Prasugrel vs. clopidogrel for cytochrome P450 2C19-genotyped subgroups: integration of the TRITON-TIMI 38 trial data.
Prasugrel is a newly marketed antiplatelet drug with improved cardiac outcomes as compared with clopidogrel for acute coronary syndromes involving percutaneous coronary intervention (PCI). Analysis of a subset of the TRITON-TIMI 38 trial demonstrated that cytochrome P450 2C19 (CYP2C19) reduced-function genotypes are associated with differential clinical responses to clopidogrel, but not prasugrel. Whether the CYP2C19 genotype has the potential to influence clinical choice of these drugs prior to PCI for individuals with unstable angina or non-ST segment elevation myocardial infarction is currently uncertain. An exploratory, secondary analysis was undertaken to estimate the clinical benefit of prasugrel over clopidogrel in subgroups defined by CYP2C19 genotype, by integrating the published results of the genetic substudy and the overall TRITON-TIMI 38 trial. Individuals with a CYP2C19 reduced-metabolizer genotype were estimated to have a substantial reduction in the risk of the composite primary outcome (cardiovascular death, myocardial infarction, or stroke) with prasugrel as compared with clopidogrel [relative risk (RR) 0.57; 95% confidence interval (CI) 0.39-0.83]. For CYP2C19 extensive metabolizers (∼70% of the population), however, the composite outcome risks with prasugrel and clopidogrel were not substantially different (RR 0.98; 95% CI 0.80-1.20). Integration of the TRITON-TIMI 38 data suggests that the CYP2C19 genotype can discriminate between individuals who receive extensive benefit from using prasugrel instead of clopidogrel, and individuals with comparable clinical outcomes with prasugrel and clopidorel. Thus, CYP2C19 genotyping has the potential to guide the choice of antiplatelet therapy, and further research is warranted to validate this estimate.


The effect of the bisphosphonate alendronate on viability of canine osteosarcoma cells in vitro.
The objective of this study was to determine the effect of alendronate on the viability of canine osteosarcoma cells and nonneoplastic canine cells. The sample population was composed of canine osteosarcoma tumor cells. Osteosarcoma cells and canine fibroblasts were maintained in culture under standard conditions. The MTT assay for cell viability was performed after 24, 48, and 72 h of incubation with alendronate (0.001 to 1000 microM) or no drug (control). Plates were set up so that each concentration and the control had a sample number of 8. The optical density (OD) of each well was measured at 540 nm using an enzyme-linked immunosorbent assay microplate reader. The percent viability was determined for each concentration and for each incubation time. After 24 h of incubation of POS (parent osteosarcoma) and HMPOS cells with alendronate, there was no significant difference in mean OD at any drug concentration when compared with control samples. A significant concentration- and time-dependent reduction in mean OD of osteosarcoma cells was observed after 48 and 72 h of incubation, with alendronate concentrations ranging from 10 to 1000 microM. The lowest percent cell viability observed in treated cells was 35%. Conversely, alendronate did not significantly affect mean OD in fibroblasts, and the lowest percent cell viability observed was 76%. Our data indicate that alendronate may have the potential to inhibit canine osteosarcoma tumor growth. It will be important to determine the clinical relevance of these in vitro findings. If similar findings are observed in vivo, use of alendronate may also be indicated as an adjuvant to existing chemotherapeutic protocols.


Solar energy storage in a Cs2AgBiBr6 halide double perovskite photoelectrochemical cell.
Storing solar energy using a stable visible light absorbing Cs2AgBiBr6 double perovskite is achieved using a photoelectrochemical (PEC) device with cobalt complexes and methyl viologen redox mediators. Under illumination, a potential gain of nearly 500 mV is achieved for charging. The charge-discharge cycling was carried out, and using in situ emission and FTIR studies, the self-discharge and solvent crossover were investigated.


c-Fos expression in hypothalamic nuclei of food-entrained rats.
The present study aimed to identify the hypothalamic nuclei involved with food entrainment by using c-Fos-like immunoreactivity (c-Fos-IR) as a marker of functional activation. We studied rats entrained 3 wk to restricted feeding schedules (RF), their ad libitum (AL) controls, and the persistence of c-Fos-IR temporal patterns in entrained-fasted rats. In addition, we included 22-h fasting and 22-h fasting-refeeding groups as controls of fasting and refeeding acute effects. Diurnal patterns of c-Fos-IR were observed in the tuberomammilar nucleus (TM) and suprachiasmatic nucleus (SCN) in AL rats. In all nuclei, except the SCN and ventromedial nucleus (VMH), restricted feeding schedules imposed a temporal pattern of increased c-Fos-IR around mealtime. An increase in c-Fos-IR before and after meal time was observed in dorsomedial nucleus (DMH), lateral nucleus (LH), perifornical area (PeF), and TM, and a marked increase was observed in the paraventricular nucleus (PVN) after feeding. Food-entrained c-Fos-IR patterns persisted after 3 days in fasting in DMH, LH, and PeF. Present data suggest that FEO might not rely on a single nucleus and rather may be a distributed system constituted of interacting nuclei in which the PVN is mainly involved with the response to signals elicited by food ingestion and, therefore, with the entraining pathway. We can suggest that the PeF and TM may be involved with the arousal state during food anticipation and the DMH and LH with the time-keeping mechanism of FEO or its output.


Carbon dioxide, bicarbonate and carbonate ions in aqueous solutions under deep Earth conditions.
We investigate the effect of pressure, temperature and acidity on the composition of water-rich carbon-bearing fluids under thermodynamic conditions that correspond to the Earth's deep crust and upper mantle. Our first-principles molecular dynamics simulations provide mechanistic insight into the hydration shell of carbon dioxide, bicarbonate and carbonate ions, and into the pathways of the acid/base reactions that convert these carbon species into one another in aqueous solutions. At temperatures of 1000 K and higher, our simulations can sample the chemical equilibrium of these acid/base reactions, thus allowing us to estimate the chemical composition of diluted carbon dioxide and (bi)carbonate ions as a function of acidity and thermodynamic conditions. We find that, especially at the highest temperature, the acidity of the solution is essential to determine the stability domain of CO2vs. HCO3-vs. CO32-.


Construction of a single-copy promoter vector and its use in analysis of regulation of the transposon Tn10 tetracycline resistance determinant.
The construction and characterization of a promoter expression vector, lambda RS205 , is described. lambda RS205 can be used for the in vitro construction of transcriptional (operon) fusions to the lacZ gene of Escherichia coli K-12. The level of beta-galactosidase activity in lysogens of lambda RS205 fusion phages provides a quantitative measure of promoter function under single-copy conditions. The regulation of the Tn10 tetracycline resistance gene ( tetA ) and the Tn10 tet repressor gene (tetR) was examined by inserting DNA fragments that span the tetR- tetA promoter-operator region into lambda RS205 . Levels of beta-galactosidase in tetA -lacZ and tetR-lacZ fusion strains indicate that the tetA and tetR promoters are strong promoters; the tetA promoter is fourfold more active than the tetR promoter. Introduction of tetR+ plasmids into tetA -lacZ and tetR-lacZ fusion strains represses beta-galactosidase synthesis 15- to 60-fold and 6- to 15-fold, respectively. The concentration of tetracycline required to induce half-maximal beta-galactosidase synthesis in these tetR+ tet-lac strains depends on both the tetracycline resistance phenotype and the level of tetR repressor in the fusion strain. However, the induction of beta-galactosidase in isogenic tetA -lacZ and tetR-lacZ strains is coordinate. The data presented here support the current model of Tn10 tet gene organization and regulation and provide quantitative information about the regulation of tetA and tetR in vivo.


Reversible non-ischaemic cardiomyopathy and left ventricular dysfunction after liver transplantation: a single-centre experience.
Non-ischaemic cardiomyopathy (NIC) is an early complication of liver transplantation (LT). Our aims were to define the prevalence, associated clinical factors, and prognosis of this condition. A retrospective study was performed on patients undergoing LT at our institution from January 2005 to December 2012. Patients who developed NIC were identified. Data collected included demographic and clinical data. A total 1460 transplants were performed in this period and seventeen patients developed NIC. Pretransplant median QTc interval was 459 (range, 405-530), and median E/A ratio was 1 (range, 0.71-1.67). Fourteen patients (82%) were severely malnourished and required nutritional support. Thirteen patients (76%) had renal insufficiency. Median time to onset was 2 days post-transplant (range, 0-20). Echocardiograms showed global left ventricular hypokinesis and a decrease in ejection fraction (EF) from a median of 65% (range, 50-81) pretransplant to a median of 21% (range, 15-32). Median raw model for end-stage liver disease (MELD) score was 29 in patients with NIC vs. 18 in patients without cardiomyopathy (P = 0.01). There was no significant difference between recipients with NIC vs. recipients without cardiomyopathy regarding donor age, donor risk index, and cold and warm ischaemia time. Recovery of cardiac function occurred in 16 patients, with a median EF of 44% (range, 25-65%) at the time of discharge. The last echocardiogram available showed a median EF of 59% (range, 49-73%). One-year survival of NIC patients was 94.1%. Non-ischaemic cardiomyopathy is a rare complication after LT. Patients with NIC are critically ill, with high MELD score, and severe malnutrition.


Explanatory models of adult patients with type 2 diabetes mellitus from urban centers of central Ethiopia.
Type 2 diabetes, which is increasing as a public health problem in the low resource settings of Africa has been associated with the high prevalence of micro-vascular complications and increasing levels of macro-vascular complications. There is evidence from the developed world that understanding patient perceptions of chronic illness is important to design effective strategies for helping patients manage these conditions. This study utilized Kleinman's model to explore the illness perceptions of type 2 diabetes patients attending treatment in Addis Ababa and Butajira (Ethiopia) and better understand how they manage their illness. Qualitative interviews were conducted to elicit the explanatory models of purposively sampled type 2 diabetes patients attending treatment in three hospitals in central Ethiopia until saturation of key emerging themes was achieved. Analysis of interview transcripts was guided by Kleinman's model. A total of 39 participants, 24 from Addis Ababa and the rest from Butajira took part in the study. This study revealed that patients' explanatory models were informed by both the traditional and biomedical models with emotional distress evident in some of the participants. The traditional model seemed to reflect the strong religious and cultural influences for the majority of study participants. The findings also revealed that symptoms played significant roles in how patients viewed their illness including assessment of its severity. Most were uncertain about the cause of their illness, with those expressing certainty citing factors over which they believed they had little or no control. This may have contributed to the perceptions about the use of religious healing and traditional medicines in a complementary or alternative manner to the biomedical regimen which could affect their adherence to recommended regimens and their health outcomes. This study suggests the need for a strong diabetes care program that is sensitive to patients' experiences of their illness including emotional distress. Individuals providing the diabetes care should consider local and individual contexts and strive to make their approach patient-centered and engage active participation of patients. There appears to be a need for better training of health providers in different areas including health communications and the fundamentals of mental healthcare.


