Increased Bone Mineral Content During Rapid Weight Gain Therapy in Anorexia Nervosa.
Patients with anorexia nervosa (AN) are at high risk of reduced bone mass. Osteocalcin (OC), a bone formation marker, has been proposed to act as a link between bone and energy metabolism. We investigated how the 3 forms of OC respond during a 12-week intensive nutrition therapy in AN patients, in whom large changes in energy metabolism are expected.Twenty-two female AN patients, mean 20.9 years of age, with a starting mean body mass index (BMI) 15.5 kg/m2 (minimum-maximum) (13.4-17.3 kg/m2) completed the study. Biochemical markers, body composition, bone mass by DXA, and pQCT were assessed. Subjects gained in median 9.9 kg (5.5-17.0 kg), and BMI increased from median 15.4 kg/m2 (13.4-17.3 kg/m2) to 19.0 kg/m2 (16.2-20.6 kg/m2), p<0.0001. Fat mass increased from median 11.4% (4.4-24.8%) to 26.7% (16.9-39.8%). Total OC, carboxylated OC (cOC), undercarboxylated OC (ucOC), and bone-specific alkaline phosphatase (BALP) increased during the study period. No change was observed for the resorption marker carboxy-terminal cross-linking telopeptide of type I collagen (CTX). Total body bone mineral content (BMC) increased, but no changes were found for whole body or lumbar spine bone mineral density. Tibial trabecular density measured by pQCT decreased. Total OC, cOC, and ucOC were not associated with BMI, insulin or body composition parameters. This prospective study demonstrates that all 3 forms of OC (total OC, cOC, ucOC) increase during rapid weight gain. BALP increased while the resorption marker CTX was unchanged, which corroborate with the increased total body BMC.


Microencapsulated fluorescent pH probe as implantable sensor for monitoring the physiological state of fish embryos.
In vivo physiological measurement is a major challenge in modern science and technology, as is environment conservation at the global scale. Proper toxicological testing of widely produced mixtures of chemicals is a necessary step in the development of new products, allowing us to minimize the human impact on aquatic ecosystems. However, currently available bioassay-based techniques utilizing small aquatic organisms such as fish embryos for toxicity testing do not allow assessing in time the changes in physiological parameters in the same individual. In this study, we introduce microencapsulated fluorescent probes as a promising tool for in vivo monitoring of internal pH variation in zebrafish embryos. The pH alteration identified under stress conditions demonstrates the applicability of the microencapsulated fluorescent probes for the repeated analysis of the embryo's physiological state. The proposed approach has strong potential to simultaneously measure a range of physiological characteristics using a set of specific fluorescent probes and to finally bring toxicological bioassays and related research fields to a new level of effectiveness and sensitivity.


Acral fibrokeratoma of the foot treated with excision and trap door flap closure: a case report.
Acral fibrokeratoma is a relatively rare, benign lesion that is typically found on the fingers and toes but can also be found on the palms and soles. Several case reports have been published of this rare lesion, with only a few investigators describing lesions of the feet. The techniques of using skin from a portion of the mass to close the defect left from mass excision have been poorly reported. Our technique, which we have termed the "trap door flap," has design similarities to the appearance and mechanics of a trap door. We present the case of a patient with a 7-year history of a slow-growing, soft tissue mass on the plantar and medial aspect of his left hallux. The mass was subsequently excised and closed with a local flap. The flap had healed uneventfully by 2 weeks postoperatively, and the patient denied any evidence of recurrence or pain at 12 months of follow-up.


Rectal perforation secondary to transanal haemorrhoidal dearterialisation.
Haemorrhoidal artery ligation has now been established as a treatment modality for symptomatic haemorrhoids. We report a case of a fit 44-year-old male who underwent the procedure as a day case, who subsequently developed pelvic sepsis due to rectal perforation. This case is the first report of a potentially life-threatening complication resulting from this procedure, which has a previously excellent safety profile.


Real-time optical interferometric image subtraction by wave polarization.
A real-time coherent optical interferometric image addition and subtraction system based on a technique similar to an earlier work by Kunstmann and Spitschan [Opt. Commun. 4, 166 (1971)] is extensively investigated. Theoretical analysis of the system has been compared with experimental data. Distortion and resolution power of the system with respect to the size and positions of the input images have been analyzed in detail.


Culture aeration in a polythermostat.
A system for the aeration of cultures growing in a polythermostat temperature-gradient block was developed. This system provided both aeration and agitation of the cultures, while allowing the growth to be followed spectrophotometrically. The necessity for aeration of cultures growing in a polythermostat was experimentally shown.


Components of airway hyperresponsiveness and their associations with inflammation and remodeling in mice.
Pathologic changes, including inflammation and remodeling, occur in the asthmatic airway. However, their relative contribution to the components of airway hyperresponsiveness (AHR) remains unclear. Attempting to delineate AHR into discrete immune-mediated and structural remodeling components, we performed a detailed time course of the development, progression, and persistence of maximal respiratory system resistance, airway reactivity, and airway sensitivity. Mice exposed to increasing durations of persistent allergen were assessed for airway function, morphometry, and inflammation. Allergen exposure resulted in increases for all indices of AHR that persisted for at least 4 weeks after chronic allergen exposure (P < .01 for all values). Early increases in AHR were associated with increases in immune-mediated events, including airway eosinophils (P < .01), whereas sustained AHR was associated with structural remodeling events. Increased maximal respiratory system resistance, evident by 6 weeks postallergen and persisting for at least 4 weeks after 8 weeks of chronic exposure, was associated with an increase in collagen deposition (P < .01). Increased airway reactivity and sensitivity, each evident by 1 week after allergen and persisting for at least 4 weeks after 8 weeks of chronic exposure, were associated with an increase in airway smooth muscle area (P < .01). Our novel observation of distinct temporal relationships in the development, progression, and persistence of the individual indices of AHR supports our hypothesis that multiple underlying factors contribute to airway dysfunction. These findings illustrate the importance of clearly addressing specific components of airway dysfunction to provide greater insight into specific pathophysiologic mechanisms in airway disease.


Physical stability of coconut oil lotions formulated using hydrophile-lipophile balance system of various emulsifier pairs.
The aim of this study was to prepare coconut oil lotions using a hydrophile-lipophile balance system to calculate the proportion of each nonionic surfactant used. The effects of emulsifier pairs in the formulations on physical properties (i.e., appearance, emulsion type, pH, flow type, viscosity) were investigated. The physical stability of the lotions was determined at ambient temperature (approximatley 30 degrees C) after the lotions were kept for 30 as well as 60 days and in accelerated conditions (6 freeze-thaw cycles). It was found that the formulations most tolerant to such harsh conditions were F1 and F2, o/w lotions containing 40% w/w coconut oil, 50% w/w water and 10% w/w of the mixed emulsifier of a low hydrophile-lipophile balance surfactant (sorbitan monostearate) and a high hydrophile-lipophile balance surfactant (either polyoxyethylene [20] sorbitan monooleate or polyethylene [20] sorbitan monolaurate).


Functions of type II pneumocyte-derived vascular endothelial growth factor in alveolar structure, acute inflammation, and vascular permeability.
Vascular endothelial growth factor-A (VEGF) is a potent regulator of vascular permeability, inflammatory response, and cell survival in the lung. To explore the functions of VEGF produced locally in type II pneumocytes, we generated mice with a conditional deletion of VEGF-A using Cre recombinase driven by the human surfactant protein C (SPC) promoter. In 7- to 10-week-old VEGF-knockout (SPC-VEGF-KO) mice, lung histology and physiology were essentially normal, except for higher dynamic lung compliance and lower pulmonary vascular permeability. Emphysema was seen in 28- to 32-week-old animals. To investigate the role of type II pneumocyte-derived VEGF in acute lung injury, we challenged 7- to 10-week-old SPC-VEGF-KO mice and their wild-type littermates with intestinal ischemia-reperfusion. Bronchoalveolar lavage fluid total cell count, pulmonary permeability, and lung injury score were significantly attenuated, and total lung VEGF levels were significantly lower in SPC-VEGF-KO mice compared with wild-type controls. In SPC-VEGF-KO mice, activated caspase 3-positive type II epithelial cells were increased after intestinal ischemia-reperfusion, even though there was no significant difference in the total number of cells positive for terminal deoxynucleotidyl transferase dUTP nick-end labeling. We conclude that VEGF in type II cells helps protect alveolar epithelial cells from caspase-dependent apoptosis. However, VEGF produced from type II cells may contribute to increased vascular permeability during acute lung injury.


Conformational rearrangements upon start codon recognition in human 48S translation initiation complex.
Selection of the translation start codon is a key step during protein synthesis in human cells. We obtained cryo-EM structures of human 48S initiation complexes and characterized the intermediates of codon recognition by kinetic methods using eIF1A as a reporter. Both approaches capture two distinct ribosome populations formed on an mRNA with a cognate AUG codon in the presence of eIF1, eIF1A, eIF2-GTP-Met-tRNAiMet and eIF3. The 'open' 40S subunit conformation differs from the human 48S scanning complex and represents an intermediate preceding the codon recognition step. The 'closed' form is similar to reported structures of complexes from yeast and mammals formed upon codon recognition, except for the orientation of eIF1A, which is unique in our structure. Kinetic experiments show how various initiation factors mediate the population distribution of open and closed conformations until 60S subunit docking. Our results provide insights into the timing and structure of human translation initiation intermediates and suggest the differences in the mechanisms of start codon selection between mammals and yeast.


