Reduced retinal neovascularization, vascular permeability, and apoptosis in ischemic retinopathy in the absence of prolyl hydroxylase-1 due to the prevention of hyperoxia-induced vascular obliteration.
Prolyl hydroxylases (PHDs) are oxygen sensors that stabilize hypoxia-inducible factors (HIFs) to induce proinflammatory, vasopermeability, and proapoptotic factors. These may be potential targets to reduce the complications of ischemic retinopathies. Oxygen-induced ischemic retinopathy (OIR) was generated as a model for retinopathy of prematurity (ROP) by placing 7-day-old mice in 75% oxygen for 5 days and returning them to the relative hypoxia of room air for 5 days. Neovascularization (NV) and avascular areas were assessed on retinal flat-mounts by image analysis. Blood-retinal barrier breakdown was assessed using ³H-mannitol as a tracer. Apoptosis was detected with TUNEL staining. HIF-1α and VEGF were quantified using Western blot analysis and ELISA. PHD1-deficient mice demonstrated reduced hyperoxia-associated vascular obliteration during oxygen-induced ischemic retinopathy. This was associated with subsequent reduced avascularity, vascular leakage, and pathologic NV during the hypoxic phase, which could be accounted for by a reduced expression of HIF-1α and VEGF. Apoptosis in the retina was also reduced in PHD1-depleted mice after 2 days in hyperoxia. PHD1 deficiency is associated with a reduction of ischemia-induced retinal NV. The regulatory mechanism in this model appears to be: PHD1 depletion prevents HIF-1α degradation in hyperoxia, which induces VEGF, thus preventing hyperoxia-related vessel loss. Without a vessel deficiency, there would not be relative hypoxia when the mice are returned to room air and there would be no need to initiate angiogenesis signaling. Blocking PHD1 may be beneficial for ischemic retinopathies and inflammatory and neurodegenerative disorders.


Successful conservative management of hereditary hemorrhagic telangiectasia in pregnancy. A case report.
Hereditary hemorrhagic telangiectasia (HHT) is a condition characterized by multiple telangiectases and arteriovenous malformations. Women with HHT may develop life-threatening complications in pregnancy. In particular, death from pulmonary hemorrhage has been reported. Consequently, these women are often advised not to conceive or to terminate their pregnancies. We report a case of conservative management of HHT in pregnancy with a good outcome. This case demonstrates that in carefully selected cases, women with HHT who are managed conservatively may have good pregnancy outcomes. A diagnosis of HHT alone is not reason to advise women against pregnancy, nor should these women routinely be advised to undergo pregnancy termination.


NPY modulates neurotransmission of CGRP-containing vasodilator nerves in rat mesenteric arteries.
The effect of neuropeptide Y (NPY) in neurotransmission of calcitonin gene-related peptide (CGRP)-containing vasodilator nerves was investigated in rats. In perfused mesenteric vascular beds with active tone, perivascular nerve stimulation (PNS; 1-8 Hz) caused a frequency-dependent vasodilator response, which was abolished by 300 nM tetrodotoxin (TTX), 500 nM capsaicin, 1 microM human CGRP-(8-37), or cold storage denervation (4 degrees C for 72 h). NPY (5, 10, and 50 nM) concentration dependently inhibited the vasodilator response to PNS, whereas NPY had little effect on vasodilation induced by exogenous CGRP (10 and 100 pmol) or 1 nmol acetylcholine (ACh). NPY (10 nM) inhibited the neurogenic release of CGRP-like immunoreactivity induced by PNS (4 and 8 Hz), which was abolished by 300 nM TTX and the removal of Ca2+ from the medium. Combined perfusion with 5 nM NPY and 10 nM norepinephrine additively inhibited the vasodilator response to PNS but not to exogenous CGRP and ACh. Immunohistochemistry showed the distinct distribution of CGRP- and NPY-like immunoreactivity-containing fibers in rat mesenteric arteries. These results suggest that NPY modulates presynaptically the release of CGRP from CGRP-containing vasodilator nerves in rat mesenteric arteries.


Changes in Albuminuria But Not GFR are Associated with Early Changes in Kidney Structure in Type 2 Diabetes.
In type 1 diabetes, changes in the GFR and urine albumin-to-creatinine ratio (ACR) are related to changes in kidney structure that reflect disease progression. However, such changes have not been studied in type 2 diabetes. Participants were American Indians with type 2 diabetes enrolled in a clinical trial of losartan versus placebo. We followed a subset who underwent kidney biopsy at the end of the 6-year trial, with annual measurements of GFR (by urinary clearance of iothalamate) and ACR. Participants had a second kidney biopsy after a mean follow-up of 9.3 years. We used quantitative morphometric analyses to evaluate both biopsy specimens. Baseline measures for 48 participants (12 men and 36 women, mean age 45.6 years) who completed the study included diabetes duration (14.6 years), GFR (156 ml/min), and ACR (15 mg/g). During follow-up, glomerular basement membrane (GBM) width, mesangial fractional volume, and ACR increased, and surface density of peripheral GBM and GFR decreased. After adjustment for sex, age, ACR, and each morphometric variable at baseline, an increase in ACR during follow-up was significantly associated with increases in GBM width, mesangial fractional volume, and mean glomerular volume, and a decrease in surface density of peripheral GBM. Decline in GFR was not associated with changes in these morphometric variables after additionally adjusting for baseline GFR. In American Indians with type 2 diabetes and preserved GFR at baseline, increasing ACR reflects the progression of earlier structural glomerular lesions, whereas early GFR decline may not accurately reflect such lesions.


Engineering the Mycomembrane of Live Mycobacteria with an Expanded Set of Trehalose Monomycolate Analogues.
Mycobacteria and related organisms in the Corynebacterineae suborder are characterized by a distinctive outer membrane referred to as the mycomembrane. Biosynthesis of the mycomembrane occurs through an essential process called mycoloylation, which involves antigen 85 (Ag85)-catalyzed transfer of mycolic acids from the mycoloyl donor trehalose monomycolate (TMM) to acceptor carbohydrates and, in some organisms, proteins. We recently described an alkyne-modified TMM analogue (O-AlkTMM-C7) which, in conjunction with click chemistry, acted as a chemical reporter for mycoloylation in intact cells and allowed metabolic labeling of mycoloylated components of the mycomembrane. Here, we describe the synthesis and evaluation of a toolbox of TMM-based reporters bearing alkyne, azide, trans-cyclooctene, and fluorescent tags. These compounds gave further insight into the substrate tolerance of mycoloyltransferases (e.g., Ag85s) in a cellular context and they provide significantly expanded experimental versatility by allowing one- or two-step cell labeling, live cell labeling, and rapid cell labeling via tetrazine ligation. Such capabilities will facilitate research on mycomembrane composition, biosynthesis, and dynamics. Moreover, because TMM is exclusively metabolized by Corynebacterineae, the described probes may be valuable for the specific detection and cell-surface engineering of Mycobacterium tuberculosis and related pathogens. We also performed experiments to establish the dependence of probe incorporation on mycoloyltransferase activity, results from which suggested that cellular labeling is a function not only of metabolic incorporation (and likely removal) pathway(s), but also accessibility across the envelope. Thus, whole-cell labeling experiments with TMM reporters should be carefully designed and interpreted when envelope permeability may be compromised. On the other hand, this property of TMM reporters can potentially be exploited as a convenient way to probe changes in envelope integrity and permeability, facilitating drug development studies.


Enhanced in-cell folding of reversibly cationized transcription factor using amphipathic peptide.
The intracellular delivery of functionally active transcription factor proteins is emerging as a promising technique for artificial regulation of cellular functions. However, in addition to the cell membrane, which acts as a barrier to macromolecules, the aggregation-favored properties of structurally flexible transcription factor proteins limit the application of this method. In-cell folding technique can be used to overcome these issues. This technique solubilizes denatured protein by reversible alkyl-disulfide cationization (S-cationization), and simultaneously endows efficient intracellular delivery and folding to the biologically active conformation in the reducing environment of the cytosol. Because cationized protein is internalized into cells by adsorption-mediated endocytosis, endosomal escape is crucial for this technique. In this study, we utilized a sensitive luciferase reporter gene assay to quantitatively evaluate in-cell folding of the artificial transcription factor GAL4-VP16. Although the cationic moiety of S-cationized protein was slightly affected, co-transduction of amphipathic peptide Endo-PORTER dramatically improved in-cell folding efficiency. Live cell imaging of fluorescent-labeled GAL4-VP16 revealed that some of the proteins diffused into the cytosol and nucleus through co-transduction with Endo-PORTER. Real-time monitoring of light output of luciferase revealed the kinetics of in-cell folding, supporting that endosomal-release assisted by Endo-PORTER was stimulated by endosome acidification. Because this method can transduce proteins uniformly and repeatedly into living cells, S-cationized transcription factor proteins are widely applicable for the artificial regulation of cellular functions.


Lasers--panacea or paradox?.
The first lasers were quite unsuitable for dental use, but the last 30 years have seen the development of a variety of lasers which allow dentists to cut soft tissue without bleeding, remove caries, cut cavities in teeth (usually without pain), cure composite resins rapidly, weld metals with precision, desensitize teeth, sterilize exposed pulps and tissue surfaces and treat skin lesions. Future research may explain why 'soft' lasers appear to promote healing and alleviate joint and muscle pain. There are now many published reports in refereed dental journals covering the various aspects of laser dentistry. Dentists interested in lasers, or who are considering purchasing one should read the relevant literature and consult their Association and colleagues before making a purchase.


The effect of anoxia on cerebral acid hydrolases in the five-day-old rat.
1. Five-day-old anaesthetized rats subjected to slow, prolonged asphyxia (50-55 min) were either allowed to die or resuscitated when at the point of death. Activities of various cerebral acid hydrolases known to be associated with lysosomes were determined in these animals and in littermate controls. 2. Asphyxia to death resulted in a significant increase in the activities of acid phosphatase, cathepsin (pH5.0) and beta-glucuronidase in whole-brain homogenates. 3. The effect of asphyxia on beta-glucuronidase activity was not apparent when the assay was performed in the presence of Triton X-100 (0.1%, v/v). 4. In resuscitated animals whole-brain-homogenate beta-glucuronidase activity showed the greatest increase (31%) 15 min after recovery. After a 60 min recovery period differences between control and asphyxiated animals were no longer apparent. 5. In animals anoxiated to death activities of acid phosphatase and beta-N-acetylglucosaminidase in brain high-speed supernatants were significantly higher than in controls. Acid phosphatase activity was similarly increased in asphyxiated animals resuscitated for 5 or 60 min. 6. It is suggested that the response of the immature rat brain to asphyxia involves a disruption or increased fragility of lysosomal particles.


