-glutaryl coenzyme A reductase exhibited lower expression in old leaves compared to other tissues. Farnesyldiphosphate synthase, squalene synthase, and 1-deoxy-D-xylulose-5-phosphate reductoisomerase showed only modest variation in expression in the different tissues, while expression of 1-deoxy-D-xylulose-5-phosphate synthase was 7-8-fold higher in flower buds and young leaves compared to old leaves. Four genes of artemisinin biosynthesis were highly expressed in flower buds and young leaves (tissues showing a high density of glandular trichomes). The expression of dihydroartemisinic aldehyde reductase has been suggested to have a negative effect on artemisinin production through reduction of dihydroartemisinic aldehyde to dihydroartemisinic alcohol. However, our results show that this enzyme is expressed only at low levels in tissues producing artemisinin and consequently its effect on artemisinin production may be limited. Finally, squalene synthase but not other sesquiterpene synthases appears to be a significant competitor for farnesyl diphosphate in artemisinin-producing tissues.


Ionic Liquids: Simple or Complex Electrolytes?
Ionic liquids belong to the most investigated systems in the recent years and this field is still significantly growing with an increased focus on developing ionic liquids for specific applications, along with fundamental research. In the present featured paper the similarity and differences between common, "classical" electrolytes and common surfactants and ionic liquids along with the surface-active ionic liquids are discussed in order to stress their significance and point out on their weak points. A short survey of the literature data reveals namely that ionic liquids in solutions behave like "classical" electrolytes and can be described by existing models in the range of their validity. There is still a lack of models describing well the concentrated electrolyte solutions and here ionic liquids, less limited by solubility as common electrolytes, can serve as model systems. The micellization of surface-active ionic liquids in aqueous solutions can be described in the same way as for common surfactants, but surface-active ionic liquids offer more possibilities to study specific ion and isomer effects. They are also quite promising systems to study the aggregation processes in non-aqueous solutions.


Study of Treatment and Reproductive Outcomes Among Reproductive-Age Women With HIV Infection in the Southern United States: Protocol for a Longitudinal Cohort Study.
Nearly a quarter of the 1.1 million individuals with HIV in the United States are women. Racial and ethnic minority women in the Southern United States are disproportionately impacted. Reproductive-age women with HIV are prone to poor HIV outcomes but remain underrepresented in HIV research. We will answer contemporary questions related to the health outcomes in this population by enrolling a prospective cohort of reproductive-age women with and without HIV in the Southern United States. The Study of Treatment and Reproductive Outcomes (STAR) will enroll and retain 2000 reproductive-age women with and without HIV. The STAR will leverage the infrastructure of the US-based Multicenter AIDS Cohort Study (MACS)/Women's Interagency HIV Study (WIHS) Combined Cohort Study, comprising the WIHS (a cohort of women with and at risk for HIV, which began in 1993), and the MACS (a cohort of gay and bisexual men with and at risk for HIV, which began in 1984). Although the advancing age of the participants enrolled in the MACS/WIHS Combined Cohort Study provides an opportunity to address the questions related to HIV and aging, the research questions pertinent to the reproductive years must also be addressed. The STAR will conduct high-priority scientific research in key areas with the overall aim of addressing the unique needs of reproductive-age women with HIV. The STAR is a prospective, observational cohort study that will be conducted at 6 sites in the United States-Atlanta, Georgia; Birmingham, Alabama; Jackson, Mississippi; Chapel Hill, North Carolina; Miami, Florida; and Washington, District of Columbia. Visits will occur semiannually for 2 years, with additional visits for up to 5 years. At each visit, the participating women will complete a structured interview for collecting key demographic, psychosocial, and clinical variables, and undergo biospecimen collection for laboratory testing and repositing (blood, urine, hair, vaginal, anal, and oral specimens). Pregnant women and infants will undergo additional study assessments. The initial scientific focus of the STAR is to understand the roles of key social determinants of health, depression, reproductive health, and oral health on HIV and pregnancy outcomes across the reproductive life span. Enrollment in the STAR commenced in February 2021 and is ongoing. Through in-depth, longitudinal data and biospecimen collection, the newly initiated STAR cohort will create a platform to answer scientific questions regarding reproductive-age women with and without HIV. STAR will be uniquely positioned to enable investigators to conduct high-impact research relevant to this population. Building on the legacy of the MACS and WIHS cohorts, the STAR is designed to foster multidisciplinary collaborations to galvanize scientific discoveries to improve the health of reproductive-age women with HIV and ameliorate the effects of the HIV epidemic in this population in the United States.


