Axon hyperexcitability in the contralateral projection following unilateral optic nerve crush in mice.
Optic neuropathies are characterized by degeneration of retinal ganglion cell axonal projections to the brain, including acute conditions like optic nerve trauma and progressive conditions such as glaucoma. Despite different aetiologies, retinal ganglion cell axon degeneration in traumatic optic neuropathy and glaucoma share common pathological signatures. We compared how early pathogenesis of optic nerve trauma and glaucoma influence axon function in the mouse optic projection. We assessed pathology by measuring anterograde axonal transport from retina to superior colliculus, current-evoked optic nerve compound action potential and retinal ganglion cell density 1 week following unilateral optic nerve crush or intraocular pressure elevation. Nerve crush reduced axon transport, compound axon potential and retinal ganglion cell density, which were unaffected by intraocular pressure elevation. Surprisingly, optic nerves contralateral to crush demonstrated 5-fold enhanced excitability in compound action potential compared with naïve nerves. Enhanced excitability in contralateral sham nerves is not due to increased accumulation of voltage-gated sodium channel 1.6, or ectopic voltage-gated sodium channel 1.2 expression within nodes of Ranvier. Our results indicate hyperexcitability is driven by intrinsic responses of αON-sustained retinal ganglion cells. We found αON-sustained retinal ganglion cells in contralateral, sham and eyes demonstrated increased responses to depolarizing currents compared with those from naïve eyes, while light-driven responses remained intact. Dendritic arbours of αON-sustained retinal ganglion cells of the sham eye were like naïve, but soma area and non-phosphorylated neurofilament H increased. Current- and light-evoked responses of sham αOFF-sustained retinal ganglion cells remained stable along with somato-dendritic morphologies. In retinas directly affected by crush, light responses of αON- and αOFF-sustained retinal ganglion cells diminished compared with naïve cells along with decreased dendritic field area or branch points. Like light responses, αOFF-sustained retinal ganglion cell current-evoked responses diminished, but surprisingly, αON-sustained retinal ganglion cell responses were similar to those from naïve retinas. Optic nerve crush reduced dendritic length and area in αON-sustained retinal ganglion cells in eyes ipsilateral to injury, while crush significantly reduced dendritic branching in αOFF-sustained retinal ganglion cells. Interestingly, 1 week of intraocular pressure elevation only affected αOFF-sustained retinal ganglion cell physiology, depolarizing resting membrane potential in cells of affected eyes and blunting current-evoked responses in cells of saline-injected eyes. Collectively, our results suggest that neither saline nor sham surgery provide a true control, chronic versus acute optic neuropathies differentially affect retinal ganglion cells composing the ON and OFF pathways, and acute stress can have near-term effects on the contralateral projection.


THE SCALING OF PLANT AND ANIMAL BODY MASS, LENGTH, AND DIAMETER.
The interspecific scaling exponents of body mass M and diameter D with respect to length L were determined to evaluate the predictions of three scaling hypotheses (geometric, stress, and elastic similitude). The relation between M and L was determined for data from a total of 133 aquatic and terrestrial species (66 plant and 67 animal species); the relation between D and L was determined independently for a total of 753 aquatic and terrestrial species (667 plant and 86 animal species). Organisms were crudely classified as to their geometry (spheres, spheroids, cylinders) and shape (defined as the body slenderness factor, L/D) to examine whether geometry and shape evinced size-dependent variations. Regression indicated M = 1.29L2.95 (r2 = 0.91, N = 133; αRMA = 3.09 ± 0.088). The stress and elastic similitude (which respectively predict αRMA = 5 and αRMA = 4) were rejected; geometric similitude was not (αRMA = 3). For animals and plants, αRMA = 2.81 ± 0.061 (r2 = 0.98), and αRMA = 2.95 ± 0.093 (r2 = 0.94), respectively. For aquatics and terrestrial organisms, αRMA = 2.82 ±0.134 (r2 = 0.97, N = 36), and αRMA = 3.08 ±0.111 (r2 = 0.89, N = 97), respectively. These results were interpreted to support the hypothesis of geometric similitude. For the pooled plant and animals data, D = 0.05L1.00 (r2 = 0.95; αRMA = 1.03 ± 0.009), which was compatible with the hypothesis of geometric similitude. For plants, D = 0.05L1.06 (r2 = 0.95; αRMA = 1.09). For animals, D = 0.29L0.98 (r2 = 0.95; αRMA = 1.01 ± 0.025). Also, for aquatics, αRMA = 0.951 ± 0.151, whereas for terrestrial plants and animals, αRMA = 1.03 ± 0.089. Although the scaling exponent for D differed among individual groupings of animals and plants, the results of regression analyses were interpreted to indicate that, on the average, body diameter scaled isometrically with respect to length as predicted by geometric similitude. For the pooled data set, organic shape varied over 3 orders of magnitude; L varied over 9 orders of magnitude reflecting 22 orders of magnitude of M. In terms of body geometry and the absolute numbers of species in the total data set: spherical shaped species (L = D) < unassigned species < prolate spheroidal species < cylindrical (squat < slender) species. The largest organisms in the data set were slender (L/D > 20) cylindrical plants; the smallest organisms were spherical plants and animals. Although not subject to statistical inference, these data were interpreted to indicate that organic shape and geometry evince size-dependent variations. These variations as well as size-dependent changes in bulk density are hypothesized to account for the scaling exponents of M and D determined for individual plant and animal clades and grades.


Dog-assisted therapy for older people with dementia: a review.
This review summarises and critiques the published literature regarding dog therapy for older people with dementia living in residential aged care facilities. Nine studies were identified for inclusion and although the methodological variability of studies makes it difficult to draw firm conclusions, research suggests that dog therapy is beneficial for people with dementia. The most frequently reported findings were an increase in social behaviour and a decrease in agitated behaviour during dog contact. Improvement in social behaviour was found to be unrelated to the severity of dementia. Various improvements on measures of global function were also reported. No study adopted a randomised controlled trial design and a number of potentially important factors were not controlled for, including halo effects of animals on caregivers that may bias caregivers' responses when acting as proxies for their relatives or residents. The premorbid relationship with dogs may be an important variable influencing outcomes.


Process and impact of an advance care planning intervention evaluated by bereaved surrogate decision-makers of dialysis patients.
Few studies have examined the process and impact of an advance care planning intervention experienced by surrogate decision-makers of dialysis patients. To explore the perspectives of the bereaved surrogates of dialysis patients on the process and impact of an advance care planning intervention and to compare the perceived impacts of the intervention between African Americans and Whites. Qualitative interviews and thematic analysis. 24 bereaved surrogates of patients from outpatient dialysis centers were interviewed. Both patients and surrogates had been participants in a larger efficacy study and had received an advance care planning intervention, SPIRIT (Sharing Patient's Illness Representations to Increase Trust). Two themes related to the process of SPIRIT were as follows: (1) SPIRIT provided a welcome opportunity to think about and discuss topics that had been avoided and (2) SPIRIT helped patients and surrogates to share their feelings. Four themes of the SPIRIT's impact were as follows: (1) SPIRIT was an eye-opening experience, acquiring knowledge and understanding of the patient's illness and end-of-life care, (2) SPIRIT helped strengthen relationships between patients and surrogates, (3) SPIRIT helped surrogates feel prepared during the time leading up to end-of-life decision-making, and (4) SPIRIT helped surrogates have peace of mind during and after actual end-of-life decision-making. Themes related to SPIRIT's impact on feeling prepared for end-of-life decision-making and the actual decision-making experience more frequently occurred in African Americans than in Whites. Our data may help explain the beneficial effects of SPIRIT on surrogates, but future trials should include data on control surrogates' perspectives.