Agent replication dynamics in a long incubation period model of mouse scrapie.
The dynamics of scrapie agent replication in brain and spleen following intracerebral or intraperitoneal infection were investigated in a model with particularly long incubation periods [IM mice (Sincp7) infected with 87V scrapie]. In contrast to other mouse scrapie models previously investigated, there was no delay in onset of replication ('zero phase') in spleen using either route of infection, For both routes infectivity levels reached a plateau in spleen, which was maintained for at least 250 days in intraperitoneally infected mice. An infectivity plateau was also apparent in brain following intracerebral infection, suggesting that there may be constraints on agent replication in brain during the later part of the incubation period. In addition, the efficiency of the intraperitoneal route to produce disease was found to be particularly low for this model.


Financial Analysis of Closed Femur Fractures in 3- to 6-Year-Olds Treated With Immediate Spica Casting Versus Intramedullary Fixation.
Health care in America continues to place more importance on providing value-based medicine. Medicare reimbursements are increasingly being tied to this and future policy changes are expected to reinforce these trends. Recent literature has shown pediatric femur fractures in preschool-age children have equivalent clinical and radiographic outcomes when treated with spica casting or flexible intramedullary nails (IMN). We compared hospital care statistics including charges for nonoperative versus operative treatment for closed femur fractures in 3- to 6-year-olds. An IRB-approved retrospective chart review was performed of 73 consecutive 3- to 6-year-olds treated at a regional level 1 pediatric hospital from January 1, 2009 to December 31, 2013 with an isolated, closed femoral shaft fracture. Exclusion criteria included open fractures, bilateral injury, and polytrauma. Immediate spica casting was performed in the Emergency Department or Anesthesia Procedure Unit versus IMN in the operating room. A total of 41 patients were treated with spica casting and 32 patients were treated operatively with flexible IMNs; 3 patients failed nonoperative care. After analysis of final treatment groups, significant differences included age at injury: 3.7 years for cast versus 5.3 years for IMN (P<0.001), time to discharge 21 versus 41 hours (P<0.001), 3.2 versus 4.4 clinic visits (P<0.001), follow-up 3.5 versus 9.4 months (P<0.001). Orthopedic surgeon charges were $1500 for casted patients versus $5500 for IMN (P<0.001). Total hospital charges were $19,200 for cast versus $59,700 for IMN (P<0.001). No difference was found between clinic charges or number of radiographs between groups. In total, 76% of cast group were discharged <24 hours from admission versus 8.6% in operative group. In the operative group, 83% had implant removal with no statistically significant charge difference between those who had implant removal versus retention. Treatment of pediatric femur fractures in 3- to 6-year-olds with IMN is associated with longer hospital stays, significantly greater hospital charges, longer follow-up and more clinic visits compared with spica casting. These findings are at odds with previous literature showing shorter hospital stays and decreased cost with nailing compared to traction and casting. This shows a clear difference between 2 treatments that yield similar clinical and radiographic outcomes. Level III-retrospective comparative study.


Reclassification of Subtercola pratensis Behrendt et al. 2002 as Agreia pratensis comb. nov.
Comparative analysis of 16S rDNA sequences revealed a close phylogenetic relationship (99.6 % similarity) between Subtercola pratensis Behrendt et al. 2002 and Agreia bicolorata Evtushenko et al. 2001. The two species were found to share genus-specific chemotaxonomic characteristics such as the occurrence of D-ornithine and L-2,4-diaminobutyric acid in the peptidoglycan and the profile of cellular fatty acids and 1,1-dimethoxy-alkanes. DNA-DNA relatedness of only 47.8 % and differences in phenotypic features such as the menaquinone profile and oxidase and Voges-Proskauer reactions confirmed the distinct species status of S. pratensis and A. bicolorata. On the basis of the data from phylogenetic and phenotypic analyses, the reclassification of S. pratensis as Agreia pratensis comb. nov. is proposed. As a result of this reclassification, the two genera are coherent, in that the cell wall composition and 1,1-dimethoxy-alkane spectrum are significant genus-specific characteristics.


Cloning, expression, and type II collagenolytic activity of matrix metalloproteinase-13 from human osteoarthritic cartilage.
Proteolysis of triple-helical collagen is an important step in the progression toward irreversible tissue damage in osteoarthritis. Earlier work on the expression of enzymes in cartilage suggested that collagenase-1 (MMP-1) contributes to the process. Degenerate reverse transcription polymerase chain reaction experiments, Northern blot analysis, and direct immunodetection have now provided evidence that collagenase-3 (MMP-13), an enzyme recently cloned from human breast carcinoma, is expressed by chondrocytes in human osteoarthritic cartilage. Variable levels of MMP-13 and MMP-1 in cartilage was significantly induced at both the message and protein levels by interleukin-1 alpha. Recombinant MMP-13 cleaved type II collagen to give characteristic 3/4 and 1/4 fragments; however, MMP-13 turned over type II collagen at least 10 times faster than MMP-1. Experiments with intact type II collagen as well as a synthetic peptide suggested that MMP-13 cleaved type II collagen at the same bond as MMP-1, but this was then followed by a secondary cleavage that removed three amino acids from the 1/4 fragment amino terminus. The expression of MMP-13 in osteoarthritic cartilage and its activity against type II collagen suggest that the enzyme plays a significant role in cartilage collagen degradation, and must consequently form part of a complex target for proposed therapeutic interventions based on collagenase inhibition.


Cellulose nanofibrils and silver nanowires active coatings for the development of antibacterial packaging surfaces.
An active ink composed of cellulose nanofibrils and silver nanowires was deposited on flexible and transparent polymer films using the bar coating process, achieving controlled thicknesses ranging from 200 nm up to 2 μm. For 350 nm thick coating on polyethylene terephthalate films, high transparency (75.6% transmittance) and strong reduction of bacterial growth equal to 89.3% and 100% was noted respectively against Gram-negative Escherichia Coli and Gram-positive Staphylococcus Aureus bacteria using AATCC contact active standard test. Retained antibacterial activity was found with films produced by reverse gravure roll-to-roll process, showing the promising capability of this antibacterial solution to be deployed industrially. Finally, the same ink was also deposited on polylactic acid substrate to investigate barrier properties: for 350 nm thick coating, a reduction of 49% of oxygen transmission rate (dry conditions) and 47% reduction of water vapor transmission rate was noted, proving the enhanced barrier properties of the coatings.


Translocation, accumulation and bioindication of trace elements in wetland plants.
This study aimed to shed further light on the capacity of macrophytes to translocate, accumulate and bioindicate the levels of trace elements present in contaminated water and sediments. Specifically, this study aimed to find evidence whether translocation, accumulation and bioindication are dependent on the kind of trace element and plant species. To investigate the correlation between trace elements in plants and in the environment, the concentrations of As, Cd, Cr, Cu, Hg, Mn, Ni, Pb, and Zn were analyzed in twenty different wetland plants, and in water and sediments from a wetland area affected by urban and industrial pollutants. Results showed that wetland plants share some common characteristics such as high tolerance to toxic element levels, capacity of phytostabilization and different element concentrations in the various organs. Moreover, element translocation from sediments to roots seems more influenced by the kind of plant species and trace element, whereas translocation across the various organs seems mainly species-specific. No clear patterns of trace element translocation were identified according to plant life forms.


Phenylalanine-544 plays a key role in substrate and inhibitor binding by providing a hydrophobic packing point at the active site of insulin-regulated aminopeptidase.
Inhibitors of insulin-regulated aminopeptidase (IRAP) improve memory and are being developed as a novel treatment for memory loss. In this study, the binding of a class of these inhibitors to human IRAP was investigated using molecular docking and site-directed mutagenesis. Four benzopyran-based IRAP inhibitors with different affinities were docked into a homology model of the catalytic site of IRAP. Two 4-pyridinyl derivatives orient with the benzopyran oxygen interacting with the Zn(2+) ion and a direct parallel ring-stack interaction between the benzopyran rings and Phe544. In contrast, the two 4-quinolinyl derivatives orient in a different manner, interacting with the Zn(2+) ion via the quinoline nitrogen, and Phe544 contributes an edge-face hydrophobic stacking point with the benzopyran moiety. Mutagenic replacement of Phe544 with alanine, isoleucine, or valine resulted in either complete loss of catalytic activity or altered hydrolysis velocity that was substrate-dependent. Phe544 is also important for inhibitor binding, because these mutations altered the K(i) in some cases, and docking of the inhibitors into the corresponding Phe544 mutant models revealed how the interaction might be disturbed. These findings demonstrate a key role of Phe544 in the binding of the benzopyran IRAP inhibitors and for optimal positioning of enzyme substrates during catalysis.


Human intestinal lamina propria CD1c+ dendritic cells display an activated phenotype at steady state and produce IL-23 in response to TLR7/8 stimulation.
Intestinal dendritic cells (DCs) play key roles in mediating tolerance to commensal flora and inflammatory responses against mucosal pathogens. The mechanisms by which intestinal "conditioning" influences human DC responses to microbial stimuli remain poorly understood. Infections with viruses, such as HIV-1, that target mucosal tissue result in intestinal epithelial barrier breakdown and increased translocation of commensal bacteria into the lamina propria (LP). It is unclear whether innate LP DC responses to concurrent viral and bacterial stimuli influence mucosal HIV-1 pathogenesis. In this study, direct ex vivo phenotype and in vitro constitutive cytokine production of CD1c+ DCs in human intestinal LP were compared with those in peripheral blood (PB). To evaluate innate responses to viral and bacterial stimuli, intracellular cytokine production by LP and PB DCs following stimulation with ligands for TLRs 2, 4, 5, and 7/8 was evaluated. At steady state, LP CD1c+ DCs expressed higher levels of activation markers (CD40, CD83, CD86, HLA-DR, and CCR7) than did PB CD1c+ DCs, and higher frequencies of LP CD1c+ DCs constitutively produced IL-6 and -10 and TNF-alpha. LP DCs had blunted cytokine responses to TLR4 ligand and TLR5 ligand stimulation relative to PB DCs, yet similarly produced IL-10 in response to TLR2 ligand. Only synthetic TLR7/8 ligand, a mimic of viral ssRNA, induced IL-23 production by LP CD1c+ DCs, and this proinflammatory cytokine response was synergistically enhanced following combined TLR7/8 and TLR4 stimulation. These findings highlight a potential mechanism by which viruses like HIV-1 may subvert homeostatic mechanisms and induce inflammation in the intestinal mucosa.


