Decoupled visually-guided reaching in optic ataxia: differences in motor control between canonical and non-canonical orientations in space.
Guiding a limb often involves situations in which the spatial location of the target for gaze and limb movement are not congruent (i.e. have been decoupled). Such decoupled situations involve both the implementation of a cognitive rule (i.e. strategic control) and the online monitoring of the limb position relative to gaze and target (i.e. sensorimotor recalibration). To further understand the neural mechanisms underlying these different types of visuomotor control, we tested patient IG who has bilateral caudal superior parietal lobule (SPL) damage resulting in optic ataxia (OA), and compared her performance with six age-matched controls on a series of center-out reaching tasks. The tasks comprised 1) directing a cursor that had been rotated (180° or 90°) within the same spatial plane as the visual display, or 2) moving the hand along a different spatial plane than the visual display (horizontal or para-sagittal). Importantly, all conditions were performed towards visual targets located along either the horizontal axis (left and right; which can be guided from strategic control) or the diagonal axes (top-left and top-right; which require on-line trajectory elaboration and updating by sensorimotor recalibration). The bilateral OA patient performed much better in decoupled visuomotor control towards the horizontal targets, a canonical situation in which well-categorized allocentric cues could be utilized (i.e. guiding cursor direction perpendicular to computer monitor border). Relative to neurologically intact adults, IG's performance suffered towards diagonal targets, a non-canonical situation in which only less-categorized allocentric cues were available (i.e. guiding cursor direction at an off-axis angle to computer monitor border), and she was therefore required to rely on sensorimotor recalibration of her decoupled limb. We propose that an intact caudal SPL is crucial for any decoupled visuomotor control, particularly when relying on the realignment between vision and proprioception without reliable allocentric cues towards non-canonical orientations in space.


Crossed-immunoelectrophoretic analyses of Trypanosoma cruzi epimastigote, metacyclic, and bloodstream forms.
Sonicated suspensions of epimastigote, metacyclic, or bloodstream forms of Trypanosoma cruzi were emulsified in Freund's complete adjuvant. Rabbits immunized with epimastigotes or metacyclics received five intramuscular (i.m.) injections of 1 x 10(9) sonicated trypanosomes at weekly intervals. Immunization with bloodstream forms included three i.m. injections of 5 x 10(7) and six injections of 2 x 10(8) sonicated trypanosomes. Selected antisera from these rabbits were employed in crossed immunoelectrophoretic studies against the homologous or heterologous extracts of sonicated trypanosomes. Extracts of epimastigote, metacyclic, and trypomastigotes produced 31, 29, and 11 precipitin peaks respectively against the homologous rabbit antisera. Tandem, crossed-immunoelectrophoresis of these extracts against antiepimastigote or antimetacyclic sera revealed that epimastigotes or metacyclics may each have at least four antigens that did not appear to be shared by the other, whereas each of these forms may have at least eight or nine antigens that were not detected with extracts from trypomastigotes. Cross-absorptions of antiepimastigote or antimetacyclic sera with live trypanosomes caused marked reductions in the numbers of precipitin peaks formed against the homologous extracts, but cross-absorptions with sonicated suspensions of epimastigotes or metacyclics showed that epimastigotes or metacyclics each have at least two antigens that were not detected in extracts of the other. Differentiation appeared to be accompanied by antigenic change. More antigens appear to be shared by epimastigotes and metacyclic forms than by trypomastigotes and epimastigotes or metacyclics.


Replantation of amputated facial tissues with microvascular anastomosis.
A challenge to the microsurgeon is perfecting the technique of replantation of small pieces of facial tissue, mainly because of the extremely small size of the arteries as well as a lack of suitable veins for drainage. In the past 4 years, we have had seven cases of facial amputations, which included one scalp, two nasal tips, two ears, one lower lip, and one eyebrow. All of these patients were replanted/revascularized by microvascular anastomosis. Only two of the cases had suitable veins for anastomosis. Alternative techniques used for improving venous outflow were arterio-venous fistula, chemical leeches, and pin pricks. Four of the cases were completely successful, two cases had partial loss of the replant, and one case failed due to absence of venous drainage. In facial amputation, an aggressive microsurgical attempt will result in more tissue surviving and a better cosmetic outcome than in any other reconstructive procedures.


Treatment of C1.1 (AO-41) tibial plateau fracture: A finite element analysis of single medial, lateral and dual plating.
Bicondylar tibial plateau fractures pose many challenges in surgical treatment. The aim of the present study was to analyze three methods of reduction, single medial, single lateral, and dual plating, for the treatment of a bicondylar tibial plateau fracture, through finite element analysis (FEA). A simple metaphyseal fracture, type C1.1 (AO-41) was modeled on a CT-derived 3D model of the knee. Lateral and medial proximal tibial polyaxial plates with screws were modeled and placed accordingly for the three methods of reduction. Simulation of physiological type loading corresponding to the maximal weight acceptance phase during a slow walking gait cycle was performed using FEA. Values of stress and strain were recorded near the fracture lines. Dual plating provided a decrease of stress and strain in the tibial plateau area. However, the differences in the values among the three cases were small. The stress concentration areas were located in the vicinity of the fracture, predominantly in the area of the tibial plateau. Considering the limitations of the present study, the results revealed that dual plating leads to smaller stress and strain values near the fracture lines in the tibial plateau area. However, values obtained for single lateral plating are close in range. Considering the complexity of the surgical approach for dual plating, single lateral plating may be a solution for good reduction with fewer surgical risks and complications. Further studies on the C1.1 fracture (AO-41) are needed to analyze the complex issue of reducing and stabilizing such a fracture and to characterize the postoperative state while providing predictable parameters for an optimal result.


Respiratory weakness after mechanical ventilation is associated with one-year mortality - a prospective study.
Diaphragm dysfunction in mechanically ventilated patients is associated with poor outcome. Maximal inspiratory pressure (MIP) can be used to evaluate inspiratory muscle function. However, it is unclear whether respiratory weakness is independently associated with long-term mortality. The aim of this study was to determine if low MIP is independently associated with one-year mortality. We conducted a prospective observational cohort study in an 18-bed ICU. Adults requiring at least 24 hours of mechanical ventilation with scheduled extubation and no evidence of pre-existing muscle weakness underwent MIP evaluation just before extubation. Patients were divided into two groups: low MIP (MIP ≤30 cmH2O) and high MIP (MIP >30 cmH2O). Mortality was recorded for one year after extubation. For the survival analysis, the effect of low MIP was assessed using the log-rank test. The independent effect of low MIP on post mechanical ventilation mortality was analyzed using a multivariable Cox regression model. One hundred and twenty-four patients underwent MIP evaluation (median age 66 years (25(th)-75(th) percentile 56-74), Simplified Acute Physiology Score (SAPS) 2 = 45 (33-57), duration of mechanical ventilation 7 days (4-10)). Fifty-four percent of patients had low MIP. One-year mortality was 31 % (95 % CI 0.21, 0.43) in the low MIP group and 7 % (95 % CI 0.02, 0.16) in the high MIP group. After adjustment for SAPS 2 score, body mass index and duration of mechanical ventilation, low MIP was independently associated with one-year mortality (hazard ratio 4.41, 95 % CI 1.5, 12.9, p = 0.007). Extubation failure was also associated with low MIP (relative risk 3.0, 95 % CI 1, -9.6; p = 0.03) but tracheostomy and ICU length of stay were not. Low MIP is frequent in patients on mechanical ventilation and is an independent risk factor for long-term mortality in ICU patients requiring mechanical ventilation. MIP is easily evaluated at the patient's bedside. This study was retrospectively registered in www.clinicaltrials.gov (NCT02363231) in February 2015.


MicroRNAs in melanocyte and melanoma biology.
The importance of microRNAs as key molecular components of cellular processes is now being recognized. Recent reports have shown that microRNAs regulate processes as diverse as protein expression and nuclear functions inside cells and are able to signal extracellularly, delivered via exosomes, to influence cell fate at a distance. The versatility of microRNAs as molecular tools inspires the design of novel strategies to control gene expression, protein stability, DNA repair and chromatin accessibility that may prove very useful for therapeutic approaches due to the extensive manageability of these small molecules. However, we still lack a comprehensive understanding of the microRNA network and its interactions with the other layers of regulatory elements in cellular and extracellular functions. This knowledge may be necessary before we exploit microRNA versatility in therapeutic settings. To identify rules of interactions between microRNAs and other regulatory systems, we begin by reviewing microRNA activities in a single cell type: the melanocyte, from development to disease.


Assessment and management of the patient with Graves' ophthalmopathy.
The appropriate management of a patient with Graves' ophthalmopathy (GO) can be determined only following a careful assessment of the signs and symptoms of the disease. A detailed examination and discussion allows the clinician to determine which components of the disease are most troublesome to that particular patient. The advantages and disadvantages of the various treatment options for each disease component can be best understood in light of current concepts of the pathogenesis of GO. This paper will begin with a discussion of the histologic and gross pathologic features of the disease, followed by a brief review of pathogenic mechanisms responsible for their development. The assessment of a patient with GO will be discussed and the modes of therapy available to address each of the signs and symptoms will be reviewed. Readers interested in a more detailed discussion of the management of GO are encouraged to read the excellent recent review by Drs. Burch and Wartofsky.


The morphological and functional diversity of apical microvilli.
Sensory neurons use specialized apical processes to perceive external stimuli and monitor internal body conditions. The apical apparatus can include cilia, microvilli, or both, and is adapted for the functions of the particular cell type. Photoreceptors detect light through a large, modified cilium (outer segment), that is supported by a surrounding ring of microvilli-like calyceal processes (CPs). Although first reported 150 years ago, CPs remain poorly understood. As a basis for future study, we therefore conducted a review of existing literature about sensory cell microvilli, which can act either as the primary sensory detector or as support for a cilia-based detector. While all microvilli are finger-like cellular protrusions with an actin core, the processes vary across cell types in size, number, arrangement, dynamics, and function. We summarize the current state of knowledge about CPs and the characteristics of the microvilli found on inner ear hair cells (stereocilia) and cerebral spinal fluid-contacting neurons, with comparisons to the brush border of the intestinal and renal epithelia. The structure, stability, and dynamics of the actin core are regulated by a complement of actin-binding proteins, which includes both common components and unique features when compared across cell types. Further, microvilli are often supported by lateral links, a glycocalyx, and a defined extracellular matrix, each adapted to the function and environment of the cell. Our comparison of microvillar features will inform further research into how CPs support photoreceptor function, and also provide a general basis for investigations into the structure and functions of apical microvilli found on sensory neurons.