Inhibitory effect of arsenic trioxide on angiogenesis and expression of vascular endothelial growth factor in gastric cancer.
To investigate the inhibitory effect of As(2)O(3) on angiogenesis of tumor and expression of vascular endothelial growth factor (VEGF) in tumor cells in vivo and in vitro. The solid tumor model was formed in nude mice with the gastric cancer cell line SGC-7901. The animals were randomly divided into three groups. As(2)O(3) was injected into the arsenic-treated groups (2.5 mg/kg and 5 mg/kg) and the same volume of saline solution was injected into the control group. Microvessel density (MVD) and expression of VEGF were detected with immunofluorescence laser confocal technology. Further expression of VEGF protein and VEGF mRNA was measured with Western bloting and fluorescence quantitative RT- PCR in SGC-7901 cells treated with As(2)O(3). In nude mice, after treatment with 5 mg/kg and 2.5 mg/kg As(2)O(3) respectively, about 50% and 30% tumor growth inhibition were observed correspondingly (P<0.05, P<0.05). Decrease in MVD appeared in As(2)O(3)-treated tumors compared with control group (P<0.001, P<0.001). MVD in tumors was significantly lower in 5 mg/kg group than in 2.5 mg/kg group (P<0.01). The fluorescence intensity levels of VEGF in tumor cells were significantly lowered in the arsenic-treated groups (P<0.01, P<0.01). The fluorescence intensity level of VEGF in 5 mg/kg group was lower than that in 2.5 mg/kg group (P<0.01). In vitro, the expression of VEGF protein decreased in dose- and time-dependent manner after the treatment with As(2)O(3), but in VEGF mRNA no significant difference was found between the control group and the treated groups. As(2)O(3) can inhibit solid tumor growth by inhibiting the formation of new blood vessels. One of the mechanisms is that As(2)O(3) can inhibit VEGF protein expression.


Expression of kinin receptors on eosinophils: comparison of asthmatic patients and healthy subjects.
Eosinophils contribute to asthmatic airway inflammation by releasing cysteinyl leukotrienes (cysLT) and other inflammatory mediators, and bradykinin (BK) induces bronchoconstriction in asthmatic patients. The aims of this study were to investigate kinin receptor expression on eosinophils of asthmatic and healthy subjects and to assess the effects of kinin stimulation on eosinophils, which were isolated from peripheral blood of asthmatic (n=27) and healthy subjects (n=14). Kinin B(1) and B(2) receptors (B(1)R and B(2)R, respectively) and mRNA expression were investigated by quantitative confocal microscopy, flow cytometry, and RT-PCR. Intracellular Ca(2+) was assessed by live-cell fluorescence confocal microscopy. Production of cysLT and eosinophil migration in response to BK and Lys-des[Arg(9)]-BK were assessed. Eosinophils expressed kinin B(1)R and B(2)R mRNA and proteins. Quantitative immunofluorescence analysis indicated that expression of B(1)R and B(2)R proteins was significantly greater in eosinophils of asthmatic patients compared with those of nonasthmatic subjects. However, kinin B(1)R and B(2)R mRNA expression did not differ significantly between these groups. Expression of kinin B(1)R and mRNA was decreased in patients using high doses of inhaled corticosteroids and in eosinophils treated with a corticosteroid in vitro. Kinin B(1) and B(2) agonists up-regulated expression of their respective receptors but did not increase intracellular Ca(2+) or the production of cysLT or enhance eosinophil migration significantly. Up-regulation of kinin receptor expression in eosinophils of asthmatic patients may be a consequence of inflammation, whereby enhanced release of kinin peptides has a positive-feedback effect on kinin receptor expression. Importantly, anti-inflammatory corticosteroids down-regulated the expression of the kinin B(1)R.


Characterization of Virus Replication, Pathogenesis, and Cytokine Responses in Syrian Hamsters Inoculated with SARS-CoV-2.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel coronavirus which caused a global respiratory disease pandemic beginning in December 2019. Understanding the pathogenesis of infection and the immune responses in a SARS-CoV-2-infected animal model is urgently needed for vaccine development. Syrian hamsters (Mesocricetus auratus) were intranasally inoculated with 105, 5×105, and 106 TCID50 of SARS-CoV-2 per animal and studied for up to 14 days. Body weight, viral load and real-time PCR amplification of the SARS-CoV-2 N gene were measured. On days 3, 6 and 9, lung, blood, liver, pancreas, heart, kidney, and bone marrow were harvested and processed for pathology, viral load, and cytokine expression. Body weight loss, increased viral load, immune cell infiltration, upregulated cytokine expression, viral RNA, SARS-CoV-2 nucleoprotein, and mucus were detected in the lungs, particularly on day 3 post-infection. Extremely high expression of the pro-inflammatory cytokines MIP-1 and RANTES was detected in lung tissue, as was high expression of IL-1β, IL-6, IL-12, and PD-L1. The glutamic oxalacetic transaminase/glutamic pyruvic transaminase (GOT/GPT) ratio in blood was significantly increased at 6 days post-infection, and plasma amylase and lipase levels were also elevated in infected hamsters. Our results provide new information on immunological cytokines and biological parameters related to the pathogenesis and immune response profile in the Syrian hamster model of SARS-CoV-2 infection.


Icariin Prevents H2O2-Induced Apoptosis via the PI3K/Akt Pathway in Rat Nucleus Pulposus Intervertebral Disc Cells.
Icariin is a prenylated flavonol glycoside derived from the Chinese herb Epimedium sagittatum. This study investigated the mechanism by which icariin prevents H2O2-induced apoptosis in rat nucleus pulposus (NP) cells. NP cells were isolated from the rat intervertebral disc and they were divided into five groups after 3 passages: (A) blank control; (B) 200 μM H2O2; (C) 200 μM H2O2 + 20 μM icariin; (D) 20 μM icariin + 200 μM H2O2 + 25 μM LY294002; (E) 200 μM H2O2 + 25 μM LY294002. LY294002 is a selective inhibitor of the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway. NP cell viability, apoptosis rate, intracellular reactive oxygen species levels, and the expression of AKT, p-AKT, p53, Bcl-2, Bax, caspase-3 were estimated. The results show that, compared with the control group, H2O2 significantly increased NP cell apoptosis and the level of intracellular ROS. Icariin pretreatment significantly decreased H2O2-induced apoptosis and intracellular ROS and upregulated p-Akt and BCL-2 and downregulated caspase-3 and Bax. LY294002 abolished the protective effects of icariin. Our results show that icariin can attenuate H2O2-induced apoptosis in rat nucleus pulposus cells and PI3K/AKT pathway is at least partly included in this protection effect.


Different definitions of esophagus influence esophageal toxicity prediction for esophageal cancer patients administered simultaneous integrated boost versus standard-dose radiation therapy.
We aim to evaluate whether different definitions of esophagus (DEs) impact on the esophageal toxicity prediction for esophageal cancer (EC) patients administered intensity-modulated radiation therapy with simultaneous integrated boost (SIB-IMRT) vs. standard-dose IMRT (SD-IMRT). The esophagus for 21 patients diagnosed with primary EC were defined in the following four ways: the whole esophagus, including the tumor (ESOwhole); ESOwhole within the treatment field (ESOinfield); ESOinfield, excluding the tumor (ESOinfield-tumor) and ESOwhole, excluding the tumor (ESOwhole-tumor). The difference in the dose variation, acute esophageal toxicity (AET) and late esophageal toxicity (LET) of four DEs were compared. We found that the mean esophageal dose for ESOwhole, ESOinfield, ESOinfield-tumor and ESOwhole-tumor were increased by 7.2 Gy, 10.9 Gy, 4.6 Gy and 2.0 Gy, respectively, in the SIB-IMRT plans. Radiobiological models indicated that a grade ≥ 2 AET was 2.9%, 3.1%, 2.2% and 1.6% higher on average with the Kwint model and 14.6%, 13.2%, 7.2% and 3.4% higher with the Wijsman model for the four DEs. A grade ≥ 3 AET increased by 4.3%, 7.2%, 4.2% and 1.2%, respectively. Additionally, the predicted LET increased by 0.15%, 0.39%, 1.2 × 10-2% and 1.5 × 10-3%. Our study demonstrates that different DEs influence the esophageal toxicity prediction for EC patients administered SIB-IMRT vs. SD-IMRT treatment.


