Calcitriol and calcium regulate cytokine production and adipocyte-macrophage cross-talk.
The objective of this study was to investigate the effects of calcitriol on adipocyte and macrophage cytokine expression as well as release and on adipocyte-macrophage cross-talk in local modulation of inflammation. We investigated calcitriol modulation of the expression of macrophage inhibitory factor (MIF) and macrophage surface-specific protein CD14, two key factors in regulating macrophage function and survival, in differentiated human adipocytes. Calcitriol significantly increased MIF and CD14 expression by 59% and 33%, respectively, while calcium-channel antagonism with nifedipine completely reversed these effects, indicating that calcitriol stimulates MIF and CD14 expression via a calcium-dependent mechanism. Similar results were also found in cultured 3T3-L1 adipocytes; in addition, calcitriol also up-regulated macrophage colony-stimulating factor, macrophage inflammatory protein, interleukin-6 (IL-6) as well as monocyte chemoattractant protein-1 expression in 3T3-L1 adipocytes and stimulated tumor necrosis factor as well as IL-6 expression in RAW 264 macrophages. These effects were blocked by either a calcium-channel antagonist (nifedipine) or a mitochondrial uncoupler (dinitrophenol). Moreover, co-culture of 3T3-L1 adipocytes with RAW 264 macrophages significantly increased the expression and production of multiple inflammatory cytokines in response to calcitriol in both cell types. These data demonstrate that calcitriol regulates local inflammation via modulating the interaction between adipocytes and macrophages as well as regulating inflammatory cytokine production in each cell type via calcium-dependent and mitochondrial uncoupling-dependent mechanisms. These data provide further mechanistic explanation for our recent observations that suppression of calcitriol by dietary calcium reduces inflammatory cytokine expression and oxidative stress in adipose tissue.


Acetabular Component Removal Systems.
As primary total hip arthroplasty procedures continue to rise, acetabular revision surgery will be in increasing demand. Treatment of acetabular component failure and associated bone defects depends on patient characteristics, the degree and location of bone loss, the ability of the columns to support biologic fixation, and the presence of pelvic discontinuity. In revision total hip arthroplasty, technological advances have been made to address limitations of acetabular cup removal while preserving host bone stock. The goal of acetabular revision reconstruction should be to obtain stable fixation and restore the hip center. The various acetabular cup removal systems are discussed.


Unidirectional ablation electrode to minimize thermal injury during radiofrequency ablation: an experimental study in an ex vivo bovine liver model.
To modify the direction of radiofrequency (RF) ablation and to find optimal technical parameters using a partially insulated electrode. The authors developed a unidirectional ablation electrode (UAE) by partial insulation of a 1-cm active electrode tip. Each electrode produced 60 ablation zones using insulation of the active tip (270°, group A; 180°, group B; and noninsulated, group C). In each group, six subgroups were created by combining the RF powers (5 W, 10 W, and 20 W) and ablation duration (6 minutes and 12 minutes). Technical success was determined when a half-moon-shaped ablation zone was achieved. The volumes of the technical success zone (Vs) and the technical failure zone (Vf) were measured. Technical success was achieved in all 10 cases using a 270° insulated electrode with a 5-W/6-minute parameter. The mean Vs of group A was significantly smaller than the mean Vs of either group B or group C (P = .003 and P < .001). The mean Vf of group A was significantly smaller than the mean Vf of group B, which was smaller than that of group C (P < .001). A UAE with a partially insulated tip can modify the ablation zone, especially when using 270° insulation with a 5-W/6-minute parameter.


Design, synthesis, and biological evaluation of novel stachydrine derivatives as potent neuroprotective agents for cerebral ischemic stroke.
Stachydrine is a natural product with multiple protective biological activities, including those involved in preventing cancer, ischemia, and cardiovascular disease. However, its use has been limited by low bioavailability and unsatisfactory efficacy. To address this problem, a series of stachydrine derivatives (A1/A2/A3/A4/B1/B2/B3/B4) were designed and synthesized, and biological studies were carried out in vitro and in vivo. When compared with stachydrine, Compound B1 exhibited better neuroprotective effects in vitro, and significantly reduced infarction size in the model of the middle cerebral artery occlusion rat model. Therefore, Compound B1 was selected for further research on ischemic stroke. Graphical abstract.


Posttransplant MRD and T-cell chimerism status predict outcomes in patients who received allografts for AML/MDS.
Allogeneic stem-cell transplant allows for the delivery of curative graft-versus-leukemia (GVL) in patients with acute myeloid leukemia/myelodysplasia (AML/MDS). Surveillance of T-cell chimerism, measurable residual disease (MRD) and blast HLA-DR expression may inform whether GVL effectiveness is reduced. We report here the prognostic impact of these biomarkers in patients allografted for AML/MDS. One hundred eighty-seven patients from FIGARO, a randomized trial of reduced-intensity conditioning regimens in AML/MDS, were alive and relapse-free at the first MRD time-point and provided monitoring samples for flow cytometric MRD and T-cell chimerism, requested to month+12. Twenty-nine (15.5%) patients had at least 1 MRD-positive result posttransplant. MRD-positivity was associated with reduced overall survival (OS) (hazard ratio [HR], 2.18; P = .0028) as a time-varying Cox variable and remained significant irrespective of pretransplant MRD status in multivariate analyses (P < .001). Ninety-four patients had sequential MRD with T-cell chimerism results at months+3/+6. Patients with full donor T-cell chimerism (FDTC) had an improved OS as compared with patients with mixed donor T-cell chimerism (MDTC) (adjusted HR=0.4; P = .0019). In patients with MDTC (month+3 or +6), MRD-positivity was associated with a decreased 2-year OS (34.3%) vs MRD-negativity (71.4%) (P = .001). In contrast, in the group with FDTC, MRD was infrequent and did not affect the outcome. Among patients with posttransplant MRD-positivity, decreased HLA-DR expression on blasts significantly reduced OS, supporting this as a mechanism for GVL escape. In conclusion, posttransplant MRD is an important predictor of the outcome in patients allografted for AML/MDS and is most informative when combined with T-cell chimerism results, underlining the importance of a GVL effect in AML/MDS.


An exploration of pregnant women and mothers' attitudes, perceptions and experiences of formula feeding and formula marketing, and the factors that influence decision-making about infant feeding in South Africa.
Despite strong evidence showing the lifelong benefits of breastfeeding for mothers and children, global breastfeeding practices remain poor. The International Code of Marketing of Breastmilk Substitutes is an internationally agreed code of practice, adopted by the World Health Assembly in 1981, to regulate promotion of commercial formula, and is supported by legislation in many countries. However, marketing of formula remains widespread and contributes to mother's decisions to formula feed. We present South African data from a multi-country, mixed-methods study exploring women's decision-making about infant feeding and how this was influenced by exposure to formula marketing. Using a consumer-based marketing approach, focus group discussions (FGDs) were conducted with pregnant women and mothers of children aged between 0 and 18 months in two urban sites in South Africa. Participants were purposively selected according to their child's age, infant feeding practices and socioeconomic status. Ten FGDs were conducted during February 2020 with a total of 69 participants. Thematic analysis was used to analyse the data with NVivo v.12 software. Despite being encouraged by health professionals to breastfeed and intending to do so, many mothers chose to give formula in the early weeks and months of their child's life. Mothers reported breastfeeding challenges as the most frequent reason for initiating infant formula, stating that family members and health professionals recommended formula to solve these challenges. Although participants described few advertisements for infant formula, advertisements for 'growing-up' formulas for older children were widespread and promoted brand recognition. Mothers experienced other marketing approaches including attractive packaging and shop displays of infant formula, and obtained information from social media and online mothers' groups, which influenced their choice of formula brand. Mothers reported strong brand loyalty derived from previous experiences and recommendations. Health professionals frequently recommended formula, including recommending specific formula brands and specialist formulas. Global formula companies use multifaceted marketing methods to promote a strong narrative portraying formula feeding as a positive lifestyle choice. Positive, coordinated efforts are required to counter pro-formula messaging and change the narrative to support breastfeeding as an aspirational choice. In particular, health professionals must stop supporting the formula industry.


Saikosaponin-d impedes hippocampal neurogenesis and causes cognitive deficits by inhibiting the survival of neural stem/progenitor cells via neurotrophin receptor signaling in mice.
Neural stem/progenitor cells (NPCs) are multipotent stem cells in the central nervous system. Damage to NPCs has been demonstrated to cause adverse effects on neurogenesis and to contribute to neurological diseases. Our previous research suggested that saikosaponin-d (SSd), a cytostatic drug belonging to the bioactive triterpenoid saponins, exhibited neurotoxicity by inhibiting hippocampal neurogenesis, but the underlying mechanism remained elusive. This study was performed to clarify the role of SSd in cognitive function and the mechanism by which SSd induced damage to hippocampal neurogenesis and NPCs. Our results indicated that SSd caused hippocampus-dependent cognitive deficits and inhibited hippocampal neurogenesis by reducing the numbers of newborn neurons in mice. RNA sequencing analysis revealed that SSd-induced neurotoxicity in the hippocampus involved neurotrophin receptor-interacting MAGE (NRAGE)/neurotrophin receptor interacting factor (NRIF)/p75NTR -associated cell death executor (NADE) cell signaling activated by the p75 neurotrophin receptor (p75NTR ). Mechanistic studies showed that a short hairpin RNA targeting p75NTR intracellular domain reversed SSd-increased NRAGE/NRIF/NADE signaling and the c-Jun N-terminal kinase/caspase apoptotic pathway, subsequently contributing to the survival of NPCs, as well as cell proliferation and differentiation. The addition of recombinant brain-derived neurotrophic factor (BDNF) ameliorated the SSd-induced inhibition of BDNF/Tyrosine kinase receptor B (TrkB) neurotrophic signaling, but did not affect SSd-activated pro-BDNF/p75NTR signaling. Moreover, the SSd-induced elevation of cytosolic Ca2+ concentration was responsible for damage to NPCs. The extracellular Ca2+ chelator ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA), rather than the intracellular Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl ester) (BAPTA/AM), attenuated SSd-induced cytosolic Ca2+ dysfunction and SSd-disordered TrkB/p75NTR signaling. Overall, this study demonstrated a new mechanism for the neurotoxic effect of SSd, which has emerging implications for pharmacological research of SSd and provides a better understanding of neurotoxicity induced by cytostatic drugs.