Imaging to study solid tumour origin and progression: lessons from research and clinical oncology.
Biomedical imaging in recent decades has clarified our understanding of normal and pathological cellular processes in vivo. In particular, this approach recently provided insights into processes occurring at a molecular or genetic level rather than at the anatomical level. The evolution of this discipline by engineering have led to its integration into biomedical research to (1) increase sensitivity and resolution imaging and to (2) improve tissue and cell specificity. Currently, imaging approaches are used in three different biomedical areas: (a) identification of cellular processes in physiological and disease state; (b) in vivo single-cell imaging; and (c) identification of new prognostic and therapeutical strategies. In this review, we will focus on the state of art of biomedical imaging in cancer. Specifically, we will highlight the most important advances in imaging tools available for basic and translational cancer research, with a particular emphasis on solid tissue malignancies.


Clinical and histopathological characteristics of primary and recurrent basal cell carcinoma: a retrospective study of the patients from a tertiary clinical centre in the northern Poland.
Basal cell carcinoma (BCC) is the most common malignant neoplasm of the skin. Management of patients with recurrent BCC remains a current clinical issue. Data concerning BCC recurrence rates as well as characteristics of this group of patients in the Polish population are scarce. Identification and analysis of clinical, epidemiological and histopathological factors influencing BCC recurrence. Histopathological diagnoses of BCC patients treated by surgical methods at the Department of Dermatology, Venereology and Allergology, Medical University of Gdansk, between 2013 and 2018, were retrospectively analysed. The analysis included 1097 tumours diagnosed in 802 patients, of which 1061 were primary BCC (pBCC) and 36 - recurring BCC (rBCC). In the analysed cohort, rBCCs constituted 3.3% of cases. 49.8% of pBCCs occurred in women; while in the rBCC group - 47.2%. The most common histopathological type was infiltrative BCC, however, it was significantly more prevalent in rBCCs (36.9% and 52.8%, respectively). The average maximum size of pBCC was 12.3 ±8.8 mm, while of rBCC 18.4 ±15.1 mm (p = 0.036). The most common location of both pBCC and rBCC was the nose (tumours in this localization constituted 23.2% and 25.0%, respectively). In the analysed cohort a relatively low percentage of rBCC was found. Among analysed risk factors, the most important ones were the infiltrative histopathological type of BCC and the non-radical treatment of the primary tumour.


Effects of stage of oestrous cycle and progesterone supplementation during culture on maturation of canine oocytes in vitro.
The aim of this study was to evaluate the effect of progesterone supplementation and stage of oestrous cycle on in vitro maturation (IVM) of canine oocytes. Oocytes were cultured in medium supplemented with 0, 2000, 4000 or 8000 ng progesterone ml(-1) (Expt 1; n=274 oocytes) or 0, 20, 200 or 2000 ng progesterone ml(-1) (Expt 2; n=789 oocytes). In Expt 3, oocytes (n=1202) were cultured in a bi-phasic system of meiotic arrest followed by IVM, both in the presence of 0, 20, 200 or 2000 ng progesterone ml(-1). Rates of meiotic resumption for Expt 1 ranged from 40.0% to 58.5%; there were no significant differences among groups. In Expt 2, rate of meiotic resumption was significantly lower in the 2000 ng progesterone ml(-1) treatment (35.5%) compared with the 200 ng progesterone ml(-1) treatment (54.0%; P<0.05). There were no significant differences in rates of maturation to metaphase II among treatments in Expt 1 (1.8-8.6%) or Expt 2 (8.4-14.7%); however, oocytes collected from ovaries of bitches in oestrus and dioestrus had higher rates of maturation to metaphase II than did oocytes from bitches at pro-oestrus or anoestrus (P<0.01). In Expt 3, no differences were observed in rates of maturation among treatment groups. Rates of maturation to metaphase II of oocytes from bitches in dioestrus were significantly higher than those from bitches in pro-oestrus (P<0.01). These results indicate that supplementation of culture medium with progesterone either during maturation or during meiotic arrest before maturation does not increase the rate of IVM of canine oocytes. However, stage of oestrous cycle is a key factor in the selection criteria for meiotically competent canine oocytes for use in in vitro experiments.


Congenital corneal opacities in a cornea referral practice.
To address the primary causes of the congenital corneal opacities seen on the Cornea Service at Wills Eye Hospital (Philadelphia, PA), the associated systemic anomalies and their management. Retrospective review of the medical records of all patients with the diagnosis of congenital corneal opacities seen at the Cornea Service from January 1, 1992 to June 30, 2003. Children 12 years old or younger at the first visit to our department were included in the study. We classified the location and extent of corneal pathology. We divided the management into medical and surgical. Seventy-two eyes of 47 patients were included in the study. The mean follow-up time from the first to the last visit was 33.1 months. The most common primary cause of congenital corneal abnormalities was Peters anomaly (40.3%), followed by sclerocornea (18.1%), dermoid (15.3%), congenital glaucoma (6.9%), microphthalmia (4.2%), birth trauma, and metabolic disease (2.8%). Seven eyes (9.7%) were classified as idiopathic. Ten patients had systemic abnormalities associated with their ocular condition. The management was medical in 38 eyes (52.7%). Twenty-four eyes (32.4%) underwent only 1 penetrating keratoplasty (PK). Only 1 eye received a regraft during the follow-up period. Eight grafts failed during the follow-up period. The importance of this study is to share our experience with this rare entity, congenital corneal opacities, describing their clinical presentation and their management.


Orienting psychotherapists to better serve low income and minority patients.
As a part of a larger NIMH study, 29 psychotherapists who were beginning training at a large urban psychiatric outpatient clinic participated in a special orientation program. The orientation program had the following objectives: First, to increase the therapists' knowledge about low income and minority patients; second, to increase the therapists' sensitivity to minority patients' requests and problems; and, third, to increase the effectiveness of the therapeutic encounter. The majority of therapists acted as their own controls; they saw patients before and then after participating in the orientation program. Data indicated that therapists improved significantly in knowledge, sensitivity, and effectiveness as a result of the special therapist orientation program.


Establishment of mouse neuron and microglial cell co-cultured models and its action mechanism.
The objective of this study is to establish a co-culture model of mouse neurons and microglial cells, and to analyze the mechanism of action of oxygen glucose deprivation (OGD) and transient oxygen glucose deprivation (tOGD) preconditioning cell models. Mouse primary neurons and BV2 microglial cells were successfully cultured, and the OGD and tOGD models were also established. In the co-culture of mouse primary neurons and microglial cells, the cell number of tOGD mouse neurons and microglial cells was larger than the OGD cell number, observed by a microscope. CCK-8 assay result showed that at 1h after treatment, the OD value in the control group is lower compared to all the other three groups (P < 0.05). The treatment group exhibited the highest OD value among the four groups. The results observed at 5h were consistent with the results at 1 h. Flow cytometry results showed that at 1h after treatment the apoptosis percentages is higher in the control group compared to other three groups (P < 0.05). Mouse brain tissues were collected and primary neurons cells were cultured. In the meantime mouse BV2 microglia cells were cultured. Two types of cells were co-cultured, and OGD and tOGD cell models were established. There were four groups in the experiment: control group (OGD), treatment group (tOGD+OGD), placebo group (tOGD+OGD+saline) and minocycline intervention group (tOGD+OGD+minocycline). CCK-8 kit was used to detect cell viability and flow cytometry was used to detect apoptosis. In this study, mouse primary neurons and microglial cells were co-cultured. The OGD and tOGD models were established successfully. tOGD was able to effectively protect neurons and microglial cells from damage, and inhibit the apoptosis caused by oxygen glucose deprivation.


Analysis of non-ketoic steroids 17alpha-methylepithiostanol and desoxymethyl- testosterone in dietary supplements.
Dietary supplements containing 17alpha-methyl-2,3-epithio-5alpha-androstane-17beta-ol (17alpha-methylepithiostanol), which is a 17-methylated analogue of epithiostanol or a prodrug of desoxymethyltestosterone (17alpha-methyl-5alpha-androst-2-en-17beta-ol), have recently appeared on the Internet. 17alpha-Methylepithiostanol and desoxymethyltestosterone are classified as prohibited substances on the World Anti-Doping Agency (WADA) list. Two preparations, EPISTANE and P-PLEX, were obtained from the Internet so that their contents could be investigated. This study involved gas chromatography/mass spectrometry (GC/MS) analysis after trimethylsilyl (TMS) derivatization, liquid chromatography/mass spectrometry (LC/MS) in atmospheric pressure photoionization (APPI) mode and nuclear magnetic resonance (NMR) spectroscopy. Analysis using LC/MS in APPI mode would be a useful tool for detecting heat-labile and non-polar steroids.Although the labelling of EPISTANE indicates that it contains 17alpha-methyl-2alpha, 3alpha-epithio-5alpha-androstane-17beta-ol only, 17alpha-methyl-2beta,3beta-epithio-5alpha-androstane-17beta-ol and desoxymethyltestosterone were identified in the supplement. The results showed that P-PLEX contained desoxymethyltestosterone and its isomer 17alpha-methyl-5alpha-androst-3-en-17beta-ol. Urine samples can be screened after EPISTANE or P-PLEX administration using the normal screening procedure for anabolic steroids with GC/MS.


PREVALENCE AND ANTIMICROBIAL RESISTANCE OF SALMONELLA ISOLATED FROM CARCASSES, PROCESSING FACILITIES AND THE ENVIRONMENT SURROUNDING SMALL SCALE POULTRY SLAUGHTERHOUSES IN THAILAND.
Salmonella is a major food-borne pathogen worldwide, including Thai- land, and poultry meat plays a role as a vehicle for the spread of the disease from animals to humans. The prevalence and characteristics of Salmonella isolated from 41 small scale poultry slaughterhouses in Chiang Mai, Thailand were determined during July 2011 through May 2012. Salmonella's prevalence in live poultry, car- casses, waste water, and soil around processing plants were 3.2%, 7.3%, 22.0% and 29.0%, respectively. Eighteen different serotypes were identified, the most common being Corvallis (15.2%), followed by Rissen (13.9%), Hadar (12.7%), Enteritidis (10.1%), [I. 4,5,12:i:-] (8.8%), Stanley (8.8%), and Weltevreden (8.8%). Antimicrobial susceptibility tests revealed that 68.4% of the Salmonella spp were resistant to at least one antimicrobial while 50.6% showed multiple drug resis- tance (MDR). Specifically, 44.3% of Salmonella were resistant to nalidixic acid, followed by streptomycin (41.8%), ampicillin (34.2%), tetracycline (34.2%), and sulfamethoxazole/trimethoprim (20.3%). Salmonella contamination was found in processing lines, carcasses, and in the environment around the processing sta- tions. These findings indicate that improving hygiene management in small scale poultry slaughterhouses as well as prudent use of antimicrobial drugs is urgently needed if Salmonella contamination is to be reduced.