Genetic studies at the receptor level: investigations in human twins and experimental animals.
In receptors, as in enzymes, quantitative as well as qualitative genetic variation may exist. Studies in inbred strains of mice have shown for various receptors that the receptor density as determined by Bmax values is under genetic control. In healthy adult twins we have shown that the density of alpha-adrenoceptors on platelets is also influenced by genetic factors, since monozygotic twins were much more similar to one another than dizygotic twins. However, Bmax values are up-regulated and down-regulated by endogenous neurotransmitters and pharmacologically active agents. Thus, receptor densities are under considerable regulatory influences. Bmax values therefore reflect regulatory mechanisms rather than innate characteristics of the receptor protein. In another twin study we failed to find evidence for a genetic influence on the density of imipramine-binding sites on platelets. Since qualitative variation (polymorphism) is well known in enzymes, it may also apply to receptors. Qualitative differences in the receptor protein within one species would be of particular interest because of possible functional implications. As a first approach we examined central benzodiazepine receptors by photoaffinity labelling and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. A comparison of fish, frog, chicken, mouse, rat and calf led to the detection of variation between species. Investigations in five inbred mouse and rat strains have not so far revealed genetic variation in benzodiazepine receptors. Nevertheless variation may be detectable by more sensitive methods such as peptide mapping after limited proteolysis or two-dimensional electrophoresis.


Isolation of Campylobacter jejuni and coliform bacilli from bile and liver obtained from slaughter cattle in Western Japan.
A total of 290 bile samples from 143 Japanese Brown, 97 Japanese Black, and 50 Holstein cattle, and a total of 148 liver samples from 81 Japanese Brown, 49 Japanese Black, and 18 Holstein cattle were examined for the presence of Campylobacter jejuni by direct plating. The bile samples were also subjected to enumeration of coliform bacilli. Sixty-eight (23%) bile samples and 2 (1.4%) liver samples were positive for C. jejuni. A significantly higher isolation rate was observed from bile samples from Holstein (50%) than from Japanese Black (22%) and Japanese Brown (15%) cattle. C. jejuni was isolated from 52 of 232 bile samples that contained < 30 CFU/ml (under the detection threshold) of coliform bacilli. The presence of C. jejuni from bile was observed throughout the year. Fifty-four of the 68 bile isolates were serologically typed into eight groups. Serotypes O:4 complex (28 isolates) and O:2 (11 isolates), which were commonly isolated from human patients in Japan, accounted for 57% of the isolates. These observations suggest that bile can be a cause of contamination with C. jejuni even though it contains only a low number of coliforms.


The social construction of illness experience: families with a chronically-ill child.
This paper examines how parents constructed the illness of a child with chronic sickness. The focus is on eliciting the parents' explanatory model of illness, that is, how they understood and described their child's illness, and how they managed their interactions with the child. The data were collected by using an ethnographic approach. In addition to obtaining parents' accounts, naturally occurring interactions between parents and children were observed and tape recorded. The paper discusses the parents' semantic definition of the "child as normal'. In actual practice, however, the child's life was regulated by the parents' understanding of the biomedical model, and included time schedules dictated by the sickness and seclusion from other children. Adjustment in family lifestyle was seen as necessary to accommodate the child's health problem. It is argued that the parents' interactions with the child contain the essential feature of paradoxical communication. An ethnographically based theoretical model of the social construction of sickness careers is introduced. The paper gives verbatim accounts of naturally occurring interactions so that the reader has access to the data upon which the author's interpretations are based. The implications for nursing practice are discussed.


Transplantation of fetal cardiomyocytes into infarcted rat hearts results in long-term functional improvement.
Studies have demonstrated the feasibility of transplanting cardiomyocytes after myocardial infarction (MI). However, persistence and effects on left ventricular (LV) function have not been elucidated in long-term studies. Ventricular fetal cardiomyocytes from embryos of both sexes were injected into marginal regions of MI 4 weeks after suture occlusion of the left anterior descending artery in adult female rats. Two and 6 months after transplantation (Tx), engrafted cells were traced by immunohistochemical in situ hybridization for Y chromosomes or bromodeoxyuridine (BrdU) staining, LV dimensions and function were assessed by echocardiography, and LV pressure was assessed ex vivo in a Langendorff perfusion system. Immunohistochemistry for alpha-sarcomeric actin and Y chromosomes revealed the presence of transplanted cells in infarcted host myocardium at both 2 and 6 months. End-diastolic LV diameter markedly decreased after Tx and fractional shortening gradually increased after Tx (31.3 +/- 4.5% before Tx, 45.4 +/- 4.2% at 6 months; p<0.005). Wall area fraction and MI size were unaffected by Tx. In hearts with MI, but not in normal hearts, Tx led to the development of higher pressures (87 +/- 18 versus 38 +/- 8 mmHg, 6 months post-Tx versus nontreated). After catecholamine stimulation, both infarcted and normal hearts developed higher pressures after Tx (p<0.005), ultimately associated with reduced mortality after Tx versus nontreated. Transplanted cardiomyocyte-rich graft cells persist in host myocardium and mediate continuous improvement of LV function and survival in a rat model of MI even during long-term follow-up, possibly involving a catecholamine-sensitive mechanism.


Delayed approach of a penile fracture with encapsulated hematoma.
Penile fracture (PF) is defined as the rupture of the tunica albuginea (TA) of the corpora cavernosa (CC) caused by trauma to the erect penis. We present a case and clinical evolution of the delayed approach of PF. Physical examination showed a ventral rounded mass in the middle surface of the penile shaft, associated with mild discoloration and edema. Surgery was performed with a vertical penoscrotal incision. We found an encapsulated hematoma on the right ventral mid penile shaft connected at its base to an approximate 1 cm transverse defect on the TA and we performed debridement and excision of the hematoma. Tunical defect was repaired with PDS 3/0 simple suture. The patient had a great postoperative evolution without local complications. The early diagnosis and surgical treatment reaches better functional results, with maintenance of erectile function in patients with penile fracture.


Methadone treatment during pregnancy.
Methadone maintenance has been used for decades to treat opioid-dependent pregnant women. The outcomes of pregnancies thus treated are vastly improved over the outcomes of pregnancies complicated by street drug use. Despite its long history of successful use during pregnancy, little is known about the long-term effects of methadone on the fetus and the newborn. Studies done in animals suggest there may be subtle effects on brain and behavior. Only recently have other treatments for opioid dependency during pregnancy been investigated. There is increasing evidence that altering the traditional methadone maintenance protocols may be beneficial, and that tapered withdrawal can be safely achieved under some circumstances.


Screening for disruptive behavior of institutionalized juvenile offenders.
Case records of 70 male adjudicated juvenile offenders were used to examine the utility of various demographic, intelligence, and personality variables in identifying youths likely to display disruptive/violent behavior while institutionalized. Stepwise discriminant function analyses revealed that variable combinations from the Psychological Screening Inventory and Shipley -Hartford Scale were generally the most effective in identifying disruptive youths, while Jesness Inventory Subscales added little to the classification process. Implications for the use of screening measures in identifying potentially disruptive youths at admission are discussed.


The hypothalamus-pituitary-adrenal axis does not influence the protective effects of nociceptin/orphanin FQ on the rat gastric mucosa.
The participation of hypothalamus-pituitary-adrenal axis in the gastroprotective effects of nociceptin/orphanin FQ (N/OFQ) has been investigated. Gastric mucosal lesions were induced by intragastric administration of 50% ethanol, 1 ml/rat. Rats received N/OFQ either by the intracerebroventricular (icv) route, at 3 microg/rat, or by the intraperitoneal (ip) route, at 10 microg/kg, 30 min before ethanol administration. The protective effect of icv and ip administered N/OFQ was assessed in adrenalectomized rats and in rats pretreated with the glucocorticoid receptor antagonist, mifepristone, or with the CRF receptor antagonist, alpha-helical CRF(9-41). The damaging effect of ethanol was apparently not influenced by adrenalectomy. N/OFQ markedly reduced macroscopically and histologically assessed gastric mucosal damage. The extent of reduction by N/OFQ was comparable in adrenalectomized and in sham-operated rats, with either icv or ip route of administration. Pretreatment with mifepristone, both icv (80 microg/rat) and ip (10 mg/kg) injected, did not modify the response to icv and ip N/OFQ. Pretreatment with alpha-helical CRF(9-41) (25 microg/rat icv or 250 microg/kg ip), had no effect on the reduction of gastric damage produced by icv or ip N/OFQ. Present findings suggest that the gastroprotective effects of N/OFQ on ethanol-induced damage do not involve the endocrine pathway through the HPA axis.