Structural and biosynthetic studies of a principal bile alcohol, 27-nor-5beta-cholestane-3alpha,7alpha,12alpha,24,25-pentol, in human urine.
The stereochemistry at C-24 and C-25 of 27-nor-5beta-cholestane-3alpha,7alpha,12alpha,24 ,25-pentol, a principal bile alcohol in human urine, and its biosynthesis are studied. Four stereoisomers of the C(26)-24,25-pentols were synthesized by reduction with LiAlH(4) of the corresponding epoxides prepared from (24S)- or (24R)-27-nor-5beta-cholest-25-ene-3alpha, 7alpha,12alpha,24-tetrol. The stereochemistries at C-25 were deduced by comparison of the C(26)-24,25-pentols with the oxidation products of (24Z)-27-nor-5beta-cholest-24-ene-3alpha,7alpha, 12alpha-triol with osmium tetraoxide. On the basis of this assignment, the principal bile alcohol excreted into human and rat urine was determined to be (24S,25R)-27-nor-5beta-cholestane-3alpha,7alpha, 12alpha,24,25-pentol, accompanied by a lesser amount of (24R, 25R)-isomer. To elucidate the biosynthesis of the C(26)-24,25-pentol, a putative intermediate, 3alpha,7alpha, 12alpha-trihydroxy-27-nor-5beta-cholestan-24-one, derived from 3alpha,7alpha, 12alpha-trihydroxy-24-oxo-5beta-cholestanoic acid by decarboxylation during the side-chain oxidation of 3alpha,7alpha, 12alpha-trihydroxy-5beta-cholestanoic acid, was incubated with rat liver homogenates. The 24-oxo-bile alcohol could be efficiently reduced to yield mainly (24R)-27-nor-5beta-cholestane-3alpha,7alpha, 12alpha,24-tetrol. If a 25R-hydroxylation of the latter steroid occurs, it should lead to formation of (24S,25R)-C(26)-24,25-pentol. Now it has appeared that a major bile alcohol excreted into human urine is (24S,25R)-27-nor-5beta-cholestane-3alpha,7alpha, 12alpha, 24, 25-pentol, which might be derived from 3alpha,7alpha, 12alpha-trihydroxy-27-nor-5beta-cholestan-24-one via (24R)-27-nor-5beta-cholestane-3alpha, 7alpha,12alpha,24-tetrol.


Radical or not radical: compared structures of metal (M = Ni, Au) bis-dithiolene complexes with a thiazole backbone.
A complete series of dianionic, monoanionic, and neutral dithiolene complexes formulated as [Ni(Et-thiazdt)2](n), with n = -2, -1, 0, and Et-thiazdt: N-ethyl-1,3-thiazoline-2-thione-4,5-dithiolate, is prepared using an optimized procedure described earlier for the N-Me derivatives. Electrochemical and spectroscopic properties confirm the electron-rich character of the Et-thiazdt dithiolate ligand. The three complexes are structurally characterized by single-crystal X-ray diffraction. The paramagnetic anionic complex [Ni(Et-thiazdt)2](-1), as Ph4P(+) salt, exhibits side-by-side lateral interactions leading to a Heisenberg spin chain behavior. The solid-state structure of the neutral, diamagnetic [Ni(Et-thiazdt)2](0) complex shows a face-to-face organization with a large longitudinal shift, at variance with the structure of its radical and neutral gold dithiolene analogue described earlier and formulated as [Au(Et-thiazdt)2](•). Comparison of the two structures, and those of the other few structurally characterized pairs of Ni/Au dithiolene complexes, demonstrates the important role played by overlap interactions between gold dithiolene radical species. Despite its closed-shell character, the neutral nickel complex [Ni(Et-thiazdt)2](0) exhibits a semiconducting behavior with a room-temperature conductivity σRT ≈ 0.014 S cm(-1).


The structure of critical care transfer networks.
Moving patients from low-performing hospitals to high-performing hospitals may improve patient outcomes. These transfers may be particularly important in critical care, where small relative improvements can yield substantial absolute changes in survival. To characterize the existing critical care network in terms of the pattern of transfers. In a retrospective cohort study, the nationwide 2005 Medicare fee-for-service claims were used to identify the interhospital transfer of critically ill patients, defined as instances where patients used critical care services in 2 temporally adjacent hospitalizations. We measured the characteristics of the interhospital transfer network and the extent to which intensive care unit patients are referred to each hospital in that network--a continuous quantitative measure at the hospital-level known as centrality. We evaluated associations between hospital centrality and organizational, medical, surgical, and radiologic capabilities. There were 47,820 transfers of critically ill patients among 3308 hospitals. 4.5% of all critical care stays of any length involved an interhospital critical care transfer. Hospitals transferred out to a mean of 4.4 other hospitals. More central hospital positions were associated with multiple indicators of increased capability. Hospital characteristics explained 40.7% of the variance in hospitals' centrality. Critical care transfers are common, and traverse an informal but structured network. The centrality of a hospital is associated with increased capability in delivery of services, suggesting that existing transfers generally direct patients toward better resourced hospitals. Studies of this network promise further improvements in patient outcomes and efficiency of care.


Analysis of BAP1 Germline Gene Mutation in Young Uveal Melanoma Patients.
To evaluate the prevalence of BAP1 germline mutations in a series of young patients with uveal melanoma (UM), diagnosed before age 30. The study was carried out on 14 young uveal melanoma patients (average age 21.4 years, range 3 months to 29 years). Germline DNA was extracted from peripheral blood. BAP1 sequencing was carried out using direct sequencing of all exons and adjacent intronic sequences. We also tested for germline mutations in additional melanoma-associated candidate genes CDKN2A and CDK4 (exon 4). We identified one patient with a pathogenic mutation (c. 1717delC, p.L573fs*3) in BAP1. This patient was diagnosed with UM at age 18 years and had a family history of a father with UM and a paternal grandfather with cancer of unknown origin. One additional patient had an intronic variant of uncertain significance (c.123-48T > G) in BAP1 while the remaining 12 patients had no alteration. None of the patients had CDKN2A or CDK4 (Exon 4) mutations. Family history was positive for a number of additional malignancies in this series, in particular for cutaneous melanoma, prostate, breast and colon cancers. There were no families with a history of mesothelioma or renal cell carcinoma. This study suggests that a small subset of patients with early onset UM has germline mutation in BAP1. While young patients with UM should be screened for germline BAP1 mutations, our results suggest that there is a need to identify other candidate genes which are responsible for UM in young patients.


Optimal lens design and use in laser-scanning microscopy.
In laser-scanning microscopy often an off-the-shelf achromatic doublet is used as a scan lens which can reduce the available diffraction-limited field-of-view (FOV) by a factor of 3 and introduce chromatic aberrations that are scan angle dependent. Here we present several simple lens designs of superior quality that fully make use of high-NA low-magnification objectives, offering diffraction-limited imaging over a large FOV and wavelength range. We constructed a two-photon laser-scanning microscope with optimized custom lenses which had a near diffraction limit point-spread-function (PSF) with less than 3.6% variation over a 400 µm FOV and less than 0.5 µm lateral color between 750 and 1050 nm.


Knowledge-based systems in laboratory medicine and pathology. A review and survey of the field.
Knowledge-based systems are computer systems designed to handle knowledge-intensive tasks, usually involving reasoning and inference. They are increasingly being applied to problems in laboratory medicine and pathology. In this article we provide a brief introduction to the basic concepts of knowledge-based systems, review some of their published applications, and report on an informal survey of specialists in laboratory medicine and pathology. The survey, sent to 102 individuals, indicated that 24% were involved in developing knowledge-based systems, with most systems at an early stage of development. Recent advances in knowledge-based systems research as well as survey responses suggest that this technology will have increasing value in laboratory medicine and pathology.


Minimally invasive lateral transpsoas approach for lumbar corpectomy and stabilization.
Here, we present our experience with the minimally invasive (MI) transpsoas approach for lumbar corpectomy and stabilization. Transpsoas approach accesses the lumbar spine and includes both the direct lateral interbody fusion and extreme lateral interbody fusion techniques. Both procedures utilize a tubular retractor system which facilitates adequate retraction and direct visualization of the target, while supposedly reducing soft tissue trauma. We evaluated two patients, one with a traumatic L2 wedge compression fracture and the other with an L3 pathological compression fracture due to multiple myeloma. Both patients underwent MI transpsoas lumbar corpectomy, anterior column reconstruction with an expandable cage, and posterior pedicle screw instrumentation to correct a kyphotic deformity. Both patients were mobilized on the 1st postoperative day and experienced significant postoperative pain relief. In two cases involving L2 and L3 compression fractures, MI transpsoas lumbar corpectomy was safely performed, with reduced perioperative and postoperative morbidity. Here, the transpsoas approach also allowed for early mobilization, adequate postoperative biomechanical stability, and resulted in immediate good outcomes.


Relationship of cognitive functioning, whole brain volumes, and T2-weighted hyperintensities in neurofibromatosis-1.
Using quantitative magnetic resonance imaging morphometry, we report that the whole brain volumes of patients with neurofibromatosis-1 are significantly larger than normal, confirm the prevalence of macrocephaly as about 50%, and report that macrocephaly in patients with neurofibromatosis-1 does not appear to be related to the familial or sporadic origin of the neurofibromatosis-1 or to the presence or absence of T2-weighted hyperintensities. No strong relationship emerged between the extent of neurofibromatosis-1-associated impairment of cognitive functions and degree of macrocephaly; however, the macrocephalic neurofibromatosis-1 group did have a significant verbal impairment relative to the nonmacrocephalic neurofibromatosis-1 group in vocabulary (P < .009).


[Chief risk factors of ischemic heart disease and mortality of the middle-aged male population of Kaunas].
A prospective study, averaging 11 years, of a representative sample of 2455 male residents of Kaunas between 45 and 59 years of age has explored relationships between coronary risk factors and mortality due to coronary heart disease (CHD), cancer, injuries, as well as total mortality rates. An original analytical approach has been developed for the analysis of associations between individual risk factors and mortality that allows to exclude the effects of age and other examined factors. A marked relationship has been demonstrated between: coronary mortality and age, arterial blood pressure, cholesterol, glucose tolerance, smoking, body weight, the presence and clinical form of CHD; cancer-related mortality, and age or smoking; traumatic mortality and smoking; total mortality and age, arterial blood pressure, glucose tolerance, body weight, smoking, and the presence and type of CHD.


Paying your respects: Transgender women and detransitioning after death.
This paper asks how do the deaths and the postmortem detransitioning (the verbal, visual, and material rejection of a person's gender identity) of transgender women impact trans activism? After analyzing the case studies of Jennifer Gable and Leelah Alcorn, I outline how the contentious memorialization of transgender women and the disenfranchized grief of survivors influence trans activism. I conclude that activism is characterized by respecting the wishes of the deceased, by preventing the violence which transgender women experience, by advocating for trans elder care, by educating about end-of-life issues, and by lobbying for laws that protect transgender men and women after death.


