Miracle-dz/UniFAR-FTUs huggingface dataset:
Repression of the NF1 gene by Tax may expain the development of neurofibromas in human T-lymphotropic virus type 1 transgenic mice

In familial neurofibromatosis type 1 (NF1), individuals with a germ line-transmitted NF1 mutation develop multiple neurofibromas. To explain the observation that transgenic mice expressing the human T-lymphotropic virus type 1 (HTLV-1) tax gene under the control of the viral regulatory element also develop multiple neurofibromas, we demonstrate that the Tax trans-regulator can functionally repress NF1 gene expression through a cis-acting element located immediately upstream of its transcriptional start site, thereby allowing the development of benign neurofibromas without the need for direct mutations in NF1. We propose that such a mechanism would suffice to epigenetically alter NF1 gene expression. The fact that transgenic animals have localized rather than diffuse neurofibroma formation, however, suggests that additional genetic or epigenetic events may be required for neurofibroma formation.
Nuclear Localization of Type I Parathyroid Hormone/Parathyroid Hormone‐Related Protein Receptors in Deer Antler Osteoclasts: Evidence for Parathyroid Hormone‐Related Protein and Receptor Activator of NF‐κB‐Dependent Effects on Osteoclast Formation in Regenerating Mammalian Bone

Parathyroid hormone‐related protein (PTHrP) is not required for osteoclastogenesis during embryonic development; however, after birth it has been shown to regulate osteoclast formation during tooth eruption. Our study explores the hypothesis that PTHrP also may regulate osteoclast differentiation in the regenerating skeletal tissues of deer antlers, bones capable of complete regeneration. Osteoclast‐like multinucleated cells (MNCs) formed spontaneously in micromass cultures derived from antler cartilage and these cells had the phenotypic characteristics of osteoclasts. PTHrP and receptor activator of NF‐κB ligand (RANKL) stimulated antler osteoclast formation although the effect of RANKL was less marked than that of PTHrP. The addition of osteoprotegerin (OPG) only partially decreased (by ∼65%) the number of osteoclasts in PTHrP‐treated cultures. To determine whether PTHrP also potentially could have direct effects on antler osteoclasts, we studied, by confocal microscopy, the expression of the type I PTH/PTHrP receptor (PTH1R) in MNCs cultured on glass and found the receptor protein to have a nuclear localization. In situ hybridization showed that antler MNCs also expressed PTH1R and PTHrP messenger RNAs (mRNAs). PTHrP was immunolocalized in MNCs cultured on glass but was undetectable in cells resorbing a dentine substrate. In tissue sections of antler cartilage, PTHrP and PTH1R were expressed in vitronectin receptor‐positive (VNR+) osteoclast‐like cells localized in the perivascular stroma. Thus, these data show that PTHrP plays a role in the regulation of osteoclast differentiation in regenerating skeletal tissues and that PTHrP can have effects on osteoclastogenesis that are independent of RANKL synthesis. Ours is the first study to describe the expression of the type I PTH/PTHrP receptor in mammalian osteoclasts at a protein and mRNA level, which indicates that PTHrP also may have a direct effect on osteoclasts. This also is the first study to show a nuclear localization of the PTHIR in cells of the osteoclast lineage, although the functional significance of this observation has yet to be established.
Freeze-fracture Cytochemistry: A New Method Combining Immunocytochemistry and Enzyme Cytochemistry on Replicas

We describe a new freeze-fracture cytochemical technique consisting of combined immunocytochemistry and enzyme cytochemistry. This technique reveals the relationship between molecules in biological membranes by double labeling with two different cytochemical markers (i.e., immunogold probes and cerium). In this method, antigens were detected with specific primary antibodies and appropriate secondary immunoprobes. Subsequently, alkaline phosphatase activity was detected with cerium as the capture agent on the same replicas. Octyl-glucoside (OG) digestion before the cytochemical reactions was crucial to the success of this combined method. OG is an efficient detergent and OG digestion can preserve both immunocytochemical antigenicity and enzyme activity on replicas. As an initial examination, we applied this technique to the study of glycosyl-phosphatidyl-inositol-anchored proteins and adhesion molecules in human neutrophils. The method described here should serve as a unique additional approach for the study of topology and dynamics of molecules in biomembranes.
Identification of novel genomic islands coding for antigenic pilus‐like structures in Streptococcus agalactiae