The psychological experience of nursing mothers upon learning of a toxic substance in their breast milk.
This study explores the psychological reactions of a group of nursing mothers whose breast milk was inadvertently contaminated with a toxic fire-retardant chemical, polybrominated bihenyl. In particular, the extent to which the 97 mothers in the study denied the presence and possible effects of the chemical in their offspring is examined. Corresponding psychological defenses and reactions such as guilt or ambivalence in the nursing relationship and efforts at mastery of the situation are described with accompanying clinical illustrations. Insofar as the presence of a variety of toxic chemicals in the environment has been reported, the psychological reactions discussed here may have widespread application.


Association between Ocular Pseudoexfoliation and Sensorineural Hearing Loss.
Background. Our study aimed to investigate an association between ocular pseudoexfoliation (PXF) and sensorineural hearing loss (SNHL) and to compare them with age and sex matched controls without pseudoexfoliation. Method. This was a case-control study of 123 patients which included 68 cases with PXF (at least one eye) and 55 controls without pseudoexfoliation. Pure-tone audiometry (PTA) was done for these patients at sound frequencies taken as important for speech comprehension, that is, 250 Hertz (Hz), 500 Hz, 1000 Hz, and 2000 Hz. Results. There were 41 patients with pseudoexfoliation syndrome (PXE) and 27 with pseudoexfoliative glaucoma (PXEG). The majority of patients with hearing loss (60%; n = 51) were PXF patients and the remaining 40% (n = 34) were controls. Below average hearing thresholds were significantly higher in the pseudoexfoliation group compared to the control group (P = 0.01; odds ratio (OR), 3.00; 95% confidence interval (CI), 1.25-7.19). However, there was no significant difference in the mean hearing threshold levels between the three groups (PXE, PXEG, and controls) in either ear (ANOVA, right ear: P = 0.46 and left ear P = 0.36). Conclusion. Our study found an association between PXF and SNHL, confirming that PXF can involve organs in the body other than the eye.


Tropicalization and kelp loss shift trophic composition and lead to more winners than losers in fish communities.
Climate-mediated species redistributions are causing novel interactions and leading to profound regime shifts globally. For species that expand their distribution in response to warming, survival depends not only on their physiological capacity, but also on the ability to coexist or be competitive within the established community. In temperate marine reefs from around the world, the range expansion of tropical species, known as 'tropicalization', has been linked to the disappearance of temperate habitat-forming kelps and shifts to dominance by low-biomass turfing algae. The consequences of these range expansions and habitat changes on resident fish communities are, however, unclear. Here, we use data derived from baited remote underwater video (BRUV) surveys to analyse changes in diversity and abundance of marine fishes over a 17-year period in warming reefs that have experienced kelp loss (occurring c. 2009). Despite the loss of kelp, we found that species richness and overall abundance of fishes (measured as probability of occurrence and relative abundance), including both tropical and temperate species, increased through time. We also found dramatic shifts in the trophic composition of fish assemblages. Tropical herbivorous fish increased most markedly through time, and temperate-associated planktivores were the only group that declined, a potential consequence of tropicalization not previously identified. At the species level, we identified 22 tropical and temperate species from four trophic guilds that significantly increased in occurrence, while only three species (all temperate associated) declined. Morphological trait space models suggest increases in fish diversity and overall occurrence are unlikely to be driven by uniqueness of traits among tropical range expanders. Our results show more winners than losers and suggest that pathways of energy flow will change in tropicalized systems, as planktonic inputs become less important and a higher proportion of algal productivity gets consumed locally by increasingly abundant herbivores.