Pathogenic Events in a Nonhuman Primate Model of Oral Poliovirus Infection Leading to Paralytic Poliomyelitis.
Despite a great deal of prior research, the early pathogenic events in natural oral poliovirus infection remain poorly defined. To establish a model for study, we infected 39 macaques by feeding them single high doses of the virulent Mahoney strain of wild type 1 poliovirus. Doses ranging from 107 to 109 50% tissue culture infective doses (TCID50) consistently infected all the animals, and many monkeys receiving 108 or 109 TCID50 developed paralysis. There was no apparent difference in the susceptibilities of the three macaque species (rhesus, cynomolgus, and bonnet) used. Virus excretion in stool and nasopharynges was consistently observed, with occasional viremia, and virus was isolated from tonsils, gut mucosa, and draining lymph nodes. Viral replication proteins were detected in both epithelial and lymphoid cell populations expressing CD155 in the tonsil and intestine, as well as in spinal cord neurons. Necrosis was observed in these three cell types, and viral replication in the tonsil/gut was associated with histopathologic destruction and inflammation. The sustained response of neutralizing antibody correlated temporally with resolution of viremia and termination of virus shedding in oropharynges and feces. For the first time, this model demonstrates that early in the infectious process, poliovirus replication occurs in both epithelial cells (explaining virus shedding in the gastrointestinal tract) and lymphoid/monocytic cells in tonsils and Peyer's patches (explaining viremia), extending previous studies of poliovirus pathogenesis in humans. Because the model recapitulates human poliovirus infection and poliomyelitis, it can be used to study polio pathogenesis and to assess the efficacy of candidate antiviral drugs and new vaccines.IMPORTANCE Early pathogenic events of poliovirus infection remain largely undefined, and there is a lack of animal models mimicking natural oral human infection leading to paralytic poliomyelitis. All 39 macaques fed with single high doses ranging from 107 to 109 TCID50 Mahoney type 1 virus were infected, and many of the monkeys developed paralysis. Virus excretion in stool and nasopharynges was consistently observed, with occasional viremia; tonsil, mesentery lymph nodes, and intestinal mucosa served as major target sites of viral replication. For the first time, this model demonstrates that early in the infectious process, poliovirus replication occurs in both epithelial cells (explaining virus shedding in the gastrointestinal tract) and lymphoid/monocytic cells in tonsils and Peyer's patches (explaining viremia), thereby supplementing historical reconstructions of poliovirus pathogenesis. Because the model recapitulates human poliovirus infection and poliomyelitis, it can be used to study polio pathogenesis, candidate antiviral drugs, and the efficacy of new vaccines.


Cystic hygroma of the head and neck. A review of 37 cases.
37 children with cystic hygroma of the head and neck are reviewed. 92 per cent of the children required surgical excision of the cystic hygroma and one-third of these children suffered a permanent nerve palsy. Spontaneous resolution of the cystic hygroma occurred in 8 per cent of the children over a period of follow-up lasting on average 7 years 2 months (range 3 months-20 years). 41 per cent of the children suffered significant upper airway and feeding problems.


Mammalian mitochondria have the innate ability to import tRNAs by a mechanism distinct from protein import.
Mitochondrial genomes generally encode a minimal set of tRNAs necessary for protein synthesis. However, a number of eukaryotes import tRNAs from the cytoplasm into their mitochondria. For instance, Saccharomyces cerevisiae imports cytoplasmic tRNA(Gln) into the mitochondrion without any added protein factors. Here, we examine the existence of a similar active tRNA import system in mammalian mitochondria. We have used subcellular RNA fractions from rat liver and human cells to perform RT-PCR with oligonucleotide primers specific for nucleus-encoded tRNA(CUG)(Gln) and tRNA(UUG)(Gln) species, and we show that these tRNAs are present in rat and human mitochondria in vivo. Import of in vitro transcribed tRNAs, but not of heterologous RNAs, into isolated mitochondria also demonstrates that this process is tRNA-specific and does not require the addition of cytosolic factors. Although this in vitro system requires ATP, it is resistant to inhibitors of the mitochondrial electrochemical gradient, a key component of protein import. tRNA(Gln) import into mammalian mitochondria proceeds by a mechanism distinct from protein import. We also show that both tRNA(Gln) species and a bacterial pre-tRNA(Asp) can be imported in vitro into mitochondria isolated from myoclonic epilepsy with ragged-red fiber cells if provided with sufficient ATP (2 mM). This work suggests that tRNA import is more widespread than previously thought and may be a universal trait of mitochondria. Mutations in mitochondrial tRNA genes have been associated with human disease; the tRNA import system described here could possibly be exploited for the manipulation of defective mitochondria.


Lysine deacetylase inhibition prevents diabetes by chromatin-independent immunoregulation and β-cell protection.
Type 1 diabetes is due to destruction of pancreatic β-cells. Lysine deacetylase inhibitors (KDACi) protect β-cells from inflammatory destruction in vitro and are promising immunomodulators. Here we demonstrate that the clinically well-tolerated KDACi vorinostat and givinostat revert diabetes in the nonobese diabetic (NOD) mouse model of type 1 diabetes and counteract inflammatory target cell damage by a mechanism of action consistent with transcription factor--rather than global chromatin--hyperacetylation. Weaning NOD mice received low doses of vorinostat and givinostat in their drinking water until 100-120 d of age. Diabetes incidence was reduced by 38% and 45%, respectively, there was a 15% increase in the percentage of islets without infiltration, and pancreatic insulin content increased by 200%. Vorinostat treatment increased the frequency of functional regulatory T-cell subsets and their transcription factors Gata3 and FoxP3 in parallel to a decrease in inflammatory dendritic cell subsets and their cytokines IL-6, IL-12, and TNF-α. KDACi also inhibited LPS-induced Cox-2 expression in peritoneal macrophages from C57BL/6 and NOD mice. In insulin-producing β-cells, givinostat did not upregulate expression of the anti-inflammatory genes Socs1-3 or sirtuin-1 but reduced levels of IL-1β + IFN-γ-induced proinflammatory Il1a, Il1b, Tnfα, Fas, Cxcl2, and reduced cytokine-induced ERK phosphorylation. Further, NF-κB genomic iNos promoter binding was reduced by 50%, and NF-κB-dependent mRNA expression was blocked. These effects were associated with NF-κB subunit p65 hyperacetylation. Taken together, these data provide a rationale for clinical trials of safety and efficacy of KDACi in patients with autoimmune disease such as type 1 diabetes.


A comprehensive review of the literature and data analysis on hypo-hyperdontia.
The objectives of this study were i) to conduct a comprehensive review of the literature on hypo-hyperdontia, and ii) to categorize the reported cases based on occurrence. An extensive search of the literature covering the period from January 1966 to January 2014 was conducted using the Embase, Google Scholar, Medline, and PubMed databases. The key words used in the search strategy were "concomitant", "agenesis", "hypodontia", "hyperdontia", "supernumerary teeth", "syndromes", and "hypohyperdontia" in various combinations. The retrieved data were analyzed based on gender, occurrence, and occurrence of both hypodontia and hyperdontia. Descriptive statistics were carried out using the chi-squared test. For hypo-hyperdontia overall, seven case studies and 40 cases involving 103 patients were reported. Our comprehensive review revealed that hypo-hyperdontia occurs most commonly in males (P < 0.05), and the bimaxillary type (65%) was reported most commonly in comparison with the maxillary, pre-maxillary and mandibular types (P < 0.001). Twenty syndromic cases and two case studies on familial occurrence of hypohyperdontia were reported. Almost 57% of cases affected the anterior region, whereas 43% of cases affected the anteroposterior region. The anterior region was most commonly affected by hyperdontia whereas the posterior region was most commonly affected by hypodontia. Hypo-hyperdontia is commonly seen in males, and the most common type is bimaxillary occurrence. G/BBB syndrome is commonly associated with hypo-hyperdontia, being relatively frequent in the anterior region, where mesiodens is frequently seen, and second premolars are the most commonly missing teeth.


Intratubular, Intracellular, and Mitochondrial Angiotensin II/AT1 (AT1a) Receptor/NHE3 Signaling Plays a Critical Role in Angiotensin II-Induced Hypertension and Kidney Injury.
Hypertension is well recognized to be the most important risk factor for cardiovascular diseases, stroke, and end-stage kidney failure. A quarter of the world's adult populations and 46% of the US adults develop hypertension and currently require antihypertensive treatments. Only 50% of hypertensive patients are responsive to current antihypertensive drugs, whereas remaining patients may continue to develop cardiovascular, stroke, and kidney diseases. The mechanisms underlying the poorly controlled hypertension remain incompletely understood. Recently, we have focused our efforts to uncover additional renal mechanisms, pathways, and therapeutic targets of poorly controlled hypertension and target organ injury using novel animal models or innovative experimental approaches. Specifically, we studied and elucidated the important roles of intratubular, intracellular, and mitochondrial angiotensin II (Ang II) system in the development of Ang II-dependent hypertension. The objectives of this invited article are to review and discuss our recent findings that (a) circulating and intratubular Ang II is taken up by the proximal tubules via the (AT1) AT1a receptor-dependent mechanism, (b) intracellular administration of Ang II in proximal tubule cells or adenovirus-mediated overexpression of an intracellular Ang II fusion protein selectively in the mitochonria of the proximal tubules induces blood pressure responses, and (c) genetic deletion of AT1 (AT1a) receptors or the Na+/H+ exchanger 3 selectively in the proximal tubules decreases basal blood pressure and attenuates Ang II-induced hypertension. These studies provide a new perspective into the important roles of the intratubular, intracellular, and mitochondrial angiotensin II/AT1 (AT1a) receptor signaling in Ang II-dependent hypertensive kidney diseases.