Intracorporeal suturing and knot tying broadens the clinical applicability of laparoscopy.
As surgeons become more experienced with basic laparoscopic procedures like cholecystectomy, they are able to expand this approach to less common operations. However, without laparoscopic suturing skills, like those obtained with Nissen fundoplication, many operations cannot be completed laparoscopically. We present a series of 10 patients with less common surgical illnesses who were successfully treated with minimal access techniques and intracorporeal suturing. Over a 6-month period at 2 medical centers, 10 patients underwent operations with laparoscopic intracorporeal suturing and knot tying. Diagnoses included bowel obstruction due to gallstone ileus (n=1), perforated uterus from an intrauterine device (n=1), urinary bladder diverticulum (n=1), bleeding Meckel's diverticulum (n=3), and perforated duodenal ulcer (n=4). Each patient was treated with standard surgical interventions performed entirely laparoscopically with intracorporeal suturing. No morbidity or mortality occurred in any patient due to the operation. Although each of these operations has been previously reported, as a series, they point out the importance of mastering laparoscopic suturing. Although devices are commercially available to facilitate certain suturing scenarios, we encourage residents and fellows to sew manually. We believe that none of these operations could have been completed as effectively by using a suture device. The ability to suture laparoscopically markedly broadens the number of clinical scenarios in which minimal access techniques can be used.


Factors affecting eco-efficiency of municipal waste services in Tuscan municipalities: An empirical investigation of different management models.
The efficient provision of municipal solid waste (MSW) services is essential from an economic and environmental perspective. This paper investigates the effect of ownership type on the eco-efficiency of MSW management in Tuscan municipalities. In the first stage of the analysis, the authors use a data envelopment analysis metafrontier approach with the integration of unsorted waste as an undesirable output. Three different clusters of municipalities are created based on the ownership type of the municipal waste operators: public, private, and mixed. In the second stage of analysis, the paper investigates factors affecting eco-efficiency in order to provide new knowledge that can be used by policy and decision makers to improve eco-efficiency. The results show that eco-efficiency is higher for municipalities that entrust the delivery of waste services to publicly owned firms compared to municipalities with delivery by mixed and private firms. Moreover, eco-efficiency in MSW management is higher for municipalities with younger residents, a larger population, and an overall higher population density. Smaller municipalities and those with less tourism manage their waste services more eco-efficiently than do bigger municipalities and those with more tourism. The results indicate that policy-makers should improve communication and engagement activities with older residents in smaller, less densely populated areas. In municipalities featuring heavy tourism, eco-taxes could be used to support these activities, avoiding any impact on residents.


Mutual interplay between cognitive offloading and secondary task performance.
Various modern tools, such as smartphones, allow for cognitive offloading (i.e., the externalization of cognitive processes). In this study, we examined the use and consequences of cognitive offloading in demanding situations in which people perform multiple tasks concurrently-mimicking the requirements of daily life. In a preregistered study, we adapted the dual-task paradigm so that one of the tasks allowed for cognitive offloading. As a primary task, our participants (N = 172) performed the pattern copy task-a highly demanding working memory task that allows for offloading at various degrees. In this task, we manipulated the temporal costs of offloading. Concurrently, half of the participants responded to a secondary N-back task. As our main research question, we investigated the impact of offloading behavior on secondary task performance. We observed that more pronounced offloading in the condition without temporal costs was accompanied by a more accurate performance in the N-back task. Furthermore, the necessity to respond to the N-back task increased offloading behavior. These results suggest an interplay between offloading and secondary task performance: in demanding situations, individuals increasingly use cognitive offloading, which releases internal resources that can then be devoted to improving performance in other, concurrent tasks.


Expansion of a single transposable element family is associated with genome-size increase and radiation in the genus Hydra.
Transposable elements are one of the major contributors to genome-size differences in metazoans. Despite this, relatively little is known about the evolutionary patterns of element expansions and the element families involved. Here we report a broad genomic sampling within the genus Hydra, a freshwater cnidarian at the focal point of diverse research in regeneration, symbiosis, biogeography, and aging. We find that the genome of Hydra is the result of an expansion event involving long interspersed nuclear elements and in particular a single family of the chicken repeat 1 (CR1) class. This expansion is unique to a subgroup of the genus Hydra, the brown hydras, and is absent in the green hydra, which has a repeat landscape similar to that of other cnidarians. These features of the genome make Hydra attractive for studies of transposon-driven genome expansions and speciation.


TNF-α is responsible for the contribution of stromal cells to osteoclast and odontoclast formation during orthodontic tooth movement.
Compressive force during orthodontic tooth movement induces osteoclast formation in vivo. TNF-α plays an important role in mouse osteoclast formation and bone resorption induced by compressive force during orthodontic tooth movement. Stromal cells, macrophages and T cells take part in TNF-α-induced osteoclast formation in vitro. Root resorption caused by odontoclasts is a major clinical problem during orthodontic tooth movement. In this study, we determined the cell type targeted by TNF-α during compressive-force-induced osteoclast and odontoclast formation to elucidate the mechanism of bone and root resorption in vivo. An orthodontic tooth movement mouse model was prepared with a nickel-titanium closed coil spring inserted between the maxillary incisors and the first molar. Using TNF receptor 1- and 2-deficient (KO) mice, we found that osteoclast and odontoclast formation was mediated by TNF-α in orthodontic tooth movement. We generated four types of chimeric mice: wild-type (WT) bone marrow cells transplanted into lethally irradiated WT mice (WT>WT), KO bone marrow cells transplanted into lethally irradiated WT mice (KO>WT), WT bone marrow cells transplanted into lethally irradiated KO mice (WT>KO), and KO marrow cells transplanted into lethally irradiated KO mice (KO>KO). Using anti-CD4 and anti-CD8 antibodies, T cells were eliminated from these mice. We subjected these chimeric mice to orthodontic tooth movement. Orthodontic tooth movement was evaluated and tartrate-resistant acid phosphatase-positive cells along the alveolar bone (osteoclasts) and along the tooth root (odontoclasts) were counted after 12 days of tooth movement. The amount of orthodontic tooth movement, and the number of osteoclasts and odontoclasts on the compression side were significantly lower in WT>KO and KO>KO mice than in WT>WT and KO>WT mice. According to these results, we concluded that TNF-α-responsive stromal cells are important for osteoclast and odontoclast formation during orthodontic tooth movement.


Improved Functional Outcome After Peripheral Nerve Stimulation of the Impaired Forelimb Post-stroke.
Lack of blood flow to the brain, i.e., ischemic stroke, results in loss of nerve cells and therefore loss of function in the effected brain regions. There is no effective treatment to improve lost function except restoring blood flow within the first several hours. Rehabilitation strategies are widely used with limited success. The purpose of this study was to examine the effect of electrical stimulation on the impaired upper extremity to improve functional recovery after stroke. We developed a rodent model using an electrode cuff implant onto a single peripheral nerve (median nerve) of the paretic forelimb and applied daily electrical stimulation. The skilled forelimb reaching test was used to evaluate functional outcome after stroke and electrical stimulation. Anterograde axonal tracing from layer V pyramidal neurons with biotinylated dextran amine was done to evaluate the formation of new neuronal connections from the contralesional cortex to the deafferented spinal cord. Rats receiving electrical stimulation on the median nerve showed significant improvement in the skilled forelimb reaching test in comparison with stroke only and stroke with sham stimulation. Rats that received electrical stimulation also exhibited significant improvement in the latency to initiate adhesive removal from the impaired forelimb, indicating better sensory recovery. Furthermore, axonal tracing analysis showed a significant higher midline fiber crossing index in the cervical spinal cord of rats receiving electrical stimulation. Our results indicate that direct peripheral nerve stimulation leads to improved sensorimotor recovery in the stroke-impaired forelimb, and may be a useful approach to improve post-stroke deficits in human patients.


Formamide as the main building block in the origin of nucleic acids.
The simplest molecules grouping the four most common elements of the universe H,C,O and N (with the exception of the biologically inert He) are isocyanate HNCO and formamide H2NCOH. Reasons for the availability of formamide on prebiotic Earth are presented. We review evidence showing that formamide in the presence of largely available catalysts and by moderate heating yields the complete set of nucleic bases necessary for the formation of nucleic acids. Formamide also favours the formation of acyclonucleosides and the phosphorylation and trans-phosphorylation of nucleosides, thus providing a plausible chemical frame for the passage from a simple one-carbon compound to nucleic polymers. Physico-chemical conditions exist in which formamide favours the stability of the phosphoester bonds in nucleic polymers relative to that of the same bonds in monomers. Starting from a formamide-laden environment subject only to the laws of chemistry, a hypothesis is outlined sketching the passage towards an aqueous world in which Darwinian rules apply.


Genetic alterations and their relationship in the phosphatidylinositol 3-kinase/Akt pathway in thyroid cancer.
To investigate the overall occurrence and relationship of genetic alterations in the phosphatidylinositol 3-kinase (PI3K)/Akt pathway in thyroid tumors and explore the scope of this pathway as a therapeutic target for thyroid cancer. We examined collectively the major genetic alterations and their relationship in this pathway, including PIK3CA copy number gain and mutation, Ras mutation, and PTEN mutation, in a large series of primary thyroid tumors. Occurrence of any of these genetic alterations was found in 25 of 81 (31%) benign thyroid adenoma (BTA), 47 of 86 (55%) follicular thyroid cancer (FTC), 21 of 86 (24%) papillary thyroid cancer (PTC), and 29 of 50 (58%) anaplastic thyroid cancer (ATC), with FTC and ATC most frequently harboring these genetic alterations. PIK3CA copy gain was associated with increased PIK3CA protein expression. A mutual exclusivity among these genetic alterations was seen in BTA, FTC, and PTC, suggesting an independent role of each of them through the PI3K/Akt pathway in the tumorigenesis of the differentiated thyroid tumors. However, coexistence of these genetic alterations was increasingly seen with progression from differentiated tumor to undifferentiated ATC. Their coexistence with BRAF mutation was also frequent in PTC and ATC. The data provide strong genetic implication that aberrant activation of PI3K/Akt pathway plays an extensive role in thyroid tumorigenesis, particularly in FTC and ATC, and promotes progression of BTA to FTC and to ATC as the genetic alterations of this pathway accumulate. Progression of PTC to ATC may be facilitated by coexistence of PI3K/Akt pathway-related genetic alterations and BRAF mutation. The PI3K/Akt pathway may thus be a major therapeutic target in thyroid cancers.