Structural and ultrastructural study of rat liver influenced by electromagnetic radiation.
Mobile communication systems are undoubtedly an environmental source of electromagnetic radiation (EMR). There is an increasing concern regarding the interactions of EMR with the humans. The aim of this study was to examine the effects of EMR on Wistar rat liver. Mature rats were exposed to electromagnetic field of frequency 2.45 GHz and mean power density of 2.8 mW/cm2 for 3 h/d for 3 wk. Samples of the liver were obtained 3 h after the last irradiation and processed histologically for light and transmission electron microscopy. Data demonstrated the presence of moderate hyperemia, dilatation of liver sinusoids, and small inflammatory foci in the center of liver lobules. Structure of hepatocytes was not altered and all described changes were classified as moderate. Electron microscopy of hepatocytes revealed vesicles of different sizes and shapes, lipid droplets, and proliferation of smooth endoplasmic reticulum. Occasionally necrotizing hepatocytes were observed. Our observations demonstrate that EMR exposure produced adverse effects on rat liver.


Strain differences in rat brain epinephrine synthesis: regulation of alpha-adrenergic receptor number by epinephrine.
Inbred tht strains Fischer 344 (F344) and Buffalo (BUF) differ in serveral physiological and behavioral measures. It was found that the activity of adrenomedullary and regional brain phenylethanolamine N-methyltransferase is at least four times higher in F344 rats than in BUF rats; these strain-dependent differences corresponded directly with the epinephrine content of the medulla-pons and hypothalamus. Conversely, alpha-adrenergic receptor density in brain regions containing phenylethanolamine N-methyltransferase is two to three times lower in F344 rats than in BUF rats; alpha-receptors in frontal cortex (a brain region lacking phenylethanolamine N-methyltransferase activity and epinephrine) are similar in both strains. These findings suggest that strain-dependent differences in alpha-receptors are regulated by inherited differences in presynaptic adrenergic neuronal function in different brain regions.


[Idiopathic recurrent ventricular tachycardia in children: characteristics and long-term prognosis].
The aim of this study was to assess the prognosis and the clinical course of incessant idiopathic ventricular tachycardia (VT) with left bundle branch morphology, in a group of 14 children consecutively evaluated in our institutions between 1983 and 1994. Mean age of these patients was 8 +/- 2 years, and they were followed for 56 +/- 32 months. In all the children, VT was present for more than 80% of the day, in non sustained runs. The mean rate of VT was of 142 +/- 30 beats/min. Four children were symptomatic. The evaluation of these children was obtained by treadmill stress test (12/14), cardiac catheterization (13/14), electrophysiologic study (10/14), nuclear magnetic resonance (7/14), and serial echocardiographic and Holter examinations. Right ventricular dysplasia was suspected in 2 cases on the basis of nuclear magnetic resonance and angiographic data. In both these cases VT was not suppressed by exercise. Medical treatment was started in 9/14 patients (69%), it was completely effective in 4 (44%) and partially effective in 2 further cases (22%). Thirteen out of fourteen patients had a regular follow-up. No patient died. All patients remained asymptomatic and in 6 (46%) VT disappeared. In 4 of these last 6 patients therapy has been previously effective. Two children, one with right ventricle dysplasia, continue antiarrhythmic treatment. Incessant VT is well tolerated in pediatric age. The frequency of spontaneous remissions is relatively high (46% of cases). In few patients this arrhythmia is associated with evident cardiac abnormalities. In our opinion, medical treatment is necessary in symptomatic patients and in those with heart abnormalities.


Single-cell RNA-seq reveals ectopic and aberrant lung-resident cell populations in idiopathic pulmonary fibrosis.
We provide a single-cell atlas of idiopathic pulmonary fibrosis (IPF), a fatal interstitial lung disease, by profiling 312,928 cells from 32 IPF, 28 smoker and nonsmoker controls, and 18 chronic obstructive pulmonary disease (COPD) lungs. Among epithelial cells enriched in IPF, we identify a previously unidentified population of aberrant basaloid cells that coexpress basal epithelial, mesenchymal, senescence, and developmental markers and are located at the edge of myofibroblast foci in the IPF lung. Among vascular endothelial cells, we identify an ectopically expanded cell population transcriptomically identical to bronchial restricted vascular endothelial cells in IPF. We confirm the presence of both populations by immunohistochemistry and independent datasets. Among stromal cells, we identify IPF myofibroblasts and invasive fibroblasts with partially overlapping cells in control and COPD lungs. Last, we confirm previous findings of profibrotic macrophage populations in the IPF lung. Our comprehensive catalog reveals the complexity and diversity of aberrant cellular populations in IPF.


Multiple tissue-specific promoters control expression of the murine tartrate-resistant acid phosphatase gene.
Tartrate-resistant acid phosphatase (TRAP) is highly expressed in osteoclasts and in a subset of tissue macrophages and dendritic cells. It is expressed at lower levels in the parenchymal cells of the liver, glomerular mesangial cells of the kidney and pancreatic acinar cells. We have identified novel TRAP mRNAs that differ in their 5'-untranslated region (5'-UTR) sequence, but align with the known murine TRAP mRNA from the first base of Exon 2. The novel 5'-UTRs represent alternative first exons located upstream of the known 5'-UTR. A similar genomic structure exists for the human TRAP gene with partial conservation of the exon and promoter sequences. Expression of the most distal 5'-UTR (Exon 1A) is restricted to adult bone and spleen tissue. Exon 1B is expressed primarily in tissues containing TRAP-positive non-haematopoietic cells. The known TRAP 5'-UTR (Exon 1C) is expressed in tissues characteristic of myeloid cell expression. In addition the Exon 1C promoter sequence is shown to comprise distinct transcription start regions, with an osteoclast-specific transcription initiation site identified downstream of a TATA-like element. Macrophages are shown to initiate transcription of the Exon 1C transcript from a purine-rich region located upstream of the osteoclast-specific transcription start point. The distinct expression patterns for each of the TRAP 5'-UTRs suggest that TRAP mRNA expression is regulated by the use of four alternative tissue- and cell-restricted promoters.


Involvement of a central alpha-adrenoceptor system in antidepressant potentiation of hyperthermia induced by thyrotropin releasing hormone.
Thyrotropin releasing hormone (TRH) causes hyperthermia in mice which is potentiated by tricyclic antidepressants (nortriptyline, imipramine, clomipramine, amitriptyline), the monoamine oxidase inhibitor, tranylcypromine, and various other antidepressants (maprotiline, nomifensin, viloxazine). Only iprindole is ineffective. The effect of mianserin, itself hypothermic, could not be interpreted. A property shared by the potentiating substances seems to be activation of a central adrenoceptor system. The potentiation of TRH-induced hyperthermia which seems to be specific to antidepressants might be used in the selection of antidepressants.


Advances in Cardiovascular Health in Women over the Past Decade: Guideline Recommendations for Practice.
Cardiovascular disease (CVD) remains the number one cause of death in women. It is estimated that 44 million women in the United States are either living with or at risk for heart disease. This article highlights the recent significant progress made in improving care, clinical decision-making, and policy implications for women with CVD. We provide our perspective supported by evidence-based advances in cardiovascular research and clinical care guidelines in seven areas: (1) primary CVD prevention and community heart care, (2) secondary prevention of CVD, (3) stroke, (4) heart failure and cardiomyopathies, (5) ischemia with nonobstructive coronary artery disease, (6) spontaneous coronary artery dissection, and (7) arrhythmias and device therapies. Advances in these fields have improved the lives of women living with and at risk for heart disease. With increase awareness, partnership with national organizations, sex-specific research, and changes in policy, the morbidity and mortality of CVD in women can be further reduced.


Modern diagnostics and treatment of Merkel cell carcinoma.
Merkel cell carcinoma (MCC) is a rare, aggressive skin cancer with epithelial and neuroendocrine differentiation, the incidence of which has increased substantially during the last decades. Risk factors include advanced age, fair skin type, UV exposure, and immunosuppression. Pathogenetically, a type caused by the Merkel cell polyomavirus is distinguished from a UV-induced type with a high tumor mutational burden. Clinically, MCC presents as a mostly painless, rapidly growing, reddish-violet tumor with a shiny surface, which is preferentially localized in the head-neck region and at the distal extremities. A reliable diagnosis can only be made based on histological and immunohistochemical features. At initial diagnosis, 20-26% of patients show locoregional metastases and 8-14% distant metastases, making staging examinations indispensable. If there is no clinical evidence of metastases, a sentinel lymph node biopsy is recommended. Essential columns of therapy are surgery, adjuvant or palliative radiotherapy and, in advanced inoperable stages, medicamentous tumor therapy. The introduction of immune checkpoint inhibitors has led to a paradigm shift, as they provide a considerably longer duration of response and better survival rates than chemotherapy. The PD-L1 inhibitor avelumab is approved for treatment of metastatic MCC in Germany, but the PD-1 antibodies pembrolizumab and nivolumab are also used with success. Adjuvant and neoadjuvant treatment concepts, immune combination therapies and targeted therapies as monotherapy or in combination with immune checkpoint inhibitors are in the clinical trial phase.


Health Behaviors Before and After the Implementation of a Health Community Organization: Gangwon's Health-Plus Community Program.
Community organization is a resident-led movement aimed at creating fundamental social changes in the community by resolving its problems through the organized power of its residents. This study evaluated the effectiveness of health community organization (HCO), Gangwon's Health-Plus community program, implemented from 2013 to 2019 on residents' health behaviors. This study had a before-and-after design using 2011-2019 Korea Community Health Survey data. To compare the 3-year periods before and after HCO implementation, the study targeted areas where the HCO had been implemented for 4 years or longer. Therefore, a total of 4512 individuals from 11 areas with HCO start years from 2013 to 2016 were included. Complex sample multi-logistic regression analysis adjusting for demographic characteristics (sex, age, residential area, income level, education level, and HCO start year) was conducted. HCO implementation was associated with decreased current smoking (adjusted odds ratio [aOR], 0.73; 95% confidence interval [CI], 0.57 to 0.95) and subjective stress recognition (aOR, 0.79; 95% CI, 0.64 to 0.97). Additionally, the HCO was associated with increased walking exercise practice (aOR, 1.39; 95% CI, 1.13 to 1.71), and attempts to control weight (aOR, 1.36; 95% CI, 1.12 to 1.64). No significant negative changes were observed in other health behavior variables. The HCO seems to have contributed to improving community health indicators. In the future, a follow-up study that analyzes only the effectiveness of the HCO through structured quasi-experimental studies will be needed.