Domoic acid impairment of cardiac energetics.
Excitatory mediated neuronal injury has been shown to involve a complex cascade of events. However, the associated cardiac damage reported in humans and marine animals following exposure to excitotoxins has not been well characterized. We hypothesized that the excitotoxin domoic acid can traverse cardiac cell membranes and elicit a deleterious effect on cardiac mitochondrial energetics. Domoic acid (0.05-0.25 microM; 10 min) treatment of isolated rat cardiac mitochondria produced a marked decrease of both mitochondrial flavin adenine dinucleotide (FAD)- and nicotinamide adenine linked respiratory control indices (p < 0.001). Enzymatic assays of the mitochondrial electron transport chain (complexes I-V) and the mitochondrial matrix marker enzyme citrate synthase, showed marked concentration-dependent impairment in activity and integrity following exposure to domoic acid (p < 0.01). Similar mitochondrial effects were seen following exposure to the glutamic acid analog, kainic acid (0.5-2 microM). Domoic acid (0.05-10 microM; 40 min) was shown by competitive enzyme-linked immunosorbent assay to traverse the cellular membrane of H9c2 rat cardiac myoblasts. Exposure of intact H9c2 cells to domoic acid (10 microM; 24 h) impaired complex II-III activity but did not compromise cellular viability as assessed using cell quantification or lactate dehydrogenase leakage assays. Assessment of reactive oxygen species (superoxide and hydrogen peroxide) production in both isolated cardiac mitochondria and H9c2 cardiomyocytes failed to show any significant differences following exposure to domoic acid (0.05-5 microM). This is the first study to demonstrate a direct effect of domoic acid on cardiac mitochondrial energetics. However, the absence of substantial damage to intact cardiomyocytes raises questions regarding direct toxicological effects on cardiac energetics or viability under conditions of natural domoic acid exposure.


Effect of unitization on associative recognition in amnesia.
We examined how associative recognition performance in amnesic patients is mediated by use of a unitized (i.e., holistic) encoding strategy, and the degree to which the unitization effect is related to sparing of familiarity-based recognition. Participants studied word pairs as either separate lexical units in sentences (i.e., nonunitized) or as compounds (unitized). Under standard recognition instructions, normal controls and patients with left-temporal lobe damage (previously determined to have impairments in both recollection and familiarity) showed no difference for unitized and nonunitized pairs, whereas hypoxics (previously determined to have impaired recollection but relatively preserved familiarity) showed an advantage of unitized over nonunitized pairs. This effect was reproduced in normal healthy participants under instructions to restrict responses to judgments of familiarity. The results indicate that unitization may mediate the degree of associative recognition impairment exhibited by some amnesic patients, and that the effect is related to preserved familiarity capacity. The relevance of the results to the debate over the importance of the hippocampus in memory for associations is discussed.


Comparison of phenolic content and antioxidant activities of millet varieties grown in different locations in Sri Lanka.
Soluble and bound phenolic compounds were extracted from different varieties of millet types namely, finger millet, foxtail, and proso millet cultivated at dry and intermediate climatic zones in Sri Lanka. The extracts were examined for their total phenolic content (TPC), total flavonoid content (TFC), and proanthocyanidin content (PC). The antioxidant activities were meassured by reducing power (RP), trolox equivalent antioxidant capacity (TEAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion chelating ability (FICA), and using a β carotene linoleate model system. The ferulic acid content of extracts were determined using high-performance liquid chromatoghraphy (HPLC). Finger millet showed the highest phenolic content and antioxidant activities compared to proso and foxtail millets. The phenolic content as well as antioxidant activites of soluble and bound phenolic extracts of millets were affected by variety and cultivated location. The highest phenolic content and antioxidant activites were reported for millet samples cultivated in areas belonging to the dry zone in Sri Lanka.


Cutaneous cladosporiosis due to Cladosporium trichoides.
A case of cutaneous cladosporiosis due to Cladosporium trichoides, manifesting as a chronic discharging ulcer near the wrist of the left arm in a farmer with ascites and hypoalbuminaemia is described. The isolate was neurotropic in experimentally infected mice. The present case constitutes one of the few reports of cutaneous cladosporiosis.


Cast-saw burns: evaluation of skin, cast, and blade temperatures generated during cast removal.
The use of an oscillating saw for cast removal creates a potential for iatrogenic injury and patient discomfort. Burns and abrasions can occur from the heat created by frictional forces and direct blade contact. With use of a cadaver model system, skin temperature measurements were recorded during cast removal with an oscillating saw. Casts of uniform thickness were applied to cadavers equilibrated to body temperature. The casts were removed by a single individual while simultaneously measuring temperatures at the skin-padding interface, cast-padding interface, and the blade. Variables tested include two removal techniques, two casting materials (fiberglass and plaster), and two cast-padding thicknesses. A poor removal technique (the cast saw blade never leaving the cast material during cutting), fiberglass casting material, and thinner cast padding resulted in significantly higher skin temperatures. The poor technique increased skin temperatures by an average of 5.0 degrees C (p < 0.05). Fiberglass casting materials increased skin temperatures by an average of 7.4 degrees C (p < 0.05). Four layers of cast padding compared with two layers decreased skin temperatures by 8.0 degrees C (p < 0.05). The highest skin temperatures were recorded for fiberglass casts with two layers of padding. The lowest skin temperatures were recorded for plaster casts with four layers of padding. Four layers of cast padding compared with two layers significantly reduced skin temperatures for both plaster and fiberglass casts.


Comparison of two chair-side tests for enumeration of Mutans Streptococci in saliva.
To compare the prevalence and levels of salivary Mutans Streptococci (MS) assessed with two commercial chair-side methods based on culture growth or monoclonal antibodies, respectively. The study group consisted of a convenience sample of 89 adults, 23-72 years of age, referred to a maxillofacial hospital clinic with a caries history. Stimulated whole saliva samples were collected and the number of MS was assessed with the Dentocult-SM Strip Mutans (DSM) and the Saliva-Check Mutans (SCM). The outcome was compared with conventional anaerobic laboratory cultivation on selective MSB agar. The prevalence of ≥ 5 × 10(5) CFU was 21%, 62%, and 73% with the SCM, DSM and MSB-cultivation, respectively. The correlation between DSM and SCM tests was r = 0.49 (p < 0.05) but the agreement on the high levels was fair. In comparison with the MSB agar, DSM provided acceptable levels of sensitivity and specificity while the SCM displayed a poor sensitivity but a perfect specificity. Both chair-side methods were significantly related to the prevalence of active root caries lesions (p < 0.05). The two chair-side salivary tests were significantly correlated and due their high specificity, they may be sensitive enough for screening purposes and for patient-centered promotion of oral health.


Multi-component hybrid organic-inorganic particles with highly dispersed silver nanoparticles in the external shell.
Complex hybrid organic-"multi inorganic" particles SiO2 PDCL-ZrO2 (Ag SiO2) (PDCL = poly(N-dicarbazolyl-lysine)) were synthesized stepwise from silica nanoparticles. First, the zirconia units were synthesized by sol-gel processing and anchored to the polyCOOH poly(dicarbazole-lysine) shell of pre-formed silica particles through metal coordination on residual COOH groups. Investigation of the particle morphology indicated a core-shell structure. A second sol-gel reaction was then carried out at the surface of the SiO2 PDCL-ZrO2 particles using a mixture of alkoxysilanes including an amino-substituted silane capable of chelating silver cations. Once the outer shell made of organically substituted silica was deposited, silver particles were generated by UV treatment, thus leading to an external layer of homogeneously dispersed silver nanoparticles in silica.


Pharmacokinetics of oral fluconazole when used for prophylaxis in bone marrow transplant recipients.
The pharmacokinetics of fluconazole was investigated in 20 bone marrow transplant patients following oral administration of 200 mg of this drug. Blood samples were collected from each patient at different time intervals within 48 h after the first dose, and fluconazole was measured in plasma by high-performance liquid chromatography with UV detection. Urine was collected from 14 of these patients and analyzed similarly. The plasma concentration-time data exhibited the characteristics of the one-compartment model with first-order absorption quite well. The means +/- standard deviations of half-lives for absorption and elimination, peak concentration, time to peak, mean residence time, apparent volumes of distribution, area under the curve, and apparent oral clearance observed in these patients were 2.84 +/- 1.34 h, 19.94 +/- 18.7 h, 4.45 +/- 1.86 microg/ml, 8.34 +/- 5.97 h, 39.57 +/- 20.5 h, 0.874 +/- 0.48 liter/kg, 156.0 +/- 60.6 microg x h/ml, and 0.0256 +/- 0.0138 liter/h x kg, respectively. The amount of fluconazole excreted in urine in 24 h was 67.1 +/- 83 mg, which represents 33.55% +/- 41.6% of the dose administered. Patients who developed hemorrhagic cystitis excreted significantly (P < or = 0.0094) more fluconazole in 24 h than did those who did not.


Neurosecretory axo-axonic synapses in the median eminence of the turtle, Geoclemys reevesii.
Neurosecretory axo-axonic synapses were found in the median eminence of the turtle. Most of the presynaptic axons contain granules approximately 95 nm in diameter, and the postsynaptic elements have granules approximately 110 nm in diameter. The functional significance of axo-axonic synapses in the median eminence is discussed in relation to the discharge of releasing hormones from the axon terminals.


Bis-diamine administration during pregnancy induces developmental and reproductive toxicities in rats.
Bis-diamine was developed as amebicidal and male contraceptive agents; however, it is also reported to induce characteristic congenital heart defects especially in the cardiac conotruncal area of rats. Because of its characteristic congenital heart defects, bis-diamine-induced animal models can be used for studying congenital heart defects. However, comprehensive toxicological information regarding bis-diamine-induced congenital heart defects in this animal model is not available. In this study, we investigated and characterized an animal model for bis-diamine-induced congenital heart defects. A single dose of 200-mg bis-diamine was administered by oral gavage to pregnant rats on gestation day 10, and then observed the representative toxicological endpoints for general systemic health of pregnant rats, embryo-fetal development, and parturition. Characteristic congenital heart defects and other birth defects similar to DiGeorge syndrome were observed in bis-diamine-administered pregnant rats. In addition, developmental and reproductive toxicity findings, including increased postimplantation loss, decreased fetal weight, increased perinatal death, and increased gestation period, were observed in bis-diamine-administered pregnant rats. In particular, these developmental and reproductive toxicities were observed without maternal toxicity findings. These results will be useful to use this animal model for further studies in congenital heart defects, cardiovascular defects, and understanding their mechanisms.