Discharging patients receiving enteral nutrition.
When a patient is receiving enteral nutrition in hospital and is otherwise medically fit (or soon to become fit) for discharge back to the home and into community institutions such as nursing homes and hospices, home enteral nutrition (HEN) is usually considered. This article describes the process used at Birmingham Heartlands Hospital for the discharge into the community of patients receiving enteral tube feeding.


Arabinoxylo-Oligosaccharides and Inulin Impact Inter-Individual Variation on Microbial Metabolism and Composition, Which Immunomodulates Human Cells.
Fecal batch fermentations coupled to cocultures of epithelial cells and macrophages were used to compare how arabinoxylo-oligosaccharides (AXOS) and inulin modulate gut microbial activity and composition of three different human donors and subsequently the epithelial permeability and immune response. Both inulin and AXOS decreased the pH during incubation (-1.5 pH units), leading to increased productions of acetate, propionate, and butyrate. Differences in terms of metabolites production could be linked to specific microbial alterations at genus level upon inulin/AXOS supplementation (i.e., Bifidobacterium, Bacteroides, Prevotella and unclassified Erysipelotrichaceae), as shown by 16S-targeted Illumina sequencing. Both products stimulated gut barrier and immune function with increases in TEER, NF-KB, IL-10, and IL-6. Ingredients with different structures selectively modulate the microbiota of a specific donor leading to differential changes at metabolic level. The extent of this effect is donor specific and is linked to a final specific modulation of the host's immune system.


Exploring the parameters of intensity, frequency, and duration within the constraint induced movement therapy published research: A content analysis.
Neurological rehabilitation specialists must determine appropriate dosage, consisting of frequency, intensity, and duration of specified treatments. The objective of this study was to perform a content analysis of the current literature related to dosage (duration, frequency and intensity) for constraint-induced movement therapy (CIMT) in the adult population. A content analysis was conducted which yielded 62 scholarly articles. The frequency of CIMT ranged from 1 to 7 days per week with the average frequency being 4.98 days. The duration of CIMT, ranged from 2 to 10 weeks with the average duration 3.14 weeks. All three components of dosage were reported collectively in outcomes studies conducted in inpatient settings and rehabilitation clinics. The findings provide a groundwork for evidence based practice for clinician in the application of CIMT dosage with consideration of settings and CIMT components.


Brief Report: Comparing Sexual Risk Behavior in a High-Risk Group of Men Who Have Sex With Men and Transgender Women in Lima, Peru.
Transgender women (TW) and men who have sex with men (MSM) are often conflated in HIV research and prevention programs, despite clear differences that exist in culture and behavior. We examined baseline data from a large treatment-as-prevention study among TW and MSM in Lima, Peru, to assess differences in risk behavior. Baseline assessment included HIV testing and a questionnaire including sociodemographics, sexual behavior, social venue attendance, and drug and alcohol use. Poisson regression with robust standard errors was used to calculate prevalence ratios adjusted for confounding variables [adjusted prevalence ratio (aPR)] and 95% confidence intervals (CIs) comparing the prevalence of covariates related to HIV risk in MSM and TW. Overall, 310 TW and 2807 MSM participated between July 2013 and September 2015 and were included in this analysis. TW engaged in some protective sexual health practices more than MSM, including HIV testing in the last year (aPR = 1.62; 95% CI: 1.42 to 1.84) and condom use at the last sexual encounter (aPR = 1.20; 95% CI: 1.06 to 1.36). TW were more likely to have sex while using alcohol (aPR 1.15, 95% CI: 1.01 to 1.31) or drugs (aPR 2.24, 95% CI: 1.47 to 3.41), have alcohol dependency (aPR 1.38, 95% CI: 1.15 to 1.66), engage in receptive anal sex (aPR 1.31, 95% CI: 1.26 to 1.36), and have received money, gifts, or favors in exchange of anal sex (1.96, 95% CI: 1.74 to 2.20). TW and MSM exhibited distinct risk profiles, suggesting that interventions specifically targeted to each group may provide new opportunities for more effective HIV prevention programs.


Children's Oncology Group's 2023 blueprint for research: Pharmacy.
Children's Oncology Group (COG) pharmacists and pharmacy technicians from more than 200 COG-member institutions comprise the COG Pharmacy Discipline. Discipline members serve an essential role in the design and execution of COG clinical trials. Core activities include study drug management, study drug access, clinical trial operations, protocol harmonization, and direct patient care. Discipline members are also actively involved in continuing education, membership engagement, and research across other COG committees/domains. Future areas of committed growth for the discipline include pharmacogenomics, pharmacokinetics, pharmacoeconomics, pharmaceutics, and implementation science.


The future of drug eluting stents.
In-stent restenosis (ISR) is the major drawback of percutaneous coronary interventions, occurring in 10-40% of patients. Drug eluting stents (DES) are successful in a large majority of patients in preventing restenosis for the first year after implantation. Recently, new stents have emerged that are loaded with anti-inflammatory, antimigratory, antiproliferative, or pro-healing drugs. These drugs are supposed to inhibit inflammation and neointimal growth and subsequently ISR. The future of DES lies in the development of better stents with new stent designs, better polymers including biological polymers and biological biodissolvable stent coatings, and new, better drugs.


Establishment and Characterization of a New Triple Negative Breast Cancer Cell Line from an Iranian Breast Cancer Tissue.
Breast cancer is the most common malignancy and the leading cause of cancer-related death among women worldwide. The underlying mechanisms for breast cancer development, especially in young women, are not completely understood. Although there are several experimental models to understand the biology of breast cancer such as immortalized cell lines, many of these cell lines have been in culture for decades and most of them have been derived from Caucasians or African-Americans. So, it is required to establish a new cell line derived from primary tumors and Asian women. In this study Pari-Institute for Cancer Research (Pari-ICR) was derived from the primary breast tumor of a 36-years old patient with invasive ductal carcinoma. We characterized the cell line by examining morphology, expression of different markers, and functional profile. Immunocytochemistry showed that this cell line does not express estrogen and progesterone receptors as well as human epidermal growth factor receptor 2 (HER2). Pari-ICR cell line expresses high levels of Vimentin, Ezrin, and S100 but does not express EpCAM, Cytokeratin19, Pan-cytokeratin, Nestin, and Desmin. Its doubling time of Pari-ICR was about 22h and was able to grow as colonies in soft agar. It displayed a higher ability of migration and invasion in comparison with MCF-7 cell line. This breast cancer cell line can serve as a model for understanding the molecular mechanisms of breast carcinogenesis. Moreover, it can be used as an appropriate resource to find novel biomarkers or assess new drugs.


Chronic postoperative pain: recent findings in understanding and management.
Chronic postoperative pain is a poorly recognized potential outcome from surgery. It affects millions of patients every year, with pain lasting for months to years, resulting in patient suffering and ensuing economic consequences. The operations with the highest incidence of chronic postoperative pain are amputations, thoracotomies, cardiac surgery, and breast surgery. Other risk factors include preoperative pain, psychological factors, demographics, and the intensity of acute postoperative pain. Attempts to prevent chronic postoperative pain have often led to debatable results. This article presents data from recently published studies examining the incidence, risk factors, mechanisms, treatment options, and preventive strategies for chronic postoperative pain in adults. In summary, many of the previously identified risk factors for chronic postoperative pain have been confirmed and some novel ones discovered, such as the importance of the trajectory of acute pain and the fact that catastrophizing may not always be predictive. The incidence of chronic postoperative pain hasn't changed over time, and there is limited new information regarding an effective preventive therapy. For example, pregabalin may actually cause more harm in certain surgeries. Further research is needed to demonstrate whether multimodal analgesic techniques have the best chance of significantly reducing the incidence of chronic postoperative pain and to determine which combination of agents is best for given surgical types and different patient populations.