Self-immolative polyprodrug-based tumor-specific cascade amplificated drug release nanosystem for orchestrated synergistic cancer therapy.
Reactive oxygen species (ROS)-activated prodrugs can potentially improve the selectivity of chemotherapeutics. However, the inability to release sufficient drugs at tumor sites due to the paucity of ROS, which is required for prodrug activation usually limits the antitumor potency. Herein, a delivery nanosystem with self-amplifiable drug release pattern is constructed by encapsulating a tumor specificity ROS inducer NAD(P)H: quinone oxidoreductase-1 (NQO1)-responsive hemicyanine fluorescent dye (NCyNH2) in a ROS-responsive self-immolative polyprodrug nanoparticle for orchestrated oxidation-chemotherapy. In response to ROS stimulation, the self-immolative polyprodrug can degrade and release doxorubicin (DOX) through a domino-like fragmentation, which can impart advanced attributes of this nanosystem such as minimum cleavage events required and maximum cleavage speed for disintegration. Thus, the NCyNH2-loaded self-immolative polyprodrug nanoparticle (SIPN) could be dissociated in response to endogenous ROS, triggering the release of DOX and NCyNH2. Subsequently, the NCyNH2 could be activated by intratumoral overexpressed NQO1 to generate additional ROS, which further induces the amplifiable degradation of self-immolative polyprodrug to release sufficient drugs. The in vitro and in vivo studies consistently demonstrate that SIPN amplifies the drug release efficiency of ROS-responsive polyprodrug by specifically upregulating intratumoral ROS levels, resulting in significant antitumor efficacy with minimal side effects.


Effects of combined THC and heroin vapor inhalation in rats.
Opioids are effective medications, but they have several key limitations including the development of tolerance, establishment of dependence, diversion for non-medical use, and the development of addiction. Therefore, any drugs which act in an additive or synergistic fashion with opioids to address medical applications have the potential to reduce opioid-related harms. To determine if heroin and Δ9-tetrahydrocannabinol (THC) interact in an additive or independent manner to alter nociception, body temperature, and spontaneous locomotor activity when inhaled or injected. Groups of female and male rats, implanted with radiotelemetry transmitters, were exposed to vapor generated from heroin (50 mg/mL in propylene glycol vehicle; PG), THC (50 mg/mL), or the combination for assessment of effects on temperature and activity. Thermal nociception was assessed with a warm water tail-withdrawal assay. Heroin inhalation increased temperature and activity whereas THC inhalation decreased temperature and activity in both female and male Sprague-Dawley rats. Effects of combined inhalation were in opposition, and additional experiments found the same outcome for the injection of heroin (0.5 mg/kg, s.c.) and THC (10 mg/kg, i.p.) alone and in combination. In contrast, the co-administration of heroin and THC by either inhalation or injection produced additive effects on thermal nociception in both male and female Sprague-Dawley and Wistar rats. This study shows that additive effects of THC with an opioid on a medical endpoint such as analgesia may not generalize to other behavioral or physiological effects, which may be a positive outcome for unwanted side effects.


Hapl-o-Mat: open-source software for HLA haplotype frequency estimation from ambiguous and heterogeneous data.
Knowledge of HLA haplotypes is helpful in many settings as disease association studies, population genetics, or hematopoietic stem cell transplantation. Regarding the recruitment of unrelated hematopoietic stem cell donors, HLA haplotype frequencies of specific populations are used to optimize both donor searches for individual patients and strategic donor registry planning. However, the estimation of haplotype frequencies from HLA genotyping data is challenged by the large amount of genotype data, the complex HLA nomenclature, and the heterogeneous and ambiguous nature of typing records. To meet these challenges, we have developed the open-source software Hapl-o-Mat. It estimates haplotype frequencies from population data including an arbitrary number of loci using an expectation-maximization algorithm. Its key features are the processing of different HLA typing resolutions within a given population sample and the handling of ambiguities recorded via multiple allele codes or genotype list strings. Implemented in C++, Hapl-o-Mat facilitates efficient haplotype frequency estimation from large amounts of genotype data. We demonstrate its accuracy and performance on the basis of artificial and real genotype data. Hapl-o-Mat is a versatile and efficient software for HLA haplotype frequency estimation. Its capability of processing various forms of HLA genotype data allows for a straightforward haplotype frequency estimation from typing records usually found in stem cell donor registries.


Hair can be a good biomarker of occupational exposure to mercury vapor: simulated experiments and field data analysis.
Generally, urine mercury (U-Hg) is widely used for assessment of inorganic mercury (I-Hg) exposure in humans. The reliability of scalp hair as an indicator of mercury vapor exposure is contentious. However, significant correlations were found between hair total mercury (T-Hg) and U-Hg and between hair I-Hg and total gaseous mercury (TGM) in ambient air in our previous studies. Simulated experiments were designed to assess the contribution of direct absorption/adsorption of mercury vapor in the hair. Results indicated that the increases of hair T-Hg concentrations were less than 1 μg/g, which was negligible compared with hair T-Hg concentrations in occupationally exposed workers. The β-mercaptoethanol washing can remove 30% of mercury (Hg) in the exposed hair samples. The inhaled Hg constituted the major fraction (97.4%) of I-Hg exposure for the artisanal Hg mining workers. From the simulated experiments and field data analysis, we can conclude that hair I-Hg can be a useful tool for monitoring occupational exposure to Hg vapor.


External cephalic version from 34 weeks under tocolysis: factors influencing success.
External cephalic version (ECV) was performed on 72 patients from 34 weeks of gestation under tocolysis, with ultrasound control and cardiotocograph surveillance. Successful ECV was associated with multiparity, decreased lower segment caesarean section (LSCS) rate, and earlier discharge home. ECV was successful in 51.4% of patients overall, with the success rate being 35% in primigravidas and 71.4% in multiparas (p < 0.005). The LSCS rate was reduced from 91.4% to 0% (p < 0.0001) on comparing the unsuccessful and the successful ECV groups, while breech presentation was reduced from 94.3% to 0% (p < 0.0001). There were no significant fetal complications.


Fabrication of activated carbon supported modified with bimetallic-platin ruthenium nano sorbent for removal of azo dye from aqueous media using enhanced ultrasonic wave.
Herein, activated carbon supported modified with bimetallic-platin ruthenium nano sorbent (PtRu@AC) was synthesized by a thermal decomposition process and used in the removal of methylene blue (MB) from aqueous solutions. The synthesized nano sorbents were characterized by X-ray Diffraction (XRD), Transmission Electron Microscopy (TEM), and X-ray Photoelectron Spectroscopy (XPS) spectroscopic techniques. The data obtained from characterization studies showed that PtRu@AC nano sorbent was highly crystalline and in a form of PtRu alloy with a monodispersed composition. The results indicated that the maximum adsorption capacity (qemax) for the removal of MB with PtRu@AC under optimum conditions was detected to be 1.788 mmol/g (569.4 mg/g). The experimental kinetic results of the study revealed that the adsorption of methylene blue was found to be more compatible with the false second-order model compared to some tested models. Calculations for thermodynamic functions including enthalpy change (ΔHo), entropy change (ΔSo), and Gibbs free energy change (ΔGo) values were performed to get an idea about the adsorption mechanism. As a result, the synthesized PtRu@AC nano adsorbent was detected as a highly effective adsorbent material in the removal of MB from aquatic mediums.


Impact of public support and government's policy on climate change in China.
Climate change is a global environmental issue that would damage the natural and biological systems. Although there are many controversies about climate change, the temperature rise has become more and more obvious in the world since the 1990s. It is worthwhile to understand whether the public supports the government's policy on climate change and how public support affects climate change. A case study of the first-tier cities in China is done to explore these questions through questionnaire surveys. 3468 valid questionnaires from four first-tier cities in China were screened out are used for individual behavior analysis. The results show that the respondents' perceptions towards climate change in China are laxer than those in other countries, meanwhile, they hope that the government would respond to climate change actively. The influencing factors mainly include the demographic characteristics and their perception of climate change. Demographic characteristics are about gender, income, marital status, age, and whether they have children. Moreover, respondents' perception of climate change has a significant impact on their attitudes towards the government's policies for mitigating climate change. The conclusions are drawn based on the comparative analysis of the survey results and suggestions are put forward for making climate change policies.


Effect of Time of Administration of Teriparatide on Bone Mineral Density in Glucocorticoid-Induced Osteoporosis.
Teriparatide (TPTD) (recombinant DNA origin human parathormone [1-34]) is approved for the treatment of glucocorticoid-induced osteoporosis (GIO). There are reports of factors that affect the response to TPTD in GIO treatment. This work describes the case of a 71-yr-old woman diagnosed with lupus nephropathy treated with 40 mg/d of meprednisone, and who suffered multiple vertebral fractures. Despite treatment with a single 5 mg dose of zoledronic acid, the patient continued to have vertebral fractures. Treatment with 20 µg/d of subcutaneous TPTD (PTH1-34, Forteo; Eli Lilly Co., Indianapolis, IN) was initiated. Nine months after the onset of treatment, bone mineral density (BMD) assessment showed a 5% decrease in lumbar spine BMD. Factors potentially affecting the results were analyzed. The patient reported injecting TPTD at night and was instructed to inject TPTD in the morning before breakfast. After changing the time of TPTD administration and 22 mo after initiating treatment, BMD assessment was repeated and showed an 18% increase at the lumbar spine and no new vertebral fractures. The time of TPTD administration might affect the response to TPTD in GIO treatment.


Placental morphology and the prediction of underlying cardiovascular risk factors.
Pre-eclampsia is associated with an increased risk of future cardiovascular disease. Maladaptive placentation or malperfusion, as well as predisposing cardiovascular or metabolic risk for endothelial dysfunction, contribute to the systemic inflammatory response that establishes the origins of the disease. The purpose of this study was to investigate the relationship between placental size and cardiovascular risk when assessed at six months postpartum in women who experienced pre-eclampsia. Maternal clinical and biochemical cardiovascular risk factors were used to categorize preeclamptic women into high vs. low lifetime cardiovascular disease risk profiles at six months postpartum. A multivariable logistic regression model was then used to identify the association between placental weight to birth weight ratio and high lifetime cardiovascular disease risk, adjusting for maternal age, pre-pregnancy BMI, and severity of pre-eclampsia. A p-value of < 0.05 was deemed statistically significant. 186/216 women with pre-eclampsia who attended the Maternal Health Clinic met inclusion criteria. No significant differences were observed for placental morphometric measurements between women who screened as having a high vs. low lifetime risk profile for cardiovascular disease at six months postpartum. However, using multivariable modelling that controlled for maternal age, pre-pregnancy body mass index, gestational age at delivery, and severity of pre-eclampsia, a low placenta to birth weight ratio (<15%) was associated with an increased odds of high lifetime cardiovascular disease risk (p < 0.009). The findings of the current study identify clinical measurements that can be collected at the time of delivery which may help identify specific women who may benefit most from postpartum cardiovascular risk screening and intervention.