Evaluation of cytotoxic potential of latex of Calotropis procera and podophyllotoxin in Allium cepa root model.
In the present study we have utilized the Allium cepa root tip meristem model to evaluate the cytotoxic and anti-mitotic activities of latex of Calotropis procera (DL) and podophyllotoxin. Standard cytotoxic drug cyclophosphamide and non-cytotoxic drugs cyprohcptadine and aspirin served as controls. Like cyclophosphamide, both DL and podophyllotoxin significantly inhibited the growth of roots and mitotic activity in a dose-dependent manner. However, podophyllotoxin was more potent in this regard and produced root decay. Cyproheptadine and aspirin, on the other hand, showed a marginal effect on the root growth and mitotic activity at much higher concentrations.


Biallelic Variants in the COLGALT1 Gene Causes Severe Congenital Porencephaly: A Case Report.
We describe a third patient with brain small vessel disease 3 (BSVD3), being the first with a homozygous essential splice site variant in the COLGALT1 gene, with a more severe phenotype than the 2 children reported earlier. Analysis of whole exome sequencing (WES) data of the child and parents was performed. We validated the missplicing of the homozygous variant using reverse transcription PCR and Sanger sequencing of the mRNA in a lymphocyte culture. The patient presented antenatally with porencephaly on ultrasound and MRI. Postnatally, he showed a severe developmental delay, refractory epilepsy, spastic quadriplegia, and a progressive hydrocephalus. WES revealed a homozygous canonical splice site variant NM_024656.3:c.625-2A>C. PCR and Sanger sequencing of the mRNA demonstrated that 2 cryptic splice sites are activated, causing a frameshift in the major transcript and in-frame deletion in a minor transcript. We report a third patient with biallelic pathogenic variants in COLGALT1, confirming the role of this gene in autosomal recessive BSVD3.


In vivo nonlinear spectral imaging in mouse skin.
We report on two-photon autofluorescence and second harmonic spectral imaging of live mouse tissues. The use of a high sensitivity detector and ultraviolet optics allowed us to record razor-sharp deep-tissue spectral images of weak autofluorescence and short-wavelength second harmonic generation by mouse skin. Real-color image representation combined with depth-resolved spectral analysis enabled us to identify tissue structures. The results show that linking nonlinear deep-tissue imaging microscopy with autofluorescence spectroscopy has the potential to provide important information for the diagnosis of skin tissues.


Functional assessment of patients with spinal cord injury: measured by the motor score and the Functional Independence Measure.
There is some information about the Functional Independence Measure (FIM) score of patients with spinal cord injury (SCI), but there are a few publications dealing with the relationship between the FIM score and the motor score of the American Spinal Injury Association (ASIA). We have studied the relationship of all FIM items with the motor score, and reviewed the disability of patients with spinal cord injury in greater detail. The purpose of this study was to describe the characteristics of impairment and disability in patients with SCI, using the FIM and motor score of the ASIA. The subjects were 100 inpatients with SCI (Frankel A, B). Neurological level, days from the onset, and the FIM were examined. In addition to these items, the ASIA motor scores were calculated for 22 tetraplegic patients. We investigated the relationships among these various respects. We also examined the changes of the physical items of the FIM score (physical FIM) over time for 18 patients. The mean FIM scores of those with tetraplegia with C4, C5, C6, C7, C8 lesions, and those with paraplegia with above T5 levels, and those below T6 were 35, 61, 82, 90, 116, 114 and 114 respectively. The FIM score reached the plateau in approximately 10 months, 6 months and 3 months post-injury, in tetraplegia, paraplegia above T5 and that below T6 respectively. The FIM scores in C6 patients were widely distributed from 56 to 104. On the other hand, the ASIA motor score could subdivide C6 patients and related well to the FIM score. The mean FIM scores for each neurological level were similar to those previously reported, thus they appeared to be plateau scores. With regard to the motor score, we feel that it could reflect the disability of the patients better than considering the neurological levels alone. Also considering the changes in the physical FIM score over time within a year from the onset of the injury, there were differences in the ADL improvement patterns among patients with different neurological levels. It appears that timing of the highest physical FIM improvement for each neurological level can exist. Thus it is important not to delay the start of the rehabilitation of patients with spinal cord injury in proper time.


[Brachioplasty. New technique].
A new brachioplasty technique is proposed, designed to correct ptosis without excision. It is based on rolling a de-epithelialised flap around from behind and sliding it underneath the anterior margin of the incision of the arm, recreating the roundness of the arm and a firmer consistency. This technique has been used in 12 patients so far, with a follow-up ranging from 8 months to 4 years. No cases of postoperative edema have been observed, but one case of hyperesthesia appeared for a period of 2 months. The main preoccupation is scar sequelae, which must limit the operation to patients clearly aware of the possibility of healing complications.


Hypoxemic Respiratory Failure Secondary to Zika Virus Infection.
An 80-year-old man experienced general weakness, myalgias, arthralgias, fever, chills, and diarrhea for one week. He had hypotension and tachycardia. He also had leukocytosis, thrombocytopenia, increased creatinine levels, elevated liver enzymes, elevated creatine phosphokinase (CPK) levels, and metabolic acidosis with hypoxemia, for which he was admitted to the Intensive Care Unit (ICU). His chest x-ray showed decreased lung volumes. Ceftriaxone and levofloxacin were empirically started to cover leptospirosis and community acquired pneumonia, respectively. The patient continued with clinical deterioration and the antibiotic therapy was changed to linezolid, cefepime, and doxycycline. He required endotracheal intubation and mechanical ventilation support due to progressive hypoxemic respiratory failure. A bronchoscopy showed no evidence of bacterial infectious process. The patient developed clinical improvement with successful extubation afterwards (4 days after initial intubation). He was later discharged home with physical therapies. A serum specimen was tested with real-time polymerase chain reaction (RT-PCR) technique, producing a positive result only for Zika virus. Confirmatory molecular diagnostic testing was performed at the Center for Disease Control (CDC).


Differences between local and orbital dynamic stability during human walking.
Currently there is no commonly accepted way to define, much less quantify, locomotor stability. In engineering, "orbital stability" is defined using Floquet multipliers that quantify how purely periodic systems respond to perturbations discretely from one cycle to the next. For aperiodic systems, "local stability" is defined by local divergence exponents that quantify how the system responds to very small perturbations continuously in real time. Triaxial trunk accelerations and lower extremity sagittal plane joint angles were recorded from ten young healthy subjects as they walked for 10 min over level ground and on a motorized treadmill at the same speed. Maximum Floquet multipliers (Max FM) were computed at each percent of the gait cycle (from 0% to 100%) for each time series to quantify the orbital stability of these movements. Analyses of variance comparing Max FM values between walking conditions and correlations between Max FM values and previously published local divergence exponent results were computed. All subjects exhibited orbitally stable walking kinematics (i.e., magnitudes of Max FM < 1.0), even though these same kinematics were previously found to be locally unstable. Variations in orbital stability across the gait cycle were generally small and exhibited no systematic patterns. Walking on the treadmill led to small, but statistically significant improvements in the orbital stability of mediolateral (p = 0.040) and vertical (p = 0.038) trunk accelerations and ankle joint kinematics (p = 0.002). However, these improvements were not exhibited by all subjects (p < or = 0.012 for subject x condition interaction effects). Correlations between Max FM values and previously published local divergence exponents were inconsistent and 11 of the 12 comparisons made were not statistically significant (r2 < or = 19.8%; p > or = 0.049). Thus, the variability inherent in human walking, which manifests itself as local instability, does not substantially adversely affect the orbital stability of walking. The results of this study will allow future efforts to gain a better understanding of where the boundaries lie between locally unstable movements that remain orbitally stable and those that lead to global instability (i.e., falling).


PBMCs are additional sites of productive infection of bovine papillomavirus type 2.
Bovine papillomavirus type 2 (BPV-2) is an oncogenic virus infecting both epithelial and mesenchymal cells. Its life cycle, similar to other papillomaviruses (PVs), appears to be linked to epithelial differentiation. Human and bovine PVs have been known to reside in a latent, episomal form in PBMCs; therefore, it is believed that blood cells, like all mesenchymal cells, function as non-permissive carriers. Here, for the first time in veterinary and comparative medicine, the BPV-2 E5 oncoprotein and the major structural L1 capsid protein, known to be expressed only in productive infections, were shown to occur in defined subsets of PBMCs. E5 oncoprotein was detected in sorted T- and B-cells as well as in monocytes by flow cytometry and Western blot analysis. However, CD4(+) and CD8(+) lymphocytes appeared to be the main circulating targets of the virus, thus possibly representing the most important reservoir of active BPV-2 in blood. L1 protein was identified by flow cytometry in a population of blood cells recognized as lymphocytes by morphological scatter properties. Western blot analysis was performed on lysates obtained from the sorted subpopulations of PBMCs and detected L1 protein in CD4(+) and CD8(+) cells only. Thus, this study showed that CD4(+) and CD8(+) lymphocytes are permissive for BPV-2 and are new, hitherto unknown sites of productive PV infection. In light of these observations, the life cycle of PVs needs to be revisited to gain novel insights into the epidemiology of BPV infection and the pathogenesis of related diseases.


Quantifying archaeo-organic degradation - A multiproxy approach to understand the accelerated deterioration of the ancient organic cultural heritage at the Swedish Mesolithic site Ageröd.
Despite a growing body of evidence concerning accelerated organic degradation at archaeological sites, there have been few follow-up investigations to examine the status of the remaining archaeological materials in the ground. To address the question of archaeo-organic preservation, we revisited the Swedish, Mesolithic key-site Ageröd and could show that the bone material had been subjected to an accelerated deterioration during the last 75 years, which had destroyed the bones in the areas where they had previously been best preserved. To understand why this has happened and to quantify and qualify the extent of the organic degradation, we here analyse the soil chemistry, bone histology, collagen preservation and palaeobotany at the site. Our results show that the soil at Ageröd is losing, or has already lost, its preservative and buffering qualities, and that pH-values in the still wet areas of the site have dropped to levels where no bone preservation is possible. Our results suggest that this acidification process is enhanced by the release of sulphuric acid as pyrite in the bones oxidizes. While we are still able to find well-preserved palaeobotanical remains, they are also starting to corrode through re-introduced oxygen into the archaeological layers. While some areas of the site have been more protected through redeposited soil on top of the archaeological layers, all areas of Ageröd are rapidly deteriorating. Lastly, while it is still possible to perform molecular analyses on the best-preserved bones from the most protected areas, this opportunity will likely be lost within a few decades. In conclusion, we find that if we, as a society, wish to keep this valuable climatic, environmental and cultural archive, both at Ageröd and elsewhere, the time to act is now and if we wait we will soon be in a situation where this record will be irretrievably lost forever.