[Utilization rate of fertilizer N and dynamic changes of soil NO3(-)-N in summer maize field in semi-humid area of Northwest China].
A field experiment on manual loessial soil was conducted to study the dynamic changes of NO3(-)-N in soil profile, utilization rate of fertilizer N, and relationships between N application rate and soil residual N accumulation during the growth period of summer maize under different N application rates (0, 45, 90, 135, and 180 kg hm(-2)). The results showed that in the whole growth period of summer maize, the NO3(-)-N concentration in the soil profile was the highest in 0-20 cm layer, and increased with increasing N application rate. The NO3(-)-N concentration in 0-60 cm soil layer changed significantly, but no significant change was observed in 60-100 cm soil layer. In the growth season of summer maize, soil NO3(-)-N accumulation presented a fluctuated decreasing trend due to the N uptake by crop and the precipitation. The N utilization rate (NUR) increased with increasing N application rate when the application rate was less than 135 kg hm(-2), but tended to decrease when the application rate exceeded 135 kg hm(-2). With the increase of N application rate, the N agronomy efficiency (NAE) decreased but the N physiology efficiency (NPE) increased. There was a significant positive correlation between soil residual N accumulation and N application rate (R2 = 0.957**, n = 5). The grain yield under N application was significantly higher than that without N application (P <0.05), and there existed a significant positive correlation between grain yield and N application rate (R2 = 0.934**, n = 5). In our experiment, the optimal application rate of fertilizer N was 135 kg hm(-2), which could harmonize the relationship between economic benefits and environment.


Investigation of the allene oxide pathway in the coral Plexaura homomalla: formation of novel ketols and isomers of prostaglandin A2 from 15-hydroxyeicosatetraenoic acid.
Prostaglandin A2 is a major constituent of the gorgonian Plexaura homomalla, and there is evidence that its biosynthesis involves a noncyclooxygenase pathway. The coral contains an 8(R)-lipoxygenase and an allene oxide synthase; from arachidonic acid, the sequential action of these enzymes gives an allene epoxide, the cyclization of which forms an analogue of prostaglandin A2 (PGA2) with no 15-hydroxyl group. In this study we examined the metabolic fate of 15-hydroxyeicosatetraenoic acid (15-HETE), which via analogous reactions could lead to PGA2. The 8(R)-lipoxygenase metabolized preferentially the 15(R) enantiomer of 15-HETE, and this reaction was stimulated fivefold by including 1 M NaCl in the incubation. Further enzymic steps were detected by comparing the metabolic profiles of the 8(R)-hydroperoxy-15(R)-hydroxy intermediate with that of its 8(S),15(S) enantiomer. Two main products were formed exclusively from the 8(R),15(R) enantiomer: an allene epoxide and the comparatively stable epoxide, 8,9-epoxy-10,15-dihydroxyeicosa-5,11,14-trienoic acid. Formation of the allene oxide was inferred from detection of its hydrolysis and cyclization products. It cyclized to give two isomers of PGA2 which have a "cis" arrangement of the side chains. The main hydrolysis product (8,15-dihydroxy-9-ketoeicosa-5,11,13-trienoic acid) was unstable and prone to oxygenation, giving 8,14,15-trihydroxy-9-ketoeicosa-5,10,12-trienoic acids after reduction of the 14-hydroperoxide. We conclude that metabolism of a 15-hydroxy eicosanoid is a potential route to the A series prostaglandins, although the low yield and lack of stereochemical control suggest that this is not the natural pathway of biosynthesis in P. homomalla. Unexpectedly, the major end products of the pathway are trihydroxy ketols and the single diastereomer of a stable epoxyalcohol.


Dual function of the nuclear export signal of the Borna disease virus nucleoprotein in nuclear export activity and binding to viral phosphoprotein.
Borna disease virus (BoDV), which has a negative-sense, single-stranded RNA genome, causes persistent infection in the cell nucleus. The nuclear export signal (NES) of the viral nucleoprotein (N) consisting of leucine at positions 128 and 131 and isoleucine at positions 133 and 136 overlaps with one of two predicted binding sites for the viral phosphoprotein (P). A previous study demonstrated that higher expression of BoDV-P inhibits nuclear export of N; however, the function of N NES in the interaction with P remains unclear. We examined the subcellular localization, viral polymerase activity, and P-binding ability of BoDV-N NES mutants. We also characterized a recombinant BoDV (rBoDV) harboring an NES mutation of N. BoDV-N with four alanine-substitutions in the leucine and isoleucine residues of the NES impaired its cytoplasmic localization and abolished polymerase activity and P-binding ability. Although an alanine-substitution at position 131 markedly enhanced viral polymerase activity as determined by a minigenome assay, rBoDV harboring this mutation showed expression of viral RNAs and proteins relative to that of wild-type rBoDV. Our results demonstrate that BoDV-N NES has a dual function in BoDV replication, i.e., nuclear export of N and an interaction with P, affecting viral polymerase activity in the nucleus.


Circulating levels of insulin-like growth factor binding protein-1 (IGFBP-1) and hepatic mRNA are increased in the small for gestational age (SGA) fetal rat.
Insulin-like growth factors (IGFs) circulate in association with a family of specific binding proteins (BPs). Recently, we reported that circulating levels of IGFBP-1 and IGFBP-2 are increased in streptozotocin-diabetic adult rats and are differentially regulated in accordance with insulin and metabolic status. Since IGF BPs appear to be important modulators of IGF bioactivity in post-natal life, we asked whether serum levels of IGF BPs also might be altered in utero when the delivery of maternal nutrients is restricted and fetal growth is impaired. Bilateral uterine artery ligation or sham surgery was performed on maternal rats on d 19 of gestation (term 21.5 d). One day after ligation (d 20), fetuses were (SGA) compared to shams (3.1 +/- 1 vs 3.7 +/- 0.2 g, p less than .02) and serum levels of glucose (70 +/- 5 vs 96 +/- 6 mg/dL, p less than .01) and insulin (62 +/- 4 vs 138 +/- 14 microU/mL) also were reduced. In contrast, serum [125I]IGF-I binding activity was markedly increased in SGA litters compared to sham (65 +/- 5% maximum binding with 2.5 microL/mL SGA serum vs 14 +/- 3% for sham serum, p less than .001), and correlated with fetal weight (r = -0.539, p less than .05) and insulin (r = -0.622, p less than .05). Ligand blotting with [125I]IGF-I revealed that serum levels of IGFBP-1 (32 K) were greater in SGA than shams, while immunoblotting with specific antiserum demonstrated that levels of IGFBP-2 (34 K), the major fetal rat IGF BP, were similar in serum from SGA and shams litters. Affinity labeling and immunoprecipitation confirmed that IGF binding activity is increased in SGA, due largely to increased availability of IGFBP-1. In addition, Northern analysis of hepatic RNA revealed that the abundance of IGFBP-1 mRNA is increased in the SGA fetal rat, while hepatic mRNA for IGFBP-2 is similar in SGA and sham-operated litters. We conclude that circulating levels of IGFBP-1 and the abundance of hepatic mRNA are increased in the SGA fetal rat. IGFBP-1 may be an important modulator of IGF bioactivity and somatic growth in utero.