Basal cell pseudopodia in cervical intraepithelial neoplasia; progressive reduction of number with severity: a morphometric quantification.
An application of a morphometric model is used to study the stroma epithelial interface in human cervical epithelium. This model derived by Okagaki and associates estimates from two dimensional transmission electron photomicrographs the number of basal cell pseudopodia per basal cell. The investigation of the full spectrum of cervical intraepithelial neoplasia (CIN) focusing on the basal cell-stroma interface with the "multiple cell-single section" model is discussed. Significant decreases in histopathologic grades of CIN are noted in the number of pseudopodia per basal cell (N) and the ratio (F) of the area of the stems of pseudopodia to the area of the base of the basal cell.


A revision of the conductive hearing loss in Cranium 4 from the Middle Pleistocene site of Sima de los Huesos (Burgos, Spain).
Pathological conditions have been previously documented in the Middle Pleistocene Sima de los Huesos hominins from northern Spain, and several of these have clear behavioral implications. Within this fossil assemblage, Cranium 4 shows bilateral external auditory exostoses which have been preliminarily interpreted as causing a significant hearing loss in this individual. If confirmed, this would be the oldest recorded case of deafness in human history and could have important implications for the antiquity of this condition, as well as social interactions. To further investigate this case, the current study presents 3D reconstructions of the entire outer and middle ear, based on computed tomography scans of both temporal bones in Cranium 4. We established the degree of stenosis in both external auditory canals, showing that in both cases the degree of stenosis is less than 52% of the original cross-sectional area of each canal. Based on clinical studies in living humans, the buildup of wax due to the degree of stenosis in Cranium 4 is unlikely to have caused frequent external ear infections. In addition, we estimated the pattern of sound power transmission up to 5 kHz in both ears relying on a comprehensive model developed in the bioengineering literature and which has been applied previously to the Sima de los Huesos hominins. The model was modified to account for the peculiar shape of the pathological external ear canals in Cranium 4. The results show that this pathology had little to no influence on the sound power transmission in this individual. Thus, we conclude that the exostoses present in both ears of Cranium 4 did not significantly affect their hearing.


Glibenclamide modulates microglial function and attenuates Aβ deposition in 5XFAD mice.
Severe neuroinflammation is known as a main pathology of neurodegenerative disorders such as Alzheimer's disease (AD) and Parkinson's disease (PD). In these diseases, excessive microglial activation is one of the main causes of inflammation in the central nervous system. Therefore, inhibition of activated microglia may be suggested as a treatment for neuroinflammatory diseases. Glibenclamide, known as a therapeutics for type 2 diabetes in clinical trials has been shown to be effective in the inhibiting inflammatory conditions of various diseases. However, studies on the effects of glibenclamide for improving AD pathologies are little known. In this study, we tested glibenclamide on microglial cell line BV2 and 5XFAD mice. We found that glibenclamide significantly inhibited nitric oxide (NO) at 10 μM and 40 μM in BV2 cells induced by lipopolysaccharide (LPS) stimulation. In addition, we confirmed that 40 μM of glibenclamide reduced pro-inflammatory cytokines and proteins in the LPS-stimulated microglial cells. The anti-inflammatory effect of glibenclamide was further tested in APP/PS1 transgenic mouse. Although further analysis would be needed to confirm whether glibenclamide affects behavioral performance, our data suggests that glibenclamide may be a therapeutic option for AD treatment.


Gender-dependent hepatic alterations in H-ras12V transgenic mice.
Although it has been proposed that Ras and related signal pathways play important roles in hepatocarcinogenesis, appropriate in vivo models are lacking. Two hepatocellular carcinoma lines were established using pronuclear microinjection techniques to create an insertion of the H-ras12V transgene under the control of the albumin enhancer/promoter. The resulting phenotypes and related molecular events were then examined. Male (but not female) transgenic mice older than 2 months showed hepatic alterations with a high degree of reproducibility, as compared to the wild-type mice. The liver/body-weight ratios were lower for the females than for the males. The transgene-carrying line 28 was investigated extensively with respect to molecular differences between the genders. Male hepatocytes showed higher Ras activity and higher reactive oxygen species (ROS) levels than female hepatocytes. The female hepatocytes showed higher expression levels of p53 and p21Waf1/Cip1, enhanced cytochrome c release, which correlated with cell cycle arrest, and higher levels of hypodiploid cell formation, as compared to the male hepatocytes. The gender-related differences in molecular responses to activated Ras may have implications for the prevalence of hepatic alterations in males. Our transgenic mice represent a potentially valuable animal model for future investigations.


Long-term prognosis after a normal exercise stress Tc-99m sestamibi SPECT study.
Patients with a normal stress technetium 99m sestamibi study were shown to have a favorable outcome at intermediate-term follow-up. However, long-term survival has not been studied. The aim of this study was to evaluate the incidence and predictors of mortality and cardiac events at long-term follow-up after a normal exercise stress sestamibi study. Methods and results We studied 218 patients (mean age, 53 +/- 10 years, 108 men) who had normal myocardial perfusion assessed by Tc-99m sestamibi single photon emission computed tomography at rest and during symptom-limited bicycle exercise stress test. Endpoints during a follow-up period of 7.4 +/- 1.8 years were hard cardiac events (cardiac death and nonfatal myocardial infarction) and all-cause mortality. During follow-up, 13 patients died of various causes (cardiac death in 1 patient). Ten patients had nonfatal myocardial infarction (a total of 11 hard cardiac events). By multivariate analysis, independent predictors of cardiac events were history of coronary artery disease (chi(2) = 5, P =.03) and lower exercise heart rate (chi(2) = 12, P =.001). Independent predictors of all-cause mortality were age (chi(2) = 4, P =.05) and exercise heart rate (chi(2) = 5, P =.03). The annual mortality rate was 0.6% in the first 5 years and 1.8% between the sixth and eighth years. The annual hard cardiac event rate was 0.7% in the first 5 years and 1.5% between the sixth and eighth years. Receiver operating characteristic curves identified an exercise heart rate lower than 130 beats/min as the cutoff value that separated patients with regard to their risk for mortality and hard cardiac events. It is concluded that the annual mortality and cardiac event rate is less than 1% during 5-year follow-up after a normal exercise sestamibi study. Therefore repeated testing would not be required unless there is a change in symptoms. Follow-up should be closer in patients with a history of coronary artery disease and in those who fail to achieve an exercise heart rate of 130 beats/min or greater.


Why the term 'persistent therapy' is not worse than the term 'medical futility'.
The discussion around the use of the term 'medical futility' began in the late 1980s. The Polish Working Group on End-of-Life Ethics (PWG) joined this discussion in 2008. They offered their own approach to the issues regarding medical futility based on the category of persistent therapy. According to the PWG, 'persistent therapy is the use of medical procedures to maintain the life function of the terminally ill in a way that prolongs their dying, introducing excessive suffering or violating their dignity'. In this paper I attempt to show that the term 'persistent therapy' is neither worse nor better than the term 'medical futility', but it captures different aspects and nuances. Additionally, the Polish social and religious background plays a significant role in shaping the category of persistent therapy.


Cytokine storms are primarily responsible for the rapid death of ducklings infected with duck hepatitis A virus type 1.
Duck hepatitis A virus type 1 (DHAV-1) is one of the most harmful pathogens in the duck industry. The infection of adult ducks with DHAV-1 was previously shown to result in transient cytokine storms in their kidneys. To understand how DHAV-1 infection impacts the host liver, we conducted animal experiments with the virulent CH DHAV-1 strain and the attenuated CH60 commercial vaccine strain. Visual observation and standard hematoxylin and eosin staining were performed to detect pathological damage in the liver, and viral copy numbers and cytokine expression in the liver were evaluated by quantitative PCR. The CH strain (108.4 copies/mg) had higher viral titers than the CH60 strain (104.9 copies/mg) in the liver and caused ecchymotic hemorrhaging on the liver surface. Additionally, livers from ducklings inoculated with the CH strain were significantly infiltrated by numerous red blood cells, accompanied by severe cytokine storms, but similar signs were not observed in the livers of ducklings inoculated with the CH60 strain. In conclusion, the severe cytokine storm caused by the CH strain apparently induces hemorrhagic lesions in the liver, which might be a key factor in the rapid death of ducklings.


[Laughter: gender differences].
Laughter is associated to many physiological and psychological benefits. Although women laugh more than men do, the daily frequency of laughter does not seem to differ. Laughter in all its forms and manifestations is an indicator of family vitality and healthy couples. Laughter is very attractive at the interpersonal level, especially for women. Men use humor much more and laughter when it comes to discussing sensitive health issues. In women, laughter would be more associated with greater social support in relationships and as a tool to cope with stress. Inviting laughter in the doctor's office may be very useful when directing certain messages on therapeutic management. Taking into account possible gender differences in the use of humor and laughter may help to improve the relationship with the patient and optimize the clinical application of laughter in health care and education setting.


An enhanced prognostic score for overall survival of patients with cancer derived from a large real-world cohort.
By understanding prognostic biomarkers, we gain insights into disease biology and may improve design, conduct, and data analysis of clinical trials and real-world data. In this context, we used the Flatiron Health Electronic Health Record-derived deidentified database that provides treatment outcome and biomarker data from >280 oncology centers in the USA, organized into 17 cohorts defined by cancer type. In 122 694 patients, we analyzed demographic, clinical, routine hematology, and blood chemistry parameters within a Cox proportional hazard framework to derive a multivariable prognostic risk model for overall survival (OS), the 'Real wOrld PROgnostic score (ROPRO)'. We validated ROPRO in two independent phase I and III clinical studies. A total of 27 variables contributed independently and homogeneously across cancer indications to OS. In the largest cohort (advanced non-small-cell lung cancer), for example, patients with elevated ROPRO scores (upper 10%) had a 7.91-fold (95% confidence interval 7.45-8.39) increased death hazard compared with patients with low scores (lower 10%). Median survival was 23.9 months (23.3-24.5) in the lowest ROPRO quartile Q1, 14.8 months (14.4-15.2) in Q2, 9.4 months (9.1-9.7) in Q3, and 4.7 months (4.6-4.8) in Q4. The ROPRO model performance indicators [C-index = 0.747 (standard error 0.001), 3-month area under the curve (AUC) = 0.822 (0.819-0.825)] strongly outperformed those of the Royal Marsden Hospital Score [C-index = 0.54 (standard error 0.0005), 3-month AUC = 0.579 (0.577-0.581)]. We confirmed the high prognostic relevance of ROPRO in clinical Phase 1 and III trials. The ROPRO provides improved prognostic power for OS. In oncology clinical development, it has great potential for applications in patient stratification, patient enrichment strategies, data interpretation, and early decision-making in clinical studies.