Improved Detection of Molecular Markers of Atherosclerotic Plaques Using Sub-Millimeter PET Imaging.
Since atherosclerotic plaques are small and sparse, their non-invasive detection via PET imaging requires both highly specific radiotracers as well as imaging systems with high sensitivity and resolution. This study aimed to assess the targeting and biodistribution of a novel fluorine-18 anti-VCAM-1 Nanobody (Nb), and to investigate whether sub-millimetre resolution PET imaging could improve detectability of plaques in mice. The anti-VCAM-1 Nb functionalised with the novel restrained complexing agent (RESCA) chelator was labelled with [18F]AlF with a high radiochemical yield (>75%) and radiochemical purity (>99%). Subsequently, [18F]AlF(RESCA)-cAbVCAM1-5 was injected in ApoE-/- mice, or co-injected with excess of unlabelled Nb (control group). Mice were imaged sequentially using a cross-over design on two different commercially available PET/CT systems and finally sacrificed for ex vivo analysis. Both the PET/CT images and ex vivo data showed specific uptake of [18F]AlF(RESCA)-cAbVCAM1-5 in atherosclerotic lesions. Non-specific bone uptake was also noticeable, most probably due to in vivo defluorination. Image analysis yielded higher target-to-heart and target-to-brain ratios with the β-CUBE (MOLECUBES) PET scanner, demonstrating that preclinical detection of atherosclerotic lesions could be improved using the latest PET technology.


Learning from cerebrospinal fluid drug-resistant HIV escape-associated encephalitis: a case report.
In the era of antiretroviral therapy (ART), central nervous system (CNS) complications in patients with human immunodeficiency virus (HIV) infection are sometimes associated with cerebrospinal fluid (CSF) viral escape. Here, we reported a case of persistent CNS viral escape with recurrent symptomatic encephalitis, which had ultimate stabilization achieved by a combination of ART adjustment and corticosteroids. A 27-year-old man with HIV infection complained of recurrent headaches during the last year. His magnetic resonance imaging (MRI) presented diffused bilateral white matter lesions, and laboratory tests confirmed elevated CSF protein level, lymphocytic pleocytosis, and detectable CSF HIV RNA (774 copies/mL). Plasma HIV RNA was well suppressed with tenofovir, lamivudine, and lopinavir/ritonavir. Prednisone 60 mg once daily was initiated to reduce intracranial inflammation, followed by a good clinical response, with CSF HIV RNA still detectable (31.1 copies/mL). During the gradual tapering of prednisone, his headache relapsed, and booming viral loads were detected in both CSF (4580 copies/mL) and plasma (340 copies/mL) with consistent drug-resistant mutations. Thereupon, prednisone was resumed and the ART regimen was switched to zidovudine, lamivudine, and dolutegravir according to drug resistance tests. Persistent clinical recovery of symptoms, neuroimaging, and laboratory abnormalities were observed in the follow-up visits. CSF and plasma HIV RNA and further drug resistance tests should be monitored in HIV-infected patients with neurologic symptoms, as opportunistic infections or tumors can be ruled out. ART optimization using a sensitive regimen may be crucial for addressing CSF viral escape and the related encephalitis.


The association between coronary microvascular dysfunction and carotid intima media thickness in patients with cardiac syndrome X.
The aim of this study was to evaluate in patients with cardiac syndrome X (CSX), using validated angiography indices, coronary blood flow and myocardial perfusion of the microcirculation to assess whether there is greater microvascular dysfunction in patients with increase of carotid intima media thickness (C-IMT), compared to those who do not have. Our study was performed on a population 124 patients with CSX that underwent coronary angiography and carotid ultrasound. We divided the sample into two categories: patients with increase of C-IMT and those without increase. We calculated Gibson and Yusuf indices for each patient based on angiographic images, including TIMI Frame Count (TFC), Myocardial Blush Grade (MBG) and Total Myocardial Blush Score (TMBS). Our sample compared two groups: patients with increase of C-IMT (n-63) and patients without increase of C-IMT (n-61). We showed that patients with increase C-IMT had a longest TFC of three major coronary arteries (TFC LAD 44.7±12.5; TFC RCA 26.2±6.9; TFC CX 27±5.9), than control group. We found lower MBG on three coronary arteries (MBG LAD 2.5±0.3; MBG RCA 2.3±0.3; MBG CX 2.1±0.32) in patients with increase of C-IMT than control group, with good statistical significance. Analysis of microcirculation trough angiography indexes in patients with CSX with increase of C-IMT and without has led to asses that patients with increase of C-IMT population has a greater involvement of microcirculation than patients without.


Immunocytochemical localization of beta-hexosaminidase and electron-microscopic characterization of human fibroblasts following treatment with monensin and nigericin.
Immunocytochemical localization of beta-hexosaminidase in cultured human skin fibroblasts was performed in the presence or absence of the Na+/K+ ionophores monensin and nigericin. In the presence of monensin, beta-hexosaminidase accumulated in the periphery of swollen vesicles in the paranuclear region of fibroblasts from normal individuals and from patients with mucolipidosis II. Nigericin-treated cells had more extensive vacuolization of the cytoplasm and the localization of the enzyme was more diffuse within these vacuoles. Morphological studies at the ultrastructral level indicated that a perturbation of the Golgi region occurred during ionophore treatment. These findings suggest that beta-hexosaminidase in ionophore-treated fibroblasts is trapped in a time- and dose-dependent manner in the paranuclear region presumed to be the swollen cisternae of the Golgi region, or adjacent vesicles derived from the Golgi region. Furthermore, fibroblasts are more sensitive to perturbation by nigericin than by monensin at similar concentrations and exposure times. These data support biochemical findings that the two ionophores differentially inhibit the transport of lysosomal enzymes in the Golgi region.


Lack of chemoprevention of dietary Agaricus blazei against rat colonic aberrant crypt foci.
The mushroom Agaricus blazei (Ab) has been widely used in folk medicine to treat various diseases including cancer. No information is available on its possible protective effects on the development of colon cancer. The potential blocking effect of Ab intake on the initiation stage of colon carcinogenesis was investigated in a short-term (4-week) bioassay using aberrant crypt foci (ACF) as biomarker. Male Wistar rats were given four subcutaneous injections of the carcinogen 1,2-dimethylhydrazine (DMH, 40 mg/kg bw, twice a week), during 2 weeks to induce ACF. The diet containing Ab at 5% was given 2 weeks before and during carcinogen treatment to investigate the potential beneficial effects of this edible mushroom on DMH-induced ACF. All groups were killed at the end of the fourth week. The colons were analyzed for ACF formation in 1% methylene blue whole-mount preparations and for cell proliferation in histological sections immunohistochemically stained for the proliferating cell nuclear antigen (PCNA). All DMH-treated rats developed ACF mainly in the middle and distal colon. Agaricus blazei intake at 5% did not alter the number of ACF induced by DMH or the PCNA indices in the colonic mucosa. Thus, the results of the present study did not confirm a chemopreventive activity of Ab on the initiation stage of rat colon carcinogenesis.


A Novel ASK Inhibitor AGI-1067 Inhibits TLR-4-Mediated Activation of ASK1 by Preventing Dissociation of Thioredoxin from ASK1.
The cell type that normally limits the inflammatory and atherosclerotic process is the vascular endothelial cell (EC) that can be regulated by proinflammatory and various stresses. Toll-like receptor-4 (TLR4) plays an important role in the pathogenesis of atherosclerosis, in part, by activating apoptosis signal-regulating kinase 1 (ASK1) to initiate the activation of MAP kinases pathways and the expression of inflammatory genes. In the present study, we test the hypothesis that AGI-1067 acts as an anti-inflammatory agent by inhibiting the activation of ASK1 in human EC. Pretreatment of human aortic endothelial cells with AGI-1067 inhibits TLR4 ligand (LPS)-induced activation of ASK1 and the downstream p38 and c-Jun N-terminal kinase (JNK) MAP kinases. LPS dissociates two endogenous inhibitors thioredoxin-1 (Trx1) and 14-3-3 from ASK1, leading to ASK1 autoactivation. Interestingly, AGI-1067 inhibits the dissociation of Trx1, but not 14-3-3, from ASK1. However, inhibition of Trx1 dissociation from ASK1 by AGI-1067 is sufficient to suppress LPS-mediated phosphorylation of the transcription factors c-Jun and activating transcription factor 2, and inhibit LPS-induced inflammatory genes including vascular cell adhesion molecule 1, E-selectin, IL-6 and monocyte chemoattractant protein 1. Our findings suggest that AGI-1067 as a unique ASK1 inhibitor to inhibit TLR4-mediated ASK1 activation, contributing to its anti-inflammatory properties.


Milk production in sows from a teat in second parity is influenced by whether it was suckled in first parity.
The impacts of a teat being suckled or not in first parity on its development, gene expression, and milk yield in the next parity were studied. Forty-seven first-parity sows (Sus scrofa) were divided into 2 groups: i) the same teats suckled in 2 subsequent lactations (controls, CTL; n = 22); and ii) different teats suckled in 2 subsequent lactations (treated, TRT; n = 25). In the first lactation, over half of the teats (Teats 1, 2, 5, 6, and 7 from 1 side of the udder, and Teats 3, 4, and 7 from the other side) were sealed with tape so that they were nonfunctional. During the next lactation, the CTL group had the same teats sealed as in the first lactation, whereas the opposite teats were sealed for the TRT group. In both parities, litters were standardized to 7 piglets around birth and to 6 piglets (1 piglet per available teat) at 48 h postpartum. During the second lactation, piglets were weighed at birth and on d 2, 4, 7, 14, 21, 35, and 56 postpartum. Weaning was at 17 ± 1 d of age. Behavioral measures were obtained (using 24-h video recording) on d 3 and d 10 of lactation on 15 sows per treatment to evaluate satiety of piglets, using aggressiveness and nursing behavior as indicators. At weaning in the second lactation, 16 sows per treatment were slaughtered and 4 functional mammary glands were collected for compositional analyses and parenchyma from 2 nonfunctional glands was collected to measure mRNA abundance for selected genes. Piglets from CTL sows weighed 1.12 kg more than piglets from TRT sows (P < 0.05) on d 56, and functional mammary glands from CTL sows contained more parenchymal tissue, more DNA, and more RNA (P < 0.01) than those from TRT sows. The relative mRNA abundance of prolactin in parenchymal tissue tended to be greater in CTL than TRT sows (P < 0.10). Behavioral measures indicated a greater hunger level for piglets using teats that were not previously suckled. Current findings clearly show that teats that were suckled in first lactation produce more milk and have a greater development in the second lactation than nonsuckled teats.


Antigenic competition: cellular or humoral.
The injection of one antigen into mice inhibited the response to a second when 1 to 10 days separated the two injections. When the same type of inhibition was attempted in gamma-irradiated mice reconstituted with normal spleen cells, the inhibition was greater in mice receiving 50 million spleen cells than in those receiving 10 million. The results are interpreted as favoring a humoral mechanism of inhibition.