Antibodies against a peptide of cholera toxin differing in cross-reactivity with the toxin differ in their specific interactions with the peptide as observed by 1H NMR spectroscopy.
The interactions between the aromatic residues of the monoclonal antibody TE34, and its peptide antigen CTP3, have been studied by 2D TRNOE difference spectroscopy. The sequence of CTP3 corresponds to residues 50-64 of the B subunit of cholera toxin (VEVPGSQHIDSQKKA). Unlike two previously studied anti-CTP3 antibodies (TE32 and TE33), the TE34 antibody does not bind the toxin. The off-rate of CTP3 from TE34 was found to be too slow to measure strong TRNOE cross-peaks between the antibody and the peptide. Much faster off-rates, resulting in a strong TRNOE, were obtained for two peptide analogues: (a) CTP3 with an amide in the C-terminus (VEVPGSQHIDSQKKA-NH2) and (b) a truncated version of the peptide (N-acetyl-IDSQKKA). These modifications do not interfere significantly either with the interactions of the unmodified part of the peptide with the antibody or with intramolecular interactions occurring in the epitope recognized by the antibody. The combined use of these peptides allows us to study the interactions between the antibody and the whole peptide. Two tyrosine residues and one or more tryptophan and phenylalanine residues have been found to interact with histidine-8, isoleucine-9, aspartate-10, lysine-13 and/or lysine-14, and alanine-15 of the peptide. In the bound peptide, we observe interactions of a lysine residue with aspartate-10 beta protons. While the peptide epitope recognized by TE34 is between histidine-8 and the negatively charged C-terminus, that recognized by TE32 and TE33 is between residues 3 and 10 of the peptide.(ABSTRACT TRUNCATED AT 250 WORDS)


Hypothermia for the treatment of neonatal ischemic encephalopathy: is the genie out of the bottle?
Evidence suggests that hypothermia for hypoxic ischemic encephalopathy in the term neonate may decrease the risk of death or neurodevelopmental impairment. The objective of this study was to determine how hypothermia has been incorporated into practice. An anonymous survey was sent to medical directors of United States neonatal intensive care units (NICUs) in October 2005. We received completed surveys from 441 (54.5%) of 809 of NICUs. Only 6.4% of respondents used hypothermia. The most common method was total body cooling (64.3%) compared with head cooling (25%) or both (10.7%). At centers that did not offer hypothermia, 29% transferred infants to an institution that did. Centers that offered hypothermia were more likely at academic institutions (76.9%) compared with private practices (11.5%; p < 0.001). Hypothermia was more likely offered at institutions that offered extracorporeal membrane oxygenation (ECMO; 57%) than centers where ECMO was not offered (43%; p < 0.001). There has not been widespread use of hypothermia. There are a variety of protocols used. As results of further outcome studies become available, educational efforts and national practice guidelines will be essential.


Prolonged-release fampridine for the treatment of myoclonus after cervical myelitis: a case report.
Prolonged-release fampridine (PR-FAM), a potassium channel blocker, is approved for improving walking ability in patients with multiple sclerosis (MS). Beyond this, positive effects on other MS symptoms like fatigue, cognition, and tremor have been described. To our knowledge, a positive effect of PR-FAM on spinal myoclonus has not been described so far. Here, we report a 32-year-old female with myoclonus after cervical myelitis affecting both hands which markedly improved after administration of PR-FAM. Treatments used before such as carbamazepine or levetiracetam had to be withdrawn because of intolerable side effects or lack of efficacy. The positive effect of PR-FAM could be confirmed by transient suspension. PR-FAM may be considered as a treatment option in refractory spinal myoclonus after myelitis in selected cases.


Effect of electrolyte concentration on the viscosity and voltammetry of supercritical solutions.
The viscosity of a supercritical electrolyte solution is measured for the first time using a modified quartz crystal microbalance, and it is shown that ionic solvation leads to a significant structuring of the solvent and an appreciable increase in solution viscosity. Voltammetric investigations in the electrolyte solutions are used to confirm the magnitude of the viscosity changes, and these account for the appreciably lower than expected peak currents.


Severe facial and ocular injuries from a potato gun.
Potato guns or spud guns are homemade firearms built primarily for recreational use. Information on how to make these can be found easily by searching the topic on the Internet. There is a surplus of Web sites dedicated to providing information to anyone looking to make one of their own. We present an interesting case that illustrates the extent of trauma, which can be caused by these dangerous devices. Despite the growing information available for consumers of any age, there is little information for clinicians regarding these devices and the serious threat of injury that they pose. The current trauma and injury databases, unfortunately, do not gather data pertaining specifically to these devices. In addition, because they are homemade and primarily built for recreational purposes, their use is not controlled or regulated by the government. It is important for clinicians to be aware of homemade firearms such as potato guns, be prepared to manage injury from these devices, and as with other weapons or firearms provide patients and families with the appropriate anticipatory guidance.


Identifying Major Health-System Challenges in Developing Countries Using PERs: Equity is the Elephant in the Room.
Despite an unprecedented increase in official development assistance to health in the last 25 years, there is no systematic way to assess dominant patterns in health-system challenges and opportunities in developing countries. Developing a new global instrument for and by donors and development partners would be resource-intensive and cumbersome. In this article, we demonstrate that Public Expenditure Reviews (PERs) can be used to reveal such patterns. PERs are analytical reports financed and conducted by the World Bank that have been used for years to identify and prioritize country-specific health sector reform needs. In order to extend their use beyond the country level, a reading instrument is developed in the form of a questionnaire to systematically identify the different themes addressed in each PER. All PERs published over a period of ten years are reviewed for health sector content. A new database is created with data on 70 PERs, spanning 61 countries. Analysis of the data reveals dominant themes globally, patterns across development levels, and some regional variations. Our main finding is that issues related to equity strongly dominate and are relevant across all regions and income groups. In addition, the article highlights the usefulness of PERs beyond providing country-specific information. Without losing the country-focus and flexibility of PERs, thoughtful and minor investments in how Health PERs are conducted can create a relatively cheap and strongly operational instrument for building global knowledge bases on health sector needs and challenges.


Update on sutureless thyroidectomy.
The Harmonic scalpel ("Focus" and the new version "Focus+") is one of the first devices for surgical simultaneous cutting and tissue coagulation which allows to obtain dissection and hemostasis by direct application of ultrasound and allows minimally invasive surgical procedures with minimal lateral thermal spread and, thus, minimal adjacent tissue destruction. The aim of the study is to complete the previous study that we made in 2014, based on the TT performed between January 2008 and December 2013, with new data about TT performed in our Surgical Division between January 2014 and December 2016 and compare the outcome using the Ultrasonic scalpel versus the device Ligasure in term of safety, operative time, overall drainage volume, complications, hospital stay. 250 patients were randomized into two groups: Group A where Ultracision were used and Group B where the Ligasure device was used. The results of the group A and of the Group B 2014-2016 have been compared with the results of the previous study and we found that they are similar, but asymptomatic hypocalcaemia increased in the patients of the new study. We found that the use of Ultrasonic scalpel and Ligasure is effective both in the hemostasis of all vessels and in dissection of tissues and confirm the results of our first study without significant difference in the rate of post-operative morbidity with these two different energy based devices used.


Sensory Drive, Color, and Color Vision.
Colors often appear to differ in arbitrary ways among related species. However, a fraction of color diversity may be explained because some signals are more easily perceived in one environment rather than another. Models show that not only signals but also the perception of signals should regularly evolve in response to different environments, whether these primarily involve detection of conspecifics or detection of predators and prey. Thus, a deeper understanding of how perception of color correlates with environmental attributes should help generate more predictive models of color divergence. Here, I briefly review our understanding of color vision in vertebrates. Then I focus on opsin spectral tuning and opsin expression, two traits involved in color perception that have become amenable to study. I ask how opsin tuning is correlated with ecological differences, notably the light environment, and how this potentially affects perception of conspecific colors. Although opsin tuning appears to evolve slowly, opsin expression levels are more evolutionarily labile but have been difficult to connect to color perception. The challenge going forward will be to identify how physiological differences involved in color vision, such as opsin expression levels, translate into perceptual differences, the selection pressures that have driven those differences, and ultimately how this may drive evolution of conspecific colors.