The Urine Albumin-to-Creatinine Ratio and Kidney Function after Nephrectomy.
Patients with kidney cancer are at risk for chronic kidney disease after radical and partial nephrectomy. We determined if the urine albumin-to-creatinine ratio is independently associated with progressive chronic kidney disease after nephrectomy. We performed a cohort study based within a large, integrated health care system. We identified patients who underwent radical or partial nephrectomy from 2004 to 2014 with urine albumin-to-creatinine ratio measured in the 12 months before surgery. We fit multivariable models to determine if the urine albumin-to-creatinine ratio was associated with the time to chronic kidney disease progression (defined as reaching stage 4 or 5 chronic kidney disease, estimated glomerular filtration rate less than 30 ml/minute/1.73 m2). We performed a parallel analysis measuring the time to stage 3b, 4 or 5 chronic kidney disease (estimated glomerular filtration rate less than 45 ml/minute/1.73 m2) among patients with normal or near normal preoperative kidney function (estimated glomerular filtration rate 60 ml/minute/1.73 m2 or greater). We also examined the association between urine albumin-to-creatinine ratio and survival. A total of 1,930 patients underwent radical or partial nephrectomy and had preoperative urine albumin-to-creatinine ratio and preoperative and postoperative estimated glomerular filtration rate. Of these patients 658 (34%) and 157 (8%) had moderate (urine albumin-to-creatinine ratio 30 to 300 mg/gm) or severe albuminuria (urine albumin-to-creatinine ratio greater than 300 mg/gm), respectively. Albuminuria severity was independently associated with progressive chronic kidney disease after radical (moderate albuminuria HR 1.7, 95% CI 1.4-2.2; severe albuminuria HR 2.3, 95% CI 1.7-3.1) and partial nephrectomy (moderate albuminuria HR 1.8, 95% CI 1.2-2.7; severe albuminuria HR 4.3, 95% CI 2.7-7.0). Albuminuria was also associated with survival following radical and partial nephrectomy. In patients undergoing radical or partial nephrectomy the severity of albuminuria can stratify risk of progressive chronic kidney disease.


Standardizing central venous catheter care by using observations from patients with cancer.
To understand the vulnerability of patients with cancer to central line-associated bloodstream infections related to tunneled central venous catheters (CVCs), patients were asked to describe their line care at home and in clinic and to characterize their knowledge and experience managing CVCs. Forty-five adult patients with cancer were recruited to participate. Patients were interviewed about the type of line, duration of use, and observations of variations in line care. They also were asked about differences between line care at home and in the clinic, precautions taken when bathing, and their education regarding line care. Demographic information and primary cancer diagnosis were taken from the patients' medical records. Patients with hematologic and gastrointestinal malignancies were heavily represented. The majority had tunneled catheters with subcutaneous implanted ports. Participants identified variations in practice among nurses who cared for them. Although many participants expressed confidence in their knowledge of line care, some were uncertain about what to do if the dressing became loose or wet, or how to recognize an infection. Patients seemed to be astute observers of their own care and offered insights into practice variation. Their observations show that CVC care practices should be standardized, and educational interventions should be created to address patients' knowledge deficits.


Fluoxetine-induced alteration of murine gut microbial community structure: evidence for a microbial endocrinology-based mechanism of action responsible for fluoxetine-induced side effects.
Depression and major depressive disorder affect 25% of the population. First line treatment utilizing selective serotonin reuptake inhibitors (SSRIs) have met with limited success due to well-recognized negative side effects which include weight gain or loss. This inability to control unwanted side effects often result in patients stopping their antidepressant medications. The mechanisms underlying the failure of SSRIs are incompletely understood. Male CF-1 mice (5 weeks of age, N = 10 per group) were per orally administered fluoxetine (20 mg per kg body weight) or diluent daily for 29 days. During this time fecal specimens were collected at three defined time points (0, 15 and 29 days). At the conclusion of the 29-day dosing regimen, animals were subjected to two behavioral assessments. For bacterial identification of the microbiota, 16S rRNA gene sequencing was performed on 60 fecal specimens (three specimens per mouse time course, N = 20 mice) using Illumina MiSeq. Analysis of community sequence data was done using mothur and LEfSe bioinformatic software packages. Daily per oral administration of fluoxetine for 29 days to male mice resulted in a significant, time dependent, alteration in microbial communities accompanying changes in body weight. The calculated species richness and diversity indicators of the murine fecal microbial communities were inconsistent and not significantly different between the groups. Among the phylotypes decreased in abundance due to fluoxetine administration were Lactobacillus johnsonii and Bacteroidales S24-7 which belong to phyla associated with regulation of body mass. The observed changes in body weight due to fluoxetine administration mimicked the dramatic shifts in weight gain/loss that has been observed in humans. Further, at the conclusion of the 29-day dosing regimen fluoxetine-dosed animals evidenced a mild anxiogenic-like behavior. We report that the most widely used antidepressant, fluoxetine, which is an SSRI-type drug, results in the selective depletion of gut microbiota, specifically the Lactobacilli which are involved in the regulation of body weight. Concomitantly, fluoxetine administration increases the abundance of phylotypes related to dysbiosis. Since Lactobacilli have been previously shown to possess a known biogenic amine transporter that regulates the uptake of fluoxetine, it is proposed that a microbial endocrinology-based mechanistic pathway is responsible for the ability of SSRIs to selectively negatively impact beneficial microbiota. The results of this study therefore suggest that the negative clinical side effects due to fluoxetine administration may be due to alterations in gut microbiota. Further, the data also suggests that supplementation of bacterial genera directly affected by fluoxetine administration may prove useful in ameliorating some of the well-known side effects of chronic fluoxetine administration such as weight alterations.


Distinct changes in chronic pain sensitivity and oxytocin receptor expression in a new rat model (Wisket) of schizophrenia.
Clinical studies have shown that schizophrenia is accompanied by hypoalgesia. Accordingly, we have previously reported that a chronic schizophrenia-related rat substrain (Wisket) showed decreased acute heat pain sensitivity. The aim of the present study was to determine the mechanical pain sensitivity and the effects of opioid ligands in a chronic osteoarthritic pain model generated using Wisket rats. Our previous molecular biological studies indicated that the impairment in opioid and cannabinoid receptor functions observed in these animals did not explain their altered pain sensitivity. Therefore, we aimed to investigate another endogenous antinociceptive system, i.e., the oxytocinergic system (which is also implicated in schizophrenia) via the determination the brain-region specific oxytocin receptor mRNA expression in Wisket rats. Osteoarthritis was induced in male adult control Wistar rats without any interventions and in Wisket rats after juvenile social isolation and ketamine treatment. The degree of allodynia and the effects of systemic morphine or intrathecal endomorphin-1 administration were determined. Furthermore, the expression of the oxytocin receptor mRNA was assessed in different brain structures (prefrontal cortex, striatum, diencephalon, brainstem, and olfactory bulb). A lower degree of allodynia was observed in the Wisket group compared with control animals 1 and 2 weeks after the induction of osteoarthritis, which was accompanied by a comparable degree of edema. Systemically or intrathecally applied opioids caused similar time-response curves in both groups, with apparently shorter effects in Wisket animals. The expression of the oxytocin receptor mRNA was lower in most of the brain regions (with the exception of the diencephalon) investigated in Wisket rats vs. the control animals. In summary, both acute and chronic hypoalgesia (as nonspecific symptoms in patients with schizophrenia) can be simulated in Wisket animals as endophenotypes despite the impairment of the endogenous antinociceptive systems evaluated. Thus, this model might be an appropriate tool for further investigation of the molecular basis of altered pain perception in schizophrenia.


Towards practical sample preparation in point-of-care testing: user-friendly microfluidic devices.
Microfluidic technologies offer a number of advantages for sample preparation in point-of-care testing (POCT), but the requirement for complicated external pumping systems limits their wide use. To facilitate sample preparation in POCT, various methods have been developed to operate microfluidic devices without complicated external pumping systems. In this review, we introduce an overview of user-friendly microfluidic devices for practical sample preparation in POCT, including self- and hand-operated microfluidic devices. Self-operated microfluidic devices exploit capillary force, vacuum-driven pressure, or gas-generating chemical reactions to apply pressure into microchannels, and hand-operated microfluidic devices utilize human power sources using simple equipment, including a syringe, pipette, or simply by using finger actuation. Furthermore, this review provides future perspectives to realize user-friendly integrated microfluidic circuits for wider applications with the integration of simple microfluidic valves.


Neonatal apnea associated with maternal clonazepam therapy: a case report.
A 2750-g female infant was born at 36 weeks' gestation to a 40-year-old woman treated with clonazepam throughout her pregnancy. The infant developed apnea, cyanosis, and hypotonia within a few hours of birth. The mother's serum clonazepam level at delivery was 32 ng/mL; the cord blood level was 19 ng/mL. The infant had no congenital malformations, evidence of infection, or seizures. Clinical episodes ceased by ten days of age. The woman elected to breastfeed; breast milk clonazepam levels were between 11 and 13 ng/mL. She was discharged with a cardiorespiratory monitor. The authors suggest that infants of mothers receiving this agent during pregnancy or while nursing have serum levels measured. Additionally, these infants should be monitored for central nervous system depression or apnea.


Rapid and Scalable Halosulfonylation of Strain-Release Reagents.
Sulfonylated aromatics are commonplace motifs in drugs and agrochemicals. However, methods for the direct synthesis of sulfonylated non-classical arene bioisosteres, which could improve the physicochemical properties of drug and agrochemical candidates, are limited. Here we report a solution to this challenge: a one-pot halosulfonylation of [1.1.1]propellane, [3.1.1]propellane and bicyclo[1.1.0]butanes that proceeds under practical, scalable and mild conditions. The sulfonyl halides used in this chemistry feature aryl, heteroaryl and alkyl substituents, and are conveniently generated in situ from readily available sulfinate salts and halogen atom sources. This methodology enables the synthesis of an array of pharmaceutically and agrochemically relevant halogen/sulfonyl-substituted bioisosteres and cyclobutanes, on up to multidecagram scale.