An Integrative Multi-Omics Workflow to Address Multifactorial Toxicology Experiments.
Toxicology studies can take advantage of omics approaches to better understand the phenomena underlying the phenotypic alterations induced by different types of exposure to certain toxicants. Nevertheless, in order to analyse the data generated from multifactorial omics studies, dedicated data analysis tools are needed. In this work, we propose a new workflow comprising both factor deconvolution and data integration from multiple analytical platforms. As a case study, 3D neural cell cultures were exposed to trimethyltin (TMT) and the relevance of the culture maturation state, the exposure duration, as well as the TMT concentration were simultaneously studied using a metabolomic approach combining four complementary analytical techniques (reversed-phase LC and hydrophilic interaction LC, hyphenated to mass spectrometry in positive and negative ionization modes). The ANOVA multiblock OPLS (AMOPLS) method allowed us to decompose and quantify the contribution of the different experimental factors on the outcome of the TMT exposure. Results showed that the most important contribution to the overall metabolic variability came from the maturation state and treatment duration. Even though the contribution of TMT effects represented the smallest observed modulation among the three factors, it was highly statistically significant. The MetaCore™ pathway analysis tool revealed TMT-induced alterations in biosynthetic pathways and in neuronal differentiation and signaling processes, with a predominant deleterious effect on GABAergic and glutamatergic neurons. This was confirmed by combining proteomic data, increasing the confidence on the mechanistic understanding of such a toxicant exposure.


The relative strengths of rapid and delayed density dependence acting on a terrestrial herbivore change along a pollution gradient.
Animal populations vary in response to a combination of density-dependent and density-independent forces, which interact to drive their population dynamics. Understanding how abiotic forces mediate the form and strength of density-dependent processes remains a central goal of ecology, and is of increasing urgency in a rapidly changing world. Here, we report for the first time that industrial pollution determines the relative strength of rapid and delayed density dependence operating on an animal population. We explored the impacts of pollution and climate on the population dynamics of an eruptive leafmining moth, Phyllonorycter strigulatella, around a coal-fired power plant near Apatity, north-western Russia. Populations were monitored at 14 sites over 26 years. The relative strengths of rapid and delayed density dependence varied with distance from the power plant. Specifically, the strength of rapid density dependence increased while the strength of delayed density dependence decreased with increasing distance from the pollution source. Paralleling the increasing strength of rapid density dependence, we observed declines in the densities of P. strigulatella, increases in predation pressure from birds and ants, and declines in an unknown source of mortality (perhaps plant antibiosis) with increasing distance from the power plant. In contrast to the associations with pollution, associations between climate change and leafminer population densities were negligible. Our results may help to explain the outbreaks of insect herbivores that are frequently observed in polluted environments. We show that they can result from the weakening of rapid (stabilizing) density dependence relative to the effects of destabilizing delayed density dependence. Moreover, our results may explain some of the variation reported in published studies of animal populations in polluted habitats. Variable results may emerge in part because of the location of the study sites on different parts of pollution gradients. Finally, in a rapidly changing world, effects of anthropogenic pollution may be as, or more, important than are effects of climate change on the future dynamics of animal populations.


Survival of Burkholderia pseudomallei and Pathogenic Leptospira in Cola, Beer, Energy Drinks, and Sports Drinks.
Burkholderia pseudomallei and pathogenic Leptospira in contaminated drinking water can cause melioidosis and leptospirosis, respectively. Here, we evaluated their survival in beverages. We mixed six isolates (three isolates per organism) in four beverages (Coca-Cola®, Red Bull®, Singha® beer, and Gatorade®) and distilled water as the control at two final concentrations (1 × 107 colony-forming units [CFU]/mL and 1 × 103 CFU/mL). The solution was kept at two temperatures (37°C and 4°C). At 4°C and at the high concentration, pathogenic Leptospira survived in Coca-Cola® up to 3 minutes and in Singha, Red Bull®, and Gatorade up to 15 minutes, whereas B. pseudomallei survived in these beverages up to 8 hours, and 14, 14, and 28 days, respectively. The survival time of both organisms was shorter at 37°C (P = 0.01) and at the lower concentration (P = 0.001). In conclusion, Leptospira can survive in some beverages for up to 15 minutes, whereas B. pseudomallei can survive in some beverages for up to 4 weeks.


Yeast as a drug discovery platform in Huntington's and Parkinson's diseases.
The high degree of conservation of cellular and molecular processes between the budding yeast Saccharomyces cerevisiae and higher eukaryotes have made it a valuable system for numerous studies of the basic mechanisms behind devastating illnesses such as cancer, infectious disease, and neurodegenerative disorders. Several studies in yeast have already contributed to our basic understanding of cellular dysfunction in both Huntington's and Parkinson's disease. Functional genomics approaches currently being undertaken in yeast may lead to novel insights into the genes and pathways that modulate neuronal cell dysfunction and death in these diseases. In addition, the budding yeast constitutes a valuable system for identification of new drug targets, both via target-based and non-target-based drug screening. Importantly, yeast can be used as a cellular platform to analyze the cellular effects of candidate compounds, which is critical for the development of effective therapeutics. While the molecular mechanisms that underlie neurodegeneration will ultimately have to be tested in neuronal and animal models, there are several distinct advantages to using simple model organisms to elucidate fundamental aspects of protein aggregation, amyloid toxicity, and cellular dysfunction. Here, we review recent studies that have shown that amyloid formation by disease-causing proteins and many of the resulting cellular deficits can be faithfully recapitulated in yeast. In addition, we discuss new yeast-based techniques for screening candidate therapeutic compounds for Huntington's and Parkinson's diseases.


Pupil Mimicry is the Result of Brightness Perception of the Iris and Pupil.
Recent scientific investigations suggest that people automatically mimic each other's pupil sizes during interaction. However, instead of being a social mimicry effect, it could also be the result of brightness perception. When observers look at individuals with dilated pupils, little of the brighter iris is visible, leading to the perception of a relatively low-illuminated eye region. In the current study we tested whether pupil mimicry remains present when pupils and irises are equalized for luminance values across pupil sizes. We tested several stimulus sets, including faces with static pupils that varied in size across images and dynamic pupils that changed in size over time in videos. Results showed that for traditional, not-luminance-equalized videos, participants' pupil sizes adapted to the observed pupils, showing a pattern that is roughly in line with pupil mimicry. However, no such pupil response in line with mimicry was seen for static images (regardless of whether they were equalized for luminance) nor for luminance-equalized videos. These findings suggest that only salient, dynamic stimuli attract enough attention to the luminance in the eye region to evoke a pupillary response. However, although such responses suggest pupil mimicry, the underlying factor is the change in brightness within the eye as a function of pupil size rather than social mimicry.


[Acute lesions of the gastric mucosa (LAMG)].
Fifty-four patients with acute lesions of the gastric mucosa (ALGM) were studied, specially from the clinical point of view and of the gastric secretion. It was found that the ALGM were fundamentally located in the gastric body and that in the majority of cases the maximum acid output obtained through the Histamine stimulation, was either increased or normal. Even though the dispeptic symptoms predominated, an important group of cases had ulcer symptoms.


Radiofrequency bipolar coagulation for radical hysterectomy: technique, feasibility and complications.
This study describes the surgical technique and intra- and postoperative complications associated with the use of a radiofrequency bipolar coagulator in a series of 18 Piver type III-IV radical hysterectomies performed in cervical cancer patients. Preliminary vessel-by-vessel dissection of the lateral parametria was possible in 17 out of 18 (94%) cases, and a direct application of a radiofrequency bipolar coagulation instrument was performed to coagulate the posterior and anterior parametrial tissues in all cases. We were able to easily coagulate isolated vessels up to 5 mm of maximal diameter. In no case were clamps or hemoclips necessary to complete hemostasis. We did not observe any parametrial vessel damage or heat-related injury of the surrounding normal tissue. The median size of the parametria removed was 44 mm (range 31-58) and nodes were detected in 15 cases (83%). Median operative time and estimated blood loss for the whole procedure including systematic pelvic and aortic lymphadenectomy was 250 min (range 200-410) and 550 ml (range 400-2500), respectively. Median follow-up time was 9 months (range 5-13). No complications specifically related to the use of radiofrequency coagulation were found. In conclusion the radio-frequency coagulation with this instrument appears to be a safe technique that is particularly useful in reducing blood loss and operative time without affecting radicality in patients undergoing radical hysterectomy.


Functional Rating Index: literature review.
In 1999, a new self-report outcome measure, the Functional Rating Index (FRI), was developed and tested. This measure demonstrated reasonable reliability, validity and responsiveness. Since the publication of the original testing, numerous independent research teams have examined the psychometric qualities of the FRI and published their findings. The aim of this study is to review the psychometric properties of the FRI as reported by published studies. Researchers independently searched the literature for relevant studies using MEDLINE and Index to Chiropractic Literature from January 2001 to August 2009. Descriptive and outcome data was extracted using standardized forms. The psychometric properties of the FRI were analyzed with established criteria and compared head-to-head with other outcome measures. The search yielded ten studies that provided psychometric data. Reliability: Test-retest: Intraclass correlation coefficient and Cronbach's alpha were good, and FRI is equivalent to the other measures. the FRI has good convergent validity with pain and function self-report scales and a weaker correlation with items that measure different constructs. Responsiveness: FRI was similar to the comparative measures for standardized response mean, effect size and receiver operating curve statistics. Patient acceptability: time required by the patient and staff averaged 78 seconds per administration, and there were few missing responses. The FRI demonstrates favorable measurement properties of reliability, validity and responsiveness. It provides an alternative to other self-report measures, because it is quicker for a patient to complete and can be used for patients with neck and back pain.


Structure, Function and Pharmacology of SLC7 Family Members and Homologues.
Amino acids are essential components of all living cells serving as building blocks of proteins, as energy source, and as precursors of metabolites and signaling molecules. Amino acid transporters are membrane proteins that mediate the transfer of amino acids across the plasma membrane, and between compartments in cells, different cells and organs. The absence, overexpression or malfunction of specific amino acid transporters have been associated with human disease. One of the projects within the Swiss National Centre of Competence in Research (NCCR) TransCure was directed at SLC7 family amino acid transporters, with a particular focus on the heteromeric amino acid transporters 4F2hc-LAT1 (SLC3A2-SLC7A5) and 4F2hc-LAT2 (SLC3A2-SLC7A8), and the bacterial homologue AdiC. The project addressed questions of basic research (function and structure), pharmacology (identification of potent inhibitors and activators), and pre-clinical medicine (e.g., physiological role in the placenta) and disease models (e.g., tumor progression) of specific SLC7 family amino acid transporters. This review presents, summarizes and discusses selected main results obtained in this NCCR TransCure project.