Raising the Bar in Aromatic Donor-Acceptor Interactions with Cyclic Trinuclear Gold(I) Complexes as Strong π-Donors.
Aromatic donor-acceptor interactions are of high importance in supramolecular chemistry, materials science and biology. Compared to other noncovalent interactions, such as hydrogen bonding, the binding is often weak. Here we show that strong donor-acceptor interactions between planar aromatics with binding free energies down to -10.1 kcal mol-1 and association constants of up to 2.34 × 107 L mol-1 for 1:1 complexes can be realized using cyclic trinuclear complexes of gold(I) with pyridinate, imidazolate, or carbeniate ligands. Data were obtained through NMR and UV/vis absorption spectroscopic studies and supported by quantum chemical calculations for a variety of acceptors. By using a specifically designed bridged naphthalene diimide-based acceptor with only one binding site, we furthermore show that a 1:2 (donor:acceptor) binding model is best suited to quantify the donor and acceptor/complex equilibrium. Scanning electron microscopy on selected donor-acceptor pairs shows crystalline supramolecular assemblies. We anticipate this study to be relevant for the future design of supramolecular systems and chemical sensors and the determination of binding energies between planar donors and acceptors.


The fate of Baker's cyst after total knee arthroplasty.
It is widely held that most Baker's cysts resolve after treatment of the intra-articular knee pathology. The present study aimed to evaluate the fate of Baker's cysts and their associated symptoms after total knee arthroplasty (TKA). In this prospective cohort study, 102 patients with (105 were included, however three were lost to follow-up) an MRI-verified Baker's cyst, primary osteoarthritis and scheduled for TKA were included. Ultrasound was performed to evaluate the existence and the gross size of the cyst before and at one year after TKA. Additionally, associated symptoms of Baker's cyst were recorded pre- and post-operatively. After one year, a Baker's cyst was still present in 87 patients (85%). There was a significant reduction in associated symptoms arising from the Baker's cyst before (71%) to after surgery (31%). No patients developed a new Baker's cyst associated symptoms. However, of the 72 patients who had reported Baker's cyst associated symptoms pre-operatively, 32 patients (44%) still complained of such symptoms one year after surgery. Baker's cysts had resolved in only a small number of patients (15%) one year after TKA and symptoms from the cysts persisted in 31%. Cite this article: Bone Joint J 2016;98-B:1185-8.


Threshold calcium levels for lamellar body exocytosis in type II pneumocytes.
Pulmonary surfactant is secreted via exocytosis of lamellar bodies (LBs) by alveolar type II cells. Here we analyzed the dependence of LB exocytosis on intracellular Ca(2+) concentration ([Ca(2+)](i)). In fura 2-loaded cells, [Ca(2+)](i) was selectively elevated by flash photolysis of a cell-permeant caged Ca(2+) compound (o-nitrophenyl EGTA-AM) or by gradually enhancing cellular Ca(2+) influx. Simultaneously, surfactant secretion by single cells was analyzed with the fluorescent dye FM 1-43, enabling detection of exocytotic events with a high temporal resolution (T. Haller, J. Ortmayr, F. Friedrich, H. Volkl, and P. Dietl. Proc. Natl. Acad. Sci. USA 95: 1579-1584, 1998). Exocytosis was initiated at a threshold concentration near 320 nmol/l with both instantaneous or gradual [Ca(2+)](i) elevations. The exocytotic response to flash photolysis was highest during the first minute after the rise in [Ca(2+)](i) and thus almost identical to purinoceptor stimulation by ATP. Correspondingly, the effects of ATP on initial secretion could be sufficiently explained by its ability to mobilize Ca(2+). This was further demonstrated by the fact that exocytosis is significantly blocked by suppression of the ATP-induced Ca(2+) signal below approximately 300 nmol/l. Our results suggest a highly Ca(2+)-sensitive step in LB exocytosis.


[Nursing Education Strategies During the COVID-19 Epidemic].
The ongoing new coronavirus disease (COVID-19) pandemic, which arose at the end of 2019, poses a severe challenge to world public health systems. Frontline medical staffs bear a great burden to provide health care services. The Taiwan government has taken rapid and decisive actions to reduce the risk of community transmission and campus cluster infection. Nursing education includes both classroom teaching and clinical practicum components. In preparing for their practicum, students must learn not only fundamental nursing care knowledge but also basic knowledge on emerging infectious diseases. All schools nationwide have complied with the Ministry of Education order to postpone the opening of the fall semester in response to the rapid evolution of the COVID-19 pandemic. Campus epidemic prevention measures were implemented through student counseling networks, and flexible teaching strategies, including online teaching and distance teaching, were implemented to protect the learning rights of students. This paper explores the strategies implemented in response to emerging infectious diseases in nursing education based on the core values of professional nursing. Examining the precautions taken at campuses and teaching strategies adopted in response to the COVID 19 pandemic may provide valuable insights that may be applied to the future development of nursing education.


Equivalent physical models and formulation of equivalent source layer in high-resolution EEG imaging.
In high-resolution EEG imaging, both equivalent dipole layer (EDL) and equivalent charge layer (ECL) assumed to be located just above the cortical surface have been proposed as high-resolution imaging modalities or as intermediate steps to estimate the epicortical potential. Presented here are the equivalent physical models of these two equivalent source layers (ESL) which show that the strength of EDL is proportional to the surface potential of the layer when the outside of the layer is filled with an insulator, and that the strength of ECL is the normal current of the layer when the outside is filled with a perfect conductor. Based on these equivalent physical models, closed solutions of ECL and EDL corresponding to a dipole enclosed by a spherical layer are given. These results provide the theoretical basis of ESL applications in high-resolution EEG mapping.


Bracon brevicornis Genome Showcases the Potential of Linked-Read Sequencing in Identifying a Putative Complementary Sex Determiner Gene.
Bracon brevicornis is an ectoparasitoid of a wide range of larval-stage Lepidopterans, including several pests of important crops, such as the corn borer, Ostrinia nubilalis. It is also one of the earliest documented cases of complementary sex determination in Hymenoptera. Here, we present the linked-read-based genome of B. brevicornis, complete with an ab initio-derived annotation and protein comparisons with fellow braconids, Fopius arisanus and Diachasma alloeum. We demonstrate the potential of linked-read assemblies in exploring regions of heterozygosity and search for structural and homology-derived evidence of the complementary sex determiner gene (csd).


The somatosensory thalamus of the raccoon: properties of single neurons responsive to light mechanical stimulation of the forepaw.
These studies were undertaken to characterize the discharge properties of single neurons of the raccoon thalamic ventrobasal complex (VB) that respond to light mechanical stimulation of the glabrous surfaces of the forepaw. Microelectrodes were used to record the extracellular activity of 146 cells in anesthetized raccoons, and all neurons were histologically verified as falling within or along the boundaries of VB. Sixty-one neurons were tested for activation by electrical stimulaton of primarily somatosensory cortex. Of these, 88% were antidromically activated, 5% were synaptically activated, and the remaining 7% were unresponsive. Out of the total sample of 146 neurons, 136 had peripheral receptive fields (RFs) that were restricted to glabrous skin and revealed properties of modality and place-specificity predictable through knowledge of properties of primary mechanoreceptive afferents. Rapidly adapting (RA) neurons accounted for 77% of this modality-place-specific sample, while 19% were slowly adapting (SA), and 4% revealed properties indicative of input from Pacinian afferents (Pc). Absolute displacement thresholds were comparable for RA and SA neurons (range = 6-415 micron). Palmar RF areas (range = 3.3-328 mm2) were significantly larger than digital RF areas (range = 0.5-98.2 mm2). As defined by exponents (b) of power functions relating instantaneous discharge frequency to displacement ramp velocity, SA neurons formed a single, homogeneous group (range of values of b = 0.633-0.720). However, RA neurons fell into three distinct groups: those showing relatively steep functions (b = 0.559-0.938), those showing relatively flat functions (b = 0.146-0.334), and those showing discontinuous, or step, functions. A small number of neurons (7% of total sample) revealed "complex" properties, not predictable from knowledge of properties of primary afferents. These included five neurons whose RFs encompassed both glabrous and hairy skin, and several linear orientation, or "tactile edge," detectors. The present results, in conjunction with those of earlier studies of the raccoon dorsal column-medial lemniscal system, lead to the conclusion that different types of information transformation are emphasized at different levels of the system. Intramodality convergences (increases in RF area) occur primarily within the cuneothalamic relay, while changes in the coding of quantitative information are primarily a function of VB neurons. The appearance of linear orientation detectors--a type of tactile "feature detector"--indicates that the synthesis of information regarding complex spatial properties of stimuli has its beginnings within the somatosensory thalamus.


Slip-shod or safely shod: the bighorn sheep as a natural model for research.
Over a million injuries caused by slipping of footwear are believed to require treatment by doctors every year in the United Kingdom and many domestic animals are injured by slipping. Recent research has revealed that surface roughness of solings and floors is an important determinant of grip on lubricated surfaces and it is also known that soling friction is affected by hardness. The bighorn sheep (Ovis canadensis) an animal species which has adapted to a slippery environment, was studied to elucidate optimum roughness and hardness and other features which influence grip. Four adult ewes were examined in the London Zoo. The cloven hooves of this species are very mobile and the cranial tips of the hooves are the first parts to make contact with the ground. A very small contact area ensures penetration of a film of water. Mean roughness of the contact area was found to be 53 microns Rtm and the mean hardness 63 Shore A. These characteristics appear to facilitate an excellent grip on wet slippery rock but not on smooth ice. Further studies of the feet of wild species could contribute to an understanding of the factors which determine the safety of solings and floors.


Dynamical properties of an harmonic oscillator impacting a vibrating wall.
The dynamics of a spring-mass system under repeated impact with a vibrating wall is investigated using the static wall approximation. The evolution of the harmonic oscillator is described by two coupled difference equations. These equations are solved numerically, and in some cases exact analytical expressions have also been found. For a periodically vibrating wall, Fermi acceleration is only found at resonance. There, the average rebounding velocity increases linearly with the number of collisions. Near resonance, the average rebounding velocity grows initially with the number of collisions and eventually reaches a plateau. In the vicinity of resonance, the motion of the oscillator exhibits scaling properties over a range of frequency ratios. The presence of dissipation at resonance destroys the Fermi-acceleration process and induces scaling behavior similar to that at near resonance. For a moving wall with a random amplitude at collisions, Fermi acceleration is observed independently of the ratio between the wall and oscillator frequencies. In this case the average rebounding velocity grows with the square root of the number of collisions with the wall. Also, in this latter case, dissipation suppresses the Fermi-acceleration mechanism and induces a scaling behavior with the same universality class as that of the dissipative bouncing ball model with random external perturbations.