The activity of 2',3'-cyclic nucleotide 3'-phosphodiesterase in rat tissues.
The activity of the myelin-associated enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) was measured in 14 rat tissues and in subcellular fractions of rat liver by a sensitive fluorometric method, using cyclic NADP as substrate. CNP activity in brain (339 mumol/h/mg protein) was fourfold that of the sciatic nerve. The activities in tissues outside the nervous system ranged from a low of 0.42 mumol/h/mg protein in the unwashed red blood cell to a high of 9.96 in the spleen. The activity was highest in tissues containing cells with membranes capable of undergoing transformation and elaboration (spleen and thymus) and low in those in which the cell membranes are morphologically stable (muscle and red cell). The enzyme was found in all major liver subfractions, with the highest activities in the microsomal and nuclear fractions. Despite the large difference in the maximal velocities of CNP in brain and liver, the affinity of the liver enzyme for the substrate (km) was similar to that of brain enzyme. Brain CNP was stable over a 48-h postmortem period.


SREBP2 gene therapy targeting striatal astrocytes ameliorates Huntington's disease phenotypes.
Brain cholesterol is produced mainly by astrocytes and is important for neuronal function. Its biosynthesis is severely reduced in mouse models of Huntington's disease. One possible mechanism is a diminished nuclear translocation of the transcription factor sterol regulatory element-binding protein 2 (SREBP2) and, consequently, reduced activation of SREBP2-controlled genes in the cholesterol biosynthesis pathway. Here we evaluated the efficacy of a gene therapy based on the unilateral intra-striatal injection of a recombinant adeno-associated virus 2/5 (AAV2/5) targeting astrocytes specifically and carrying the transcriptionally active N-terminal fragment of human SREBP2 (hSREBP2). Robust hSREBP2 expression in striatal glial cells in R6/2 Huntington's disease mice activated the transcription of cholesterol biosynthesis pathway genes, restored synaptic transmission, reversed dopamine receptor D2 (Drd2) transcript levels decline, cleared mutant huntingtin aggregates and attenuated behavioural deficits. We conclude that glial SREBP2 participates in Huntington's disease brain pathogenesis in vivo and that AAV-based delivery of SREBP2 to astrocytes counteracts key features of the disease.


Characterization of monoclonal antibodies to terminal and internal O-antigen epitopes of Francisella tularensis lipopolysaccharide.
The lipopolysaccharide (LPS) of Francisella tularensis (Ft), the Gram negative bacterium that causes tularemia, has been shown to be a main protective antigen in mice and humans; we have previously demonstrated that murine anti-Ft LPS IgG2a monoclonal antibodies (MAbs) can protect mice against otherwise lethal intranasal infection with the Ft live vaccine strain (LVS). Here we show that four IgG2a anti-LPS MAbs are specific for the O-polysaccharide (O-antigen [OAg]) of Ft LPS. But whereas three of the MAbs bind to immunodominant repeating internal epitopes, one binds to a unique terminal epitope of Ft OAg. This was deduced from its even binding to both long and short chains of the LPS ladder in Western blots, its rapid decrease in ELISA binding to decreasing solid-phase LPS concentrations, its inability to compete for LPS binding with a representative of the other three MAbs, and its inability to immunoprecipitate OAg despite its superior agglutination titer. Biacore analysis showed the end-binding MAb to have higher bivalent avidity for Ft OAg than the internal-binding MAbs and provided an immunogenicity explanation for the predominance of internal-binding anti-Ft OAg MAbs. These findings demonstrate that non-overlapping epitopes can be targeted by antibodies to Ft OAg, which may inform the design of vaccines and immunotherapies against tularemia.


Phantom limb pain, cortical reorganization and the therapeutic effect of mental imagery.
Using functional MRI (fMRI) we investigated 13 upper limb amputees with phantom limb pain (PLP) during hand and lip movement, before and after intensive 6-week training in mental imagery. Prior to training, activation elicited during lip purse showed evidence of cortical reorganization of motor (M1) and somatosensory (S1) cortices, expanding from lip area to hand area, which correlated with pain scores. In addition, during imagined movement of the phantom hand, and executed movement of the intact hand, group maps demonstrated activation not only in bilateral M1 and S1 hand area, but also lip area, showing a two-way process of reorganization. In healthy participants, activation during lip purse and imagined and executed movement of the non-dominant hand was confined to the respective cortical representation areas only. Following training, patients reported a significant reduction in intensity and unpleasantness of constant pain and exacerbations, with a corresponding elimination of cortical reorganization. Post hoc analyses showed that intensity of constant pain, but not exacerbations, correlated with reduction in cortical reorganization. The results of this study add to our current understanding of the pathophysiology of PLP, underlining the reversibility of neuroplastic changes in this patient population while offering a novel, simple method of pain relief.


Ciprofloxacin for treatment of drug-resistant epilepsy.
To investigate the efficacy of short-term treatment with ciprofloxacin in alteration of gut microbiota pattern and reduction of seizure frequency in adult patients with drug-resistant epilepsy. In a prospective study, we investigated the effect of a 5-day course of treatment with ciprofloxacin on gut microbiota pattern and seizure frequency of 23 adults with drug-resistant epilepsy. Fecal samples were collected before and after treatment and were analyzed for microbial load and species. Changes in seizure frequency were registered for 12 weeks. Responders were defined as patients who experienced ≥50 % seizure reduction in comparison to baseline. Outcome measures were specified as alteration in fecal microbial burden in days 5-7 and responder rate in 4th and 12th weeks. The mean baseline frequency of seizures was5.6 ±7.7 per week. All patients were on polytherapy with a mean of 3 ± 1.2 anti-seizure medications. Microbial analysis showed a considerable increase in Bacteroidetes/Firmicutes ratio after treatment. Seizure frequency significantly decreased at the end of first week and the therapeutic effect continued to week 12 (P < 0.001). The responder rate at 4th and 12th weeks were 69.6 % and 73.9 % respectively with a more prominent response in patients with symptomatic generalized epilepsy (P:0.06). Alteration of abnormal gut microbiota pattern by methods such as short-course antibiotic therapy, prescription of probiotics and fecal microbiota transplant might be effective in treatment of drug-resistant epilepsy.


Malignant lymphomas of follicular center cell origin in man. II. Ultrastructural and cytochemical studies.
Tissues from malignant lymphomas with both nodular and diffuse growth patterns, thought by light microscopy to be composed of cells of follicular center cell (FCC) origin, Were examined by electron microscopy; the tumor cells were similar to lymphoid cells found in reactive follicular centers. Tumor cells from neoplasms thought to be composed of cleaved FCC often had more pronounced nuclear folding than did cleaved FCC of reactive follicles, whereas cells in tumors of noncleaved FCC type were indistinguishable from their presumed counterparts in reactive follicles. Large cell noeplasms, previously classified as "histiocytic" lymphomas were composed of cells with ultrastructural characteristics of transformed lymphocytes; they showed neither ultrastructural nor cytochemical features of mononuclear phagocytes. These findings support the concept that a major group of lymphomas arises from lymphocytes of follicular centers.


Evaluation of photopolymer resins for dental prosthetics fabricated via the stereolithography process at different polymerization temperatures-Part I: Conversion rate and mechanical properties.
Improvement in the mechanical properties of 3-dimensional (3D) printed dental prostheses is necessary to prevent wear caused by an antagonist or fracture. However, how different printing temperatures affect their mechanical properties is unclear. The purpose of this in vitro study was to evaluate the mechanical properties of 3D printed parts fabricated at different printing temperatures. Photopolymer specimens were fabricated at 3 different temperatures (room temperature, 50 °C, and 70 °C) using a stereolithography 3D printer. After rinsing to remove the residual monomer, the specimens were divided into 2 groups: with or without postprocessing. The viscosity of the photopolymerization resin was measured while the temperature was increased. Furthermore, the double-bond conversion (DBC) of the printed part was evaluated (n=3). Mechanical properties were investigated via dynamic mechanical analysis (n=1) and tensile testing (n=5). Statistical comparisons were performed via 1-way analysis of variance, followed by the Tukey honestly significant difference test (α=.05). The DBC rates of the green condition group increased from 66.67% to 86.33% with increasing temperature. In addition, these specimens exhibited improved mechanical properties and reduced residual monomer levels. Specimens fabricated at a temperature of 70 °C exhibited mechanical properties suitable for clinical application.


Acceptability of an "on-demand" pre-exposure HIV prophylaxis trial among men who have sex with men living in France.
Although predictors of willingness to take daily, self-administered pre-exposure HIV prophylaxis (PrEP) for men who have sex with men (MSM) have been studied in the context of several PrEP trials internationally, little is known about MSM interested in participating in a trial on the use of PrEP on an "on -demand" basis, i.e., taking a first dose of combined tenofovir/emtricitabine a few hours before possible HIV sexual exposure and a second dose a few hours afterwards. A double-blind placebo randomized PrEP trial will soon begin in France to evaluate the effectiveness of PrEP in terms of reducing HIV infection rates, among MSM self-administering "on-demand" PrEP. To assess potential participants' characteristics associated with willingness to participate in the trial and identify barriers and facilitators to implementation, MSM completed a self-administered questionnaire, distributed via gay venues and community websites. Among the 443 respondents who reported being HIV-negative, 40% reported being interested in participating. Factors independently associated with interest included: reporting lower educational level, more than 20 male sexual partners in the previous year, reporting unprotected anal sex with casual partners and preferring PrEP follow-up visits in a devoted area within a hospital. There is great interest in participating in a future "on-demand" PrEP trial among HIV-negative MSM and particularly in those at potentially high risk of HIV exposure. Providing confidentiality and tailored counseling during PrEP follow-up are important issues.