We have recently reported the presence of covalently linked pilus‐like structures in the human pathogen, Group B Streptococcus (GBS). The pilus operon codes for three proteins which contain the conserved amino acid motif, LPXTG, associated with cell wall‐anchored proteins together with two genes coding for sortase enzymes. Analysis of the eight sequenced genomes of GBS has led to the identification of a second, related genomic island of which there are two variants, each containing genes coding for proteins with LPXTG motifs and sortases. Here we show that both variant islands also code for pilus‐like structures. Furthermore, we provide a thorough description and characterization of the genomic organization of the islands and the role of each protein in the assembly of the pili. For each pilus, polymerization of one of the three component proteins is essential for incorporation of the other two proteins into the pilus structure. In addition, two sortases are required for complete pilus assembly, each with specificity for one of the pilus components. A component protein of one of the newly identified pili is also a previously identified protective antigen and a second component of this pilus is shown to confer protection against GBS challenge. We propose that pilus‐like structures are important virulence factors and potential vaccine candidates.
Expression of Heteropolymeric Ferritin Improves Iron Storage in Saccharomyces cerevisiae

ABSTRACT Saccharomyces cerevisiae was engineered to express different amount of heavy (H)- and light (L)-chain subunits of human ferritin by using a low-copy integrative vector (YIp) and a high-copy episomal vector (YEp). In addition to pep4::HIS3 allele, the expression host strain was bred to have the selection markers leu2− and ura3− for YIplac128 and YEp352, respectively. The heterologous expression of phytase was used to determine the expression capability of the host strain. Expression in the new host strain (2805-a7) was as high as that in the parental strain (2805), which expresses high levels of several foreign genes. Following transformation, Northern and Western blot analyses demonstrated the expression of H- and L-chain genes. The recombinant yeast was more iron tolerant, in that transformed cells formed colonies on plates containing more than 25 mM ferric citrate, whereas none of the recipient strain cells did. Prussian blue staining indicated that the expressed isoferritins were assembled in vivo into a complex that bound iron. The expressed subunits showed a clear preference for the formation of heteropolymers over homopolymers. The molar ratio of H to L chains was estimated to be 1:6.8. The gel-purified heteropolymer took up iron faster than the L homopolymer, and it took up more iron than the H homopolymer did. The iron concentrations in transformants expressing the heteropolymer, L homopolymer, and H homopolymer were 1,004, 760, and 500 μg per g (dry weight) of recombinant yeast cells, respectively. The results indicate that heterologously expressed H and L subunits coassemble into a heteropolymer in vivo and that the iron-carrying capacity of yeast is further enhanced by the expression of heteropolymeric isoferritin.
Prenatal Iron Supplementation Reduces Maternal Anemia, Iron Deficiency, and Iron Deficiency Anemia in a Randomized Clinical Trial in Rural China, but Iron Deficiency Remains Widespread in Mothers and Neonates.