Outcomes After Rotator Cuff Repair With Transverse Scapular Ligament Release in Patients With Severe Fatty Degeneration of the Infraspinatus.
In some large to massive rotator cuff tears (RCTs), fatty degeneration (FD) is more severe in the infraspinatus than the supraspinatus muscle, and in such cases, suprascapular neuropathy is highly suspected. Nerve release at the suprascapular notch might alleviate this problem. To evaluate the effects of the transverse scapular ligament (TSL) release in patients with large to massive RCTs with more severe FD of the infraspinatus than the supraspinatus. Cohort study; Level of evidence, 3. Between September 2017 and January 2022, arthroscopic TSL release with rotator cuff repair was performed in patients with large to massive RCTs and more severe FD of the infraspinatus muscle than the supraspinatus muscle (TSL group). Cuff integrity, FD, and atrophy of cuff muscles were evaluated using preoperative and 1-year postoperative magnetic resonance imaging. In addition, results were compared with those of patients who did not undergo TSL release during arthroscopic large to massive rotator cuff repair (NTSL group). A total of 103 patients-20 in the TSL group and 83 in the NTSL group-were included. Group preoperative characteristics, including tear size and supraspinatus FD, were not significantly different, but infraspinatus FD (TSL vs NTSL; grade, 0-4, 0/0/5/10/5 vs 1/33/42/4/3) and atrophy (grade, 1-3. 3/9/8 vs 56/20/7) differed significantly (P < .001). Healing failure occurred in 13 of 20 (65%) patients in the TSL group and 30 of 83 (36%) patients in the NTSL group, which was a statistically significant difference (P = .019). Postoperatively, infraspinatus FD and atrophy were more severe in the TSL group than in the NTSL group (P < .001), and supraspinatus FD was more severe in the TSL group (P = .029). Seven patients in the TSL group achieved healing, but FD and atrophy of the supraspinatus and the infraspinatus showed no improvement in this group (all, P > .05). In patients with more FD in the infraspinatus than the supraspinatus muscle, TSL release appeared to have no benefit for cuff healing or FD reversal in cuff muscles. The possibility of suprascapular nerve entrapment remains in patients with more FD in the infraspinatus than the supraspinatus, and this potential nerve problem is not properly addressed by TSL release alone.


Analysis of carbon emission performance and regional differences in China's eight economic regions: Based on the super-efficiency SBM model and the Theil index.
China's carbon emission performance has significant regional heterogeneity. Identified the sources of carbon emission performance differences and the influence of various driving factors in China's eight economic regions accurately is the premise for realizing China's carbon emission reduction goals. Based on the provincial panel data from 2005 to 2017, the super-efficiency SBM model and Malmquist model are constructed in this paper to measure regional carbon emission performance's static and dynamic changes. After that, the Theil index is used to distinguish the impact of inter-regional and intra-regional differences on different regions' carbon emissions performance. Finally, by introducing the Tobit model, the effect of various driving factors on carbon emission performance differences is analyzed quantitatively. The results show that: (1) There are significant differences in different regions' carbon emission performance, but the overall carbon emission performance presents an upward fluctuation trend. Malmquist index decomposition results show substantial differences in technology progress index and technology efficiency index in different regions, leading to significant carbon emission performance differences. (2) Overall, inter-regional differences contribute the most to the overall carbon emission performance, up to more than 80%. Among them, the inter-regional and intra-regional differences in ERMRYR contributed significantly. (3) Through Tobit regression analysis, it is found that residents' living standards, urbanization level, ecological development degree, and industrial structure positively affect carbon emission performance. On the contrary, energy intensity presents an apparent negative correlation on carbon emission performance. Therefore, to improve the carbon emission performance, we should put forward targeted suggestions according to the characteristics of different regional development stages, regional carbon emission differences, and influencing driving factors.


[Research Progress of Post-Transplant Cyclophosphamide Combined with Antithymocyte Globulin in Preventing GVHD after Hematopoietic Stem Cell Transplantation --Review].
Graft-versus-host disease (GVHD) is one of the major complications after allogeneic hematopoietic stem cell transplantation (allo-HSCT), which seriously affects the prognosis of patients. At present, a new regimen of post-transplantation cyclophosphamide (PTCy) combined with antithymocyte globulin (ATG) has been used to prevent GVHD, indicating that PTCy combined with ATG may have a good effect on the prevention of GVHD in different types of transplantation. However, the mechanism of this regimen, its effect on immune reconstitution and viral reactivation still needs to be further studied. Therefore, this article briefly reviews the research progress of PTCy combined with ATG in preventing GVHD after HSCT.


Association Between Community Social Capital and Hospital Readmission Rates.
Hospital readmissions remain frequent, and are partly attributable to patients' social needs. The authors sought to examine whether local community levels of social capital are associated with hospital readmission rates. Social capital refers to the connections among members of a society that foster norms of reciprocity and trust, which may influence the availability of support for postdischarge recovery after hospitalization. Associations between hospital-wide, risk-stratified readmission rates for hospitals in the United States (n = 4298) and levels of social capital in the hospitals' service areas were examined. Social capital was measured by an index of participation in associational activities and civic affairs. A multivariate linear regression model was used to adjust for hospital and community factors such as hospital financial performance, race, income, and availability of heath care services. Results showed that higher social capital was significantly associated with lower readmission rates (P < .01), a finding that held across income-stratified analyses as well as sensitivity analyses that included hospital performance on process quality measures and hospital community engagement activities. A hospital is unlikely to be able to influence prevailing levels of social capital in its region, but in areas of low social capital, it may be possible for public or philanthropic sectors to buttress the types of institutions that address nonmedical causes of readmission.


Risk Factors of Extensively Drug Resistant Typhoid Fever Among Children in Karachi: Case-Control Study.
Extensively drug resistant typhoid fever (XDR-TF) has been responsible for an ongoing outbreak in Pakistan, which began in November 2016. This study aimed to determine the risk factors associated with XDR-TF. This age- and sex-matched case-control study was conducted during May-October 2018 in Karachi. All patients with XDR-TF were identified from the laboratory-based surveillance system data. Cases included patients aged <15 years living in Karachi with culture-positive Salmonella enterica serovar Typhi with resistance to chloramphenicol, ampicillin, trimethoprim/sulfamethoxazole, fluoroquinolones, and third-generation cephalosporins. Age- and sex-matched controls included children free from the symptoms of TF, aged under 15 years, and residing in Karachi. All controls were recruited from among those who attended outpatient clinics. A total of 75 cases and 75 controls were included in this study. On univariate analysis, the odds of having XDR-TF were 13-fold higher among participants who used piped municipal water than among those who did not (odds ratio [OR] 12.6, 95% CI 4.1-38.6). The use of bore water was significantly associated with XDR-TF (OR 5.1, 95% CI 1.4-19.0). Cases were more likely to report eating French fries with sauce (OR 13.5, 95% CI 3.9-47.0) and poppadum (OR 3.4, 95% CI 1.7-6.7) from street vendors than controls. Boiling water at home was negatively associated with XDR-TF (OR 0.3, 95% CI 0.2-0.7). On multivariate analysis, 2 factors were independently associated with XDR-TF. Using piped municipal water (OR 10.3, 95% CI 3.4-30.4) and eating French fries with sauce from street vendors (OR 8.8, 95% CI 2.1-36.2) were significantly associated with an increased odds of XDR-TF. Community water supply and street food eating habits were implicated in the spread of the superbug S typhi outbreak, which continues to grow in Karachi. Therefore, it is recommended to improve the community water supply to meet recommended standards and to develop a policy to improve the safety of street food. In addition, health authorities are required to conduct mass vaccination for TF among high-risk groups.


The heterogeneity of the cigarette price effect on body mass index.
Previous studies estimate the average effect of cigarette price on body mass index (BMI), with recent research showing that their different methodologies all point to a negative effect after several years. This literature, however, ignores the possibility that the effect could vary throughout the BMI distribution or across socioeconomic and demographic groups due to differences in underlying obesity risks or preferences for health. We evaluate heterogeneity in the long-run impact of cigarette price on BMI by performing quantile regressions and stratifying the sample by race, education, age, and sex. Cigarette price has a highly heterogeneous negative effect that is more than three times as strong at high BMI levels--where weight loss is most beneficial for health--than at low levels. The effects are also strongest for blacks, college graduates, middle-aged adults, and women. We also assess the implications for disparities, conduct robustness checks, and evaluate potential mechanisms.


Branded prescription drug spending: a framework to evaluate policy options.
High drug spending is a concern for policy makers due to limits on access for patients. Numerous policies have been proposed to address high drug spending. The existence of multifarious proposals makes it difficult for policy makers to consider all the alternatives. We developed an approach to select the most viable options to present to policy makers. We identified 41 different proposals in the peer-reviewed literature to reduce the level of spending or change the incentives for branded prescription drugs; ten of which we identified as promising proposals. Based on criterion used to assess various legislative proposals regarding branded pharmaceuticals we developed a framework to evaluate the ten promising proposals. We then used a modified Delphi technique to iteratively evaluate these ten proposals starting with the initial criterion. During each iteration, five researchers independently evaluated the ten policies based on available criterion and assessed how to modify the criterion to achieve consensus on what attributes the criterion were intended to measure. We highlight areas of disagreement to show where modifications to existing criterion are needed. We found general agreement for most policy-criterion combinations after three iterations. Areas with the greatest remaining disagreement include possible unintended consequences, the concept of value implied by many of the policies, and secondary effects by the pharmaceutical industry, insurers, and the FDA. Our analysis provides an approach that can be applied to evaluate policy proposals. It also suggests factors that policy analysts and researchers should consider when they propose policy options and where additional research is needed to assess policy impacts. Developing an objective approach to compare alternatives may facilitate the adoption of policies for branded prescription drugs in the U.S. by allowing policy makers to focus on the most viable options.