Diagnosis, Treatment Response, and Prognosis: The Role of 18F-DOPA PET/CT in Children Affected by Neuroblastoma in Comparison with 123I-mIBG Scan: The First Prospective Study.
Our purpose was to evaluate the diagnostic role of 18F-3,4-dihydroxyphenylalanine (DOPA) PET/CT at the time of staging in children with neuroblastoma and to investigate its ability to assess treatment response. We also investigated the prognostic value of 18F-DOPA PET/CT at the same time points. Methods: We enrolled children with neuroblastoma at onset. Before and after induction chemotherapy, all patients underwent 18F-DOPA PET/CT and 123I-metaiodobenzylguanidine (MIBG) scanning plus SPECT/CT. 18F-DOPA PET/CT results were compared with those of 123I-MIBG whole-body scanning (WBS). For each modality, patient-based analysis and lesion-based analysis were performed and sensitivity was calculated. We applied scoring systems to 123I-MIBG scanning and 18F-DOPA PET/CT (i.e.,123I-MIBG WBS score and whole-body metabolic burden [WBMB], respectively) and evaluated the association between these parameters, the principal neuroblastoma risk factors, and outcome. Results: We enrolled 16 high-risk and 2 intermediate-risk neuroblastoma patients. On patient-based analysis, sensitivity in detecting primary tumors, soft-tissue metastases, and bone or bone-marrow metastases was 83%, 50%, and 92%, respectively, for 123I-MIBG WBS versus 94%, 92%, and 100%, respectively, for 18F-DOPA PET/CT. On lesion-based analysis, the sensitivity of 18F-DOPA PET/CT in detecting soft-tissue and bone or bone-marrow metastases was 86% and 99%, respectively-significantly higher than that of 123I-MIBG WBS, at 41% and 93%, respectively. After therapy, on patient-based analysis, the sensitivity in detecting primary tumors, soft-tissue metastases, and bone or bone-marrow metastases was 72%, 33%, and 38%, respectively, for 123I-MIBG WBS versus 83%, 75% and 54%, respectively, for 18F-DOPA PET/CT. On lesion-based analysis, the sensitivity of 18F-DOPA PET/CT in detecting soft-tissue and bone or bone-marrow metastases was 77% and 86%, respectively-significantly higher than that of 123I-MIBG WBS, at 28% and 69%, respectively. During follow-up, 8 cases of disease progression and 5 deaths occurred. On multivariate analysis, only posttherapeutic 18F-DOPA WBMB (>7.5) was associated with progression-free survival. Conclusion:18F-DOPA PET/CT is more sensitive than 123I-MIBG WBS in staging neuroblastoma patients and evaluating disease persistence after chemotherapy. In a time-to-event analysis, posttherapeutic 18F-DOPA WBMB remained the only risk factor associated with disease progression.


Antibacterial susceptibility of anaerobes by agar dilution method.
Of the 9 antibacterials, vancomycin alone at its break-point (optimum blood level) revealed highest (93.3%) efficacy on Gram-positive cocci. The remarkable activity of carbenicillin, chloramphenicol and vancomycin on Gram-positive cocci (95.4 to 96.9%), carbenicillin and chloramphenicol on Gram-positive bacilli (94.8% to 98.2%) and chloramphenicol on Gram-negative bacilli (95.4%) was noted by increasing their concentrations 1.5 to 2.5 times above their break-points. These (and even higher) concentration, however, showed relatively lower efficacy of metronidazole, rifampicin, penicillin, clindamycin and erythromycin on Gram-positive cocci (76 to 84.1%), rifampicin, tetracycline, erythromycin, clindamycin and vancomycin on Gram-positive bacilli (81 to 87.9%) and clindamycin, metronidazole, rifampicin and carbenicillin in Gram-negative bacilli (72.7 to 84%). By and large, rifampicin, chloramphenicol and carbenicillin are also quite effective (72.1 to 75.1%) at lower concentration.


Hydroxyurea interaction with α7 nicotinic acetylcholine receptor can underlie its therapeutic efficacy upon COVID-19.
In this paper the authors provide evidence that hydroxyurea (hydroxycarbamide) interacts with α7 nicotinic acetylcholine receptor, exerts anti-inflammatory and pro-survival effect, prevents α7 nicotinic receptor interaction with angiotensin-converting enzyme-2 and stimulates IgM to IgG class switch upon immunization with SARS spike protein fragment 674-685. Hydroxyurea shifts immunoglobulin glycosylation profile to anti-inflammatory phenotype and prevents the appearance of anti-idiotypic α7(179-190)-specific antibodies, as well as memory impairment. According to these results, interaction with α7 nicotinic acetylcholine receptor may underlie positive therapeutic effects of hydroxyurea upon SARS-Cov-2 infection by interfering with virus penetration into the cell and providing anti-inflammatory and immunomodulatory effects.


MMP20 hemopexin domain mutation in amelogenesis imperfecta.
Proteolytic enzymes serve important functions during dental enamel formation, and mutations in the kallikrein 4 (KLK4) and enamelysin (MMP20) genes cause autosomal-recessive amelogenesis imperfecta (ARAI). So far, only 1 KLK4 and 3 MMP20 mutations have been reported in ARAI kindreds. To determine whether ARAI in a family with a hypomaturation-type enamel defect is caused by mutations in the genes encoding enamel proteolytic enzymes, we performed mutational analysis on candidate genes. Mutational and haplotype analyses revealed an ARAI-causing point mutation (c.910G>A, p.A304T) in exon 6 of MMP20 that results in a single amino acid substitution in the hemopexin domain. Western blot analysis showed decreased expression of the mutant protein, but zymogram analysis demonstrated that this mutant was a functional protein. The proband and an affected brother were homozygous for the mutation, and both unaffected parents were carriers. The enamel of newly erupted teeth had normal thickness, but was chalky white and became darker with age.


An integrative model of ion regulation in yeast.
Yeast cells are able to tolerate and adapt to a variety of environmental stresses. An essential aspect of stress adaptation is the regulation of monovalent ion concentrations. Ion regulation determines many fundamental physiological parameters, such as cell volume, membrane potential, and intracellular pH. It is achieved through the concerted activities of multiple cellular components, including ion transporters and signaling molecules, on both short and long time scales. Although each component has been studied in detail previously, it remains unclear how the physiological parameters are maintained and regulated by the concerted action of all components under a diverse range of stress conditions. In this study, we have constructed an integrated mathematical model of ion regulation in Saccharomyces cerevisiae to understand this coordinated adaptation process. Using this model, we first predict that the interaction between phosphorylated Hog1p and Tok1p at the plasma membrane inhibits Tok1p activity and consequently reduces Na(+) influx under NaCl stress. We further characterize the impacts of NaCl, sorbitol, KCl and alkaline pH stresses on the cellular physiology and the differences between the cellular responses to these stresses. We predict that the calcineurin pathway is essential for maintaining a non-toxic level of intracellular Na(+) in the long-term adaptation to NaCl stress, but that its activation is not required for maintaining a low level of Na(+) under other stresses investigated. We provide evidence that, in addition to extrusion of toxic ions, Ena1p plays an important role, in some cases alongside Nha1p, in re-establishing membrane potential after stress perturbation. To conclude, this model serves as a powerful tool for both understanding the complex system-level properties of the highly coordinated adaptation process and generating further hypotheses for experimental investigation.


Benzylidene-6-hydroxy-3,4-dihydronaphthalenone chalconoids as potent tyrosinase inhibitors.
Tyrosinase enzyme has a key role in melanin biosynthesis by converting tyrosine into dopaquinone. It also participates in the enzymatic browning of vegetables by polyphenol oxidation. Therefore, tyrosinase inhibitors are useful in the fields of medicine, cosmetics, and agriculture. Many tyrosinase inhibitors having drawbacks have been reported to date; so, finding new inhibitors is a great need. A variety of 6-hydroxy-3,4-dihydronaphthalenone chalcone-like analogs (C1-C10) have been synthesized by aldol condensation of 6-hydroxy tetralone and differently substituted benzaldehydes. The compounds were evaluated for their inhibitory effect on mushroom tyrosinase by a spectrophotometric method. Moreover, the inhibition manner of the most active compound was determined by Lineweaver-Burk plots. Docking study was done using AutoDock 4.2. The drug-likeness scores and ADME features of the active derivatives were also predicted. Most of the compounds showed remarkable inhibitory activity against the tyrosinase enzyme. 6-Hydroxy-2-(3,4,5-trimethoxybenzylidene)-3,4-dihydronaphthalen-1(2H)-one (C2) was the most potent derivative amongst the series with an IC50 value of 8.8 μM which was slightly more favorable to that of the reference kojic acid (IC50 = 9.7 μM). Inhibitory kinetic studies revealed that C2 behaves as a competitive inhibitor. According to the docking results, compound C2 formed the most stable enzyme-inhibitor complex, mainly via establishing interactions with the two copper ions in the active site. In silico drug-likeness and pharmacokinetics predictions for the proposed tyrosinase inhibitors revealed that most of the compounds including C2 have proper drug-likeness scores and pharmacokinetic properties. Therefore, C2 could be suggested as a promising tyrosinase inhibitor that might be a good lead compound in medicine, cosmetics, and the food industry, and further drug development of this compound might be of great interest.


Monocyte Polarization is Altered by Total-Body Irradiation in Male Rhesus Macaques: Implications for Delayed Effects of Acute Radiation Exposure.
Radiation-induced fibrosis (RIF) is a common delayed effect of acute ionizing radiation exposure (DEARE) affecting diverse tissues including the heart, lungs, liver and skin, leading to reduced tissue function and increased morbidity. Monocytes, which may be classified into classical (CD14++, CD16-), intermediate (CD14++, CD16+) and non-classical (CD14+/low, CD16++) subtypes in humans and non-human primates (NHPs), and monocyte-derived macrophages may play an integral role in the pathogenesis of RIF. We tested the hypothesis that moderate to high levels of total-body exposure to radiation would alter monocyte polarization and produce phenotypes that could promote multi-organ fibrosis in a wellestablished NHP model of DEARE. Subjects were 16 young adult male rhesus macaques, ten of which were exposed to high-energy, 4 Gy X-ray total-body irradiation (TBI) and six that received sham irradiation (control). Total monocytes assessed by complete blood counts were 89% depleted in TBI animals by day 9 postirradiation (P < 0.05), but recovered by day 30 postirradiation and did not differ from control levels thereafter. Monocytes were isolated from peripheral blood mononuclear cells (PBMCs) and sorted into classical, intermediate and non-classical subsets using fluorescence-activated cell sorting (FACS) prior to and at 6 months post-TBI. At 6 months postirradiation, monocyte polarization shifted towards lower classical (92% → 86%) and higher intermediate (7% → 12%) and non-classical monocyte subsets (0.6% → 2%) (all P < 0.05) in TBI animals compared to baseline. No change in monocyte subsets was observed in control animals. Transcriptional profiles in classical and intermediate monocyte subsets were assessed using RNAseq. Classical monocyte gene expression did not change significantly over time or differ cross-sectionally between TBI and control groups. In contrast, significant numbers of differentially expressed genes (DEGs) were detected in intermediate monocyte comparisons between the TBI animals and all animals at baseline (304 DEGs), and in the TBI versus control animals at 6 months postirradiation (67 DEGs). Intermediate monocytes also differed between baseline and 6 months in control animals (147 DEGs). Pathway analysis was used to identify genes within significant canonical pathways, yielding 52 DEGs that were specific to irradiated intermediate monocytes. These DEGs and significant canonical pathways were associated with pro-fibrotic and anti-inflammatory signaling pathways that have been noted to induce M2 macrophage polarization. These findings support the hypothesis that TBI may alter monocyte programming and polarization towards a profibrotic phenotype, providing a novel target opportunity for therapies to inhibit or prevent RIF.