Parents' perceptions on policies of early childhood care and education programmes in selected states of Malaysia.
The 2030 agenda for sustainable development proposed global equitable quality education and lifelong learning opportunities for all children . The quality of early childhood care and education (ECCE) programs helps shape children's minds, attitudes and behaviors, and has short and long-term effects on a child, a family and a country. In Malaysia, the government has formulated some policies and laws to protect children's rights. However, ECCE is facing some challenges. The purpose of this study is to investigate parents' perceptions of the quality of ECCE programs implemented by Malaysian government. A mixed method was used to collect data on parents' perceptions of ECCE policies in selected states in Malaysia. The questionnaires, (P1/POL) from the research project "Development of a Comprehensive and Integrated Model of Quality Malaysian ECCE", were distributed among 629 respondents who have a child in a preschool, and 22 participants were randomly selected to take part in five focus group interviews Results: The key findings of the study revealed 68% parents were not familiar with ECCE Malaysian government policy, however 84.3% stressed it is important for the government to educate them about ECCE. Thus findings indicated that the majority of parents lack awareness of the ECCE policies and quality of early childhood care and education programs related to the policies remain the issue. While interviewing the focus group ,most of them were not aware of ECCE and pointed out parents are stressing children's academic learning in particular preschools. It is concluded that parents' awareness regarding the ECCE program must be part of the policies and needs to improve. It is recommended that the government of Malaysia should supply more information on ECCE policies to parents and focus on policy implementation. Moreover, the quality of ECCE programs should be improved based on the parents' perceptions.


Lentinus edodes: a macrofungus with pharmacological activities.
Lentinus edodes is the first medicinal macrofungus to enter the realm of modern biotechnology. It is the second most popular edible mushroom in the global market which is attributed not only to its nutritional value but also to possible potential for therapeutic applications. Lentinus edodes is used medicinally for diseases involving depressed immune function (including AIDS), cancer, environmental allergies, fungal infection, frequent flu and colds, bronchial inflammation, heart disease, hyperlipidemia (including high blood cholesterol), hypertension, infectious disease, diabetes, hepatitis and regulating urinary inconsistencies. It is the source of several well-studied preparations with proven pharmacological properties, especially the polysaccharide lentinan, eritadenine, shiitake mushroom mycelium, and culture media extracts (LEM, LAP and KS-2). Antibiotic, anti-carcinogenic and antiviral compounds have been isolated intracellularly (fruiting body and mycelia) and extracellularly (culture media). Some of these substances were lentinan, lectins and eritadenine. The aim of this review is to discuss the therapeutic applications of this macrofungus. The potential of this macrofungus is unquestionable in the most important areas of applied biotechnology.


Fine structure of cells infected with human cytomegalovirus after treatment with 9-(1,3-dihydroxy-2-propoxymethyl)guanine.
Infection with human cytomegalovirus (CMV) is characterized by cytological changes which are readily visualized by electron microscopy using ultrathin sections of infected cells. Treatment of such cells with 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG), a potent inhibitor of CMV, is effective when initiated at early or late times after infection and the response to such treatment has been studied by fine structural analysis. Inhibition of viral DNA synthesis by DHPG treatment (50 microM) late in virus infection resulted in a cessation of virus growth accompanied by a lack of development and possible regression in skein-like intranuclear inclusions together with a depletion in cytoplasmic dense bodies. Such changes were accompanied by the appearance of nuclear dense bodies. These were also present when virus growth was reduced (5 microM-DHPG) rather than completely inhibited (50 microM-DHPG) by treatment initiated from the time of infection. The nuclear bodies were predominantly of a reticular type structure after the early treatment but mainly of a homogeneous form when virus growth was interrupted at late times. Their presence appeared to be connected with the ability of infected cells to initiate the synthesis of late proteins and their morphology may relate to the extent of such protein synthesis. Unlike cytoplasmic dense bodies, provisional findings on the characterization of the nuclear bodies suggested that the 69K matrix protein was not present in abundance.