Vaccinia virus-mediated cancer immunotherapy: cancer vaccines and oncolytics.
Cancer vaccines and oncolytic immunotherapy are promising treatment strategies with potential to provide greater clinical benefit to patients with advanced-stage cancer. In particular, recombinant vaccinia viruses (VV) hold great promise as interventional agents. In this article, we first summarize the current understanding of virus biology and viral genes involved in host-virus interactions to further improve the utility of these agents in therapeutic applications. We then discuss recent findings from basic and clinical studies using VV as cancer vaccines and oncolytic immunotherapies. Despite encouraging results gleaned from translational studies in animal models, clinical trials implementing VV vectors alone as cancer vaccines have yielded largely disappointing results. However, the combination of VV vaccines with alternate forms of standard therapies has resulted in superior clinical efficacy. For instance, combination regimens using TG4010 (MVA-MUC1-IL2) with first-line chemotherapy in advanced-stage non-small cell lung cancer or combining PANVAC with docetaxel in the setting of metastatic breast cancer have clearly provided enhanced clinical benefits to patients. Another novel cancer vaccine approach is to stimulate anti-tumor immunity via STING activation in Batf3-dependent dendritic cells (DC) through the use of replication-attenuated VV vectors. Oncolytic VVs have now been engineered for improved safety and superior therapeutic efficacy by arming them with immune-stimulatory genes or pro-apoptotic molecules to facilitate tumor immunogenic cell death, leading to enhanced DC-mediated cross-priming of T cells recognizing tumor antigens, including neoantigens. Encouraging translational and early phase clinical results with Pexa-Vec have matured into an ongoing global phase III trial for patients with hepatocellular carcinoma. Combinatorial approaches, most notably those using immune checkpoint blockade, have produced exciting pre-clinical results and warrant the development of innovative clinical studies. Finally, we discuss major hurdles that remain in the field and offer some perspectives regarding the development of next generation VV vectors for use as cancer therapeutics.


Comparison between an African town and a neighbouring village shows delayed, but not decreased, sleep during the early stages of urbanisation.
The well-established negative health outcomes of sleep deprivation, and the suggestion that availability of electricity may enable later bed times without compensating sleep extension in the morning, have stimulated interest in studying communities whose sleep pattern may resemble a pre-industrial state. Here, we describe sleep and activity in two neighbouring communities, one urban (Milange) and one rural (Tengua), in a region of Mozambique where urbanisation is an ongoing process. The two communities differ in the amount and timing of daily activity and of light exposure, with later bedtimes (≈1 h) associated with more evening and less daytime light exposure seen in the town of Milange. In contrast to previous reports comparing communities with and without electricity, sleep duration did not differ between Milange (7.28 h) and Tengua (7.23 h). Notably, calculated sleep quality was significantly poorer in rural Tengua than in Milange, and poor sleep quality was associated with a number of attributes more characteristic of rural areas, including more intense physical labour and less comfortable sleeping arrangements. Thus, whilst our data support the hypothesis that access to electricity delays sleep timing, the higher sleep quality in the urban population also suggests that some aspects of industrialisation are beneficial to sleep.


The plasma free amino acid dose-response technique: A proposed methodology for determining lysine relative bioavailability of rumen-protected lysine supplements.
Estimates of Lys bioavailability of rumen-protected Lys (RP-Lys) supplements are often obtained using in vitro or 2-step in situ techniques, with little to no data determining efficacy and bioavailability in vivo. The objective of this study was to further evaluate and refine the use of the plasma free AA dose-response technique as a method for determining Lys relative bioavailability of RP-Lys supplements. Thirteen dose-response Latin square studies using 87 lactating, ruminally cannulated multiparous Holstein cows (days in milk from 55 to 315 and milk yield from 12 to 62 kg/d at the start of the studies) were conducted to measure the relative bioavailability of RP-Lys supplements. Intestinal (1 study) and abomasal (12 studies) infusions of Lys ranged from 0 to 84 g/d, and experimental periods ranged from 4 to 21 d. Basal diets were formulated to be adequate in metabolizable Met, but varied in predicted metabolizable Lys (5.04 to 6.81% of metabolizable protein). One to 4 daily blood samples were taken from the coccygeal vessels for 1 to 3 consecutive days in each period. Plasma Lys concentration in cows assigned to the control treatment (0 g/d Lys) ranged from 1.83 to 5.21% of total plasma AA, whereas that from cows duodenally or abomasally infused with Lys ranged from 2.53 to 7.51% of total plasma AA. Results from studies involving more than 2 amounts of infused Lys confirmed linearity of response. The following variables were regressed against the plasma Lys dose-response slopes generated from the Lys infusion treatments to examine their effects on the magnitude of the slopes: plasma Lys concentration of the control diet, plasma Lys concentration at the greatest amount of infused Lys, net energy of lactation and metabolizable protein balances, metabolizable protein supply, days in milk, milk yield, milk concentrations of fat, true protein, and lactose, milk true protein yield, and dry matter intake. The variable having the greatest effect on the magnitude of the dose-response slope was the plasma Lys concentration at the greatest amount infused. The relative bioavailability of evaluated RP-Lys supplements using the plasma free AA dose-response technique ranged from 5 to 87%. It was concluded that plasma free Lys increases in a linear fashion to increasing amounts of absorbed Lys and that the dose-response technique is an appropriate technique for evaluating RP-Lys supplements.


Prognostic value of cancer stem cell marker CD133 expression in pancreatic ductal adenocarcinoma (PDAC): a systematic review and meta-analysis.
CD133 is one of the most commonly used markers of pancreatic cancer stem cells (CSCs), which are characterized by their ability for self-renewal and tumorigenicity. Although the expression of CD133 has been reported to correlate with poor prognosis of PDAC in most literatures, some controversies still exist. In this study, we aimed to investigate the correlation between CD133 expression and prognosis and clinicopathological features in PDAC. A search in the Medline, EMBASE and Chinese CNKI (China National Knowledge Infrastructure) database (up to 1 March 2015) was performed using the following keywords pancreatic cancer, CD133, AC133, prominin-1 etc. Data from eligible studies were extracted and included into meta-analysis using a random effects model. Outcomes included overall survival and various clinicopathological features. We performed a final analysis of 723 patients from 11 evaluable studies for prognostic value and 687 patients from 12 evaluable studies for clinicopathological features. Our study shows that the pooled hazard ratio (HR) of overexpression CD133 for overall survival in PDAC was 0.58 (95% confidence interval (CI): 0.49-0.67) by univariate analysis and 0.73 (95% CI: 0.52-1.03) by multivariate analysis. With respect to clinicopathological features, CD133 overexpression by immunohistochemistry (IHC) method was closely correlated with clinical TNM stage (TNM stage III+IV, OR=0.32, 95% CI: 0.19-0.54), tumor differentiation (poor differentiation, OR=0.56, 95% CI: 0.37-0.83), and lymph node metastasis (N1, 3.15, 95% CI: 1.56-6.36) in patients with PDAC. Our meta-analysis results suggest that CD133 is an efficient prognostic factor in PDAC. Overexpression of CD133 was significantly associated with clinical TNM stage, tumor differentiation and lymph node metastasis.


Combined cognitive and vocational interventions after mild to moderate traumatic brain injury: study protocol for a randomized controlled trial.
A considerable proportion of patients with mild to moderate traumatic brain injury (TBI) experience long-lasting somatic, cognitive, and emotional symptoms that may hamper their capacity to return to work (RTW). Although several studies have described medical, psychological, and work-related factors that predict RTW after TBI, well-controlled intervention studies regarding RTW are scarce. Furthermore, there has traditionally been weak collaboration among health-related rehabilitation services, the labor and welfare sector, and workplaces. This study protocol describes an innovative randomized controlled trial in which we will explore the effect of combining manualized cognitive rehabilitation (Compensatory Cognitive Training [CCT]) and supported employment (SE) on RTW and related outcomes for patients with mild to moderate TBI in real-life competitive work settings. The study will be carried out in the southeastern region of Norway and thereby be performed within the Norwegian welfare system. Patients aged 18-60 years with mild to moderate TBI who are employed in a minimum 50% position at the time of injury and sick-listed 50% or more for postconcussive symptoms 2 months postinjury will be included in the study. A comprehensive assessment of neurocognitive function, self-reported symptoms, emotional distress, coping style, and quality of life will be performed at baseline, immediately after CCT (3 months after inclusion), following the end of SE (6 months after inclusion), and 12 months following study inclusion. The primary outcome measures are the proportion of participants who have returned to work at 12-month follow-up and length of time until RTW, in addition to work stability as well as work productivity over the first year following the intervention. Secondary outcomes include changes in self-reported symptoms, emotional and cognitive function, and quality of life. Additionally, a qualitative RTW process evaluation focused on organizational challenges at the workplace will be performed. The proposed study will combine cognitive and vocational rehabilitation and explore the efficacy of increased cross-sectoral collaboration between specialized health care services and the labor and welfare system. If the intervention proves effective, the project will describe the cost-effectiveness and utility of the program and thereby provide important information for policy makers. In addition, knowledge about the RTW process for persons with TBI and their workplaces will be provided. ClinicalTrials.gov, NCT03092713 . Registered on 10 March 2017.


Clear rules for clear fluids fasting in children.
Preoperative fasting guidelines published in 2022 by the European Society of Anaesthesiology and Intensive Care represent a paradigm shift in the preoperative preparation of children undergoing general anaesthesia. Schmitz and colleagues report the results from a multi-institutional prospective cohort study to determine if application of the recent guidelines increased the risk of regurgitation and pulmonary aspiration. This study provides support for the concept of reducing real fasting times by allowing clear fluids until 1 h before induction of anaesthesia. Although the study cohort was large, further prospective multicentre studies with even greater sample sizes are warranted to provide definitive evidence for the safety of the new fasting rules.


[Molecular mechanisms in liver ischemic-reperfusion injury and ischemic preconditioning].
Ischemia-reperfusion (IR) liver injury is associated with temporary clamping of hepatoduodenal ligament during liver surgery, hypoperfusion shock and graft failure after liver transplantation. Mechanisms of IR liver injury include: i) loss of calcium homeostasis, ii) reactive oxygen and nitrogen species generation, iii) changes in microcirculation, iv) Kupffer cell activation, and (v) complement activation. Pre-exposure of the liver to transient ischemia increases the tolerance to IR injury, a phenomenon known as hepatic ischemic preconditioning (IP). IP involves: i) recovery of the energy supply and calcium, sodium and pH homeostasis, ii) enhancement in the antioxidant potential, and iii) expression of multiple stress-response proteins, including acute phase proteins, heat shock proteins, and heme oxygenase. These observations and preliminary studies in humans give a rationale for the assessment of IP in minimizing or preventing IR injury during surgery and non surgical conditions of tissue hypoperfusion.