Calcium requirement of uterine contraction induced by PGE1: importance of intracellular calcium stores.
The contraction of the rat uterus in response to PGE1 in high K+ medium and in Ca-free solution which contained EDTA has been investigated in order to examine whether excitation-contraction coupling involves the release of Ca from an intracellular store. In uterus maximally contracted by K+, cumulative concentrations of PGE1 (1.25 - 20 ng/ml) caused maintained concentration-dependent contraction. PGE1 induced sustained contraction of rat uterus in Ca-free medium after incubation with 3mM EDTA for 50 min. In these conditions the involvement of extracellular Ca is highly unlikely. The PGE1-induced contraction could be repeated without exposure to external Ca ions and with only slight reduction in magnitude. The PGE1 concentrations required to elicit uterine contraction in Ca-free solution were about 1000 times higher than the effective doses in KCl-depolarized uterus. In conclusion, the present investigation shows that Ca influx is not essential for PGE1-induced contraction of rat uterus, although extracellular Ca enhances it presumably by increasing the free Ca levels in the cytosol.


Crossing the Golden Training Divide: The Science and Practice of Training World-Class 800- and 1500-m Runners.
Despite an increasing amount of research devoted to middle-distance training (herein the 800 and 1500 m events), information regarding the training methodologies of world-class runners is limited. Therefore, the objective of this review was to integrate scientific and best practice literature and outline a novel framework for understanding the training and development of elite middle-distance performance. Herein, we describe how well-known training principles and fundamental training characteristics are applied by world-leading middle-distance coaches and athletes to meet the physiological and neuromuscular demands of 800 and 1500 m. Large diversities in physiological profiles and training emerge among middle-distance runners, justifying a categorization into types across a continuum (400-800 m types, 800 m specialists, 800-1500 m types, 1500 m specialists and 1500-5000 m types). Larger running volumes (120-170 vs. 50-120 km·week-1 during the preparation period) and higher aerobic/anaerobic training distribution (90/10 vs. 60/40% of the annual running sessions below vs. at or above anaerobic threshold) distinguish 1500- and 800-m runners. Lactate tolerance and lactate production training are regularly included interval sessions by middle-distance runners, particularly among 800-m athletes. In addition, 800-m runners perform more strength, power and plyometric training than 1500-m runners. Although the literature is biased towards men and "long-distance thinking," this review provides a point of departure for scientists and practitioners to further explore and quantify the training and development of elite 800- and 1500-m running performance and serves as a position statement for outlining current state-of-the-art middle-distance training recommendations.


Heterozygous effects of x-ray induced mutations on viability of Drosophila melanogaster.
Drosophila second chromosomes that had received 1000 R of X-irradiation mainly in the mature sperm stage were tested for heterozygous effects on viability. When the genetic background was heterozygous the viability was reduced by about 1.5 percent; in a nearly homozygous background the viability was enhanced by about the same amount. In a partially inbred background the results were intermediate. The results are therefore in agreement with earlier experiments of Wallace and Mukai.


Clinical measurements of hard palate and implications for subepithelial connective tissue grafts with suggestions for palatal nomenclature.
The objectives of the present study were to 1) identify a reliable measuring technique for a palatal graft, 2) observe the patterns of the neurovascular bundle, and 3) identify the morphology of the hard palate. Subepithelial connective tissue grafts are performed at an increasing rate to improve esthetics and oral health. Palatal graft techniques began in 1963, and today the subepithelial connective tissue graft is the most widely accepted technique. The greater palatine nerve and artery are critical neurovascular structures to identify. Their pattern and palate morphology are not well defined. Anatomy texts, atlases, and specialty texts were analyzed. We dissected 17 palates (17 left and right halves) from embalmed human cadavers, implementing a measuring technique to locate the most coronal structure of the greater palatine artery and greater palatine nerve bundle, observing the patterns and palatal morphology. Electronic digital calipers and a periodontal probe were used for data collection. The dissection results revealed a reliable measuring technique, a common pattern of the bundle, and osseous palatal landmarks not clearly defined in contemporary texts. The dissections also demonstrated a medial and lateral groove, along with a crest in the palatine process of the maxillary bone. The greater palatine artery traversed the lateral groove, and the greater palatine nerve traversed the medial groove consistently. The crest was located anteroposteriorly between the grooves. The results of our study suggest a useful measurement technique, a consistent neurovascular pattern, and the need to reconsider the palatal nomenclature.


Membranous obstruction of the inferior vena cava in a patient with factor V Leiden: evidence for a post-thrombotic aetiology.
Membranous obstruction of the inferior vena cava is a rare cause of hepatic venous outflow obstruction in Caucasians. There has been much debate in the literature about its aetiology. We describe a Caucasian with hepatic venous outflow obstruction due to an inferior vena cava web, who was found to have hypercoagulability due to factor V Leiden. Following balloon rupture of the membrane and anticoagulation, his symptoms resolved and he has remained well for a year. The age at presentation in this patient, the presence of hypercoagulability and the excellent response to membrane rupture and anticoagulation suggest that in this case the membrane may have been derived from organised thrombus. Balloon rupture of the membrane and anticoagulation appears to be an effective treatment in such cases.


Pyrosequencing of clinically relevant polymorphisms.
The data generated from the Human Genome Project has led to an explosion of technology for low-, medium-, and high-throughput genotyping methods. Pyrosequencing is a genotyping assay based on sequencing by synthesis. Short runs of sequence around each polymorphism are generated, allowing for internal controls for each sample. Pyrosequencing can also be used to identify tri-allelic, indel, and short-repeat polymorphisms, as well as determining allele percentages for methylation or pooled sample assessment. Assays details for Pyrosequencing of clinically relevant polymorphisms are described in this chapter.


Retinal anomalies in trisomy 18.
The eyes of three patients with trisomy 18, the second most common human autosomal trisomy, were examined histopathologically. In the posterior retina transmission and scanning electron microscopic examinations reveal cytological details characteristic of immature neural retinas. We are able to confirm the report that the posterior retinal pigment epithelium in trisomy 18 has a paucity of mature melanosomes and, in fact, resembles human albino retinal pigment epithelium. The asociation of hypopigmentation and retinal immaturity suggests pigmentation plays a role in the control of the maturation of the neural retina.


Interleukin 1 enhances the development of spontaneous arthritis in MRL/lpr mice.
We previously reported that treatments with human recombinant interleukin-1 beta (rIL-1 beta) in DBA/1 mice which were suboptimally immunized with native chick type II collagen (NcII) markedly accelerated the onset of collagen-induced arthritis (CIA). In the present study, we further characterized this IL-1-mediated enhancement of murine arthritis by examining the in vivo effects of rIL-1 beta in another arthritis model, namely, the spontaneous arthritis of the MRL/lpr mouse strain. The results of these studies demonstrated that IL-1 treatments also enhanced the onset and progression of the spontaneous arthritic disease in MRL/lpr mice. A substantial proportion of the IL-1-treated MRL/lpr mice that were between 3 and 3.5 months of age exhibited swelling in either the hind or front paws. Moreover, histopathologic studies demonstrated the presence of striking alterations within the various joints of these IL-1-treated MRL/lpr mice. Such abnormalities were not detected in the non-IL-1-treated, age-matched MRL/lpr mice. Therefore, as in the experimentally induced disease of CIA, IL-1 may also be capable of contributing to the pathogenesis of the spontaneous arthritis of MRL/lpr mice.


Evaluation of the visual system in multiple sclerosis. II. Colour vision.
A study of colour vision (CV) in 65 patients with multiple sclerosis (MS), (30 patients had had previous optic neuritis) and 51 controls was carried out with Ishihara's pseudoisochromatic plates (I-test), Farnsworth's panel D-15 test (F-test), and Lanthony's desaturated 15-hue test (L-test). CV defects were classified as to type and severity. Error scorings were calculated by Bowman's computerized method and our own simple proposal for scoring, which was found of equal value. Results were compared with pattern-reversal (black/white) visual evoked potentials (PR-VEP) (80% of eyes abnormal). The I-test (56% of eyes abnormal) was a more sensitive indicator of demyelination than the L-test (47%) and F-test (26%). In 14 eyes CV defects (10 blue-yellow, 4 red-green) were only revealed with the L-test. Abnormal CV, mostly blue-yellow defects, occurred in 16 patients (19 eyes) having normal VEP latencies; 29 patients were re-tested within one week. It is proposed that the performance of the I-test, which showed the highest reproducibility, could be improved by adding more cards, particularly blue-yellow, to the test.


Abnormal cilia in a malignant ependymoma.
Abnormal cilia were frequently exposed in a malignant ependymoma of the cerebellar vermis by an electron microscope study. Among these erratic cilia, compound cilia, huge axoneme with bizarre shapes, were repeatedly observed. These giant cilia commonly had random orientation of many microtubular doublets as well as complicated cavities in their granular ciliary matrix. Additionally, abnormal cilia of normal-sized axoneme showed diversified arrangement of peripheral doublets and central singlets. The formation process of these abnormal cilia is discussed.


Dysregulation of angiogenesis-specific signalling in adult testis results in xenograft degeneration.
Ectopic xenografting of testis is a feasible option for preservation of male fertility and angiogenesis plays a pivotal role in xenograft survival and functionality. When compared to immature testis, the adult testis is unable to establish functional xenografts due to potentially lower efficiency to induce angiogenesis. The precise molecular mechanism, however, remains elusive. In the present study, we compared adult and immature testis xenografts for survival, maturation and germ cell differentiation. Further, we evaluated differential expression of angiogenesis signalling-specific proteins in adult and immature testis and their xenografts. Results showed that adult testis xenografts degenerated whereas immature testis xenografts survived and established spermatogenesis with the production of haploid germ cells. Protein expression analysis demonstrated that immature testis xenografts were able to establish angiogenesis either through eNOS activation via VEGF and PI3K/AKT or through EGFR-mediated STAT3 pathway. The role of ERK/MAPK pathway in xenograft angiogenesis was ruled out. The absence or reduced expression of angiogenesis-specific proteins in adult testis and its xenografts possibly resulted in poor angiogenesis and in their subsequent degeneration. This study provides insight into angiogenesis mechanism that can be utilized to augment testis xenografting efficiency.