Pancreas-specific ablation of beta1 integrin induces tissue degeneration by disrupting acinar cell polarity.
Integrin contact with basement membrane is a major determinant of epithelial cell polarity. beta1 integrin heterodimers are the primary receptors for basement membrane in pancreatic acinar cells, which function to synthesize and directionally secrete digestive enzymes into a central lumen. Aberrant acinar secretion and exposure of the parenchyma to digestive enzyme activity lead to organ damage and pancreatitis. beta1 integrin conditional knockout mice were crossed to Ptf1a-Cre mice to ablate beta1 integrin in the pancreas. Histopathology of aged and cerulein-treated mice were assessed by histology and immunocytochemistry. Directional secretion was determined in vitro by FM1-43 loading with cerulein stimulation. Pancreas-specific ablation of beta1 integrin led to progressive organ degeneration, associated with focal acinar cell necrosis and ductal metaplasia along with widespread inflammation and collagen deposition. beta1 Integrin-null pancreata were highly susceptible to cerulein-induced acute pancreatitis, displaying an enhanced level of damage with no loss in regeneration. Degenerating beta1 integrin-null pancreata were marked by disruption of acinar cell polarity. Protein kinase C epsilon, normally localized apically, was found in the cytoplasm where it can lead to intracellular digestive enzyme activation. beta1 Integrin-null acinar cells displayed indiscriminate secretion to all membrane surfaces, consistent with an observed loss of basolateral membrane localization of Munc18c, which normally prevents basal secretion of digestive enzymes. Ablation of beta1 integrin induces organ atrophy by disrupting acinar cell polarity and exposing the pancreatic parenchyma to digestive enzymes.


[High risk factors of lung cancer and its early detection].
High risk factors and early detection of lung cancer were studied in a population of greater than 45 years old of 10 122 in Lu Wen District. It was shown as follows: 1. The high risk factors of lung cancer were male greater than 50 years old, and with a smoking history of greater than 400 pack year. 2. Questionnaire 70-110 mm X-ray chest film and sputum cytological examination for the suspicious lung cancer were the screen test for the high risk group. Their prevalence rate of lung cancer was 4.74/1000, 50% of them was in p-staging I, it was 6.3 times higher, than in the specialized hospital, their 5-yr survival rate was 31.5%. It was higher than that in the general population.


Antioxidant Activity and Chemical Composition of Geranium Oil and Its Synergistic Potential against Pneumococci with Various Antibiotic Combinations.
The essential oil of Pelargonium graveolens L. is valuable for its therapeutic benefits, so this study aimed to determine the synergistic effect of the combination of the essential oil of this plant with antibiotics instead of the extracts prepared with various solvents. In addition, the second goal of this study was to determine whether the essential oil combined with various antibiotics increased the overall killing activity in mouse macrophage cells with the aim of introducing an immunotherapeutic approach to the infection treatments used today. Herein, the volatile profile of Geranium oil (G.O) was analyzed using GC/MS. The current study sought to assess the synergistic characteristics of several antibiotic combinations using G.O against pneumococci, as well as the oil's antioxidant and antimicrobial activities. The major components of the oil were citronellol, geraniol, and isomenthone. In the combinations of G.O and antibiotics, the synergism of the Streptococcus pneumoniae to antibiotics advanced. When the time-kill data were evaluated, G.O + antibiotic combinations quickly diminished the viable cell count of S. pneumoniae from the 6th h. In this study, the combined use of existing antibiotics used in infection treatments with G.O could improve antibiotic effectiveness and thus prevent bacteria from developing antibiotic resistance.


Detection of Incidental Prostate Cancer or Urothelial Carcinoma Extension in Urinary Bladder Cancer Patients by Using Multiparametric MRI: A Retrospective Study Using Prostate Imaging Reporting and Data System Version 2.0.
The study aimed to investigate the role of Prostate Imaging Reporting and Data System version 2 (PI-RADS v2) in predicting incidental prostate cancer (PCa) or urothelial carcinoma (UCa) extension in urinary bladder (UB) cancer patients. A total of 72 UB cancer patients who underwent radical cystoprostatectomy and 3 Tesla multiparametric MRI before surgery were enrolled. PI-RADS v2 ratings were assigned by two independent radiologists. All prostate specimens were examined by a single pathologist. We compared the multiparametric MRI findings rated using PI-RADS v2 with the pathologic data. Of the 72 UB cancer patients, 29 had incidental PCa (40.3%) and 20 showed UCa extension (27.8%), with an overlap for 3 patients. With a score of 4 as the cut-off value for predicting incidental PCa, the diagnostic accuracy was 65.3%, specificity was 90.7%, and positive predictive value (PPV) was 66.7%. The diagnostic accuracy for incidental UCa extension was 47.2%, specificity was 92.3%, and PPV was 83.3%. Despite the low diagnostic accuracy, the PPV and specificity were relatively high. Therefore, PI-RADS v2 scores of 1, 2, or 3 may help exclude the probability of incidental PCa or UCa extension.


Minimum data set necessary to promote the care of the elderly in general practice.
A preliminary data set was designed to allow uniform information collection, to initiate structured intervention and thereby to promote anticipatory care of the elderly. A questionnaire inviting comment on the data set was sent to 116 general practitioners in training practices and 40 consultants in geriatric medicine in the northern region and also to the 18 professors in general practice in the United Kingdom. The response rates from the three groups were 76%, 55% and 67% respectively.There was good agreement between the three groups regarding the content of the data set suggesting that it may be possible to introduce uniform information collection which in turn would standardize records of the elderly and aid computerization.The need for periodic screening of the elderly was considered necessary by 37% of general practitioners, 59% of consultants and 33% of professors. The cause of such low interest in screening and its effect on computerized care and surveillance of the elderly are discussed.


Carry-over effects of early thermal conditions on somatic and germline oxidative damages are mediated by compensatory growth in sticklebacks.
Most studies of climate change impacts focus on the effects of summer temperatures, which can immediately impact fitness of breeders, but winter temperatures are expected to have a greater impact on development and growth of animals with long-lasting consequences. Exposure to warmer temperatures can increase cellular oxidative damage in ectotherms. Yet, it is unknown whether thermal stress during early life has prolonged effects on oxidative status during adulthood. In an experiment using F1 fish originated from a wild three-spined stickleback population at the southern edge of its European distribution, we examined whether experimental thermal conditions experienced in winter had carry-over effects on oxidative status and telomere length, a marker of accumulated stress, in the soma and germline during adulthood. For this, oxidative DNA damage, enzymatic antioxidant activities and telomere length were measured three months after the termination of the temperature manipulation. In addition, we tested whether such delayed effects, if any, were due to individuals' compensatory growth after experiencing unfavourable growth conditions in winter. Warm acclimation during winter induced increased levels of oxidative DNA damage in muscle and sperm and increased enzymatic antioxidant defences in muscle during the breeding season. Telomere length of adult fish was not influenced by thermal conditions experienced during early life. Winter temperature manipulation influenced fish to alter the temporal pattern of growth trajectories across the juvenile and adult stages. Fish reared in warm winter conditions grew at a slower rate than the controls during the period of temperature manipulation then accelerated body mass gain to catch up during the breeding season. Faster somatic growth during the breeding season incurred a higher cost in terms of oxidative damage in the warm-treated individuals. For the first time, we experimentally show the long-lasting detrimental effects of thermal stress on and the positive link between catch-up growth and oxidative DNA damage in the soma and germline. Winter temperature increases due to climate change can reduce fertility and survival of fish by inducing catch-up growth. The detrimental effects of winter climate change may accumulate across generations through the pre-mutagenic DNA damage in the germline.


[A new approach to research graphical performance of MRI RF coil based on the quantitative comparison method].
To compare the performance of three different types of RF coil in 3.0T MRI, the signal-to-noise ratio (SNR) and image uniformity in 3.0 Tesla magnetic resonance imaging system have been studied. In addition to the measurement of SNR and Uniformity based on the protocol recommend by AAPM, the images had also been analyzed using SPSS and MATLAB to get the SNR distribution along two vertical direction and the whole imaging plane. The SNR with T/R birdcage coil and 8-ch surface coil is seven and twelve times higher than surface coil in the region of interest (ROI), which was set in the center of the phantom images. With regard to the spatial variation of sensitivity, the uniformity of T/R birdcage coil and 8-ch surface coil are also much higher than that of surface coil. The images from T/R birdcage and phased array coils are all better than those of surface coil. The SNR of the images from phased array coil is highest while the uniformity of the images from T/R birdcage coil is highest.


Attenuation of renal ischemia-reperfusion injury by FR167653 in dogs.
Inflammatory cytokines are known to contribute to ischemia-reperfusion injury. We investigated the effect of FR167653 (FR), a suppressor of interleukin-1beta and tumor necrosis factor-alpha, on ischemia-reperfusion injury of the kidney in dogs. The left kidney was subjected to ischemia for 60 minutes followed by removal of the right kidney. A control group (n = 10) and an FR group (n = 8) were evaluated for tissue blood flow; resistive index, pulsatility index, arterial oxygen pressure, serum creatinine, blood urea nitrogen, aspartate transaminase, and alanine transaminase levels; interleukin-1beta messenger RNA expression in the peripheral blood; apoptotic index; and histopathology. The FR group showed lower creatinine, serum urea nitrogen, aspartate transaminase, and alanine transaminase levels (P <.038 each) and lower interleukin-1beta mRNA expression and apoptotic index (P <.041 each) than did the control group. Arterial oxygen pressure during the 120 minutes after reperfusion in the FR group decreased but recovered quickly (P =.024). Renal tissue damage in the FR group was less than that in the control group (P =.036). FR ameliorates ischemia-reperfusion injury of the kidney potentially by reduced production of inflammatory cytokines that may contribute to damage to the ischemic kidney and the distant organs.