The neonatal sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA1b): a neglected pump in scope.
The neonatal isoform of the sarcoplasmic/endoplasmic reticulum Ca(2+) ATPase (SERCA1b) is formed by developmental splicing and expressed fully only in developing muscle. As a major Ca(2+) pump in myotubes, SERCA1b must be detected in excitation contraction coupling or in store-operated calcium entry. The available pan SERCA1 antibodies also recognise SERCA1b but these are more frequently used to detect SERCA1a, the adult muscle-specific isoform characteristically expressed in fast fibres of skeletal muscle. In such applications, the pan SERCA1 antibodies are frequently claimed to be SERCA1a antibodies without proving it. Realistically, such an antibody cannot be made since it should recognise a single glycine at the C-terminal, the only part of SERCA1a that is different from SERCA1b. The false interpretation of the antibody specificity created inconsistence in the literature and led to false conclusions attributing features only to SERCA1a although those at least are also shared by SERCA1b. In contrast, a SERCA1b antibody has been made against the eight amino acid peptide tail that replaces the glycine of SERCA1a at the C-terminal. Therefore, the expression of SERCA1b can be specifically demonstrated, unlike that of SERCA1a, in various stages and conditions of skeletal muscle. This review argues against misbeliefs related to the distinction, expressions and functions of the two muscle-specific SERCA1 isoforms.


Cardiac titin: an adjustable multi-functional spring.
The giant elastic protein titin contains a molecular spring segment that underlies the majority of myocardial passive stiffness. The mechanical characteristics of this spring may be tuned to match changing mechanical demands placed on muscle, using mechanisms that operate on different time scales and that include post-transcriptional and post-translational processes. Recent work also suggests that titin performs roles that go beyond passive stiffness generation. In contracting myocardium, titin may modulate actomyosin interaction by a titin-based alteration of the distance between myosin heads and actin. Furthermore, novel ligands have been identified that link titin to membrane channels, protein turnover and gene expression. This review highlights that titin is a versatile and adjustable spring with a range of important functions in passive and contracting myocardium.


Successful treatment of fetal supraventricular tachycardia with maternal digoxin therapy.
In a fetus with supraventricular tachycardia (SVT) and cardiac failure, normal sinus rhythm (NSR) was restored with maternal digoxin therapy at 26 weeks' gestation. The diagnosis of cardiac failure was based on ultrasound evidence of ascites and scalp edema. Cardiac failure was attributed to the persistent SVT. The infant remained in NSR and was delivered at 36 weeks' gestation because of persistent ascites. Intracardiac anatomy was normal. This case confirms the usefulness of prenatal ultrasound examinations in the diagnosis of fetal SVT and cardiac failure and illustrates the effectiveness and safety of transplacental digoxin therapy in the management of fetal SVT.


Molecular Responses of Red Ripe Tomato Fruit to Copper Deficiency Stress.
Fruit nutritional value, plant growth, and yield can be compromised by deficient copper (Cu) bioavailability, which often appears in arable lands. This condition causes low Cu content and modifications in the ripening-associated processes in tomato fruit. This research studies the transcriptomic changes that occur in red ripe tomato fruit grown under suboptimal Cu conditions to shed light on the molecular mechanisms underlying this stress. Comparative RNA-sequencing and functional analyses revealed that Cu deficiency during cultivation activates signals for metal ion transport, cellular redox homeostasis, pyridoxal phosphate binding, and amino acid metabolism while repressing the response to phosphate starvation in harvested fruit. Transcriptomic analyses highlighted a number of novel Cu stress-responsive genes of unknown function and indicated that Cu homeostasis regulation in tomato fruit may involve additional components than those described in model plants. It also studied the regulation of high-affinity Cu transporters and a number of well-known Cu stress-responsive genes during tomato fruit ripening depending on Cu availability, which allowed potential candidates to be targeted for biotechnological improvements in reproductive tissues. We provide the first study characterizing the molecular responses of fruit to Cu deficiency stress for any fruit crop.


An Arabidopsis glutathione peroxidase functions as both a redox transducer and a scavenger in abscisic acid and drought stress responses.
We isolated two T-DNA insertion mutants of Arabidopsis thaliana GLUTATHIONE PEROXIDASE3 (ATGPX3) that exhibited a higher rate of water loss under drought stress, higher sensitivity to H(2)O(2) treatment during seed germination and seedling development, and enhanced production of H(2)O(2) in guard cells. By contrast, lines engineered to overexpress ATGPX3 were less sensitive to drought stress than the wild type and displayed less transpirational water loss, which resulted in higher leaf surface temperature. The atgpx3 mutation also disrupted abscisic acid (ABA) activation of calcium channels and the expression of ABA- and stress-responsive genes. ATGPX3 physically interacted with the 2C-type protein phosphatase ABA INSENSITIVE2 (ABI2) and, to a lesser extent, with ABI1. In addition, the redox states of both ATGPX3 and ABI2 were found to be regulated by H(2)O(2). The phosphatase activity of ABI2, measured in vitro, was reduced approximately fivefold by the addition of oxidized ATGPX3. The reduced form of ABI2 was converted to the oxidized form by the addition of oxidized ATGPX3 in vitro, which might mediate ABA and oxidative signaling. These results suggest that ATGPX3 might play dual and distinctive roles in H(2)O(2) homeostasis, acting as a general scavenger and specifically relaying the H(2)O(2) signal as an oxidative signal transducer in ABA and drought stress signaling.


Flies lacking all synapsins are unexpectedly healthy but are impaired in complex behaviour.
Vertebrate synapsins are abundant synaptic vesicle phosphoproteins that have been proposed to fine-regulate neurotransmitter release by phosphorylation-dependent control of synaptic vesicle motility. However, the consequences of a total lack of all synapsin isoforms due to a knock-out of all three mouse synapsin genes have not yet been investigated. In Drosophila a single synapsin gene encodes several isoforms and is expressed in most synaptic terminals. Thus the targeted deletion of the synapsin gene of Drosophila eliminates the possibility of functional knock-out complementation by other isoforms. Unexpectedly, synapsin null mutant flies show no obvious defects in brain morphology, and no striking qualitative changes in behaviour are observed. Ultrastructural analysis of an identified 'model' synapse of the larval nerve muscle preparation revealed no difference between wild-type and mutant, and spontaneous or evoked excitatory junction potentials at this synapse were normal up to a stimulus frequency of 5 Hz. However, when several behavioural responses were analysed quantitatively, specific differences between mutant and wild-type flies are noted. Adult locomotor activity, optomotor responses at high pattern velocities, wing beat frequency, and visual pattern preference are modified. Synapsin mutant flies show faster habituation of an olfactory jump response, enhanced ethanol tolerance, and significant defects in learning and memory as measured using three different paradigms. Larval behavioural defects are described in a separate paper. We conclude that Drosophila synapsins play a significant role in nervous system function, which is subtle at the cellular level but manifests itself in complex behaviour.


EVOLUTIONARY PATTERNS OF ALTERED BEHAVIOR AND SUSCEPTIBILITY IN PARASITIZED HOSTS.
Adaptation is the usual context for interpreting parasite-host interactions. For example, altered host behavior is often interpreted as a parasite adaptation, because in some cases it enhances parasite transmission. Resistance to parasites also has obvious adaptive value for hosts. However, it is difficult to evaluate the adaptive significance of host-parasite interactions without considering the historical context in which these traits have evolved and if they can be predicted by host (or parasite) phylogeny. We examined the influence of host phylogeny on patterns of altered behavior and resistance to parasitism in a cockroach-acanthocephalan system. A consensus cladogram for cockroach subfamilies was produced from the morphological data of McKittrick. We used this cladogram to predict patterns of altered host behavior in seven cockroach host species. Each species was experimentally infected with a single species of acanthocephalan, Moniliformis moniliformis, a parasite that is transmitted when cockroaches are eaten by rodent final hosts. Activity patterns, substrate choices, and responses to light were measured for control and infected animals. These data were recoded into a behavioral matrix of discrete characters. We determined the most parsimonious distribution of the behavioral characters on the tree obtained from McKittrick's data. We then measured the concordance between the behavioral data and the cockroach cladogram with the consistency index (CI). We compared the observed CI to the expected value based on a randomization of observed character states. For three different models of evolutionary character change, there was no evidence of strong concordance (significantly large CI) between altered host behavior and host relationships. Parsimony analysis of the interior nodes of the phylogenetic reconstruction suggested that unaltered behavior was the ancestral state for most host behaviors. We also compared host phylogeny to a data set on the susceptibility of 29 cockroach species to infection with M. moniliformis. At the species level, there was a significant concordance between susceptibility and host phylogeny. This pattern was consistent with the finding that susceptibility of species varied significantly among different subfamilies. However, at the subfamily level, susceptibility was not strongly concordant with phylogeny. We predict that, given enough time, resistance should be lost in subfamilies that are currently resistant to parasitism. In spite of the potential importance of phylogeny in the evolution of behavior and susceptibility, we found little evidence for phylogenetic effects in this system. We conclude that changes in the behavioral responses of hosts to parasites and, to a lesser extent, changes in susceptibility are more frequent than cockroach speciation events in different cockroach lineages. This finding strengthens the assertion that at least some of the altered behaviors are adaptive for host and/or parasite.