Are virtual planning and guided surgery for head and neck reconstruction economically viable?
Virtual planning and guided surgery with or without prebent or milled plates are becoming more and more common for mandibular reconstruction with fibular free flaps (FFFs). Although this excellent surgical option is being used more widely, the question of the additional cost of planning and cutting-guide production has to be discussed. In capped payment systems such additional costs have to be offset by other savings if there are no special provisions for extra funding. Our study was designed to determine whether using virtual planning and guided surgery resulted in time saved during surgery and whether this time gain resulted in self-funding of such planning through the time saved. All consecutive cases of FFF surgery were evaluated during a 2-year period. Institutional data were used to determine the price of 1 minute of operative time. The time for fibula molding, plate adaptation, and insetting was recorded. During the defined period, we performed 20 mandibular reconstructions using FFFs, 9 with virtual planning and guided surgery and 11 freehand cases. One minute of operative time was calculated to cost US $47.50. Multiplying this number by the time saved, we found that the additional cost of virtual planning was reduced from US $5,098 to US $1,231.50 with a prebent plate and from US $6,980 to US $3,113.50 for a milled plate. Even in capped health care systems, virtual planning and guided surgery including prebent or milled plates are financially viable.


Glycogen synthase kinase 3 as an anticancer drug target: novel experimental findings and trends in the design of inhibitors.
Glycogen synthase kinase 3 (GSK-3) is a ubiquitous serine/threonine protein kinase participating in numerous pathways. Very important among them are cancer-related pathways, such as Wnt pathway and nuclear factor κB pathway. The recent findings concerning possible applicability of GSK-3 inhibitors as anticancer agents are summarized in this review, and controversies in the data are highlighted. The most actual concepts of GSK-3 inhibitor design are also thoroughly discussed.


Use of immobilized Candida yeast cells for xylitol production from sugarcane bagasse hydrolysate: cell immobilization conditions.
Candida guilliermondii cells were immobilized in Ca-alginate beads and used for xylitol production from concentrated sugarcane bagasse hydrolysate. A full factorial design was employed to determine whether variations in the immobilization conditions would have any effects on the beads, chemical stability and on the xylitol production rates. Duplicate fermentation runs were carried out in 125-mL Erlenmeyer flasks maintained in a rotatory shaker at 30 degrees C and 200 rpm for 72 h. Samples were periodically analyzed to monitor xylose and acetic acid consumption, xylitol production, free cell growth, and bead solubilization. Concentrations of sodium alginate at 20.0 g/L and calcium chloride at 11.0 g/L and bead curing time of 24 h represented the most appropriate immobilization conditions within the range of conditions tested.


The PKB/FOXO switch in aging and cancer.
Aging is characterized by the general decline in tissue and body function and the increased susceptibility to age-related pathologies, such as cancer. To maintain optimal tissue and body function, organisms have developed complex mechanisms for tissue homeostasis. Importantly, it is becoming apparent that these same mechanisms when deregulated also result in the development of age-related disease. The build in fail safe mechanisms of homeostasis, which prevent skewing toward disease, themselves contribute to aspects of aging. Thus, longevity is limited by an intrinsic trade-off between optimal tissue function and disease. Consequently, aging and age-related diseases, such as cancer and diabetes are driven by the same genetic determinants. Illustrative in this respect is the insulin/IGF-1 signaling pathway acting through PI3K/PKB and FOXO. Loss of PKB signaling contributes to diabetes, whereas gain of function of PKB drives cancer. Enhanced FOXO activity, at least in model organism contributes to extended lifespan and acts as a tumor suppressive mechanism. Here, we focus on the linkage between PKB and FOXO as a central switch in contributing to tissue homeostasis and age-related diseases in particular cancer. This article is part of a Special Issue entitled: P13K-AKT-FoxO axis in cancer and aging.


N-Heterocyclic carbene-catalyzed direct cross-aza-benzoin reaction: Efficient synthesis of α-amino-β-keto esters.
An efficient catalytic synthesis of α-amino-β-keto esters has been newly developed. Cross-coupling of various aldehydes with α-imino ester, catalyzed by N-heterocyclic carbene, leads chemoselectively to α-amino-β-keto esters in moderate to good yields with high atom efficiency. The reaction mechanism is discussed, and it is proposed that the α-amino-β-keto esters are formed under thermodynamic control.


Stroke Rehabilitation.
Despite contemporary rehabilitation strategies, stroke remains a leading cause of loss of function, limited mobility, psycho-social complications, and decreased quality of life. Stroke rehabilitation is a process that aims to prevent deterioration of function, increase function, and assist the patient in achieving the highest possible level of independence physically, socially, spiritually, psychologically, vocationally, and economically. The process begins with relearning activities of daily living such as grooming, bathing, toileting, eating, and dressing. As the patient progresses, stroke rehabilitation works on instrumental activities of daily living such as housekeeping, cooking, driving, and managing financial responsibilities.


IIHS small overlap frontal crash test ratings and real-world driver death risk.
To evaluate how ratings for the Insurance Institute for Highway Safety (IIHS) driver-side small-overlap frontal crash test predict real-world driver death risk in frontal impacts. IIHS released the driver-side small-overlap frontal crash test in 2012, after manufacturers had improved vehicle designs to make good ratings in the IIHS moderate overlap frontal crash test virtually ubiquitous. In the small overlap test, the vehicle impacts a rigid barrier at 40 mph (64 km/h) with 25% of the vehicle's width overlapping the barrier. As in other IIHS tests, vehicles are rated as good, acceptable, marginal, or poor. Drivers' risk of dying in a frontal crash was estimated by dividing driver deaths by driver involvements in police-reported crashes and modeling with logistic regression to estimate the effect of crash test rating, while controlling for driver age and sex, vehicle type and curb weight, and number of vehicles in the crash. Drivers of good-rated vehicles were 12% less likely to die in frontal impacts than drivers of poor-rated vehicles. This estimate was 11% for acceptable-rated vehicles and 5% (not statistically significant) for marginal-rated vehicles, compared with vehicles rated poor. The current study demonstrates that the IIHS driver-side small-overlap crash test rating encourages vehicle designs that reduce drivers' real-world risk of dying in frontal crashes.


Does Potassium Modify the Response of Zinnia (Zinnia elegans Jacq.) to Long-Term Salinity?
Salinity is one of the major abiotic stress factors hindering crop production, including ornamental flowering plants. The present study examined the response to salt stress of Zinnia elegans 'Lilliput' supplemented with basic (150 mg·dm-3) and enhanced (300 mg·dm-3) potassium doses. Stress was imposed by adding 0.96 and 1.98 g of NaCl per dm-3 of the substrate. The substrate's electrical conductivity was 1.1 and 2.3 dS·m-1 for lower potassium levels and 1.2 and 2.4 dS·m-1 for higher potassium levels. Salt stress caused a significant and dose-dependent reduction in leaf RWC, increased foliar Na and Cl concentrations, and reduced K. About 15% and 25% of cell membrane injury at lower and higher NaCl doses, respectively, were accompanied by only slight chlorophyll reduction. Salt stress-induced proline increase was accompanied by increased P5CS activity and decreased PDH activity. More than a 25% reduction in most growth parameters at EC 1.1-1.2 dS·m-1 but only a slight decrease in chlorophyll and a 25% reduction in the decorative value (number of flowers produced, flower diameter) only at EC 2.3-2.4 dS·m-1 were found. Salt stress-induced leaf area reduction was accompanied by increased cell wall lignification. An enhanced potassium dose caused a reduction in leaf Na and Cl concentrations and a slight increase in K. It was also effective in membrane injury reduction and proline accumulation. Increasing the dose of potassium did not improve growth and flowering parameters but affected the lignification of the leaf cell walls, which may have resulted in growth retardation. Zinnia elegans 'Lilliput' may be considered sensitive to long-term salt stress.


Reference Module-Based Analysis of Ovarian Cancer Transcriptome Identifies Important Modules and Potential Drugs.
Ovarian cancer (OVC) is often diagnosed at the advanced stage resulting in a poor overall outcome for the patient. The disease mechanisms, prognosis, and treatment require imperative elucidation. A rank-based module-centric framework was proposed to analyze the key modules related to the development, prognosis, and treatment of OVC. The ovarian cancer cell line microarray dataset GSE43765 from the Gene Expression Omnibus database was used to construct the reference modules by weighted gene correlation network analysis. Twenty-three reference modules were tested for stability and functionally annotated. Furthermore, to demonstrate the utility of reference modules, two more OVC datasets were collected, and their gene expression profiles were projected to the reference modules to generate a module-level expression. An epithelial-mesenchymal transition module was activated in OVC compared to the normal epithelium, and a pluripotency module was activated in ovarian cancer stroma compared to ovarian cancer epithelium. Seven differentially expressed modules were identified in OVC compared to the normal ovarian epithelium, with five up-regulated, and two down-regulated. One module was identified to be predictive of patient overall survival. Four modules were enriched with SNP signals. Based on differentially expressed modules and hub genes, five candidate drugs were screened. The hub genes of those modules merit further investigation. We firstly propose the reference module-based analysis of OVC. The utility of the analysis framework can be extended to transcriptome data of other kinds of diseases.