Fabrication of bacterial cellulose/chitosan-MIL-100(Fe) composite for adsorptive removal of dacarbazine.
In this research, a polymeric composite based on a chitosan/bacterial cellulose (CS/BC) matrix filled with MIL-100(Fe) particles was prepared to solve the recyclability of issue MIL-100(Fe) particles and utilized as an efficient adsorbent for removing dacarbazine (DTIC) from wastewater. The adsorption capacity of the composite (CS/BC-MIL) was higher than both MIL-100(Fe) and the CS/BC polymeric matrix. The adsorption performance of the fabricated composite was evaluated through kinetics and isotherm studies. While isotherm studies revealed that the adsorption of DTIC onto the adsorbent can be well described by the Freundlich model, kinetics studies indicated that a combination of factors, rather than a single rate-limiting factor, are responsible for the adsorption rate. Thermodynamics investigation showed that the adsorption of DTIC to CS/BC-MIL composite is exothermic and occurs spontaneously. Additionally, due to the negative entropy change, it was established that the adsorption is governed by the enthalpy change. Exploring the solution chemistry revealed that the optimum pH for the adsorption process was about 4. Moreover, the CS/BC-MIL can selectively adsorb DTIC in the presence of other pharmaceuticals like doxorubicin (DOX). Furthermore, regeneration investigations disclosed that the composite holds its structural features and has an acceptable adsorption capacity after several cycles of adsorption/desorption.


Role of PDK1 in insulin-signaling pathway for glucose metabolism in 3T3-L1 adipocytes.
To investigate the role of 3-phosphoinositide-dependent protein kinase 1 (PDK1) in the insulin-signaling pathway for glucose metabolism, wild-type (wt), the kinase-dead (kd), or the plecstrin homology (PH) domain deletion (DeltaPH) mutant of PDK1 was expressed using an adenovirus gene transduction system in 3T3-L1 adipocytes. wt-PDK1 and kd-PDK1 were found in both membrane and cytosol fractions, whereas DeltaPH-PDK1, which exhibited PDK1 activity similar to that of wt-PDK1, was detected exclusively in the cytosol fraction. Insulin dose dependently activated protein kinase B (PKB) but did not change atypical protein kinase C (aPKC) activity in control cells. aPKC activity was not affected by expression of wt-, kd-, or DeltaPH-PDK1 in either the presence or the absence of insulin. Overexpression of wt-PDK1 enhanced insulin-induced activation of PKB as well as insulin-induced phosphorylation of glycogen synthase kinase (GSK)3alpha/beta, a direct downstream target of PKB, although insulin-induced glycogen synthesis was not significantly enhanced by wt-PDK1 expression. Neither DeltaPH-PDK1 nor kd-PDK1 expression affected PKB activity, GSK3 phosphorylation, or glycogen synthesis. Thus membrane localization of PDK1 via its PH domain is essential for insulin signaling through the PDK1-PKB-GSK3alpha/beta pathway. Glucose transport activity was unaffected by expression of wt-PDK1, kd-PDK1, or DeltaPH-PDK1 in either the presence or the absence of insulin. These findings suggest the presence of a signaling pathway for insulin-stimulated glucose transport in which PDK1 to PKB or aPKC is not involved.


Speakers and listeners exploit word order for communicative efficiency: A cross-linguistic investigation.
Pragmatic theories and computational models of reference must account for people's frequent use of redundant color adjectives (e.g., referring to a single triangle as "the blue triangle"). The standard pragmatic view holds that the informativity of a referential expression depends on pragmatic contrast: Color adjectives should be used to contrast competitors of the same kind to preempt an ambiguity (e.g., between several triangles of different colors), otherwise they are redundant. Here we propose an alternative to the standard view, the incremental efficiency hypothesis, according to which the efficiency of a referential expression must be calculated incrementally over the entire visual context. This is the first theoretical account of referential efficiency that is sensitive to the incrementality of language processing, making different cross-linguistic predictions depending on word order. Experiment 1 confirmed that English speakers produced more redundant color adjectives (e.g., "the blue triangle") than Spanish speakers (e.g., "el triángulo azul"), but both language groups used more redundant color adjectives in denser displays where it would be more efficient. In Experiments 2A and 2B, we used eye tracking to show that pragmatic contrast is not a processing constraint. Instead, incrementality and efficiency determine that English listeners establish color contrast across categories (BLUE SHAPES > TRIANGULAR ONE), whereas Spanish listeners establish color contrast within a category (TRIANGLES > BLUE ONE). Spanish listeners, however, reversed their visual search strategy when tested in English immediately after. Our results show that speakers and listeners of different languages exploit word order to increase communicative efficiency. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Pros and cons of catheter lock solutions.
Catheter-related bacteremia (CRB) and catheter dysfunction are major sources of morbidity and mortality when central venous catheters are used as long-term vascular access in hemodialysis patients. Attempts have been made to minimize these complications by the prophylactic use of catheter locking solutions. This review aims to describe the recent advances in the field of lock solutions and to discuss the benefits and risks associated with the routine use of antimicrobial and/or antithrombotic lock solutions. Antithrombotic lock solutions may improve patency and CRB but may be cost-prohibitive. Antimicrobial lock solutions may decrease the incidence of CRB, but their routine use is concerning for the risk of systemic toxicity and the development of resistant organisms. Preliminary results suggest a novel antimicrobial and antithrombotic lock solution may be promising in maintaining patency, while decreasing catheter-related bacteremia. The ideal catheter lock solution would be one that prevents infections and thrombosis safely and effectively, while being economically viable. Recent developments have led to improvements in the development of catheter locking solutions, but the search for the perfect 'solution' is still ongoing.


The Rapid Implementation of a Psychological Support Model for Frontline Healthcare Workers During the COVID-19 Pandemic: A Case Study and Process Evaluation.
The COVID-19 pandemic highlighted the need for psychological support initiatives directed toward frontline healthcare workers, which can be rapidly and sustainably implemented during an infectious disease outbreak. The current case study presents a comprehensive model of psychological support that was implemented at an intensive care unit (ICU) during the first wave of the COVID-19 pandemic. The psychological support model aimed at promoting a resilient stress reaction among frontline staff by protecting physical, social, and psychological resources. The initiatives, targeting different groups of workers, included education and training, peer support, psychologist-supervised and unsupervised group sessions, on-boarding for transferred staff, manager support, and individual sessions for workers experiencing strong stress reactions. The results of the process evaluation of this rapid implementation suggest that peer support initiatives as well as daily group sessions were the most appreciated forms of psychological support. Psychologists involved in organizing and providing the support highlighted several aspects of a successful implementation of the support model: offering support during work hours (preferably after shift), positive attitude of line managers that framed support initiatives as a team effort, and involvement of experienced psychologists able to quickly adjust the content of the support according to the current needs. The study also identified two main problems of the current implementation: the lack of efficient planning due to the use of volunteer work and the need for more structural resources on the organizational level to ensure long-term sustainability of the support model and its implementation among all groups of healthcare staff. The current case study highlights the importance of establishing permanent structural resources and routines for psychological support integrated in clinical practice by healthcare organizations to improve both rapid and sustainable response to future crises.


Endoscopic hemostasis techniques for upper gastrointestinal hemorrhage: A review.
Upper gastrointestinal hemorrhage (UGIH) is an urgent disease that is often encountered in daily medical practice. Endoscopic hemostasis is currently indispensable for the treatment of UGIH. Initially, when UGIH is suspected, a cause of UGIH is presumed from the medical interview and physical findings. After ample primary treatment, urgent endoscopy is performed. Many methods of endoscopic hemostasis are in wide use, including hemoclip, injection and thermo-coagulation methods. Although UGIH develops from a wide variety of diseases, such as esophageal varices and gastric and duodenal ulcer, hemostasis is almost always possible. Identification of the causative diseases, primary treatment and characteristic features of endoscopic hemostasis are needed to allow appropriate treatment.


Phosphorylation of 5-lipoxygenase at ser523 by protein kinase A determines whether pioglitazone and atorvastatin induce proinflammatory leukotriene B4 or anti-inflammatory 15-epi-lipoxin a4 production.
The 5-lipoxygenase (5LO) produces leukotriene B(4) and 15-epilipoxin-A(4) (15-epi-LXA(4)). Phosphorylation at Ser(523) by protein kinase A (PKA) prevents 5LO shift to the perinuclear membrane. Atorvastatin and pioglitazone up-regulate 15-epi-LXA(4) production in the heart. We assessed whether phosphorylation of 5LO by PKA determines whether 5LO interacts with the membranous cytosolic phospholipase A(2) (cPLA(2)) to produce leukotriene B(4) or with cyclooxygenase-2 (COX2) to produce 15-epi-LXA(4). Rats received either pioglitazone, atorvastatin, pioglitazone plus atorvastatin, vehicle, or LPS. Rat myocardial cells were incubated with pioglitazone plus atorvastatin, pioglitazone plus atorvastatin plus H-89 (PKA inhibitor), H-89, or vehicle for 8 h. Pioglitazone and atorvastatin did not affect total 5LO expression. However, both increased 5LO levels in the cytosolic fraction. H-89 caused a shift of 5LO to the membranous fraction in atorvastatin- and pioglitazone-treated rats. Pioglitazone and atorvastatin increased phospho-5LO levels. H-89 attenuated this increase. Both pioglitazone and atorvastatin increased COX2 levels in the cytosolic fraction and the membranous fraction. H-89 prevented this increase. Pioglitazone and atorvastatin increased cPLA(2) expression in the membranous fraction. This effect was not attenuated by H-89. Pioglitazone plus atorvastatin increased 15-epi-LXA(4) levels. H-89 attenuated the effect of pioglitazone plus atorvastatin. Pioglitazone plus atorvastatin plus H-89 increased leukotriene B(4) levels. Coimmunoprecipitation showed that without H-89, atorvastatin and pioglitazone induced an interaction between 5LO and COX2 in the cytosolic fraction, whereas when H-89 was added, 5LO interacted with cPLA(2) on the membranous fraction. The 5LO phosphorylation determines whether 15-epi-LXA(4) (anti-inflammatory) or leukotriene B(4) (inflammatory mediator) is produced.