Degradable Zinc-Phosphate-Based Hierarchical Nanosubstrates for Capture and Release of Circulating Tumor Cells.
Circulating tumor cells (CTCs) play a significant role in cancer diagnosis and personalized therapy, and it is still a significant challenge to efficiently capture and gently release CTCs from clinical samples for downstream manipulation and molecular analysis. Many CTC devices incorporating various nanostructures have been developed for CTC isolation with sufficient capture efficiency, however, fabricating such nanostructured substrates often requires elaborate design and complicated procedures. Here we fabricate a degradable zinc-phosphate-based hierarchical nanosubstrate (HZnPNS), and we demonstrate its excellent CTC-capture performance along with effective cell-release capability for downstream molecular analysis. This transparent hierarchical architecture prepared by a low-temperature hydrothermal method, enables substantially enhanced capture efficiency and convenient imaging. Biocompatible sodium citrate could rapidly dissolve the architecture at room temperature, allowing that 88 ± 4% of captured cells are gently released with a high viability of 92 ± 1%. Furthermore, antiepithelial cell adhesion molecule antibody functionalized HZnPNS (anti-EpCAM/HZnPNS) was successfully applied to isolate CTCs from whole blood samples of cancer patients, as well as release CTCs for global DNA methylation analysis, indicating it will serve as a simple and reliable alternative platform for CTC detection.


Highly Robust Silver Nanowire Network for Transparent Electrode.
Solution-processed silver nanowire networks are one of the promising candidates to replace a traditional indium tin oxide as next-generation transparent and flexible electrodes due to their ease of processing, moderate flexibility, high transparency, and low sheet resistance. To date, however, high stability of the nanowire networks remains a major challenge because the long-term usages of these electrodes are limited by their poor thermal and chemical stabilities. Existing methods for addressing this challenge mainly focus on protecting the nanowire network with additional layers that require vacuum processes, which can lead to an increment in manufacturing cost. Here, we report a straightforward strategy of a sol-gel processing as a fast and robust way to improve the stabilities of silver nanowires. Compared with reported nanoparticles embedded in nanowire networks, better thermal and chemical stabilities are achieved via sol-gel coating of TiO2 over the silver nanowire networks. The conformal surface coverage suppressed surface diffusion of silver atoms and prevented chemical corrosion from the environment. These results highlight the important role of the functional layer in providing better thermal and chemical stabilities along with improved electrical properties and mechanical robustness. The silver nanowire/TiO2 composite electrodes were applied as the source and drain electrodes for In2O3 thin-film transistors (TFTs) and the devices exhibited improved electrical performance annealed at 300 °C without the degradation of the electrodes. These key findings not only demonstrated a general and effective method to improve the thermal and chemical stabilities of metal nanowire networks but also provided a basic guideline toward rational design of highly efficient and robust composite electrodes.


Novel Pharmacologic Candidates for Treatment of Primary Open-Angle Glaucoma.
Primary open-angle glaucoma (OAG) affects approximately 45 million people worldwide and more than 2.5 million people aged 40 years or older in the United States. Pharmacologic treatment for glaucoma is directed towards lowering intraocular pressure (IOP) to slow disease progression and delay visual field loss. Current medical treatment options for the lowering of IOP include the following classes of topical medications: beta-adrenergic antagonists, alpha-adrenergic agonists, cholinergic agonists, carbonic anhydrase inhibitors, and prostaglandin analogs. Issues with existing drugs include failure to achieve target IOP with monotherapy, drug-related side effects, and low patient compliance with multiple daily administration of eye drops. In recent years, the scientific and medical community has seen encouraging development of novel classes of drugs for primary OAG, the majority of which lower IOP by targeting the trabecular meshwork outflow pathway to increase aqueous humor outflow. Among the most promising new pharmacologic candidates are rho kinase inhibitors including ripasudil (K-115), netarsudil (AR-13324), and AMA0076; adenosine receptor agonists including trabodenoson (INO-8875); and modified prostaglandin analogs including latanoprostene bunod (LBN, BOL-303259-X) and ONO-9054. This study aims to systematically review and summarize the most recent developments in clinical trials for new pharmacologic options for the treatment of primary open-angle glaucoma.


National Museum of Dentistry exhibition: the future is now! African Americans in dentistry.
Inspired by recently published NDA II: The Story of America's Second National Dental Association and sponsored jointly by the National Dental Association Foundation and the Colgate-Palmolive Company, an historical exhibit on dentistry in the African-American community was one of the celebrations for the Golden Anniversary of the American Academy of the History of Dentistry. This exhibit premiered on Sept. 27, 2002 in the National Museum of Dentistry located on the medical/dental campus of the University of Maryland in Baltimore. The Museum recently became an affiliate of the Smithsonian Institution. Contents of the exhibit were photographs, charts, artifacts, memorabilia, etc. These materials presented an overview of African-American activities in dental education, research, patient care, general practice, dental specialities, military service, and public health. Also included were inter-racial relationships, socioeconomic developments, and participation in civil rights endeavors that played a major role in changing out-dated accepted customs. The exhibit's purpose was to celebrate dentistry's ministrations as a health professional among African Americans in particular and the nation at large over the past two centuries. Respect for and progress of black dentists paralleled that of black physicians who were instrumental in including dentist and pharmacists as equal members in the National Medical Association since the latter's inception in 1895.


The use of animal models in research on dental implants.
The use of animal models in the study of dental implants has contributed greatly to our present understanding of the many different devices in use today. Animal testing plays a major role in assessing the safety and efficacy of dental implants. Standardized tests for determination of the toxicity and general biocompatibility responses of dental implants have been developed by the American Dental Association, the American National Standards Institute, and the Fédération Dentaire Internationale. To date, animal testing has shown the nature of soft tissue attachment to implants and the types of interfacial tissues within bone sites. There have been increased studies correlating animal tests with in vitro analysis and human studies. Additional development of testing remains to be accomplished to show the effects of function on host tissue and the long-term safety of dental implants.


Effect of Oxidation Process on Complex Refractive Index of Secondary Organic Aerosol Generated from Isoprene.
Oxidation of isoprene by hydroxyl radical (OH), ozone (O3), or nitrate radical (NO3) leads to the formation of secondary organic aerosol (SOA) in the atmosphere. This SOA contributes to the radiation balance by scattering and absorbing solar radiation. In this study, the effect of oxidation processes on the wavelength-dependent complex refractive index (RI) of SOA generated from isoprene was examined. Oxidation conditions did not have a large effect on magnitude and wavelength dependence of the real part of the RI. In the case of SOA generated in the presence of sulfur dioxide (SO2), significant light absorption at short visible and ultraviolet wavelengths with the imaginary part of the RI, up to 0.011 at 375 nm, was observed during oxidation with OH. However, smaller and negligible values were observed during oxidation with O3 and NO3, respectively. Moreover, in the absence of SO2, light absorption was not observed regardless of the oxidation process. There was an empirical correlation between the imaginary part of the RI and the average degree of unsaturation of organic molecules. The results obtained herein demonstrate that oxidation processes should be considered for estimating the radiative effect of isoprene-derived SOA.


Rapid, Low Temperature Formation of Imine-Linked Covalent Organic Frameworks Catalyzed by Metal Triflates.
Imine-linked two-dimensional covalent organic frameworks (2D COFs) are crystalline polymer networks with enhanced stability compared to boronate ester-linked systems and with broad monomer scope. They are traditionally prepared by condensing polyfunctional aldehydes and amines at elevated temperature in a mixture of organic solvents and aqueous CH3CO2H, which catalyzes imine formation and exchange. Here we employ metal triflates, which are water-tolerant Lewis acids, to accelerate 2D imine-linked COF synthesis and improve their materials quality. Low catalyst loadings provide crystalline polymer networks in nearly quantitative yields. These conditions are demonstrated for several COFs, including heteroatom-containing systems of interest for optoelectronic applications.


Adult reference levels in diagnostic and interventional radiology for temporary use in Switzerland.
This work aims at establishing a set of diagnostic reference levels (DRLs) for various types of examinations performed in diagnostic and interventional radiology. The average doses for 257 types of radiological examinations were established during the 1998 nationwide survey on the exposure of the Swiss population by radiodiagnostics. They were calculated using appropriate dosimetric models and average technical parameters. The DRLs were derived from the average doses using a multiplying factor of 1.5. The DRLs obtained were rounded and compared to the data reported in the literature. The results are in most cases comparable to the DRLs determined by the 3rd-quartile method. These discrepancies registered in some cases, particularly for complex examinations, can be explained by significant differences in the protocols and/or the technical parameters used. A set of DRLs is proposed for a large number of examinations to be used in Switzerland as temporary values until a national dosimetric database is set up.


Toxoplasma gondii Parasitophorous Vacuole Membrane-Associated Dense Granule Proteins Orchestrate Chronic Infection and GRA12 Underpins Resistance to Host Gamma Interferon.
Toxoplasma gondii evades host immunity to establish a chronic infection. Here, we assessed the role of parasitophorous vacuole (PV) membrane (PVM)- and intravacuolar network (IVN) membrane-localized dense granule (GRA) proteins in the development of acute and chronic Toxoplasma infection. Deletion of PVM-associated GRA3, GRA7, GRA8, and GRA14 or IVN membrane-associated GRA2, GRA9, and GRA12 in the low-virulence type II Prugniaud (Pru) strain induced severe defects in the development of chronic-stage cysts in vivo without affecting the parasite growth rate or the ability to differentiate into cysts in vitro Acute virulence of the PruΔgra2, PruΔgra3, and PruΔgra4 mutants was reduced but not abolished. In contrast, the PruΔgra12 mutant was avirulent in mice and PruΔgra12 parasites failed to establish a chronic infection. High-virulence type I strain RHΔgra12 parasites also exhibited a major defect in acute virulence. In gamma interferon (IFN-γ)-activated macrophages, type I RHΔgra12 and type II PruΔgra12 parasites resisted the coating of the PVM with host immunity-related GTPases as effectively as the parental type I RHΔku80 and type II PruΔku80 strains, respectively. Despite this resistance, Δgra12 PVs ultimately succumbed to IFN-γ-activated host cell innate immunity. Our findings uncover a key role for GRA12 in mediating resistance to host IFN-γ and reveal that many other IVN membrane-associated GRA proteins, as well as PVM-localized GRA proteins, play important roles in establishing chronic infection.IMPORTANCEToxoplasma gondii cysts reactivate during immune deficiency and cause fatal encephalitis. Parasite molecules that coordinate the development of acute and chronic infection are poorly characterized. Here, we show that many intravacuolar network membrane and parasitophorous vacuole membrane-associated dense granule (GRA) proteins orchestrate the development of chronic cysts in vivo A subset of these GRA proteins also modulate acute virulence, and one protein that associates with the intravacuolar network membranes, namely GRA12, was identified as a major virulence factor required for parasite resistance to host gamma interferon (IFN-γ). Our results revealed that many parasitophorous vacuole membrane and intravacuolar network membrane-associated GRA proteins are essential for successful chronic infection.