Lived experience and attitudes of people with plantar heel pain: a qualitative exploration.
Plantar heel pain is a common source of pain and disability. Evidence-based treatment decisions for people with plantar heel pain should be guided by the best available evidence, expert clinical reasoning, and consider the needs of the patient. Education is a key component of care for any patient and needs to be tailored to the patient and their condition. However, no previous work has identified, far less evaluated, the approaches and content required for optimal education for people with plantar heel pain. The aim of this study was to gather the patients' perspective regarding their lived experience, attitudes and educational needs in order to inform the content and provision of meaningful education delivery approaches. Using a qualitative descriptive design, semi-structured interviews were conducted with participants with a clinical diagnosis of plantar heel pain. A topic guide was utilised that focused on the experience of living with plantar heel pain and attitudes regarding treatment and educational needs. Interviews were audio recorded, transcribed verbatim and analysed using the Framework approach. Each transcription, and the initial findings, were reported back to participants to invite respondent validation. Eighteen people with plantar heel pain were interviewed. Descriptive analysis revealed eight themes including perceptions of plantar heel pain, impact on self, dealing with plantar heel pain, source of information, patient needs, patient unmet needs, advice to others and interest in online education. Participants revealed doubt about the cause, treatment and prognosis of plantar heel pain. They also expressed a desire to have their pain eliminated and education individually tailored to their condition and needs. Respondent validation revealed that the transcripts were accurate, and participants were able to recognise their own experiences in the synthesised themes. Plantar heel pain has a negative impact on health-related quality of life. Participants wanted their pain eliminated and reported that their expectations and needs were frequently unmet. Health professionals have an important role to be responsive to the needs of the patient to improve their knowledge and influence pain and behaviour. Our study informs the content needed to help educate people with plantar heel pain.


Differential mechanisms of posterior cingulate cortex downregulation and symptom decreases in posttraumatic stress disorder and healthy individuals using real-time fMRI neurofeedback.
Intrinsic connectivity networks, including the default mode network (DMN), are frequently disrupted in individuals with posttraumatic stress disorder (PTSD). The posterior cingulate cortex (PCC) is the main hub of the posterior DMN, where the therapeutic regulation of this region with real-time fMRI neurofeedback (NFB) has yet to be explored. We investigated PCC downregulation while processing trauma/stressful words over 3 NFB training runs and a transfer run without NFB (total n = 29, PTSD n = 14, healthy controls n = 15). We also examined the predictive accuracy of machine learning models in classifying PTSD versus healthy controls during NFB training. Both the PTSD and healthy control groups demonstrated reduced reliving symptoms in response to trauma/stressful stimuli, where the PTSD group additionally showed reduced symptoms of distress. We found that both groups were able to downregulate the PCC with similar success over NFB training and in the transfer run, although downregulation was associated with unique within-group decreases in activation within the bilateral dmPFC, bilateral postcentral gyrus, right amygdala/hippocampus, cingulate cortex, and bilateral temporal pole/gyri. By contrast, downregulation was associated with increased activation in the right dlPFC among healthy controls as compared to PTSD. During PCC downregulation, right dlPFC activation was negatively correlated to PTSD symptom severity scores and difficulties in emotion regulation. Finally, machine learning algorithms were able to classify PTSD versus healthy participants based on brain activation during NFB training with 80% accuracy. This is the first study to investigate PCC downregulation with real-time fMRI NFB in both PTSD and healthy controls. Our results reveal acute decreases in symptoms over training and provide converging evidence for EEG-NFB targeting brain networks linked to the PCC.


Hierarchical structure of the Sicilian goats revealed by Bayesian analyses of microsatellite information.
Genetic structure and relationship amongst the main goat populations in Sicily (Girgentana, Derivata di Siria, Maltese and Messinese) were analysed using information from 19 microsatellite markers genotyped on 173 individuals. A posterior Bayesian approach implemented in the program STRUCTURE revealed a hierarchical structure with two clusters at the first level (Girgentana vs. Messinese, Derivata di Siria and Maltese), explaining 4.8% of variation (amovaФ(ST) estimate). Seven clusters nested within these first two clusters (further differentiations of Girgentana, Derivata di Siria and Maltese), explaining 8.5% of variation (amovaФ(SC) estimate). The analyses and methods applied in this study indicate their power to detect subtle population structure.


Heterogeneity of tissue IL-17 and tight junction proteins expression demonstrated in patients with autoimmune thyroid diseases.
Th17 cells together with their hallmark cytokine interleukin (IL)-17 were identified as crucial contributing factors in the pathogenesis of thyroid autoimmunity. The cytokine-regulated tight junction (Tj) disruption is thought to be essential in the initiation and/or development of several diseases. Still, the role of IL-17 maintaining Tj integrity in autoimmune thyroid diseases (AITDs) has not yet been evaluated. We aimed to investigate integrity of the thyroid follicle by studying immunoexpression of cellular Tj - zonula occludens (ZO)-1 and claudin-1 proteins coupled to IL-17A and CD68 detection in AITD patients compared with controls.Thirty-five adult patients undergoing thyroidectomy and presenting 18 cases of Hashimoto thyroiditis (HT), 7 of Graves' disease (GD) as well as 10 subjects of colloid goiter without autoimmune component served as controls were enrolled in this study. An immunohistochemical analysis including IL-17A, ZO-1, claudin-1, and CD68 detection was performed in each case. The correlation of IL-17A with Tj and CD68 in patients with AITD was also analyzed.Apart from inflammatory cells, we evidenced a stronger expression level of IL17A in the thyroid follicular cells in HT patients when compared with GD or colloid goiter. A significant reduction of ZO-1 immunoreactivity was observed in the thyrocytes in HT patients, whereas no significant differences were found in claudin-1 expression in HT and GD compared with colloid goiter patients. A significantly higher number of thyroid follicles with CD68-positive cells was found in HT patients than that in patients with GD or colloid goiter. In HT patients, the expression of IL-17A in the follicular cells was positively correlated with CD68 immunopositivity, whereas no association with claudin-1 or ZO-1 expression was found. GD patients did not reveal any significant correlation of IL-17A with Tj and CD68.Strong overexpression of IL-17A observed in the thyroid epithelial cells is associated with the presence of intrafollicular CD68-positive cells in HT patients. We evidenced the changes in molecules of thyrocyte junctional complexes highlighting impairment of the thyroid follicle integrity in HT, but no association with IL-17A was found.


Metabolic remodeling: a pyruvate transport affair.
Metabolic remodeling is a major determinant for many cell fate decisions, and a switch from respiration to aerobic glycolysis is generally considered as a hallmark of cancer cell transformation. Pyruvate is a key metabolite at the major junction of carbohydrate metabolism between cytosolic glycolysis and the mitochondrial Krebs cycle. In this issue of The EMBO Journal, Bender et al show that yeast cells regulate pyruvate uptake into mitochondria, and thus its metabolic fate, by expressing alternative pyruvate carrier complexes with different activities.


Learning deep features for dead and living breast cancer cell classification without staining.
Automated cell classification in cancer biology is a challenging topic in computer vision and machine learning research. Breast cancer is the most common malignancy in women that usually involves phenotypically diverse populations of breast cancer cells and an heterogeneous stroma. In recent years, automated microscopy technologies are allowing the study of live cells over extended periods of time, simplifying the task of compiling large image databases. For instance, there have been several studies oriented towards building machine learning systems capable of automatically classifying images of different cell types (i.e. motor neurons, stem cells). In this work we were interested in classifying breast cancer cells as live or dead, based on a set of automatically retrieved morphological characteristics using image processing techniques. Our hypothesis is that live-dead classification can be performed without any staining and using only bright-field images as input. We tackled this problem using the JIMT-1 breast cancer cell line that grows as an adherent monolayer. First, a vast image set composed by JIMT-1 human breast cancer cells that had been exposed to a chemotherapeutic drug treatment (doxorubicin and paclitaxel) or vehicle control was compiled. Next, several classifiers were trained based on well-known convolutional neural networks (CNN) backbones to perform supervised classification using labels obtained from fluorescence microscopy images associated with each bright-field image. Model performances were evaluated and compared on a large number of bright-field images. The best model reached an AUC = 0.941 for classifying breast cancer cells without treatment. Furthermore, it reached AUC = 0.978 when classifying breast cancer cells under drug treatment. Our results highlight the potential of machine learning and computational image analysis to build new diagnosis tools that benefit the biomedical field by reducing cost, time, and stimulating work reproducibility. More importantly, we analyzed the way our classifiers clusterize bright-field images in the learned high-dimensional embedding and linked these groups to salient visual characteristics in live-dead cell biology observed by trained experts.


Steering droplets on substrates using moving steps in wettability.
Droplets move on substrates with a spatio-temporal wettability pattern as generated, for example, on light-switchable surfaces. To study such cases, we implement the boundary-element method to solve the governing Stokes equations for the fluid flow field inside and on the surface of a droplet and supplement it by the Cox-Voinov law for the dynamics of the contact line. Our approach reproduces the relaxation of an axisymmetric droplet in experiments, which we initiate by instantaneously switching the uniform wettability of a substrate quantified by the equilibrium contact angle. In a step profile of wettability the droplet moves towards higher wettability. Using a feedback loop to keep the distance or offset between step and droplet center constant, induces a constant velocity with which the droplet surfs on the wettability step. We analyze the velocity in terms of droplet offset and step width for typical wetting parameters. Moving instead the wettability step with constant speed, we determine the maximally possible droplet velocities under various conditions. The observed droplet speeds agree with the values from the feedback study for the same positive droplet offset.


Ginkgolic acid inhibits HIV protease activity and HIV infection in vitro.
Several HIV protease mutations, which are resistant to clinical HIV protease inhibitors (PIs), have been identified. There is a great need for second-generation PIs with different chemical structures and/or with an alternative mode of inhibition. Ginkgolic acid is a natural herbal substance and a major component of the lipid fraction in the nutshells of the Ginkgo biloba tree. The objective of this study was to determine whether ginkgolic acid could inhibit HIV protease activity in a cell free system and HIV infection in human cells. Purified ginkgolic acid and recombinant HIV-1 HXB2 KIIA protease were used for the HIV protease activity assay. Human peripheral blood mononuclear cells (PBMCs) were used for HIV infection (HIV-1SF162 virus), determined by a p24gag ELISA. Cytotoxicity was also determined. Ginkgolic acid (31.2 µg/ml) inhibited HIV protease activity by 60%, compared with the negative control, and the effect was concentration-dependent. In addition, ginkgolic acid treatment (50 and 100 µg/ml) effectively inhibited the HIV infection at day 7 in a concentration-dependent manner. Ginkgolic acid at a concentration of up to 150 µg/ml demonstrated very limited cytotoxicity. Ginkgolic acid effectively inhibits HIV protease activity in a cell free system and HIV infection in PBMCs without significant cytotoxicity. Ginkgolic acid may inhibit HIV protease through different mechanisms than current FDA-approved HIV PI drugs. These properties of ginkgolic acid make it a promising therapy for HIV infection, especially as the clinical problem of viral resistance to HIV PIs continues to grow.