BACKGROUND Previous trials of prenatal iron supplementation had limited measures of maternal or neonatal iron status. OBJECTIVE The purpose was to assess effects of prenatal iron-folate supplementation on maternal and neonatal iron status. METHODS Enrollment occurred June 2009 through December 2011 in Hebei, China. Women with uncomplicated singleton pregnancies at ≤20 wk gestation, aged ≥18 y, and with hemoglobin ≥100 g/L were randomly assigned 1:1 to receive daily iron (300 mg ferrous sulfate) or placebo + 0.40 mg folate from enrollment to birth. Iron status was assessed in maternal venous blood (at enrollment and at or near term) and cord blood. Primary outcomes were as follows: 1) maternal iron deficiency (ID) defined in 2 ways as serum ferritin (SF) <15 μg/L and body iron (BI) <0 mg/kg; 2) maternal ID anemia [ID + anemia (IDA); hemoglobin <110 g/L]; and 3) neonatal ID (cord blood ferritin <75 μg/L or zinc protoporphyrin/heme >118 μmol/mol). RESULTS A total of 2371 women were randomly assigned, with outcomes for 1632 women or neonates (809 placebo/folate, 823 iron/folate; 1579 mother-newborn pairs, 37 mothers, 16 neonates). Most infants (97%) were born at term. At or near term, maternal hemoglobin was significantly higher (+5.56 g/L) for iron vs. placebo groups. Anemia risk was reduced (RR: 0.53; 95% CI: 0.43, 0.66), as were risks of ID (RR: 0.74; 95% CI: 0.69, 0.79 by SF; RR: 0.65; 95% CI: 0.59, 0.71 by BI) and IDA (RR: 0.49; 95% CI: 0.38, 0.62 by SF; RR: 0.51; 95% CI: 0.40, 0.65 by BI). Most women still had ID (66.8% by SF, 54.7% by BI). Adverse effects, all minor, were similar by group. There were no differences in cord blood iron measures; >45% of neonates in each group had ID. However, dose-response analyses showed higher cord SF with more maternal iron capsules reported being consumed (β per 10 capsules = 2.60, P < 0.05). CONCLUSIONS Prenatal iron supplementation reduced anemia, ID, and IDA in pregnant women in rural China, but most women and >45% of neonates had ID, regardless of supplementation. This trial was registered at clinicaltrials.gov as NCT02221752.
Parent–Child Interaction Therapy

It is common practice to augment efficacious treatment protocols for special populations (Durlak & DuPre, 2008), but this is often done before establishing that standard services are not appropriate. In this randomized controlled trial with families at risk or with a history of maltreatment (N = 151), we investigated the effectiveness of standard 12-session Parent–Child Interaction Therapy (PCIT). This is in contrast to other PCIT studies with similar parents, which have allowed for longer and sometimes variable treatment length and with modifications to PCIT protocol. After treatment and compared to Waitlist, mothers reported fewer child externalizing and internalizing behaviors, decreased stress, and were observed to have more positive verbalizations and maternal sensitivity. These outcomes were equivalent or better than outcomes of our previous PCIT trial with high-risk families (Thomas & Zimmer-Gembeck, 2011) when treatment length was variable and often longer. These findings support standard protocol PCIT as an efficacious intervention for families in the child welfare system.
Intraspecies transmission of L‐type‐like bovine spongiform encephalopathy detected in Japan

It has been assumed that the agent causing BSE in cattle is a uniform strain (classical BSE); however, different neuropathological and molecular phenotypes of BSE (atypical BSE) have been recently reported. We demonstrated the successful transmission of L‐type‐like atypical BSE detected in Japan (BSE/JP24 isolate) to cattle. Based on the incubation period, neuropathological hallmarks, and molecular properties of the abnormal host prion protein, the characteristics of BSE/JP24 prion were apparently distinguishable from the classical BSE prion and closely resemble those of bovine amyloidotic spongiform encephalopathy prion detected in Italy.
The passage of protozoa from the reticulo-rumen through the omasum of sheep

1. Protozoa in rumen contents and omasal effluent of growing wethers were counted. The wethers were equipped with rumen and abomasal cannulas, and omasal sleeves attached to the omasal-abomasal orifice. Rumen fluid dilution rates were elevated by continuous infusions of hypertonic mineral solutions (34 litres/d) for 24 d. Rumen contents and omasal effluent were sampled between 9 and 21 h during the last 10 d of each experiment. 2. Protozoa1 concentrations in omasal effluent were only 0.24.3 those found in the rumen under normal conditions. The ratio of protozoal concentrations in rumen: those in omasal effluent was for small Diplodinium spp. 4.6 (SD 0.9), for Ophryoscolex spp. 4.3 (SD 1.0), for Dasytricha ruminantium 4.0 (SD 0.5), for Isotricha spp. 3.8 (SD 0.8), for Entodinium spp. 3.6 (SD 0.9) and for Polyplastron multivesiculatum 2.6 (SD 0.5). 3. Elevation of rumen fluid dilution rate by 20 and 55% respectively, increased protozoal concentrations in omasal effluents from 22 to 33% and from 31 to 47% those in rumen contents. The apparent residence times of protozoa in the rumen were decreased 50% by the infusion of a mineral-salt solution. The increase in rumen fluid dilution rate had no significant effect on concentrations of protozoa in the rumen or on the differences of the apparent residence times between different species. The apparent residence time of holotrichs remained the same before and after infusion of the mineral-salt solution. 4. Apparent residence times of individual species of protozoa in the rumen were, under normal feeding conditions, 2.55 d, and were four to six times longer than the mean residence time of CrEDTA in the rumen.
Flat frequency response in the electronic measurement of the Boltzmann constant