Plasma Lipidomics Profiling Reveals Biomarkers for Papillary Thyroid Cancer Diagnosis.
The objective of this study was to identify potential biomarkers and possible metabolic pathways of malignant and benign thyroid nodules through lipidomics study. A total of 47 papillary thyroid carcinomas (PTC) and 33 control check (CK) were enrolled. Plasma samples were collected for UPLC-Q-TOF MS system detection, and then OPLS-DA model was used to identify differential metabolites. Based on classical statistical methods and machine learning, potential biomarkers were characterized and related metabolic pathways were identified. According to the metabolic spectrum, 13 metabolites were identified between PTC group and CK group, and a total of five metabolites were obtained after further screening. Its metabolic pathways were involved in glycerophospholipid metabolism, linoleic acid metabolism, alpha-linolenic acid metabolism, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, Phosphatidylinositol signaling system and the metabolism of arachidonic acid metabolism. The metabolomics method based on PROTON nuclear magnetic resonance (NMR) had great potential for distinguishing normal subjects from PTC. GlcCer(d14:1/24:1), PE-NME (18:1/18:1), SM(d16:1/24:1), SM(d18:1/15:0), and SM(d18:1/16:1) can be used as potential serum markers for the diagnosis of PTC.


Severity of atopic disease inversely correlates with intestinal microbiota diversity and butyrate-producing bacteria.
The reports on atopic diseases and microbiota in early childhood remain contradictory, and both decreased and increased microbiota diversity have been associated with atopic eczema. In this study, the intestinal microbiota signatures associated with the severity of eczema in 6-month-old infants were characterized. Further, the changes in intestinal microbiota composition related to the improvement of this disease 3 months later were assessed. The severity of eczema correlated inversely with microbiota diversity (r = -0.54, P = 0.002) and with the abundance of butyrate-producing bacteria (r = -0.52, P = 0.005). During the 3-month follow-up, microbiota diversity increased (P < 0.001) and scoring atopic dermatitis values decreased (P < 0.001) in all infants. This decrease coincided with the increase in bacteria related to butyrate-producing Coprococcus eutactus (r = -0.59, P = 0.02). In conclusion, the high diversity of microbiota and high abundance of butyrate-producing bacteria were associated with milder eczema, thus suggesting they have a role in alleviating symptoms of atopic eczema.


[The possibilities of fractionated external beam repeat irradiation of relapsed primary brain tumours: the first Hungarian experience].
The practice of image-based three dimensional treatment planning and conformal radiotherapy techniques offer the opportunity to elaborate novel treatment forms, e.g. repeat irradiation techniques for primary brain tumours. The authors analysed the effect on survival and toxicity of fractionated external beam repeat irradiation in brain tumour patients. At the National Institute of Oncology, between 2002 and 2006, fractionated external beam repeat irradiation was performed in eleven patients with recurrent primary brain tumour, with total of 50-54 Gy or near total of 34-40 Gy doses. All patients were previously treated with total radiotherapy doses of 50-64 Gy. The intervals between radiotherapy courses were in the range of 7-30 years. All the treatments were carried out with 3D image-based conformal methods, the fractionation was conventional, with 1,8-2,0 Gy daily fractions in all cases. The repeat irradiation was tolerated well in the material. No grade 3-4 acute toxicity was detected, and serious, grade 3 mental deterioration, not related tumour progression was observed in only one case. In one case reoperation was necessary due to histologically verified radio-necrosis with mass-effect, and we believe that late neurotoxicity caused serious functional inabilities in one case. The median progression free survival was 8 (2-33) months, the median survival was 13 (4,5-33) months. Three of our patients were alive at the end of the study. Based on this experience and current knowledge, in absence of other treatment possibilities, the fractionated external beam repeat irradiation with near total doses could be a therapeutic choice in case of recurrent primary brain tumours, if having appropriate background. To define the optimal treatment strategy and regimens, further clinical trials should be carried out.


Maximum step length as a potential screening tool for falls in non-disabled older adults living in the community.
Identification of the risk of falls in a cohort of interest is a prerequisite for a targeted fall prevention study. Motor tasks are widely used as baseline assessment in such studies, but there are only a few well-evaluated tests of motor performance to predict falls prospectively. This study was conducted to find out if the potential of the maximum step length (MSL) test can predict future falls in non-disabled older persons. A modified version of the MSL test was used for baseline assessment in 56 community-dwelling, non-disabled elderly persons (mean age 67.7 yrs, SD 6 yrs; 57% women). During a follow-up of 1 year, falls were recorded in a daily calendar. During the follow-up, 30 persons (54%) fell, with no gender difference in reporting of falls between men and women. The adjusted mean valid step length and adjusted maximum valid step length were predictive of future falls with a sensitivity/specificity of 77%/62% and 70%/69%, respectively. Combining MSL test results with fall history increased sensitivity to 93% and 90%, respectively, but decreased specificity to 54% and 58%, respectively. The MSL test is a feasible tool, with low requirements in space, predicting future falls in community-dwelling older persons. In combination with history of falls, the sensitivity of the test increased considerably.


Comparison of the level(s) of DNA damage using Comet assay in bovine oocytes subjected to selected vitrification methods.
It was suggested that the cryodamage to oocytes' DNA has been responsible for the compromised developmental competence of cryopreserved oocytes. Vitrification of bovine oocytes affected not only cellular components, but also nuclear material. A significant rate of DNA fragmentation was found in bovine frozen or vitrified oocytes analysed by Comet assay regardless of cryopreservation method. Our method of vitrification using droplet system after gentle pre-equilibration treatment is one of the most effective cryopreservation methods employed for bovine oocytes so far, making it possible to develop 30% blastocyst stage embryos. In this study, the extent of DNA damage in bovine oocytes vitrified using three vitrification methods (droplet system, Open Pulled Straw and traditional vitrification in 0.25 ml insemination straws) was compared using Comet assay. Vitrification in droplet system and Open Pull Straws vitrification did not result in detectable cryoinjuries of DNA of bovine oocytes. On the contrary, DNA fragmentation was found in four of 26 oocytes vitrified in 0.25 ml straws (15.4%, p <or= 0.05 in comparison with the other vitrification methods).


Chinese herbal medicine for obesity: a randomized, double-blinded, multicenter, prospective trial.
Obesity is a serious medical problem worldwide. As a holistic therapy, traditional Chinese medicine (TCM) may have a potential in obesity management. In this controlled trial, we evaluated the safety and effectiveness of xin-ju-xiao-gao-fang (XJXGF), a TCM herbal formulation, in 140 obese subjects over a 24-week period. The XJXGF formula mainly consists of rhubarb, coptis, semen cassia, and citrus aurantium. Subjects with body mass index (BMI) 28-40 kg/m(2) were recruited at 5 centers in China. We assessed the changes in subjects' body weight, its related parameters, and the reduction of insulin resistance (IR) after administration of XJXGF formula or low-dose XJXGF (10% of the XJXGF formula, as control). After 24-week treatment, among participants in the XJXGF formula group and low-dose XJXGF group, the mean ± SE changes in the body weight were -3.58 ± 0.48 and -1.91 ± 0.38 kg, respectively (p < 0.01). The changes in the IR-index of two groups were -2.65 ± 1.04 and -1.58 ± 1.3, respectively (p < 0 .05). There were no serious adverse events reported during the 24-week trial. Participants reported 7 minor adverse events, 4 in the XJXGF formula group and 3 in the low-dose XJXGF group (p = 0.578). Future studies are needed to investigate the clinical utility of this TCM formulation in the treatment of obese subjects.


Improved Performance for PMSM Sensorless Control Based on Robust-Type Controller, ESO-Type Observer, Multiple Neural Networks, and RL-TD3 Agent.
This paper summarizes a robust controller based on the fact that, in the operation of a permanent magnet synchronous motor (PMSM), a number of disturbance factors naturally occur, among which both changes in internal parameters (e.g., stator resistance Rs and combined inertia of rotor and load J) and changes in load torque TL can be mentioned. In this way, the performance of the control system can be maintained over a relatively wide range of variation in the types of parameters mentioned above. It also presents the synthesis of robust control, the implementation in MATLAB/Simulink, and an improved version using a reinforcement learning twin-delayed deep deterministic policy gradient (RL-TD3) agent, working in tandem with the robust controller to achieve superior performance of the PMSM sensored control system. The comparison of the proposed control systems, in the case of sensored control versus the classical field oriented control (FOC) structure, based on classical PI-type controllers, is made both in terms of the usual response time and error speed ripple, but also in terms of the fractal dimension (DF) of the rotor speed signal, by verifying the hypothesis that the use of a more efficient control system results in a higher DF of the controlled variable. Starting from a basic structure of an ESO-type observer which, by its structure, allows the estimation of both the PMSM rotor speed and a term incorporating the disturbances on the system (from which, in this case, an estimate of the PMSM load torque can be extracted), four variants of observers are proposed, obtained by combining the use of a multiple neural network (NN) load torque observer and an RL-TD3 agent. The numerical simulations performed in MATLAB/Simulink validate the superior performance obtained by using properly trained RL-TD3 agents, both in the case of sensored and sensorless control.


Echinococcus granulosus sensu lato GENOTYPES IN DOMESTIC LIVESTOCK AND HUMANS IN GOLESTAN PROVINCE, IRAN.
Cystic echinococcosis (CE) is a globally parasitic zoonosis caused by larval stages of Echinococcus granulosus. This study investigated E. granulosus genotypes isolated from livestock and humans in the Golestan province, northern Iran, southeast of the Caspian sea, using partial sequencing data of the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1) mitochondrial genes. Seventy E. granulosus isolates were collected from animals in slaughterhouses: 18 isolates from sheep, 40 from cattle, nine from camels, two from buffaloes and one from a goat, along with four human isolates (formalin-fixed, paraffin-embedded tissues) from CE patients of provincial hospitals. All isolates were successfully analysed by PCR amplification and sequencing. The sequence analysis found four E. granulosus genotypes among the 74 CE isolates: G1 (78.3%), G2 (2.7%), G3 (15%) and G6 (4%). The G1-G3 complex genotype was found in all of the sheep, goat, cattle and buffalo isolates. Among the nine camel isolates, the frequency of G1-G3 and G6 genotypes were 66.7% and 33.3%, respectively. All four human CE isolates belonged to E. granulosus sensu stricto. This study reports the first occurrence of the G2 genotype in cattle from Iran and confirms the previously reported G3 genotype in camels in the same country.