Effect of root surface treatment with denusomab after delayed tooth replantation.
To evaluate denosumab, a human monoclonal antibody that mimics the effects of osteoprotegerin in bone metabolism, as a topical treatment of root surface to be used prior to delayed tooth replantation. Thirty-six rats' right incisors were used. Teeth were extracted and divided into: delayed replantation without root surface treatment (control); delayed replantation with root surface treatment with denosumab 60 mg/mL and 30 mg/mL, respectively, for 10 min both experimentals groups. After that, the root canals were filled with calcium hydroxide and replanted. After 15 and 60 days, the animals were euthanized, and the samples were collected and processed for microscopic analysis. Histological sections were performed, and stained with HE to describe the dental characteristics, measure ankylosis, replacement resorption, and dental resorption by conventional microscopy. Also, was performed Brown & Brenn staining and immunohistochemistry for RANKL, OPG, and periostin. Denosumab 60 mg/mL reducted ankylosis (p < 0.0001), replacement resorption (p < 0.0001), and tooth resorption, 60 days after replantation, compared to untreated replanted teeth (p < 0.005). Lower bacterial contamination in root surface in the denosumab treatment groups was found, regardless of the concentration used (p < 0.001). Also, denosumab treatment inhibited the expression of RANKL without modulating OPG. Periostin was observed in periodontal ligament of replanted tooth, although this labelling was absent in the ankylosis areas, in both experimental periods. Treatment of the root surface with denosumab at 60 mg/mL of rat teeth before delayed replantation reduced dental root resorption compared with the untreated teeth after 60 days. Survival of a replanted tooth has been a challenge in clinical practice. The use of a medication, such as denosumab, to limit dental root resorption represents an important therapeutical approach.


Alternative ball-milling synthesis of vanadium-substituted polyoxometalates as catalysts for the aerobic cleavage of C-C and C-O bonds.
Vanadium-substituted phosphomolybdic acids (H3+x[PMo12-xVxO40], denoted as Vx) are well-known oxidation catalysts that are generally prepared by the hydrothermal treatment of MoO3 and V2O5 in the presence of H3PO4. This synthesis procedure is highly energy consuming and the Vx yields are not always acceptable. In the present work, an alternative hybrid mechanochemical/hydrothermal synthesis of Vx is proposed, comprising the ball-milling of MoO3 and V2O5, followed by a hydrothermal attack. The resulting materials, with 2 ≤ x ≤ 3, obtained from this new route were compared, in terms of yield, energy consumption and catalytic activity, with a reference V3 sample prepared through a conventional hydrothermal treatment. The ball-milling step proved to lead not only to a shorter and far more energy-saving synthesis procedure, but also to high yields of Vx. Moreover, Vx from this alternative route proved to be generally more active than the conventionally prepared V3 in the aerobic oxidative cleavage of C-O and C-C bonds in 2-phenoxyacetophenone, used herein as a lignin model compound.


Neck arthritis pain is reduced and range of motion is increased by massage therapy.
The literature on the effects of massage therapy on neck arthritis pain is mixed depending on the dose level, and it is also based on self-report. In the present study an attempt was made to enhance the effects of weekly massage therapy by having the participants massage themselves daily. And in addition to self-reports on pain, range of motion (ROM) and the associated ROM pain were assessed before and after the first massage session and pre-post the last session one month later. Staff and faculty members at a medical school who were eligible for the study if they had neck arthritis pain were randomly assigned to a massage or a waitlist control group (N = 24 per group). The massage group received moderate pressure massages weekly by a massage therapist plus daily self-massages. The waitlist control group received the same schedule massages one month after being control subjects. The massage group showed significant short-term reductions after the first and last day massages in self-reported pain and in ROM-associated pain as well as an increase in ROM. Comparisons between the massage group (N = 23) and the control group (N = 14) on the last versus the first day data suggested significantly different changes including increased ROM and reduced ROM-associated pain for the massage group and reduced ROM and increased ROM-associated pain for the control group. These changes occurred specifically for flexion and right and left lateral flexion motions. These data highlight the importance of designing massage therapy protocols that target the most affected neck muscle groups and then assessing range of motion and related pain before and after the massage therapy. Comparisons with other studies also suggest that moderate pressure may contribute to the massage effects, and the use of daily self-massages between sessions may sustain the effects and serve as a cost-effective therapy for individuals with neck arthritis pain.


Total or partial prostate sparing cystectomy for invasive bladder cancer: long-term implications on erectile function.
To review the long-term results in patients treated with either total or partial prostate-sparing cystectomy, focusing on erectile function (EF), as en-bloc radical cystectomy (RC) with or without urethrectomy has been the method of choice for managing invasive bladder carcinoma, but has inherent risks of subsequent urinary incontinence and erectile dysfunction, with a marked effect on quality of life, especially in younger patients. Between 2003 and 2005 we assessed 21 men (mean age 56 years) who had either a prostate apex-sparing cystectomy (PASC, 15) or total prostate-sparing cystectomy (TPSC, six). The mean follow-up was 30 months for PASC and 24 months for TPSC. The evaluation before surgery included standard bladder cancer staging, prostate specific antigen level, a digital rectal examination and a complete medical history, with attention to self-reported EF before surgery and the EF domain of the International Index of EF (IIEF) after surgery. The EF domain score was 20 after PASC and 30 after TPSC; this correlates with mild to moderate ED in the PASC group vs normal erectile function in the TPSC group. After transurethral resection of the bladder tumours (TURBT) 10 of 14 in the PASC group were T1 or T2a, and in the TPSC group, five of six were T2a and one patient was T2b. From the cystectomy specimen, in the PASC group eight were understaged compared with the TURBT specimen (T2b/T4a vs T1/T2a), while in the TPSC group there was understaging two (T3a vs T2a/T2b); this was significantly different (P < 0.05). There was recurrence of urothelial carcinoma in one of 15 and one of six after PASC and TPSC, respectively. The EF domain score after PASC was 10 points lower than after TPSC, representing a 30% increase in EF by preserving the entire prostate. We conclude that in patients with invasive bladder cancer, EF can be significantly preserved by prostate-sparing cystectomy. If adequate selection criteria are applied, EF can be preserved without compromising cancer control.


Exemplary professional practice through nurse peer review.
The development and execution of a nurse peer review program to evaluate nursing practice associated with significant adverse events has resulted in systemic changes. Descriptive analyses were conducted for 23 peer-reviewed cases involving 41 RNs and 2 advanced practice nurses from 14 specialties over a 4-year period. Thematic analysis revealed four common event categories: assessment and monitoring, team communication, skin integrity, and vascular access. This approach demonstrates the impact of professional nurse accountability for improving the quality of care and may serve as an exemplar for professional practice.


Prevalence and outcomes of malnutrition among hospitalized COVID-19 patients: A systematic review and meta-analysis.
Malnutrition is inevitable in patients with Coronavirus Disease 2019 (COVID-19) due to its effect on the gastrointestinal system, immune system, and high metabolic activity. However, the prevalence of malnutrition and its outcomes is uncertain. This study aimed to investigate the prevalence and outcome of malnutrition among patients with COVID-19. A comprehensive search was conducted in PubMed/Medline; Science direct and LILACS from December 29, 2019 to September, 2020 without language restriction. All observational studies reporting the prevalence of malnutrition were included while case reports and reviews were excluded. The data were extracted with two independent authors with a customized format and the disagreements were resolved by the other authors. The methodological quality of included studies was evaluated using a standardized critical appraisal tool. A total of 511 articles were identified from different databases and 27 articles were selected for evaluation after the successive screening. Fourteen articles with 4187 participants were included. The pooled prevalence of malnutrition among hospitalized patients with COVID-19 was 49.11% (95% CI: 31.67 to 66.54). The odd of mortality among patients COVID-19 with malnutrition was 10 times more likely as compared to those who were well-nourished. The prevalence of malnutrition and mortality associated with malnutrition among COVID-19 hospitalized patients was very high which entails a mitigating strategy by different stakeholders to prevent and manage malnutrition and its outcomes. This systematic review was registered in Prospero's international prospective register of systematic reviews (CRD42020215396).


Altered heparin pharmacodynamics in patients with pulmonary embolism.
Heparin clearance and pharmacodynamic response were examined in 12 patients being treated for deep venous thrombosis (DVT, 6 patients) or pulmonary embolism (PE, 6 patients). A loading dose of 70 units/kg was administered to DVT patients and 100 units/kg to PE patients followed by an initial infusion rate of 15 or 25 units/kg/h for DVT or PE patients, respectively. Heparin clearance was determined at 4, 12, and 24 h after initiating heparin therapy. The mean heparin clearance in the DVT group was 2,164 +/- 1,024 ml/h at 4 h, 2,591 +/- 1,239 ml/h at 12 h, and 2,795 +/- 1,863 m/h at 24 h. The PE patients had clearances of 1,775 +/- 494, 2,004 +/- 321, and 2,843 +/- 1,000 ml/h at 4, 12, and 24 h, respectively. The difference between the two groups was not statistically significant (p greater than 0.50). The activated partial thromboplastin time (aPTT) was used as a measure of heparin effect. The maximum effect (EMAX) and concentration required to attain 50% of the maximum effect (EC50) were determined for each group using the Lineweaver-Burke linearization method. The mean EMAX and EC50 for the DVT patients were 130 +/- 40.99 s and 1.01 +/- 0.70 units/ml, respectively. For the PE patients, the mean EMAX was 418 +/- 200 s and the mean EC50 was 4.32 +/- 2.81 units/ml. The difference between both groups for each parameter was statistically significant (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Updated clinical pharmacologic considerations for HIV-1 protease inhibitors.
Many data associate low protease inhibitor plasma concentrations with suboptimal virologic responses, whereas fewer data associate high plasma concentrations with toxicity. Knowledge of relationships between concentrations and virologic response is important because significant variability in concentrations exists among patients. For antiretroviral-naïve patients, target trough concentrations have been suggested on the basis of retrospective associations with virologic responses. Two prospective studies demonstrated improved virologic responses when indinavir and nelfinavir doses were managed based on these troughs. Investigations among antiretroviral-experienced patients have identified a relationship between the trough concentration and the in-vitro susceptibility of the patient's virus with virologic outcome. However, differences in virologic response may further depend on other pharmacologic factors, such as protein binding, intracellular kinetics, function of drug transporters, and the activity of other drugs in the regimen. In the future, dosing strategies that accommodate the variability in pharmacokinetics and pharmacodynamics may improve virologic outcomes.