A new quantum voltage calibrated Johnson noise thermometer (JNT) was developed at NIM to demonstrate the electrical approach that determines the Boltzmann constant k by comparing electrical and thermal noise power. A measurement with an integration period of 10 hours and bandwidth of 640 kHz results in relative offset of 0.5×10-6 from the current CODATA value of k, and type A relative standard uncertainty of 23×10-6. The quadratic fitting parameters of the ratio spectrum show a flat response with respect to the measurement bandwidth. This flat response is a dramatic improvement compared to the response produced by the NIST JNT system that dominated the relative combined uncertainty of previous measurements of k.
Prevalence of Depression After Stroke: The Perth Community Stroke Study

Background The Perth Community Stroke Study (PCSS) was a population-based study of the incidence, cause, and outcome of acute stroke. Method Subjects from the study were assessed initially, by examination and interview, and at four- and 12-month follow-ups to determine differences in prevalence of depression between the sexes and between patients with first-ever and recurrent strokes. Results The prevalence of depressive illness four months after stroke in 294 patients from the PCSS was 23% (18–28%), 15% (11–19%) major depression and 8% (5–11 %) minor depression. There were no significant differences between the sexes or between patients with first-ever and recurrent strokes. With a non-hierarchic approach to diagnosis of those with depression, 26% of men and 39% of women had an associated anxiety disorder, mainly agoraphobia. Nine per cent of male and 13% of female patients interviewed had evidence of depression at the time of the stroke. Twelve months after stroke 56% of the men were still depressed (40% major and 16% minor), as were 30% of the women (12% major and 18% minor). Conclusion The prevalence of depression after stroke was comparable with that reported from other studies, and considerably less than that reported from in-patient and rehabilitation units.
Conformational selection and adaptation to ligand binding in T4 lysozyme cavity mutants

Significance Analysis of protein packing reveals the prevalence of cavities, some of which are evolutionarily conserved. Despite the fact that these packing defects are generally destabilizing, they can play important roles in facilitating functionally important protein motions and in the evolution of protein function. In this study, we used site-directed spin-labeling with electron paramagnetic resonance spectroscopy to investigate the structural and dynamical response of an enzyme to engineered ligand-binding cavities. The results show that the engineered cavities introduce conformational fluctuations between a native-like state and a new state. Ligand binding shifts the equilibrium toward the native-like state, suggesting binding via conformational selection, although additional structural adaptation is observed for one mutant. The results underscore the potential role of cavities in modulating molecular flexibility. The studies presented here explore the relationship between protein packing and molecular flexibility using ligand-binding cavity mutants of T4 lysozyme. Although previously reported crystal structures of the mutants investigated show single conformations that are similar to the WT protein, site-directed spin labeling in solution reveals additional conformational substates in equilibrium exchange with a WT-like population. Remarkably, binding of ligands, including the general anesthetic halothane shifts the population to the WT-like state, consistent with a conformational selection model of ligand binding, but structural adaptation to the ligand is also apparent in one mutant. Distance mapping with double electron-electron resonance spectroscopy and the absence of ligand binding suggest that the new substates induced by the cavity-creating mutations represent alternate packing modes in which the protein fills or partially fills the cavity with side chains, including the spin label in one case; external ligands compete with the side chains for the cavity space, stabilizing the WT conformation. The results have implications for mechanisms of anesthesia, the response of proteins to hydrostatic pressure, and protein engineering.
‘You are here’: locating ‘spirituality’ on the map of the current medical world