Upfront Use of 177Lu-Labeled PSMA Radioligand Therapy and Treatment Response Assessment in Treatment-Naive Prostate Cancer.
Prostatic malignancy is the most common type of nonskin cancer and associated with significant morbidity and mortality. Early androgen deprivation therapy (ADT) is the treatment of choice in metastatic prostate cancer, whereas ADT delays progression of disease, but it is associated with significant adverse effects and frequently impairs the quality of life. Therefore, there is growing interest in treatments to postpone ADT while achieving a good progression-free survival. We present a case of oligometastatic prostate cancer, who was treated upfront with 177Lu-labeled PSMA radioligand therapy and demonstrated excellent response to a single dose of 177Lu-PSMA-617.


Cardiomyopathies of infancy.
Cardiomyopathies in infancy constitute an ill-defined group of conditions. Recently, however, a number of specific disorders of metabolism and mitochondrial abnormalities have been described. Idiopathic or primary cardiomyopathies remain poorly defined. Endocardial fibroelastosis, the most prominent member in this group, probably represents a reaction of the endocardium to various insults. The different types of cardiomyopathy (CM) present a similar clinical picture and require the expertise of a group of specialists for proper diagnosis. Therapy is mostly supportive, as only a few cases of CM are amenable to specific treatment. Orthopic cardiac transplantation may become a therapeutic option in the future for infants unresponsive to current therapy.


[A study on the sustainable effects of reeducation on cardiopulmonary resuscitation on nurses' knowledge and skills].
This study was to analyze the sustainable effects of cardiopulmonary resuscitation (CPR) reeducation on nurses' knowledge and skills. A repeated experimental design was used for a single sample group of 47 nurses working for a general hospital. The nurses were tested on their skill of CPR 3 times at an interval of 4 months. In order to test nurses' knowledge and skills, the researcher used a CPR assessment program linked to an adult practice doll (Anne). 1) The amount of decrease of nurses' knowledge about CPR between points of time was wide between the first and second points of time but small between the second and third time owing to the effects of reeducation between the two points of time. 2) Nurses' skills between the first and second time dropped but they improved between the second and third time owing to the effects of reeducation. As confirmed by the above findings, reeducation of CPR clearly affects nurses' knowledge and skills. Given the fact that the same period of time (4 months) elapsed between the 3 tests, it could be argued that the reeducation at the second test served to maintain nurses' knowledge and enhance their skills.


Antiangiogenic potency of vitamin E.
We investigated the antiangiogenic property and mechanism of vitamin E compounds, with particular emphasis on tocotrienol (T3), a natural analogue of tocopherol (Toc). T3 inhibited both the proliferation and tube formation of bovine aortic endothelial cells, with delta-T3 appearing to have the highest activity. delta-T3 also reduced the vascular endothelial growth factor (VEGF)-stimulated tube formation by human umbilical vein endothelial cells. Moreover, delta-T3 inhibited the new blood vessel formation on the growing chick embryo chorioallantoic membrane (assay for in vivo angiogenesis). Orally administered T3 suppressed the tumor cell-induced angiogenesis in the mouse dorsal air sac assay. In contrast with T3, Toc showed very weak inhibition. Based on DNA microarray analysis, antiangiogenic effect of T3 was attributable in part to regulation of intracellular VEGF signaling (phospholipase C-gamma and protein kinase C). Our findings suggest that T3 has potential as a therapeutic dietary supplement for preventing angiogenic disorders.