The role of hydrolases in insecticide metabolism and the toxicological significance of the metabolites.
The hydrolytic enzymes involved in insecticide metabolism are the phosphorotriester hydrolases, carboxylesterases, carboxylamidases and epoxide hydrolases. The phosphorotriester hydrolases (arylesterases and DFP-ases) catalyze the P-anhydride bond cleavage of the "leaving group", a major route of detoxication of organophosphates. Carboxylesterases hydrolyze the carboethoxy group of malathion and also have hydrolytic activity toward certain synthetic pyrethroids. Carboxylamidases are involved in the hydrolysis of amide-containing phosphorothionates, N-formyl metabolites and substituted fluoroacetamides. Epoxide hydrolases hydrate certain cyclodiene insecticides and are probably involved in the metabolism of some other insecticides. Overall, the hydrolysis of insecticides increases the polarity of the metabolites and decreases their biological activity.


Switching mechanism of sensor-motor coordination through an oscillator network model.
Insects have small brains, but their behavior is highly adaptive; this leads us to conclude that their brains possess a simple adaptation mechanism. This paper focuses on the pheromone processing of crickets, varying their aggression depending on their global neural connection, and proposes a behavior selection mechanism that can be controlled by network transformation. The controller is composed of an oscillator network, and its behavior is decided by the synchrony of organic oscillations. Furthermore, every network component corresponds to a certain brain module. A model is realized by using an analog circuit, and it is applied to a simple robot that displays the behavior of a real insect.


[Selection in "hybrid" cattle populations].
Natural selection in populations of cattle was studied. Heterozygosity level, pure-blooded proportion, stabilizing selection, and directional selection II acted as quantitative traits. Changes in the proportion of pure bloods with time were found, something not taken into account in breeding practice.


Biopolyesters from microorganisms: Biochemical basis of microbial synthesis, properties and applications.
Biopolyester produced by microorganisms has many useful properties for commercial and environmental applications. Poly(3-hydroxyalkonates) [poly(HA)s] and poly (â -malate) are among the best known biopolyesters produced by microor-ganisms. Poly(HA)s having a wide variety of repeating units can be produced by bacteria and they all posses very high biodegradability and biocompatibility. Poly (â -malate) is especially of interest for biomedical applications including drug delivery systems. During the last 20 yrs a large volume of research work has focused on the physiology, biochemistry and applications of poly(HA)s. In this review, the biochemical basis of bacterial production, the biodegradation properties and applications of poly(HA)s are considered.


Design of an amphiphilic hyperbranched core/shell-type polymeric nanocarrier platform for drug delivery.
An amphiphilic core/shell-type polymer-based drug carrier system (HPAE- PCL-b -MPEG), composed of hyperbranched poly(aminoester)-based polymer (HPAE) as the core building block and poly(ethylene glycol)-b - poly(ε-caprolactone) diblock polymers (MPEG-b -PCL) as the shell building block, was designed. The synthesized polymers were characterized with FTIR, 1 H NMR, 13 C NMR, and GPC analysis. Monodisperse HPAE-PCL-b - MPEG nanoparticles with dimensions of < 200 nm and polydispersity index of < 0.5 were prepared by nanoprecipitation method and characterized with SEM, particle size, and zeta potential analysis. 5-Fluorouracil was encapsulated within HPAE-PCL-b -MPEG nanoparticles. In vitro drug release profiles and cytotoxicity of blank and 5-fluorouracil-loaded nanoparticles were examined against the human colon cancer HCT116 cell line. All results suggest that HPAE-PCL-b - MPEG nanoparticles offer an alternative and effective drug nanocarrier system for drug delivery applications.


Thought disorder in children at risk for psychosis.
The Thought Disorder Index, which has been shown to be a reliable and valid measure of the degree of thought disorder in adult psychotic patients, was used to assess thought disorder in children. Normal children aged 5 to 16 years, hospitalized psychotic children, children hospitalized for nonpsychotic behavioral problems, and children born to a psychotic mother (high-risk) were tested. In the normal sample, thought disorder decreases with age. The level of thought disorder in psychotic children and high-risk children was about three times higher than that of the normal children, whereas the level of thought disorder of the nonpsychotic hospitalized children was no different from that of the normal children. The study also detected differences in the kinds of thought disorder that characterize the psychotic and high-risk groups vs the other children. The presence of thought disorder may be thought of as a useful indicator of the diathesis for functional psychosis. The Thought Disorder Index merits selection as a tool for investigating thought disorder as a potential precursor variable in studies of children at risk for schizophrenia and manic-depressive illness.


Investigating the association of bed bugs with infectious diseases: A retrospective case-control study.
Bed bugs are common urban pests. Unlike many other blood-feeding human ectoparasites, bed bugs are not known to be vectors of human infectious diseases, but clinical and epidemiological studies to directly interrogate this link have been limited. Here, we aimed to determine whether bed bugs were associated with infectious diseases in a set of infested patients presenting to emergency departments (ED) in the greater Cleveland, OH area. We performed a retrospective case-control study involving 332 ED patients with bed bugs and 4,952 control patients, seen from February 1, 2011, through February 1, 2017. Cases and controls were matched by age, sex, and the presenting ED. Additionally, data were adjusted for ≥20 sociodemographic variables, triage data, and comorbidities in multivariable regression analyses. Seventeen laboratory values, ten different ED and inpatient diagnoses, chest radiographs, infectious disease consults, and blood cultures were examined. The odds of bed bug infestation were significantly higher for patients that had positive blood cultures, had blood cultures growing coagulase-negative Staphylococcus, were diagnosed with pneumonia, were diagnosed with cellulitis, received an infectious disease consult, received a chest radiograph, and had higher percentages of eosinophils in the blood (P < .05 for all). Additional investigations are needed to determine whether bed bugs directly contribute to disease by transmitting causative agents, whether bed bug exposure contributes secondarily contributes to infections, or whether these associations are better explained by other environmental and social determinants of health.


Overall survival in Japanese patients with ER+/HER2- advanced breast cancer treated with first-line palbociclib plus letrozole.
An open-label, single-arm, Japanese phase 2 study (J-Ph2) investigated the efficacy and safety of first-line (1L) palbociclib (PAL) + letrozole (LET) in postmenopausal Japanese women with ER+/HER2- advanced breast cancer (ABC). In the final analysis, median progression-free survival was 35.7 months (95% CI 21.7-46.7); but overall survival (OS) data were immature. Here, we report the findings from a follow-up study of J-Ph2 (NCT04735367) evaluating OS and subsequent therapy in these Japanese women. Patients (N = 42) who participated in J-Ph2 were enrolled in the OS follow-up study. The primary endpoint was OS and secondary endpoints included type and duration of subsequent therapy. Patients were a median age of 62.5 years; 48% had visceral metastases. At a median follow-up of 89.7 months, the median OS was 85.4 months (95% CI 64.3-not estimable). Median OS was longer in patients with nonvisceral versus visceral metastases (not reached vs 67.3 months), or with treatment-free interval > 12 months versus ≤ 12 months (85.4 vs 45.4 months), or with treatment duration ≥ 24 months versus < 24 months (not reached vs 47.5 months). Of patients who received a first subsequent therapy (81%), most (67%) continued endocrine-based therapy, while 7% received chemotherapy. The median duration of the first subsequent therapy was 8.3 months (95% CI 3.9-12.2), and the median chemotherapy-free survival was 69.1 months (95% CI 24.2-85.4). In this population of Japanese women with ER+/HER2- ABC, median OS was over 7 years with 1L PAL + LET, supporting the use of 1L PAL + endocrine therapy. NCT04735367.


Comparison of photocatalytic and transport properties of TiO2 and ZnO nanostructures for solar-driven water splitting.
Titanium dioxide (TiO2) and zinc oxide (ZnO) nanostructures have been widely used as photo-catalysts due to their low-cost, high surface area, robustness, abundance and non-toxicity. In this work, four TiO2 and ZnO-based nanostructures, i.e. TiO2 nanoparticles (TiO2 NPs), TiO2 nanotubes (TiO2 NTs), ZnO nanowires (ZnO NWs) and ZnO@TiO2 core-shell structures, specifically prepared with a fixed thickness of about 1.5 μm, are compared for the solar-driven water splitting reaction, under AM1.5G simulated sunlight. Complete characterization of these photo-electrodes in their structural and photo-electrochemical properties was carried out. Both TiO2 NPs and NTs showed photo-current saturation reaching 0.02 and 0.12 mA cm(-2), respectively, for potential values of about 0.3 and 0.6 V vs. RHE. In contrast, the ZnO NWs and the ZnO@TiO2 core-shell samples evidence a linear increase of the photocurrent with the applied potential, reaching 0.45 and 0.63 mA cm(-2) at 1.7 V vs. RHE, respectively. However, under concentrated light conditions, the TiO2 NTs demonstrate a higher increase of the performance with respect to the ZnO@TiO2 core-shells. Such material-dependent behaviours are discussed in relation with the different charge transport mechanisms and interfacial reaction kinetics, which were investigated through electrochemical impedance spectroscopy. The role of key parameters such as electronic properties, specific surface area and photo-catalytic activity in the performance of these materials is discussed. Moreover, proper optimization strategies are analysed in view of increasing the efficiency of the best performing TiO2 and ZnO-based nanostructures, toward their practical application in a solar water splitting device.


Effects of stroke severity and training duration on locomotor recovery after stroke: a pilot study.
Locomotor training using partial body weight-supported treadmill (BWST) walking has been widely investigated for people after stroke, yet there remains a lack of evidence concerning the optimal training duration and the effect of locomotor impairment severity. Previous protocols have not emphasized the transfer of locomotor skills from the BWST environment to overground. To assess the feasibility of a program combining locomotor training using BWST with task-specific overground training and to obtain pilot data on the effects of severity and training duration on recovery of locomotion. Seven adults with chronic poststroke hemiparesis and gait speed less than 0.8 m/s were recruited to participate in a 12-week (36 session) locomotor training program. Each session comprised 20 to 30 minutes of training using BWST with manual assistance, followed by 10 to 15 minutes of overground training to transfer the skills trained in the BWST environment. Gait speed was the primary outcome measure. Six out of the 7 enrolled individuals completed the intervention program; 1 was withdrawn due to transportation difficulties affecting compliance with the training schedule. Four of the 6 participants had a functionally significant improvement in walking speed after 36 sessions, defined as having achieved a 0.4 m/s gait speed or greater for those with initial severe gait speed impairment (<0.4 m/s) or as having achieved a 0.8 m/s gait speed or greater for persons with initial moderate gait speed impairment (>or=0.4 m/s and <0.8 m/s). All participants improved in balance and distance walked over 6 minutes, and 5 of the 6 participants showed increases in their daily home and community step activity. A locomotor training program combining walking using BWST and manual assistance with overground practice is feasible for people with chronic poststroke hemiparesis and moderate or severe gait speed impairment. This intervention shows promise for achieving functionally significant improvements in walking speed.