Purpose of review Clinical works at the intersection of ‘spirituality, religion, theology and medicine’ are studied to identify various aspects of what constitutes spirituality, what contributes to spiritual health and how to provide spiritual-healers for our current health-care system. Recent findings Spiritual care in the current medical world can be classed grossly into two departments: complementary and alternative medicine, considered as proxy variable for spirituality, and physician-initiated clinical Chaplaincy, informed by theology. The large body of research on ‘self’ as a therapeutic tool, though, falls into subtle categories: phenomenological studies, empathy, embodied care, and mindfulness-based therapies. Development in the field of ‘spiritual medicine’ has focused on spirituality-related curricula. Summary As mindfulness-based meditation programs help build deep listening skills needed to stay aware of the ‘self’, Clinical Pastoral Education trains the chaplain to transcend the ‘self’ to provide embodied care. Clinical chaplaincy is the destination for health-care professionals as well as theological/religious scholars who have patients’ spiritual health as their primary focus. Medical education curricula that train students in chaplain's model of transpersonal-mindfulness/empathy founded on neuro-physiological principles would help them gain skills in embodied care. Such education would seamlessly integrate evidence-based clinical practice and spiritual-theological concepts.
Comparison of Two Pharmacodynamic Transduction Models for the Analysis of Tumor Therapeutic Responses in Model Systems

Semi-mechanistic pharmacodynamic (PD) models that capture tumor responses to anticancer agents with fidelity can provide valuable insights that could aid in the optimization of dosing regimens and the development of drug delivery strategies. This study evaluated the utility and potential interchangeability of two transduction-type PD models: a cell distribution model (CDM) and a signal distribution model (SDM). The evaluation was performed by simulating dense and sparse tumor response data with one model and analyzing it using the other. Performance was scored by visual inspection and precision of parameter estimation. Capture of tumor response data was also evaluated for a liposomal formulation of paclitaxel in the paclitaxel-resistant murine Colon-26 model. A suitable PK model was developed by simultaneous fitting of literature data for paclitaxel formulations in mice. Analysis of the simulated tumor response data revealed that the SDM was more flexible in describing delayed drug effects upon tumor volume progression. Dense and sparse data simulated using the CDM were fit very well by the SDM, but under some conditions, data simulated using the SDM were fitted poorly by the CDM. Although both models described the dose-dependent therapeutic responses of Colon-26 tumors, the fit by the SDM contained less bias. The CDM and SDM are both useful transduction models that recapitulate, with fidelity, delayed drug effects upon tumor growth. However, they are mechanistically distinct and not interchangeable. Both fit some types of tumor growth data well, but the SDM appeared more robust, particularly where experimental data are sparse.
Divergent requirements for fibroblast growth factor signaling in zebrafish maxillary barbel and caudal fin regeneration

The zebrafish maxillary barbel is an integumentary organ containing skin, glands, pigment cells, taste buds, nerves, and endothelial vessels. The maxillary barbel can regenerate (LeClair & Topczewski 2010); however, little is known about its molecular regulation. We have studied fibroblast growth factor (FGF) pathway molecules during barbel regeneration, comparing this system to a well‐known regenerating appendage, the zebrafish caudal fin. Multiple FGF ligands (fgf20a, fgf24), receptors (fgfr1‐4) and downstream targets (pea3, il17d) are expressed in normal and regenerating barbel tissue, confirming FGF activation. To test if specific FGF pathways were required for barbel regeneration, we performed simultaneous barbel and caudal fin amputations in two temperature‐dependent zebrafish lines. Zebrafish homozygous for a point mutation in fgf20a, a factor essential for caudal fin blastema formation, regrew maxillary barbels normally, indicating that the requirement for this ligand is appendage‐specific. Global overexpression of a dominant negative FGF receptor, Tg(hsp70l:dn‐fgfr1:EGFP)pd1 completely blocked fin outgrowth but only partially inhibited barbel outgrowth, suggesting reduced requirements for FGFs in barbel tissue. Maxillary barbels expressing dn‐fgfr1 regenerated peripheral nerves, dermal connective tissue, endothelial tubes, and a glandular epithelium; in contrast to a recent report in which dn‐fgfr1 overexpression blocks pharyngeal taste bud formation in zebrafish larvae (Kapsimali et al. 2011), we observed robust formation of calretinin‐positive tastebuds. These are the first experiments to explore the molecular mechanisms of maxillary barbel regeneration. Our results suggest heterogeneous requirements for FGF signaling in the regeneration of different zebrafish appendages (caudal fin versus maxillary barbel) and taste buds of different embryonic origin (pharyngeal endoderm versus barbel ectoderm).