Predictors of access to palliative care services among patients who died at a Comprehensive Cancer Center.
Palliative care services can decrease physical and psychosocial distress in patients with advanced cancer. However, most patients with cancer die without access to palliative care services (APCS), and patterns of referral are not well understood. The purpose of this study was to determine predictors of patients' access to palliative care. We reviewed patient records from the computerized database at UT M. D. Anderson Cancer Center over 2 (2003 and 2004) to determine differences in characteristics and outcomes between patients with and without APCS. APCS was defined as a palliative care consultation and follow-up or transfer to the palliative care unit. A total of 499 of 1453 (34%) inpatients who died at our cancer center had APCS. There were no significant differences in race, age, or insurance status between the APCS groups. The two major predictors of a low rate of APCS were hematologic malignancies (20% rate of APCS versus 44% for solid tumors, p < 0.0001) and intensive care unit (ICU) admission (15% versus 52% for non-ICU admission, p < 0.0001). Patients with hematologic malignancies who were admitted to the ICU had the lowest APCS rate (10%, p < 0.0001). The median relative cost of care per patient for decedents with APCS was 0.62 compared to non-referred patients (p < 0.0001). APCS was lower among patients with hematologic malignancies and those admitted to the ICU. APCS resulted in a lower cost of care. Mortality in comprehensive cancer centers is quite variable among different primary malignancies. More research is needed to better define patterns of referral.


Two-sample tests and one-way MANOVA for multivariate biomarker data with nondetects.
Testing whether the mean vector of a multivariate set of biomarkers differs between several populations is an increasingly common problem in medical research. Biomarker data is often left censored because some measurements fall below the laboratory's detection limit. We investigate how such censoring affects multivariate two-sample and one-way multivariate analysis of variance tests. Type I error rates, power and robustness to increasing censoring are studied, under both normality and non-normality. Parametric tests are found to perform better than non-parametric alternatives, indicating that the current recommendations for analysis of censored multivariate data may have to be revised. Copyright © 2016 John Wiley & Sons, Ltd.


Antibody breadth and protective efficacy are increased by vaccination with computationally optimized hemagglutinin but not with polyvalent hemagglutinin-based H5N1 virus-like particle vaccines.
One of the challenges for developing an H5N1 influenza vaccine is the diversity of antigenically distinct isolates within this subtype. Previously, our group described a novel hemagglutinin (HA) derived from a methodology termed computationally optimized broadly reactive antigen (COBRA). This COBRA HA, when used as an immunogen, elicits a broad antibody response against H5N1 isolates from different clades. In this report, the immune responses elicited by the COBRA HA virus-like particle (VLP) vaccine were compared to responses elicited by a mixture of VLPs expressing representative HA molecules from clade 2.1, 2.2, and 2.3 primary H5N1 isolates (polyvalent). The COBRA HA VLP vaccine elicited higher-titer antibodies to a panel of H5N1 HA proteins than did the other VLPs. Both COBRA and polyvalent vaccines protected vaccinated mice and ferrets from experimental infection with highly lethal H5N1 influenza viruses, but COBRA-vaccinated animals had decreased viral replication, less inflammation in the lungs of mice, and reduced virus recovery in ferret nasal washes. Both vaccines had similar cellular responses postchallenge, indicating that higher-titer serum antibodies likely restrict the duration of viral replication. Furthermore, passively transferred immune serum from the COBRA HA VLP-vaccinated mice protected recipient animals more efficiently than immune serum from polyvalent-vaccinated mice. This is the first report comparing these two vaccine strategies. The single COBRA HA antigen elicited a broader antibody response and reduced morbidity and viral titers more effectively than a polyvalent mixture of primary H5N1 HA antigens.


Effects of zinc supplementation on metabolomic profiles in Tanzanian infants: a randomized trial.
Provision of zinc supplementation to young children has been associated with reduced infectious morbidity and better growth outcomes. However, the metabolic pathways underlying these outcomes are unclear, and metabolomic data from humans undergoing zinc supplementation, particularly infants, are generally lacking. To examine the effect of zinc supplementation on metabolic profiles in Tanzanian infants ages 6 weeks and 6 months METHODS: Blood samples were collected at age 6 weeks and 6 months from 50 Tanzanian infants who were enrolled in a randomized placebo-controlled trial of zinc supplementation (5 mg oral daily). Metabolomic analysis using an ultrahigh-performance liquid chromatography/tandem mass spectroscopy (UPLC-MS/MS) platform were performed to identify potential metabolomic profiles and biomarkers associated with zinc supplementation. Principal component analysis (PCA) was used to summarize metabolomic data from all samples. Two-way repeated measures ANOVAs with compound symmetry covariance structures were used to compare metabolome levels over time between infants in the two treatment arms. In PCA, our samples tended to be more separated by child age (6 weeks versus 6 months) than by zinc supplementation status. We found that zinc supplementation affected a variety of metabolites associated with amino acid, lipid, nucleotide, and xenobiotic metabolism, including indoleacetate in the tryptophan metabolism pathway,3-methoxytrosine and 4-hydrxoyphenylphruvate in the tyrosine pathway, eicosanedioate, 2-aminooctanoate, and N-acetyl-2-aminooctanoate in the fatty acid pathway, and N6-succinyladenosine in the purine metabolism pathway. Compared to the relatively small number of metabolites associated with zinc supplements, many infant metabolites changed significantly from age 6 weeks to 6 months. Zinc supplementation, despite having overall clinical benefits, appears to induce limited metabolomic changes in blood metabolites in young infants. Future larger studies may be warranted to further examine metabolic pathways associated with zinc supplementation.


Etiology of diarrhea in pediatric outpatient settings.
The frequency with which bacteria cause diarrhea evaluated in ambulatory settings is often unknown. We attempted to determine the microbiologic etiology of diarrhea in a private pediatric practice (site A) and a clinic serving largely immigrant children (site B) and to establish guidelines for bacterial culture. Children with diarrhea were prospectively enrolled, and their stools were examined for diarrheagenic bacteria, viruses and parasites. A total of 123 and 103 children were enrolled at sites A and B, respectively. Stools from all (100%), 126 (55.8%), 104 (46.0%) and 75 (33.2%) were tested for bacterial enteric pathogens, parasites, Clostridium difficile toxin and viruses, respectively. Of the 75 patients whose stool underwent complete testing, 36 (48%) contained at least 1 definitive or plausible pathogen. Twelve stools (5.3%) tested positive for bacteria [Campylobacter jejuni (n = 7), Yersinia enterocolitica, Shigella flexneri, Shigella sonnei, Salmonella serogroup D and Salmonella Braenderup (n = 1 each)]. One contained Blastocystis hominis, 8 contained C. difficile toxin and 16 contained viruses (9 rotavirus, 5 adenovirus and 2 astrovirus). Visible fecal blood (P = 0.029), increased stool frequency (P = 0.035), abdominal tenderness (P = 0.011) and fecal white (P < 0.001) or red blood cells (P = 0.002) were associated with bacterial infection. All children with stool yielding diarrheagenic bacteria or C. difficile toxin had at least 1 of these factors, but so did 75% of children without these agents (positive predictive value, 11%; negative predictive value, 100%; sensitivity, 100%; specificity, 25%). The bacterial diarrhea prevalence is similar to that in other ambulatory studies, although the spectrum differs. Exclusion criteria for stool testing in diarrhea remain elusive. Studies to determine the etiology of unexplained diarrhea and cost-effective algorithms for diarrhea diagnosis, are needed.


[Endocrine cells of mucosal epithelium in the distal part of the intestine of Lacerta vivipara].
The epithelium of the distal part of intestine of the lizard Lacerta vivipara has been studied by light and electron microscopy. The total number of endocrinocytes (argyrophilic cells) was found to increase from small bowel (57 +/- cell/mm2) to colon (9 +/- 69), and cloaca (99 +/- 8). Although the number of argentaffin cells increases from the small bowel to colon, cell decrease occurs from colon (42 +/- 6 cell/mm2) to cloaca (65 +/- 10 cell/mm2). On electronograms of the colon mucosal epithelium four types of endocrinocytes were identified. Type I--with secretory granules polymorphic for the size and form, with a high electron density core, and average size 206 +/- 31 nm. Type II--with secretory granules 265 +/- 20 nm in size, having spherical form and highly electronic dense contents. Type III--contains largest (350 +/- 12 nm), spherical, oval or irregularly-shaped secretory granules, with contents of various electronic density. Type IV--endocrine cells having small (176 +/- 5 nm) spherical or oval secretory granules with a highly electronic dense core. Besides, "mixed" cells were identified, whose cytoplasm contained simultaneously mucous and endocrinous granules.


High sodium intake is associated with important risk factors in a large cohort of chronic kidney disease patients.
An increased risk of mortality and cardiovascular disease (CVD) is observed in people with chronic kidney disease (CKD) even in early stages. Dietary sodium intake has been associated with important CVD and CKD progression risk factors such as hypertension and proteinuria in this population. We aimed to investigate the relationship between sodium intake and CVD or CKD progression risk factors in a large cohort of patients with CKD stage 3 recruited from primary care. A total of 1733 patients with previous estimated glomerular filtration rate (eGFR) of 30-59 ml/min/1.73m(2), with a mean age 72.9±9.0 years, were recruited from 32 general practices in primary care in England. Medical history was obtained and participants underwent clinical assessment, urine and serum biochemistry testing. Sodium intake was estimated from three early-morning urine specimens using an equation validated for this study population. Sixty percent of participants who had estimated sodium intake above recommendation (>100 mmol/day or 6 g salt/day) also had higher diastolic blood pressure, mean arterial pressure (MAP), urinary albumin-to-creatinine ratio, high-sensitive C-reactive protein and uric acid and used a greater number of anti-hypertensive drugs. In multivariable regression analysis, excessive sodium intake was an independent predictor of MAP (B=1.57, 95% confidence interval (CI) 0.41-2.72; P=0.008) and albuminuria (B=1.35, 95% CI 1.02-1.79; P=0.03). High sodium intake was associated with CVD and CKD progression risk factors in patients with predominantly early stages of CKD followed up in primary care. This suggests that dietary sodium intake could afffect CVD risk even in early or mild CKD. Intervention studies are warranted to investigate the potential benefit of dietary advice to reduce sodium intake in this population.


To tweet or not to tweet, that is the question: A randomized trial of Twitter effects in medical education.
Many medical education journals use Twitter to garner attention for their articles. The purpose of this study was to test the effects of tweeting on article page views and downloads. The authors conducted a randomized trial using Academic Medicine articles published in 2015. Beginning in February through May 2018, one article per day was randomly assigned to a Twitter (case) or control group. Daily, an individual tweet was generated for each article in the Twitter group that included the title, #MedEd, and a link to the article. The link delivered users to the article's landing page, which included immediate access to the HTML full text and a PDF link. The authors extracted HTML page views and PDF downloads from the publisher. To assess differences in page views and downloads between cases and controls, a time-centered approach was used, with outcomes measured at 1, 7, and 30 days. In total, 189 articles (94 cases, 95 controls) were analyzed. After days 1 and 7, there were no statistically significant differences between cases and controls on any metric. On day 30, HTML page views exhibited a 63% increase for cases (M = 14.72, SD = 63.68) when compared to controls (M = 9.01, SD = 14.34; incident rate ratio = 1.63, p = 0.01). There were no differences between cases and controls for PDF downloads on day 30. Contrary to the authors' hypothesis, only one statistically significant difference in page views between the Twitter and control groups was found. These findings provide preliminary evidence that after 30 days a tweet can have a small positive effect on article page views.