Palliative and prognostic approach in cancer patients identified in the multicentre NECesidades PALiativas 2 study in Argentina.
Early identification of palliative needs has proven benefits in quality of life, survival and decision-making. The NECesidades PALiativas (NECPAL) Centro Coordinador Organización Mundial de la Salud - Instituto Catalán de Oncología (CCOMS-ICO©) tool combines the physician's insight with objective disease progression parameters and advanced chronic conditions. Some parameters have been independently associated with mortality risk in different populations. According to the concept of the 'prognostic approach' as a companion of the 'palliative approach', predictive models that identify individuals at high mortality risk are needed. We aimed to identify prognostic factors of mortality in cancer in our cultural context. We assessed cancer patients with palliative needs until death using this validated predictive tool at three hospitals in Buenos Aires City. This multifactorial, quantitative and qualitative non-dichotomous assessment process combines subjective perception (the surprise question: Would you be surprised if this patient dies in the next year?) with other parameters, including the request (and need) for palliative care (PC), the assessment of disease severity, geriatric syndromes, psychosocial factors and comorbidities, as well as the use of healthcare resources. 2,104 cancer patients were identified, 681 were NECPAL+ (32.3%). During a 2-year follow-up period, 422 NECPAL+ patients died (61.9%). The mean overall survival was 8 months. A multivariate model was constructed with significant indicators in univariate analysis. The best predictors of mortality were: nutritional decline (p < 0.000), functional decline (p < 0.000), palliative performance scale (PPS) ≤ 50 (p < 0.000), persistent symptoms (p < 0.002), functional dependence (p < 0.000), poor treatment response (p < 0.000), primary cancer diagnosis (p = 0.024) and condition (in/outpatients) (p < 0.000). Only three variables remained as survival predictors: low response to treatment (p < 0.001), PPS ≤ 50 (p < 0.000) and condition (in/outpatients) (p < 0.000). This prospective model aimed to improve cancer survival prediction and timely PC referral in Argentinian hospitals.


[Beekeeping: study of organization, general hazards assessment, pre-assessment of risk for mechanical overload using a new tool for easy application].
Beekeeping is common activity in the two regions in this study, Marche and Tuscany: in both regions the numbers of beekeepers, both amateur and professional, and honey production are high. The aim was to study, through the application of simple tools, the organization of beekeeping activity so as to identify hazardous situations in the work process. We followed the production cycle of two businesses that differed in size and work organization for a period of twelve months. Subsequently each homogeneous period was assessed via increasingly complex levels of intervention which made it possible to identify the work phases where preventive measures could be applied. The results obtained made it possible to detect the presence of risk situations for the musculoskeletal system of beekeepers. Organizational analysis in the two enterprises showed that is possible to apply easy solutions to improve safety and health at the workplace.


[A special closure system of the gastrointestinal tract of the rat (author's transl)].
As a result of animal experiments, morpho-histologic and surgical findings, the presence of a new closure system of the gastrointestinal tract is described. The mode of action is different from that of all the known muscle sphincters. It is found in the ascending colon of the rat and consists of a specially arranged and adjustable mucosal folds with aunique vascularisation. A functional unit which comosed of a pair of folds and a vascular plexus constitute the valvular mechanism. We, therefore, consider appropriate to call it the Angioplical Closure System. Surgical reversion of even a very small portion of the system causes a lethal ileus.


Development and validation of a scale for the tendency to exercise in response to mood, eating, and body image cues: the Reactive Exercise Scale (RES).
Earlier work on engaging in physical exercise when experiencing negative affect demonstrated robust associations with eating disorder (ED) behaviors and attitudes; however, measurement of the behavior was primitive, relying on one yes/no question that cannot capture much variability. We report on the development of a self-report measure, the Reactive Exercise Scale (RES), that disentangles the tendency to engage in exercise in response to negative mood cues from the tendency to engage in exercise in response to eating and body image cues, which themselves may be associated with negative mood. The measure also assesses exercising in response to positive mood cues. Exploratory factor analysis (EFA) guided item and factor selection. Confirmatory factor analysis (CFA) in an independent sample tested a 3-factor solution-exercising in response to negative mood cues, eating and body image cues, and positive mood cues. Correlations with exercise attitudes, eating disorder and body image attitudes, mood, and personality were used to evaluate construct validity. Results supported the 3-factor structure and indicated that exercising in response to negative mood cues may not uniquely relate to most aspects of ED psychopathology when accounting for eating and body image cues, which themselves are associated with negative mood. The RES captures the tendency to exercise in response to negative mood, positive mood, and eating and body image cues. Together, these constructs allow researchers to examine the unique relations of negative mood cued exercise with ED constructs, while accounting for appearance-related motives for which exercise may also be used. Level III: evidence obtained from well-designed cohort or case-control analytic studies.


Sounding out stability of enteric coated dosage forms using Broadband Acoustic Resonance Dissolution Spectroscopy (BARDS).
Stability testing is essential in the pharmaceutical industry to determine product shelf- life and the conditions under which drug products should be stored. Stability testing involves a complex set of procedures, considerable cost, time, and scientific expertise to build quality, efficacy and safety in a drug formulation. This paper highlights a new complementary approach to stability testing called Broadband Acoustic Resonance Dissolution Spectroscopy (BARDS). BARDS measurements are based on reproducible changes in the compressibility of a solvent during dissolution. It is monitored acoustically via associated changes in the frequency of induced acoustic resonances. This study presents a novel approach to track the change of various drug formulations to determine the formulation's stability. Pellets, tablet and multiple-unit pellet system (MUPS) formulations were investigated to examine the effect of polymer coating and formulation core degradation over time. In combination with minimal usage of Ultra Violet - Visible Spectroscopy, BARDS can effectively track these changes. The technique offers a rapid approach to characterizing pharmaceutical formulations. BARDS can enable rapid development of solid drug formulation dissolution and disintegration testing as an In-Process Control test and drug stability analysis. The data show that a solid oral dose formulation has an intrinsic acoustic signature specific to the method of manufacture, excipient composition and elapsed time since the production of a product. BARDS data are also indicative of which aspect of a formulation may be unstable, whether a coating, sub-coating or core. It is potentially a time-efficient, cost-effective and greener approach to testing coating stability, disintegration and overall formulation stability.


Integrating the therapeutic community and work release for drug-involved offenders. The CREST Program.
As the nation's first therapeutic community (TC) and work release center for drug involved offenders, CREST combines the basic elements of both modalities into an effective agent for behavioral change. This article explores the ways in which these elements are integrated and applied, and the outcome of such treatment as determined by subsequent substance abuse and criminal activity. Clients entering the program from prison progress through several phases of counseling, group interaction, confrontation, and education before they enter the work release phase, where they gain realistic experience and can implement what they learned in the TC concerning living drug free. Follow-up data collected at 6 and 18 months after entry into the program indicate that CREST clients have significantly lower relapse and recidivism rates than a comparable comparison group. CREST has similar effects on relapse and recidivism across sexes, racial/ethnic groups, and different age categories, although length of time in treatment and whether clients graduated do impact outcome variables.


Event-related potentials as indicators of repetition priming in young and older adults: amplitude, duration, and scalp distribution.
Event-related brain potentials (ERPs) were recorded in young and older adults in an indirect repetition priming paradigm. Compared with the young adults, the older adults' ERP repetition effect was larger and of longer duration, due entirely to greater amplitude elicited by the repeated item. These data suggest that, although indirect memory (as indexed by a robust ERP repetition effect) appeared to be intact in the older adult, the possibility exists for qualitative age-related differences consistent with inefficient and/or additional processing of the repeated item.


Asperimides A-D, anti-inflammatory aromatic butenolides from a tropical endophytic fungus Aspergillus terreus.
Four new aromatic butenolides, asperimides A-D (1-4), together with a known analogue, butyrolactone I (5), were isolated from solid cultures of a tropical endophytic fungus Aspergillus terreus. The structures of these compounds were elucidated on the basis of spectroscopic methods and electronic circular dichroism calculations. Compounds 1-4 represent the first examples of butenolides with a maleimide core isolated from Aspergillus sp. Inhibitory effects of the isolated compounds on nitric oxide production were investigated in lipopolysaccaride (LPS)-mediated RAW 264.7 cells. Compounds 3 and 4 exhibited potent anti-inflammatory with IC50 values of 0.78 ± 0.06 and 1.26 ± 0.11 μM, respectively.


Globalization and local response to epidemiological overlap in 21st century Ecuador.
Third World countries are confronted by a complex overlay of two sets of health problems. Traditional maladies, including communicable diseases, malnutrition, and environmental health hazards coexist with emerging health challenges, including cardiovascular disease, cancer, and increasing levels of obesity. Using Ecuador as an example, this paper proposes a conceptual framework for linking epidemiologic overlap to emerging social structures and processes at the national and global levels. Epidemiologic trends can be seen as part of broader processes related to globalization, but this does not imply that globalization is a monolithic force that inevitably and uniformly affects nations, communities, and households in the same manner. Rather, characteristics and forms of social organization at the subnational level can shape the way that globalization takes place. Thus, globalization has affected Ecuador in specific ways and is, at the same time, intimately related to the form in which the epidemiologic transition has transpired in that country. Ecuador is among neither the poorest nor the wealthiest countries and its situation may illuminate trends in other parts of the world. As in other countries, insertion into the global economy has not taken place in a vacuum; rather, Ecuador has experienced unprecedented social and demographic change in the past several decades, producing profound transformation in its social structure. Examples of local represent alternatives to centralized health systems that do not effectively address the complex overlay of traditional and emerging health problems.


Identification of a phosphatidylinositol 4,5-bisphosphate-binding site in chicken skeletal muscle alpha-actinin.
We previously reported that phosphatidylinositol 4,5-bisphosphate (PIP2) dramatically increases the gelating activity of smooth muscle alpha-actinin (Fukami, K., Furuhashi, K., Inagaki, M., Endo, T., Hatano, S., and Takenawa, T. (1992) Nature 359, 150-152) and that the hydrolysis of PIP2 on alpha-actinin by tyrosine kinase activation may be important in cytoskeletal reorganization (Fukami, K., Endo, T., Imamura, M., and Takenawa, T. (1994) J. Biol. Chem. 269, 1518-1522). Here we report that a proteolytic fragment with lysylendopeptidase comprising amino acids 168-184 (TAPYRNVNIQNFHLSWK) from striated muscle alpha-actinin contains a PIP2-binding site. A synthetic peptide composed of the 17 amino acids remarkably inhibited the activities of phospholipase C (PLC)-gamma 1 and -delta 1. Furthermore, we detected an interaction between PIP2 and a bacterially expressed alpha-actinin fragment (amino acids 137-259) by PLC inhibition assay. Point mutants in which arginine 172 or lysine 184 of alpha-actinin were replaced by isoleucine reduced the inhibitory effect on PLC activity by nearly half. Direct interactions between PIP2 and the peptide (amino acids 168-184) or the bacterially expressed protein (amino acids 137-259) were confirmed by enzyme-linked immunosorvent assay. We also found this region homologous to the sequence of the PIP2-binding site in spectrin and the pleckstrin homology domains of PLC-delta 1 and Grb7. Synthetic peptides from the homologous regions in spectrin and PLC-delta 1 inhibited PLC activities. These results indicate that residues 168-184 comprise a binding site for PIP2 in alpha-actinin and that similar sequences found in spectrin and PLC-delta 1 may be involved in the interaction with PIP2.