Evaluation of toxicity studies of flavonoid fraction of Lithocarpus polystachyus Rehd in rodents.
The aim of the study was to evaluate the safety of flavonoid fraction of Lithocarpus polystachyus Rehd (Sweet Tea-F, ST-F) in mice and rats through acute and sub-chronic toxicity studies respectively. For acute toxicity study, a single dose of 5000 mg/kg of ST-F was given orally to healthy KM mice. The mice were observed mortality and toxic symptoms for 24 h, then once a day up to 14 days. In the sub-chronic toxicity study, ST-F was administered orally at doses of 0, 70, 140, 560 mg/kg/day to rats for 26 weeks. Body weight and food intake were recorded weekly. Hematological, biochemical, coagulation and organ parameters were analyzed at the end of 26 weeks administration. Vital organs were evaluated by histopathology. In the acute toxicity study, ST-F caused neither significant toxic symptoms, nor mortality in mice. In sub-chronic toxicity study, daily oral administration of ST-F at the dose of 70 mg/kg resulted in a significant increase (P < 0.05) in the relative body weight at the 10-week, and the same situation brought at the dose of 140 mg/kg/day at the 22-week. Hematological and biochemical showed significant changes (P < 0.01 or P < 0.05) in WBC, GLU, ALP, AST and serum electrolytes levels at the dose of 560 mg/kg/day. The amount of RBC decreased significantly (P < 0.05) while the content of PLT slightly increased (P < 0.05) at the dose of 140 mg/kg/day. In additional, no obvious histological changes were observed in vital organs of ST-F treated animals compared to control group. The ST-F may be exit slight side effects at the dose of 560 mg/kg/day in rats. Thus, the overall results show that the no-observed adverse effect level (NOAEL) of ST-F was considered to be 140 mg/kg for male SD rats.


Angiomotin-like 2 interacts with and negatively regulates AKT.
Angiomotin-like 2 (AMOTL2) is a member of Angiomotin family proteins, which are potent inhibitors for the Hippo pathway effector YAP. However, additional cellular functions of AMOTL2 besides YAP inhibition are largely unknown. Here, we reported that AMOTL2 associates with and negatively regulates AKT. Mechanistically, AMOTL2 binds to AKT pleckstrin homology domain and interrupts AKT's membrane localization. Liver-specific depletion of AMOTL2 enlarged mouse liver and activated both YAP and AKT. Downregulation of AMOTL2 is found in human liver cancers, correlating with the concomitant activation of YAP and AKT. Together, our results provide novel insights into a dual tumor suppressive function of AMOTL2 by targeting both YAP and AKT in liver size control and cancer prevention.


Immobilizing enzymes onto electrode arrays by hydrogel photolithography to fabricate multi-analyte electrochemical biosensors.
This paper describes a biomaterial microfabrication approach for interfacing functional biomolecules (enzymes) with electrode arrays. Poly (ethylene glycol) (PEG) hydrogel photopatterning was employed to integrate gold electrode arrays with the enzymes glucose oxidase (GOX) and lactate oxidase (LOX). In this process, PEG diacrylate (DA)-based prepolymer containing enzyme molecules as well as redox species (vinylferrocene) was spin-coated, registered, and UV cross-linked on top of an array of gold electrodes. As a result, enzyme-carrying circular hydrogel structures (600 microm diameter) were fabricated on top of 300 microm diameter gold electrodes. Importantly, when used with multiple masks, hydrogel photolithography allowed us to immobilize GOX and LOX molecules on adjacent electrodes within the same electrode array. Cyclic voltammetry and amperometry were used to characterize biosensor electrode arrays. The response of the biosensor array was linear for up to 20 mM glucose with sensitivity of 0.9 microA cm(-2) mM(-1) and 10 mM lactate with sensitivity of 1.1 microA cm(-2) mM(-1). Importantly, simultaneous detection of glucose and lactate from the same electrode array was demonstrated. A novel strategy for integrating biological and electrical components of a biosensor described in this paper provides the flexibility to spatially resolve and register different biorecognition elements with individual members of a miniature electrode array. Of particular interest to us are future applications of these miniature electrodes for real-time monitoring of metabolite fluxes in the vicinity of living cells.


Posttraumatic headache.
The authors review posttraumatic headache (PTH). The most common symptom following head injury, PTH is paradoxically most severe after mild head injury. Although most cases resolve within 6 to 12 months, many patients have protracted or even permanent headache. Because PTH generally has no objective findings, it is often controversial whether the symptom is "real," "psychogenic," or "fabricated." Despite persisting beliefs by physicians, attorneys, and insurers that PTH resolves upon legal settlement, recent studies have shown that "permanent" PTH is usually still present several years after a legal settlement. Often PTH affects family life, recreation, and employment. Patients require education and support as well as appropriate evaluation and treatment.


Association of Acinetobacter baumannii EF-Tu with cell surface, outer membrane vesicles, and fibronectin.
A conundrum has long lingered over association of cytosol elongation factor Tu (EF-Tu) with bacterial surface. Here we investigated it with Acinetobacter baumannii, an emerging opportunistic pathogen associated with a wide spectrum of infectious diseases. The gene for A. baumannii EF-Tu was sequenced, and recombinant EF-Tu was purified for antibody development. EF-Tu on the bacterial surface and the outer membrane vesicles (OMVs) was revealed by immune electron microscopy, and its presence in the outer membrane (OM) and the OMV subproteomes was verified by Western blotting with the EF-Tu antibodies and confirmed by proteomic analyses. EF-Tu in the OM and the OMV subproteomes bound to fibronectin as detected by Western blot and confirmed by a label-free real-time optical sensor. The sensor that originates from photonic crystal structure in a total-Internal-reflection (PC-TIR) configuration was functionalized with fibronectin for characterizing EF-Tu binding. Altogether, with a novel combination of immunological, proteomical, and biophysical assays, these results suggest association of A. baumannii EF-Tu with the bacterial cell surface, OMVs, and fibronectin.


Seven-year experience of enterocutaneous fistula with univariate and multivariate analysis of factors associated with healing: development of a validated scoring system.
The management of enterocutaneous fistulae (ECF) is complex and challenging. We examined factors associated with fistula healing at a National Intestinal Failure Centre and devised the first scoring system to predict spontaneous fistula healing prior to surgery. A retrospective audit of 177 patients (mean age 48.7 years) treated over 7 years was undertaken. Results were compared with a previously reported series from this unit. Univariate and multivariate analyses wete performed on variables to assess relationship with ECF healing. A scoring system was devised and validated on a prospective cohort. One-hundred and fifty patients underwent surgery between January 2003 and December 2009. The overall healing rate following surgery in the current series was 94.6% (82% in the previous series). Mean delay from previous surgery to the current operation was 1 year (compared with 8 months previously). Thirty-day postfistula resection mortality was 0% (compared with 3.5% previously). Twenty-seven patients underwent medical management alone with overall healing rate of 46.4% (vs 19.9%). Multivariate analysis revealed that comorbidity (P = 0.02), source of referral (P = 0.01) and aetiology (P = 0.006) had associations with healing. Almost all patients with scores of 0 and 1 healed, whereas the highest scores healed least frequently. Surgical management of ECF is safe and improving. Fistula healing is affected by aetiology, comorbidity and source of referral. The scoring system has the potential to predict ECF healing and can be a useful clinical decision-making tool.


Regulatory T cells ameliorate tissue plasminogen activator-induced brain haemorrhage after stroke.
Delayed thrombolytic treatment with recombinant tissue plasminogen activator (tPA) may exacerbate blood-brain barrier breakdown after ischaemic stroke and lead to lethal haemorrhagic transformation. The immune system is a dynamic modulator of stroke response, and excessive immune cell accumulation in the cerebral vasculature is associated with compromised integrity of the blood-brain barrier. We previously reported that regulatory T cells, which function to suppress excessive immune responses, ameliorated blood-brain barrier damage after cerebral ischaemia. This study assessed the impact of regulatory T cells in the context of tPA-induced brain haemorrhage and investigated the underlying mechanisms of action. The number of circulating regulatory T cells in stroke patients was dramatically reduced soon after stroke onset (84 acute ischaemic stroke patients with or without intravenous tPA treatment, compared to 115 age and gender-matched healthy controls). Although stroke patients without tPA treatment gradually repopulated the numbers of circulating regulatory T cells within the first 7 days after stroke, post-ischaemic tPA treatment led to sustained suppression of regulatory T cells in the blood. We then used the murine suture and embolic middle cerebral artery occlusion models of stroke to investigate the therapeutic potential of adoptive regulatory T cell transfer against tPA-induced haemorrhagic transformation. Delayed administration of tPA (10 mg/kg) resulted in haemorrhagic transformation in the ischaemic territory 1 day after ischaemia. When regulatory T cells (2 × 106/mouse) were intravenously administered immediately after delayed tPA treatment in ischaemic mice, haemorrhagic transformation was significantly decreased, and this was associated with improved sensorimotor functions. Blood-brain barrier disruption and tight junction damages were observed in the presence of delayed tPA after stroke, but were mitigated by regulatory T cell transfer. Mechanistic studies demonstrated that regulatory T cells completely abolished the tPA-induced elevation of MMP9 and CCL2 after stroke. Using MMP9 and CCL2 knockout mice, we discovered that both molecules partially contributed to the protective actions of regulatory T cells. In an in vitro endothelial cell-based model of the blood-brain barrier, we confirmed that regulatory T cells inhibited tPA-induced endothelial expression of CCL2 and preserved blood-brain barrier integrity after an ischaemic challenge. Lentivirus-mediated CCL2 knockdown in endothelial cells completely abolished the blood-brain barrier protective effect of regulatory T cells in vitro. Altogether, our studies suggest that regulatory T cell adoptive transfer may alleviate thrombolytic treatment-induced haemorrhage in stroke victims. Furthermore, regulatory T cell-afforded protection in the tPA-treated stroke model is mediated by two inhibitory mechanisms involving CCL2 and MMP9. Thus, regulatory T cell adoptive transfer may be useful as a cell-based therapy to improve the efficacy and safety of thrombolytic treatment for ischaemic stroke.