Hepatoprotective activity of cinnamon ethanolic extract against CCI4-induced liver injury in rats.
The inner bark of cinnamon (Cinnamomum zeylanicum L.) is commonly used as a spice and has also been widely employed in the treatment and prevention of disease. The aim of the present study is to evaluate the protective effect of cinnamon bark extract against carbon tetrachloride (CCl4)-induced liver damage in male Wistar rats. Administration with cinnamon extracts (0.01, 0.05 and 0.1 g/kg) for 28 days significantly reduced the impact of CCl4 toxicity on the serum markers of liver damage, aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase. In addition, treatment of cinnamon extract resulted in markedly increased the levels of superoxide dismutase and catalase enzymes in rats. The histopathological studies in the liver of rats also supported that cinnamon extract markedly reduced the toxicity of CCl4 and preserved the histoarchitecture of the liver tissue to near normal. Thus, the results suggest that cinnamon extract acts as a potent hepatoprotective agent against CCl4 induced hepatotoxicity in rats.


Screening and social prescribing in healthcare and social services to address housing issues among children and families: a systematic review.
Housing is a social determinant of health that impacts the health and well-being of children and families. Screening and referral to address social determinants of health in clinical and social service settings has been proposed to support families with housing problems. This study aims to identify housing screening questions asked of families in healthcare and social services, determine validated screening tools and extract information about recommendations for action after screening for housing issues. The electronic databases MEDLINE, PsycINFO, EMBASE, Ovid Emcare, Scopus and CINAHL were searched from 2009 to 2021. Inclusion criteria were peer-reviewed literature that included questions about housing being asked of children or young people aged 0-18 years and their families accessing any healthcare or social service. We extracted data on the housing questions asked, source of housing questions, validity and descriptions of actions to address housing issues. Forty-nine peer-reviewed papers met the inclusion criteria. The housing questions in social screening tools vary widely. There are no standard housing-related questions that clinical and social service providers ask families. Fourteen screening tools were validated. An action was embedded as part of social screening activities in 27 of 42 studies. Actions for identified housing problems included provision of a community-based or clinic-based resource guide, and social prescribing included referral to a social worker, care coordinator or care navigation service, community health worker, social service agency, referral to a housing and child welfare demonstration project or provided intensive case management and wraparound services. This review provides a catalogue of housing questions that can be asked of families in the clinical and/or social service setting, and potential subsequent actions.


Effective treatment of advanced estrogen-independent MXT mouse mammary cancers with targeted cytotoxic LH-RH analogs.
Cytotoxic agents linked to hormonal carriers provide new approaches to tumor therapy, and LH-RH receptors expressed by breast cancers can be used for targeting chemotherapeutic compounds. In the present study, large, advanced estrogen-independent MXT mouse mammary cancers were treated with cytotoxic LH-RH analog AN-152 containing doxorubicin (DOX) or AN-207 incorporating superactive derivative 2-pyrrolino-DOX (AN-201). These cytotoxic hybrid molecules were administered once i.v., close to their maximum tolerated doses, at various time intervals after transplantation of tumors. The cytotoxic LH-RH analogs and the radicals alone, given at earlier stages of tumor development, inhibited growth of MXT cancers. Cytotoxic LH-RH conjugate AN-207 had significantly stronger effect than its respective cytotoxic radical, particularly when larger tumors were treated, causing 95%, 89%, 100% and 96% tumor growth reduction when administered on days 1, 7, 10 or 14, respectively. AN-152, AN-201, and DOX, given on day 14, were virtually ineffective. Histological characteristics of tumor cell proliferation and cell death were analyzed in large MXT cancers 1-4 days after treatment with AN-207 and AN-201. AgNOR scores were decreased and apoptotic indices increased after treatment of tumors with AN-207 or AN-201, but enhanced apoptosis and decreased AgNOR numbers persisted longer in the case of AN-207. In contrast to AN-201, AN-207 also increased the extent of necrosis in tumors. In conclusion, on the basis of its powerful inhibitory effect on the aggressive MXT mouse mammary tumor, the cytotoxic LH-RH analog AN-207 could be considered for treatment of advanced human mammary carcinomas that express LH-RH receptors.


Transcriptional regulation of CYP19 gene (aromatase) expression in adipose stromal cells in primary culture.
Estrogen biosynthesis in adipose tissue increases with age and obesity, and has been implicated in the development of endometrial cancer and breast cancer. In normal human adipose tissue, expression of the CYP19 gene which encodes aromatase P450, the enzyme responsible for estrogen biosynthesis, is regulated by a distal promoter, namely promoter I.4. Stimulation of expression in adipose stromal cells by members of the type 1 cytokine family, i.e. interleukin (IL)-6, IL-11, leukemia inhibitory factor (LIF) and oncostatin M (OSM), is mediated via a Jak-STAT3 signaling pathway and a GAS element upstream of promoter I.4. In contrast, aromatase expression in breast adipose tissue proximal to tumor is increased three- to four-fold to the utilization of another promoter, namely promoter II, proximal to the translation initiation site. In the present report, we show that prostaglandin (PG) E2 is the most potent factor which stimulates aromatase expression via cyclic AMP and promoter II. PGE2 acts via EP1 and EP2 receptor subtypes to stimulate both the PKC and PKA pathways. The combined stimulation of both of these pathways results in the maximal expression of promoter II-specific CYP19 transcripts. Because PGE2 is a major secretory product both of breast tumor epithelial cells and fibroblasts, as well as of macrophages infiltrating the tumor site, then this could be the mechanism whereby estrogen biosynthesis is stimulated in breast sites adjacent to a tumor, leading in turn to increased growth and development of the tumor itself.


Protonated 2'-aminoguanosine as a probe of the electrostatic environment of the active site of the Tetrahymena group I ribozyme.
We have probed the electrostatic environment of the active site of the Tetrahymena group I ribozyme (E) using protonated 2'-aminoguanosine (), in which the 2'-OH of the guanosine nucleophile (G) is replaced by an group. At low concentrations of divalent metal ion (2 mM Mg(2+)), binds at least 200-fold stronger than G or G(NH)()2, with a dissociation constant of </=1 microM from the ribozyme. oligonucleotide substrate. complex (). This strong binding suggests that the group interacts with negatively charged phosphoryl groups within the active site. Increasing the concentration of divalent metal ion weakens the binding of to E. S more than 10(2)-fold. The Mn(2+) concentration dependence suggests that M(C), the metal ion that interacts with the 2'-moiety of G in the normal reaction, is responsible for this effect. M(C) and compete for binding to the active site; this competition could arise from electrostatic repulsion between the positively charged and M(C) and, possibly, from their competition for interaction with active site phosphoryl groups. The reactive phosphoryl group of S increases the competition between M(C) and, consistent with a network of interactions involving M(C) that help position the reactive phosphoryl group and the guanosine nucleophile with respect to one another. The chemical step with bound is at least 10(4)-fold slower than with G or G(NH)()2. These results provide additional support for an integral role of M(C) in catalysis by the Tetrahymena ribozyme, and demonstrate the utility of the moiety as an electrostatic probe within a structured RNA.


Structure and dynamics of water and lipid molecules in charged anionic DMPG lipid bilayer membranes.
Molecular dynamics simulations have been used to investigate the influence of the valency of counter-ions on the structure of freestanding bilayer membranes of the anionic 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) lipid at 310 K and 1 atm. At this temperature, the membrane is in the fluid phase with a monovalent counter-ion and in the gel phase with a divalent counter-ion. The diffusion constant of water as a function of its depth in the membrane has been determined from mean-square-displacement calculations. Also, calculated incoherent quasielastic neutron scattering functions have been compared to experimental results and used to determine an average diffusion constant for all water molecules in the system. On extrapolating the diffusion constants inferred experimentally to a temperature of 310 K, reasonable agreement with the simulations is obtained. However, the experiments do not have the sensitivity to confirm the diffusion of a small component of water bound to the lipids as found in the simulations. In addition, the orientation of the dipole moment of the water molecules has been determined as a function of their depth in the membrane. Previous indirect estimates of the electrostatic potential within phospholipid membranes imply an enormous electric field of 10(8)-10(9) V m(-1), which is likely to have great significance in controlling the conformation of translocating membrane proteins and in the transfer of ions and molecules across the membrane. We have calculated the membrane potential for DMPG bilayers and found ∼1 V (∼2 ⋅ 10(8) V m(-1)) when in the fluid phase with a monovalent counter-ion and ∼1.4 V (∼2.8 ⋅ 10(8) V m(-1)) when in the gel phase with a divalent counter-ion. The number of water molecules for a fully hydrated DMPG membrane has been estimated to be 9.7 molecules per lipid in the gel phase and 17.5 molecules in the fluid phase, considerably smaller than inferred experimentally for 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC) membranes but comparable to the number inferred for 1,2-dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE) membranes. Some of the properties of the DMPG membrane are compared with those of the neutral zwitterionic DMPC bilayer membrane at 303 K and 1 atm, which is the same reduced temperature with respect to the gel-to-fluid transition temperature as 310 K is for the DMPG bilayer membrane.


DNA sequence symmetries from randomness: the origin of the Chargaff's second parity rule.
Most living organisms rely on double-stranded DNA (dsDNA) to store their genetic information and perpetuate themselves. This biological information has been considered as the main target of evolution. However, here we show that symmetries and patterns in the dsDNA sequence can emerge from the physical peculiarities of the dsDNA molecule itself and the maximum entropy principle alone, rather than from biological or environmental evolutionary pressure. The randomness justifies the human codon biases and context-dependent mutation patterns in human populations. Thus, the DNA 'exceptional symmetries,' emerged from the randomness, have to be taken into account when looking for the DNA encoded information. Our results suggest that the double helix energy constraints and, more generally, the physical properties of the dsDNA are the hard drivers of the overall DNA sequence architecture, whereas the selective biological processes act as soft drivers, which only under extraordinary circumstances overtake the overall entropy content of the genome.