Prokaryotic Heme Biosynthesis: Multiple Pathways to a Common Essential Product.
The advent of heme during evolution allowed organisms possessing this compound to safely and efficiently carry out a variety of chemical reactions that otherwise were difficult or impossible. While it was long assumed that a single heme biosynthetic pathway existed in nature, over the past decade, it has become clear that there are three distinct pathways among prokaryotes, although all three pathways utilize a common initial core of three enzymes to produce the intermediate uroporphyrinogen III. The most ancient pathway and the only one found in the Archaea converts siroheme to protoheme via an oxygen-independent four-enzyme-step process. Bacteria utilize the initial core pathway but then add one additional common step to produce coproporphyrinogen III. Following this step, Gram-positive organisms oxidize coproporphyrinogen III to coproporphyrin III, insert iron to make coproheme, and finally decarboxylate coproheme to protoheme, whereas Gram-negative bacteria first decarboxylate coproporphyrinogen III to protoporphyrinogen IX and then oxidize this to protoporphyrin IX prior to metal insertion to make protoheme. In order to adapt to oxygen-deficient conditions, two steps in the bacterial pathways have multiple forms to accommodate oxidative reactions in an anaerobic environment. The regulation of these pathways reflects the diversity of bacterial metabolism. This diversity, along with the late recognition that three pathways exist, has significantly slowed advances in this field such that no single organism's heme synthesis pathway regulation is currently completely characterized.


Antimicrobial activities of Ferulago essential oils.
Essential oils from Ferulago asparagifolia Boiss., F. galbanifera (Miller) W. Koch, F. humilis Boiss. (Endemic), F. trachycarpa Boiss. growing in Turkey were evaluated against 15 microorganisms for their antifungal and antibacterial activity using an agar tube dilution and microdilution broth susceptibility assay, respectively. The essential oil compositions were investigated by GC/MS. Inhibitory effects against Escherichia coli, Enterobacter aerogenes, Candida albicans, Gaeumannomyces graminis var. tritici, Sclerotium rolfsii and Fusarium moniliforme were remarkable. Results are discussed in comparison with the chemical composition of the essential oils.


A key role for EZH2 in epigenetic silencing of HOX genes in mantle cell lymphoma.
The chromatin modifier EZH2 is overexpressed and associated with inferior outcome in mantle cell lymphoma (MCL). Recently, we demonstrated preferential DNA methylation of HOX genes in MCL compared with chronic lymphocytic leukemia (CLL), despite these genes not being expressed in either entity. Since EZH2 has been shown to regulate HOX gene expression, to gain further insight into its possible role in differential silencing of HOX genes in MCL vs. CLL, we performed detailed epigenetic characterization using representative cell lines and primary samples. We observed significant overexpression of EZH2 in MCL vs. CLL. Chromatin immune precipitation (ChIP) assays revealed that EZH2 catalyzed repressive H3 lysine 27 trimethylation (H3K27me3), which was sufficient to silence HOX genes in CLL, whereas in MCL H3K27me3 is accompanied by DNA methylation for a more stable repression. More importantly, hypermethylation of the HOX genes in MCL resulted from EZH2 overexpression and subsequent recruitment of the DNA methylation machinery onto HOX gene promoters. The importance of EZH2 upregulation in this process was further underscored by siRNA transfection and EZH2 inhibitor experiments. Altogether, these observations implicate EZH2 in the long-term silencing of HOX genes in MCL, and allude to its potential as a therapeutic target with clinical impact.


Does the use of bilateral mammary artery grafts compared with the use of a single mammary artery graft offer a long-term survival benefit in patients undergoing coronary artery bypass surgery?
A best evidence topic in cardiac surgery was written according to a structured protocol. The question addressed was: 'Does the use of bilateral mammary artery grafts compared with the use of a single mammary artery graft offer a long-term survival benefit in patients undergoing coronary artery bypass surgery?' Altogether 214 papers were found using the reported search, of which 13 represented the best evidence to answer the clinical question. The authors, journal, date and country of publication, patient group studied, study type, relevant outcomes and results of these papers are tabulated. All the included studies were follow-up studies; eight studies used prospective data collection, and five studies collected the study data retrospectively. No randomized controlled trials were found. Nine of the 13 included papers used a propensity-score-matched comparison of the survival of bilateral mammary artery graft [or, bilateral internal thoracic artery (BITA) graft] patients vs single mammary artery graft [or, single internal thoracic artery (SITA) graft] patients. These studies consistently showed an enhanced survival of BITA patients compared with propensity-score-matched SITA patients. Three of the 13 included papers used Cox proportional hazards regression analysis to compare survival of BITA vs SITA patients; one larger study showed better crude survival of BITA patients, but did not identify BITA grafts as independent predictor of enhanced survival. The remaining two studies also did not identify BITA grafts as independent predictor of enhanced survival. One study only presented crude survival estimates of BITA vs SITA patients and therefore was of limited informative value. We conclude that the use of BITA grafts seems to offer a long-term survival benefit compared with a SITA graft for patients undergoing coronary artery bypass grafting surgery. Although randomized evidence is lacking, observational evidence supporting this hypothesis is mounting.


The detection of double stranded DNA antibody using human metaphase chromosomes.
Double stranded native DNA (dsDNA) antibody detection is important in the diagnosis and management of patients with Systemic Lupus Erythematosus (SLE). Current radioimmunoassay and haemagglutination assays for DNA antibodies are technically complicated or show considerable inter-laboratory variation. A simple inexpensive indirect immunofluorescence method using fixed human metaphase chromosomes for DNA antibody estimation is described. The method has been compared with the Farr radioimmunoassay and Crithidia luciliae techniques. The metaphase test showed 100% correlation with a positive Farr assay in 38 of 373 routine DNA antibody estimations. Simultaneous testing using Crithidia showed 84% correlation, indicating greater sensitivity of the metaphase method. Enzyme treatment of metaphase and Crithidia substrate abolished reactivity on DNAase treated substrate alone, and DNA specific fluorescent staining further confirmed metaphase substrate specificity. The metaphase test appears simple, specific, of greater sensitivity, easily read, and can be utilized in large or small laboratories to determine DNA antibodies and their subclasses.


Frequency among Enterobacteriaceae of the DNA sequences encoding type 1 pili.
Type 1 pili, characterized by mannose-inhibitable agglutination of fowl or guinea pig erythrocytes, have been found throughout the family Enterobacteriaceae. A radiolabeled probe was prepared from a restriction endonuclease-digested fragment of the Escherichia coli pil operon and used to detect homologous DNA sequences in 236 bacteria representing 11 genera of Enterobacteriaceae. Only isolates identified as E. coli or Shigella spp. exhibited homology. In contrast, mannose-sensitive hemagglutination was observed in nine genera. Probe DNA did not hybridize to plasmid DNA, indicating a chromosomal location for the pil operon. Analysis of restriction nuclease-digested whole-cell DNA from 60 E. coli and two Shigella sp. isolates indicated that internal sequences were conserved in most strains, but that changes in flanking sequences in the chromosome were common.


Communication, swallowing and feeding in the intensive care unit patient.
It is not uncommon for patients requiring a period of time in the intensive care unit (ICU) to experience difficulties with communication and/or swallowing, either as a result of their illness or as a result of the treatments they receive. These difficulties can be both short term and long term and require timely and appropriate intervention in order to improve the patient's experience and expedite recovery/rehabilitation. The purpose of this article is to provide critical care nurses with an update on aspects of communication, swallowing and feeding in the ICU. The paper will focus on each area in relation to the current evidence base and factors of 'best practice' (as determined by expert opinion). IMPLICATIONS FOR CRITICAL CARE NURSING PRACTICE: Enabling communication can improve well-being, increase compliance and reduce length of stay; Simple modes of communication, e.g. writing/gesture/pictures can be very effective; Coded eye blinking may be unreliable because of confusion with reflexive blinking; Non-oral nutrition will generally not meet the psychological and physical needs and benefits of oral intake; Not all patients with a tracheostomy in situ will experience dysphagia; however, those considered 'at risk' should have their swallow assessed by an appropriately trained professional, e.g. speech and language therapist/dysphagia trained professional; An inflated tracheostomy cuff will not prevent aspiration. The decision to commence oral intake in the presence of an inflated cuff should be made as a team and take into consideration the patient's medical and psychological status; The use of blue dye to assess the swallow carries a high false-negative rate and cannot be relied on alone to predict either the presence or the absence of aspiration. There is still much more research to be performed and evidence to be gained regarding the input into communication, swallowing and feeding in the ICU; however, a full-team approach to these areas can have very positive effects on the patient's experience.


Chemokine Signaling Controls Integrity of Radial Glial Scaffold in Developing Spinal Cord and Consequential Proper Position of Boundary Cap Cells.
Radial glial cells are the neural progenitors of the developing CNS and have long radial processes that guide radially migrating neurons. The integrity of the radial glial scaffold, in particular proper adhesion between the endfeet of radial processes and the pial basement membrane (BM), is important for the cellular organization of the CNS, as indicated by evidence emerging from the developing cortex. However, the mechanisms underlying the maintenance of radial glial scaffold integrity during development, when the neuroepithelium rapidly expands, are still poorly understood. Here, we addressed this issue in the developing mouse spinal cord. We show that CXCR4, a receptor of chemokine CXCL12, is expressed in spinal cord radial glia. Conditional knock-out of Cxcr4 in radial glia caused disrupted radial glial scaffold with gaps at the pial endfeet layer and consequentially led to an invasion of boundary cap (BC) cells into the spinal cord. Because BC cells are PNS cells normally positioned at the incoming and outgoing axonal roots, their invasion into the spinal cord suggests a compromised CNS/PNS boundary in the absence of CXCL12/CXCR4 signaling. Both disrupted radial glial scaffold and invasion of BC cells into the CNS were also present in mice deficient in CXCR7, a second receptor of CXCL12. We further show that CXCL12 signaling promotes the radial glia adhesion to BM components and activates integrin β1 avidity. Our study unravels a novel molecular mechanism that deploys CXCL12/CXCR4/CXCR7 for the maintenance of radial glial scaffold integrity, which in turn safeguards the CNS/PNS boundary during spinal cord development.