Insulin-Like Growth Factor 2 mRNA Binding Protein 3 Promotes Cell Proliferation of Malignant Mesothelioma Cells by Downregulating p27Kip1.
Malignant mesothelioma is a tumor with a poor prognosis, mainly caused by asbestos exposure and with no adequate treatment yet. To develop future therapeutic targets, we analyzed the microarray dataset GSE 29370 of malignant mesothelioma and reactive mesothelial hyperplasia, downloaded from the Gene Expression Omnibus (GEO) database. We identified insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3) as one of the significantly upregulated genes in malignant mesothelioma. IGF2BP3 functions as an oncoprotein in many human cancers; however, to our knowledge, this is the first study on the biological function of IGF2BP3 in malignant mesothelioma cells. The knockdown of IGF2BP3 in malignant mesothelioma cells resulted in the suppression of cell proliferation with an increase in the proportion of cells in the G1 phase of the cell cycle. Furthermore, knockdown of IGF2BP3 inhibited cell migration and invasion. We focused on the cell cycle assay to investigate the role of IGF2BP3 in cell proliferation in malignant mesothelioma. Among the various proteins involved in cell cycle regulation, the expression of p27 Kip1 (p27) increased significantly upon IGF2BP3 knockdown. Next, p27 siRNA was added to suppress the increased expression of p27. The results showed that p27 knockdown attenuated the effects of IGF2BP3 knockdown on cell proliferation and G1 phase arrest. In conclusion, we found that IGF2BP3 promotes cell proliferation, a critical step in tumorigenesis, by suppressing the expression of p27 in malignant mesothelioma.


Developing a prototype digital risk mitigation pathway for children and young people admitted to acute paediatric NHS care in mental health crisis: Protocol of the Safety Assessment in Paediatric healthcare Environments (SAPhE) pathway study.
Globally, there are increasing numbers of Children and young people (CYPs) experiencing a mental health crisis requiring admission to acute paediatric inpatient care. These CYPs can often experience fluctuating emotional states accompanied by urges to self-harm or attempt to end their life, leading to reduced safety and poorer experiences. Currently, in the UK National Health Service (NHS) there are no standardised, evidence-based interventions in acute paediatric care to mitigate or minimise immediate risk of self-harm and suicide in CYP admitted with mental health crisis. To outline the protocol for the SAPhE Pathway study which aims to: 1) identify and prioritise risk mitigation strategies to include in the digital prototype, 2) understand the feasibility of implementing a novel digital risk mitigation pathway in differing NHS contexts, and 3) co-create a prototype digital risk mitigation pathway. This is a multi-centre study uses a mixed-methods design. A systematic review and exploratory methods (interviews, surveys, and focus groups) will be used to identify the content and feasibility of implementing a digital risk mitigation pathway. Participants will include healthcare professionals, digital experts and CYP with experience of mental health conditions. Data will be collected between January 2022 and March 2023 and analysed using content and thematic analysis, case study, cross-case analysis for qualitative data and descriptive statistics for quantitative data. Findings will inform the experience-based co-design workshops. The study received full ethical approval from NHS REC [Ref: 22/SC/0237 and 22/WM/0167]. Findings will be made available to all stakeholders using multiple approaches.


[Corrective possibilities by subcapital metatarsal osteotomy in the management of hallux valgus and metatarsus primus valgus].
The extent of correction achievable by inclined subcapital osteotomy of the metatarsus I. was studied in 100 cases of the deformation. The different angular values and dimensions were compared in the X-ray plates taken pre- and postoperatively and the clinical changes of the patients were evaluated on the basis of subjective complaints and objective symptoms. The angular limit values of the performability of osteotomy were determined. In deformations larger than the limit values wedge and half-wedge osteotomy, basisosteotomy, respectively, are the expedient methods.


Maternal-fetal relationships in the parathyroidectomized rat. Intestinal calcium transport, serum calcium, immunoreactive parathyroid hormone and calcitonin.
We studied the role of the parathyroids in the adaptation of intestinal Ca transport that occurs during pregnancy, and whether maternal hypoparathyroidism causes fetal hyperparathyroidism. Serum Ca of pregnant parathyroidectomized (PTX) rats was significantly greater than nonpregnant, PTX animals. Intestinal active Ca transport was increased 2.1- and 2.2-fold by pregnancy in intact and PTX rats, respectively. Serum levels of immunoreactive parathyroid hormone (PTH) were nondetectable in PTX-pregnant rats. Fetuses from PTX rats appeared grossly normal. The serum PTH was not different in fetuses from PTX compared to fetuses from intact mothers and serum Ca, Mg, and P were normal. Thus, alleviation of maternal hypocalcemia during pregnancy in PTX rats may be due to an adaptive increase in intestinal Ca transport, which does not require the parathyroids. Fetuses from PTX mothers were euparathyroid and were protected from Ca deficiency during pregnancy.


Free tissue breast reconstruction in older patients.
We present the case of a 72-year-old woman who underwent mastectomy with immediate breast reconstruction using a free transverse rectus abdominis myocutaneous (TRAM) flap, to illustrate that age is not necessarily a barrier to microvascular reconstructive techniques for breast reconstruction.


Expression of carbonic anhydrase IX in human fetal joints, ligaments and tendons: a potential marker of mechanical stress in fetal development?
Carbonic anhydrase type IX (CA9) is known to express in the fetal joint cartilage to maintain pH against hypoxia. Using paraffin-embedded histology of 10 human fetuses at 10-16 weeks of gestation with an aid of immunohistochemistry of the intermediate filaments, matrix components (collagen types I and II, aggrecan, versican, fibronectin, tenascin, and hyaluronan) and CA9, we observed all joints and most of the entheses in the body. At any stages examined, CA9-poisitive cells were seen in the intervertebral disk and all joint cartilages including those of the facet joint of the vertebral column, but the accumulation area was reduced in the larger specimens. Glial fibrillary acidic protein (GFAP), one of the intermediate filaments, expressed in a part of the CA9-positive cartilages. Developing elastic cartilages were positive both of CA9 and GFAP. Notably, parts of the tendon or ligament facing to the joint, such as the joint surface of the annular ligament of the radius, were also positive for CA9. A distribution of each matrix components examined was not same as CA9. The bone-tendon and bone-ligament interface expressed CA9, but the duration at a site was limited to 3-4 weeks because the positive site was changed between stages. Thus, in the fetal entheses, CA9 expression displayed highly stage-dependent and site-dependent manners. CA9 in the fetal entheses seemed to play an additional role, but it was most likely to be useful as an excellent marker of mechanical stress at the start of enthesis development.


Action of naloxone on the interferon-lowering activity of morphine in the mouse.
The serum titer of interferon produced by polyinosinic:polycytidylic acid (poly I:C) was studied in mice following subcutaneous injections of either morphine alone, naloxone alone, morphine and naloxone together, morphine preceded by naloxone, naloxone preceded by morphine, or a sequence of injections of naloxone, morphine, and naloxone again. Poly I:C injected without the other drugs present served as the control. Both morphine and naloxone produced a significant decrease in the ability of the mice to increase the serum titer of interferon in response to poly I:C. Naloxone appeared to function as an antagonist to morphine when administered following the morphine injection, but not when injected before or simultaneously with the opiate. When naloxone was injected both before and following the morphine injection, results were interpreted to indicate the possibility that naloxone could function as both an agonist and antagonist under this condition.


Novel insight into the role of heterotrophic dinoflagellates in the fate of crude oil in the sea.
Although planktonic protozoans are likely to interact with dispersed crude oil after a spill, protozoan-mediated processes affecting crude oil pollution in the sea are still not well known. Here, we present the first evidence of ingestion and defecation of physically or chemically dispersed crude oil droplets (1-86 μm in diameter) by heterotrophic dinoflagellates, major components of marine planktonic food webs. At a crude oil concentration commonly found after an oil spill (1 μL L(-1)), the heterotrophic dinoflagellates Noctiluca scintillans and Gyrodinium spirale grew and ingested ~0.37 μg-oil μg-C(dino)(-1) d(-1), which could represent ~17% to 100% of dispersed oil in surface waters when heterotrophic dinoflagellates are abundant or bloom. Egestion of faecal pellets containing crude oil by heterotrophic dinoflagellates could contribute to the sinking and flux of toxic petroleum hydrocarbons in coastal waters. Our study indicates that crude oil ingestion by heterotrophic dinoflagellates is a noteworthy route by which petroleum enters marine food webs and a previously overlooked biological process influencing the fate of crude oil in the sea after spills.


Physical modelling in biomechanics.
Physical models, like mathematical models, are useful tools in biomechanical research. Physical models enable investigators to explore parameter space in a way that is not possible using a comparative approach with living organisms: parameters can be varied one at a time to measure the performance consequences of each, while values and combinations not found in nature can be tested. Experiments using physical models in the laboratory or field can circumvent problems posed by uncooperative or endangered organisms. Physical models also permit some aspects of the biomechanical performance of extinct organisms to be measured. Use of properly scaled physical models allows detailed physical measurements to be made for organisms that are too small or fast to be easily studied directly. The process of physical modelling and the advantages and limitations of this approach are illustrated using examples from our research on hydrodynamic forces on sessile organisms, mechanics of hydraulic skeletons, food capture by zooplankton and odour interception by olfactory antennules.


Modification of the existing maximum residue levels and setting of import tolerances for flupyradifurone and DFA in various crops and animal commodities.
In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Bayer CropScience SA-NV submitted a request to the competent national authority in the Netherlands to modify the existing maximum residue levels (MRLs) and to set import tolerances for the active substance flupyradifurone and its metabolite difluoroacetic acid (DFA) in various plant and animal commodities. The data submitted in support of the requests were found sufficient to derive MRL proposals for flupyradifurone and/or DFA in the crops under consideration. The calculated EU livestock dietary burden indicated that for several animal matrices the EU MRLs for flupyradifurone and/or DFA would need to be modified. Adequate analytical methods for enforcement are available to control the residues of both compounds in the plant and animal commodities under consideration, and in honey. Based on the risk assessment results, EFSA concluded that the proposed and authorised uses of flupyradifurone on various crops and subsequent residues of flupyradifurone and DFA in plant and animal commodities will not result in a consumer exposure exceeding the toxicological reference values and therefore is unlikely to pose a risk to consumers' health.


[Composition of Eimeria in turkeys in Bulgaria].
Studied was the species composition of Eimeria in turkeys on a pedigree farm in the territory of the Higher Institute of Zootechnics and Veterinary Medicine. Metrical and morphologic investigations were carried out with more than 1000 sporulated oocysts isolated from parental flocks, turkey poults for fattening, clinically affected poults at various ages, and from experimentally infected poults. On the basis of the results obtained the following species were ascertained: Eimeria meleagridis Tyzzer, 1927; E. adenoeides Moore, Brown, 1951; E. meleagrimitis Tyzzer, 1929. Most widely spread proved the Eimeria meleagridis species (52 per cent), followed by E. adenoeides (28.4 per cent), and E. meleagrimitis (26.6 per cent).