Association of levels of lipoprotein Lp(a), plasma lipids, and other lipoproteins with coronary artery disease documented by angiography.
In a study of 307 white patients who underwent coronary angiography, the relationship of coronary artery disease (CAD) to plasma levels of lipoprotein Lp(a) and other lipid-lipoprotein variables was examined. Lp(a) resembles low-density lipoprotein (LDL) in several ways, but can be distinguished and quantified by electroimmunoassay. CAD was rated as present or absent and was also represented by a quantitative lesion score derived from estimates of stenosis in four major coronary vessels. Coronary lesion scores significantly correlated with Lp(a), total cholesterol, triglycerides, LDL cholesterol, and high-density lipoprotein (HDL) cholesterol levels by univariate statistical analysis. By multivariate analysis levels of Lp(a) were associated significantly and independently with the presence of CAD (p less than .02), and tended to correlate with lesion scores (p = .06). Among subgroups Lp(a) level was associated with CAD in women of all ages and in men 55 years old or younger. An apparent threshold for coronary risk occurred at Lp(a) lipoprotein mass concentrations of 30 to 40 mg/dl, corresponding to Lp(a) cholesterol concentrations of approximately 10 to 13 mg/dl. Plasma Lp(a) in white patients appears to be a major coronary risk factor with an importance approaching that of the level of LDL or HDL cholesterol.


CO2 activation by electrogenerated divalent samarium for aryl halide carboxylation.
The reductive carboxylation of aryl halides has been investigated using a samarium electrode as a sacrificial anode to yield the corresponding benzoic acids, providing a smooth strategy for CO2 activation. Carboxylation occurred after an efficient reduction of carbon dioxide mediated by an electrogenerated Sm(ii)-complex acting as a strong monoelectronic reductive reagent.


Chi3l3 induces oligodendrogenesis in an experimental model of autoimmune neuroinflammation.
In demyelinating diseases including multiple sclerosis (MS), neural stem cells (NSCs) can replace damaged oligodendrocytes if the local microenvironment supports the required differentiation process. Although chitinase-like proteins (CLPs) form part of this microenvironment, their function in this differentiation process is unknown. Here, we demonstrate that murine Chitinase 3-like-3 (Chi3l3/Ym1), human Chi3L1 and Chit1 induce oligodendrogenesis. In mice, Chi3l3 is highly expressed in the subventricular zone, a stem cell niche of the adult brain, and in inflammatory brain lesions during experimental autoimmune encephalomyelitis (EAE). We find that silencing Chi3l3 increases severity of EAE. We present evidence that in NSCs Chi3l3 activates the epidermal growth factor receptor (EGFR), thereby inducing Pyk2-and Erk1/2- dependent expression of a pro-oligodendrogenic transcription factor signature. Our results implicate CLP-EGFR-Pyk2-MEK-ERK as a key intrinsic pathway controlling oligodendrogenesis.


Health-Related Anxiety and Hypochondriacal Concerns in Medical Students: A Cross-Sectional Study From Pakistan.
Phenomenon: Transient health-related anxiety/hypochondriacal concerns in medical students are well documented. The literature suggests that after studying a particular disease, medical students are likely to consider any symptoms earlier regarded as normal to be signs of the disease they are studying. The aim of this study was to investigate the prevalence of these phenomena and their cognitive and distress aspects among medicals students in Karachi, Pakistan. This was an analytical, cross-sectional study. Self-administered questionnaires comprising demographic details, the Short Health Anxiety Inventory, Medical Students' Disease (MSD) Perception Scale, and MSD Distress Scale were distributed to 1st- through 5th-year medical students. In total, 513 medical students (66% female) participated. Their mean age was 21 ± 1.6 years. Three hundred seventy-five students (73%) reported having visited a doctor at least once in the past 6 months. Fifty students (9.9%) admitted to having addictions. The overall prevalence of significant hypochondriacal concerns was 11.9% (61 students). The presence of addiction was associated with a greater likelihood of developing significant health-related anxiety (odds ratio = 3.82, p = .003), 95% confidence interval [1.51, 7.11]. Age, gender, medical school, year of medical school, and visits to the doctor in the previous 6 months were not associated with greater likelihood of developing significant health-related anxiety. Second-year medical students experienced a significantly greater degree of worry (MSD-Distress scale) than 5th-year students (M score = 12.6 ± 4.6 vs. 10.7 ± 4.4, p = .04). Insights: The prevalence of substantial hypochondriacal concerns in medical students in Pakistan was low in comparison to similar studies published in literature. Student health physicians should be aware of the true prevalence of hypochondriacal concerns and behavior and not dismiss legitimate complaints. Educational sessions to counteract this phenomenon can be incorporated into the curriculum of undergraduate medicine. By defining heightened awareness of symptoms as a normal process, different coping techniques can be discussed to help medical students reduce their level of stress.


A principal components analysis of the abbreviated Desires for Alcohol Questionnaire (DAQ).
The aim of this study was to examine the abbreviated Desires for Alcohol Questionnaire (DAQ) with respect to component structure and concurrent validity. The DAQ was administered to 2,960 adults participating in the Collaborative Studies on the Genetics of Alcohol. Rotated principal components analysis was conducted on 1,500 subjects with an alcohol-use disorder (AUD) and on 1,460 non-AUD subjects. Total DAQ scores were compared for these two subsamples. In addition, correlations were computed between DAQ scores and the following: (1) a sum of alcohol symptoms, and (2) endorsement of a single interview craving question. Similar solutions emerged in the AUD and non-AUD subsamples, with dimensions characterized by (1) strong desires/intentions to drink, (2) negative reinforcement, and (3) positive reinforcement + ability to control drinking. Each component was significantly correlated with the alcohol symptom scale in both subsamples (r(s) = .25-.64 and .31-.40, respectively, p < .0001) and with the interview craving item in the AUD subsample (r(s) = .22-.55, p < .0001). Total DAQ score was significantly higher for AUD subjects (40.5) than for non-AUD subjects (23.1, p < .0001) and exhibited significant correlations with the alcohol symptom scale in the AUD and non-AUD subsamples (r(s) = .61 and .39, respectively, p < .0001) and with the interview craving item in the AUD subsample (r(s) = .51, p < .0001). The DAQ is an appropriate measure of alcohol craving, as demonstrated by similar component structures across two samples as well as its concurrent validity.


Proteomic profiling of membrane vesicles from Mycobacterium avium subsp. paratuberculosis: Navigating towards an insilico design of a multi-epitope vaccine targeting membrane vesicle proteins.
Bacteria typically produce membrane vesicles (MVs) at varying levels depending on the surrounding environments. Gram-negative bacterial outer membrane vesicles (OMVs) have been extensively studied for over 30 years, but MVs from Gram-positive bacteria only recently have been a focus of research. In the present study, we isolated MVs from Mycobacterium avium subsp. paratuberculosis (MAP) and analyzed their protein composition using LC-MS/MS. A total of 316 overlapping proteins from two independent preparations were identified in our study, and topology prediction showed these cargo proteins have different subcellular localization patterns. When MVs were administered to bovine-derived macrophages, significant up-regulation of pro-inflammatory cytokines was observed via qRT-PCR. Proteome functional annotation revealed that many of these proteins are involved in the cellular protein metabolic process, tRNA aminoacylation, and ATP synthesis. Secretory proteins with high antigenicity and adhesion capability were mapped for B-cell and T-cell epitopes. Antigenic, Immunogenic and IFN-γ inducing B-cell, MHC-I, and MHC-II epitopes were stitched together through linkers to form multi-epitope vaccine (MEV) construct against MAP. Strong binding energy was observed during the docking of the 3D structure of the MEV with the bovine TLR2, suggesting that the putative MEV may be a promising vaccine candidate against MAP. However, in vitro and in vivo analysis is required to prove the immunogenic concept of the MEV which we will follow in our future studies. SIGNIFICANCE: Johne's disease is a chronic infection caused by Mycobacterium avium subsp. paratuberculosis that has a potential link to Crohn's disease in humans. The disease is characterized by persistent diarrhea and enteritis, resulting in significant economic losses due to reduced milk yield and premature culling of infected animals. The dairy industry in the United States alone experiences losses of approximately USD 250 million due to Johne's disease. The current vaccine against Johne's disease is limited by several factors, including variable efficacy, limited duration of protection, interference with diagnostic tests, inability to prevent infection, and logistical and cost-related challenges. Nevertheless, a multiepitope vaccine design approach targeting M. avium subsp. paratuberculosis has the potential to overcome these challenges and offer improved protection against Johne's disease.


Bortezomib treatment and regulatory T-cell depletion enhance the antitumor effects of adoptively infused NK cells.
Ligation of inhibitory receptors renders natural killer (NK) cells inactive against autologous tumors. Recently, the proteasome inhibitor bortezomib was shown to sensitize tumors to autologous NK-cell cytotoxicity in vitro. Here, we show bortezomib augments the antitumor effects of syngeneic NK-cell infusions in tumor-bearing animals; this effect is further enhanced in regulatory T cell (Treg cell)-depleted hosts. In vitro, bortezomib-treated tumors had higher tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and perforin/granzyme-mediated caspase-8 activity, which enhanced their susceptibility to NK-cell lysis. Bioluminescence imaging of mice with established tumors showed treatment with bortezomib and syngeneic NK cells reduced tumor growth and prolonged survival compared with controls receiving bortezomib or NK cells alone. In contrast, tumor progression was not delayed when animals received bortezomib and perforin-deficient NK cells, showing drug-induced augmentation in NK-cell cytotoxicity was mediated through perforin/granzyme. Furthermore, tumor growth was slower in bortezomib-treated recipients when host Treg cells were eradicated with anti-CD25 antibody before infusing NK cells compared with mice without Treg-cell ablation (tumor doubling time, 16.7 vs 4.9 days, respectively; P = .02). These findings suggest that depletion of Treg cells followed by bortezomib-induced tumor sensitization to autologous NK cells could be used as a novel strategy to treat cancer.