Characterization and phylogenetic diversity of carboxymethyl cellulase producing bacillus species from a landfill ecosystem.
Total population of cellulose degrading bacteria was studied in a landfill ecosystem as a part of microbial diversity study. Samples were obtained from 3 and 5 feet depth of a local landfill being operated for past 10 years. Among many isolates, 22 bacterial strains were selected based on their capability to decompose carboxymethyl cellulose (CMC). These isolates were cultivated on agar medium with CMC as the carbon source. All isolates were Gram positive, endospore forming and alkalophilic bacteria with optimum growth pH 9-10. They were grouped based on the phenotypic and chemotaxonomic characters and representative strains of different groups along with high carboxymethyl cellulase (CMCase) producing strains were included for further characterization. Analysis of 16S rRNA gene indicated that these strains belong to different species of the genus Bacillus. Maximum CMCase activity of 4.8 U/ml at 50°C was obtained by strain LFC15. Results in the present study indicated the potential of waste land ecosystems such as landfill are potential source for isolation of industrially important microorganisms.


Phylogenetic relationships of Amazonian anole lizards (Dactyloa): taxonomic implications, new insights about phenotypic evolution and the timing of diversification.
The ecology and evolution of Caribbean anoles are well described, yet little is known about mainland anole species. Lack of phylogenetic information limits our knowledge about species boundaries, morphological evolution, and the biogeography of anoles in South America. To help fill this gap, we provide an updated molecular phylogeny of the Dactyloa (Dactyloidae), with emphasis on the punctata species group. By sampling understudied Amazonian taxa, we (i) assess the phylogenetic placement of the 'odd anole', D. dissimilis; (ii) infer the relationships of the proboscis-bearing D. phyllorhina, testing the hypothesis of independent nasal appendage evolution within the anole radiation; and (iii) examine genetic and dewlap color variation in D. punctata and D. philopunctata. Combining multiple nuclear loci with a review of the fossil record, we also (iv) estimate divergence times within the pleurodont iguanian clade of lizards, including Amazonian representatives of Dactyloa and Norops (Dactyloidae) and of Polychrus (Polychrotidae). We recover the five Dactyloa clades previously referred to as the aequatorialis, heteroderma, latifrons, punctata and roquet species groups, as well as a sixth clade composed of D. dissimilis and the non-Amazonian D. neblinina and D. calimae. We find D. phyllorhina to be nested within the punctata group, suggesting independent evolution of the anole proboscis. We consistently recover D. philopunctata nested within D. punctata, and report limited genetic divergence between distinct dewlap phenotypes. The most recent common ancestor of Dactyloa, Anolis and Norops dates back to the Eocene. Most Amazonian taxa within both Dactyloa and Norops diverged in the Miocene, but some diversification events were as old as the late Eocene and late Oligocene. Amazonian Polychrus diverged in the Pliocene. Our findings have broad implications for anole biogeography, disputing recent suggestions that modern dactyloid genera were present in the Caribbean region during the Cretaceous.


Triacylglycerol and very low density lipoprotein secretion into plasma of squirrel monkeys.
We determined the effects of varying the types and level of dietary fat and cholesterol on the increase in plasma total triacylglycerol concentrations after injection of Triton WR-1339, an inhibitor of lipoprotein lipase, into monkeys that had been subjected to an overnight fast. The monkeys that had been treated with Triton WR-1339 were then given a test meal by intragastric intubation. Dietary cholesterol, high levels of fat and saturated fat in the habitual diet reduced the rate of release of triacylglycerol to plasma in the fasted monkey. We also determined the changes in protein and lipid concentrations of the different lipoprotein fractions. The injection of Triton WR-1339 resulted in a linear increase with time in the concentration of protein and triacylglycerol in the very low density (chylomicron-free and d less than 1.006) lipoproteins, but there was an increase in the ratio of traicylglycerol to protein in that fraction. Most of the increase (96%) in very low density protein was in the B protein. Regardless of the habitual diet, a test meal accentuated the rate of triacylglycerol appearance in whole plasma and in the very low density lipoproteins of Triton WR-1339-treated monkeys, and the rate of increase of the protein component after feeding was slightly higher. Thus the administration of a meal to the fasted Triton WR-1339-treated squirrel monkey further increased the proportion of triacylglycerol in very low density lipoproteins. Although dietary cholesterol and saturated fat in the habitual diet depressed the rate of increase in very low density triacylglycerol during fasting, the rate of protein synthesis was not significantly affected. After administration of a test meal the rates of increase in triacylglycerol and protein in the very low density lipoproteins were similar for monkeys from the different diet groups. Triton WR-1339 administration caused a slight and progressive increase in the intermediate density (d 1.006-1.019) lipoproteins and a marked and progressive decrease in the low density (d 1.019-1.063) lipoproteins. There was an immediate (by 5 min) drop of 70% or more in high density (d 1.063-1.21) lipoprotein protein, but the lipids except triacylglycerol remained unchanged. There was a decrease in both the A (the major fraction) and C proteins. The rates of very low density B protein secretion were comparable to the rates of low density lipoprotein catabolism that had been previously demonstrated for this species.


High-purity alkaline lignin extraction from Saccharum ravannae and optimization of lignin recovery through response surface methodology.
Saccharum ravannae, known as "Ekra" in the Northeast region of India, is an elephant grass species that abundantly grows in the natural habitat of Assam. This study aims to utilize this wild grass species and extract alkaline lignin of high purity through KOH-mediated alkaline hydrothermal pretreatment using the Oil bath process. Lignin recovery was optimized using RSM (response surface methodology) combined with a central composite model. Three process parameters, namely KOH concentration (1-3 %), reaction time (50-200 min), and solid loading (5-15 %), varied to optimize the combined effect of these parameters. RSM predicted a maximum lignin recovery of 15.38 g/100 g of raw biomass at optimum conditions (2.4 % KOH, 6.41 % solid loading, 176.57 min). Three experimental runs were performed at optimum conditions, and 15.81 ± 0.32 g/100 g lignin recovery was obtained, thus verifying the predicted result. Maximum 93.7 % purity of extracted lignin was achieved in a different operating condition (3 % KOH, 10 % solid loading, 125 min). The commercial and extracted alkaline lignin with maximum purity was characterized by Nuclear Magnetic Resonance (NMR), Fourier Transform Infrared Spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). The extracted lignin shows higher phenolic content and more functional groups than commercial lignin and can be used for future applications.


Geospatial epidemiology of leprosy in northwest Bangladesh: a 20-year retrospective observational study.
Leprosy is known to be unevenly distributed between and within countries. High risk areas or 'hotspots' are potential targets for preventive interventions, but the underlying epidemiologic mechanisms that enable hotspots to emerge, are not yet fully understood. In this study, we identified and characterized leprosy hotspots in Bangladesh, a country with one of the highest leprosy endemicity levels globally. We used data from four high-endemic districts in northwest Bangladesh including 20 623 registered cases between January 2000 and April 2019 (among ~ 7 million population). Incidences per union (smallest administrative unit) were calculated using geospatial population density estimates. A geospatial Poisson model was used to detect incidence hotspots over three (overlapping) 10-year timeframes: 2000-2009, 2005-2014 and 2010-2019. Ordinal regression models were used to assess whether patient characteristics were significantly different for cases outside hotspots, as compared to cases within weak (i.e., relative risk (RR) of one to two), medium (i.e., RR of two to three), and strong (i.e., RR higher than three) hotspots. New case detection rates dropped from 44/100 000 in 2000 to 10/100 000 in 2019. Statistically significant hotspots were identified during all timeframes and were often located at areas with high population densities. The RR for leprosy was up to 12 times higher for inhabitants of hotspots than for people living outside hotspots. Within strong hotspots (1930 cases among less than 1% of the population), significantly more child cases (i.e., below 15 years of age) were detected, indicating recent transmission. Cases in hotspots were not significantly more likely to be detected actively. Leprosy showed a heterogeneous distribution with clear hotspots in northwest Bangladesh throughout a 20-year period of decreasing incidence. Findings confirm that leprosy hotspots represent areas of higher transmission activity and are not solely the result of active case finding strategies.


In vitro antibiotic susceptibility of rapidly growing nontuberculous mycobacteria isolated from patients with microbial keratitis.
This study was undertaken to determine the antibiotic susceptibility and minimum inhibitory concentrations (MIC) of amikacin, tobramycin, ciprofloxacin, gatifloxacin, azithromycin, and clarithromycin against rapidly growing nontuberculous mycobacteria isolated from patients with keratitis. A total of 15 rapidly growing nontuberculous mycobacteria isolated from corneal scrapings of keratitis patients from January 1999 through December 2007 were subjected to antimicrobial susceptibility testing by the E-Test to amikacin, tobramycin, ciprofloxacin, gatifloxacin, azithromycin, and clarithromycin. Out of 15 isolates, 13 were identified as Mycobacterium chelonae complex and 2 as Mycobacterium fortuitum complex. Based on minimum inhibitory concentration (MIC) cut off, all 15 (100%) isolates were sensitive to amikacin, azithromycin, and clarithromycin, 13 (86%) were sensitive to tobramycin, nine (60%) to gatifloxacin, and only 6 (40%) to ciprofloxacin. The MIC range was 0.25-4 microg/ml for amikacin, 0.5-1 microg/ml for azithromycin, 0.125-1 microg/ml for clarithromycin, 0.5-16 microg/ml for ciprofloxacin, and 0.25-16 microg/ml for tobramycin. MIC(90) for amikacin was 2 microg/ml, azithromycin 1 microg/ml, clarithromycin 0.75 microg/ml, ciprofloxacin 8 microg/ml, gatifloxacin 8 microg/ml, and for tobramycin it was 4 microg/ml. All the isolates were sensitive to amikacin, azithromycin, and clarithromycin, but the MIC values of clarithromycin and azithromycin were lower than amikacin. Based on in vitro susceptibility results it appears that the topical amikacin in combination with oral clarithromycin or azithromycin is the best treatment option for rapidly growing nontuberculous mycobacterial keratitis.