Biocompatible coupling of therapeutic fusion proteins to human erythrocytes.
Carriage of drugs by red blood cells (RBCs) modulates pharmacokinetics, pharmacodynamics, and immunogenicity. However, optimal targets for attaching therapeutics to human RBCs and adverse effects have not been studied. We engineered nonhuman-primate single-chain antibody fragments (scFvs) directed to human RBCs and fused scFvs with human thrombomodulin (hTM) as a representative biotherapeutic cargo (hTM-scFv). Binding fusions to RBCs on band 3/glycophorin A (GPA; Wright b [Wrb] epitope) and RhCE (Rh17/Hr0 epitope) similarly endowed RBCs with hTM activity, but differed in their effects on RBC physiology. scFv and hTM-scFv targeted to band 3/GPA increased membrane rigidity and sensitized RBCs to hemolysis induced by mechanical stress, while reducing sensitivity to hypo-osmotic hemolysis. Similar properties were seen for other ligands bound to GPA and band 3 on human and murine RBCs. In contrast, binding of scFv or hTM-scFv to RhCE did not alter deformability or sensitivity to mechanical and osmotic stress at similar copy numbers bound per RBCs. Contrasting responses were also seen for immunoglobulin G antibodies against band 3, GPA, and RhCE. RBC-bound hTM-scFv generated activated protein C (APC) in the presence of thrombin, but RhCE-targeted hTM-scFv demonstrated greater APC generation per bound copy. Both Wrb- and RhCE-targeted fusion proteins inhibited fibrin deposition induced by tumor necrosis factor-α in an endothelialized microfluidic model using human whole blood. RhCE-bound hTM-scFv more effectively reduced platelet and leukocyte adhesion, whereas anti-Wrb scFv appeared to promote platelet adhesion. These data provide a translational framework for the development of engineered affinity ligands to safely couple therapeutics to human RBCs.


Effects of sesame (Sesamum indicum L.) and bioactive compounds (sesamin and sesamolin) on inflammation and atherosclerosis: A review.
Inflammation, oxidative stress, obesity, infection, hyperlipidemia, hypertension, and diabetes are the main causes of atherosclerosis, which in the long term lead to hardening of the arteries. In the current study, we reviewed recent findings of the mechanism of sesame and its active compounds of sesamin and sesamolin regulates on atherosclerosis. Sesame can decrease the lipid peroxidation and affect the enzymes, which control the balance of oxidative status in the body. Besides modulating the inflammatory cytokines, sesame regulates the main mediators of the signaling pathways in the process of inflammation, such as prostaglandin E2 (PGE2), nuclear factor kappa light-chain enhancer of activated B cells (NF-kB) and peroxisome proliferator-activated receptor gamma (PPAR-γ). Sesame decreases the growth of different pathogens. It fights against obesity and helps to reduce weight, body mass index (BMI), waist circumference, and lipid count of serum and liver. In addition to lowering fasting blood sugar (FBS), it decreases the hemoglobin A1c (HbA1c) and glucose levels and improves insulin function. With high content of linoleic acid, α-linolenic acid, and total polyunsaturated fatty acid (PUFA), sesame efficiently controls the blood plasma lipids and changes the lipid profile. In the case of hypertension, it maintains the health of endothelium through multiple mechanisms and conserves the response of the arteries to vasodilation. PUFA in sesame suppresses blood clotting and fibrinogen activity. All the mentioned properties combat atherosclerosis and hardening of blood vessels, which are detailed in the present review for sesame.


L-DOPA-induced activation of striatal p38MAPK and CREB in neonatal dopaminergic denervated rat: relevance to self-injurious behavior.
The destruction of nigrostriatal dopaminergic neurons with 6-hydroxydopamine (6OHDA) during the neonatal period results in dopamine (DA) loss and susceptibility for self-injurious behavior (SIB) when challenged with L-dihydroxyphenylalanine (L-DOPA), via a supersensitive D1 receptor-mediated mechanism. However, there are no changes in D1 receptor binding or mRNA levels, suggesting a potential postreceptor signaling mechanism(s). Here, we examined whether L-DOPA-induced SIB is associated with altered MAPK signaling (p38MAPK, ERK1/2, and JNK) and their nuclear target, CREB. Neonatal dopaminergic lesioned animals were challenged, as adults, with L-DOPA, observed for SIB for 6 hr, and then sacrificed. The data were grouped as follows: control, lesioned rats without SIB (SIB(-)), and lesioned rats that were positive for SIB (SIB(+)). HPLC analysis of striatal extracts revealed a more significant loss of DA and an increase of serotonin in the SIB(+) than in the SIB(-) group. The striatal levels of TH protein were severely decreased, but D1 receptor levels were unaltered in the lesioned groups. These results confirm and extend previous studies indicating that SIB is associated with a near-total loss of DA and TH, an increase in serotonin, and no change in D1 receptor levels. The present studies further revealed that the levels of active phosphorylated forms of p38MAPK and CREB were significantly higher in the SIB(+) group than in the SIB(-) group in the striatum, but not in cortex or olfactory tubercle. The results indicate an induction of striatal p38MAPK and an activation of its nuclear target, CREB, as additional mechanisms in the genesis of L-DOPA-induced SIB.


Recombinant growth hormone and insulin-like growth factor I do not alter gonadotrophin stimulation of the baboon testis in vivo.
In vitro studies indicate a physiological role for insulin-like growth factor I (IGF-I) in paracrine regulation of testicular function and recent clinical studies suggest a potential role for growth hormone (GH) and/or IGF-I in the treatment of hypogonadotrophic states in males. This study aimed to examine the effects of pretreatment with recombinant human GH (rhGH) or rhIGF-I on the response to gonadotrophins of the non-human primate testis in vivo. Using a balanced Latin square design with repeated measures, six prepubertal male hamadryas baboons (Papio hamadryas hamadryas) were treated in a cross-over sequence for periods of 18 days with daily im injections of rhGH (0.4 IU.kg-1.day-1), rhIGF-I (0.1 mg.kg-1.day-1) or saline with a 2-week washout period between each treatment. A single im injection of hCG (1500 IU) increased serum testosterone (p = 0.0002) but neither rhGH nor rhIGF-I influenced the timing or magnitude of this response (p > 0.5). A single im dose of FSH (75 IU) stimulated immunoreactive inhibin (p = 0.01) but also was unaffected in magnitude or timing by pretreatment with rhGH or rhIGF-I (p > 0.2). Circulating IGF-I levels were increased independently by hCG (p = 0.01) and FSH (p < 0.0001) administration. These findings indicate that neither GH nor IGF-I pre-treatment enhance acute gonadal responses to gonadotrophin stimulation of the prepubertal non-human primate testis in vivo. These findings suggest that GH or IGF-I treatment of hypogonadotrophic men without somatotrophin deficiency is unlikely to be beneficial.


Endovascular treatment of malignant superior vena cava syndrome: is bilateral wallstent placement superior to unilateral placement?
To report our experience with unilateral versus bilateral stent placement in the treatment of malignant superior vena cava syndrome (SVCS). The records and films of 84 consecutive patients (69 men; mean age 64+/-10 years, range 39-79) referred for stent placement in malignant SVCS were reviewed for venous compromise, technical and clinical success, complications, and reocclusions. Wallstents were placed covering the SVC and both (bilateral technique) brachiocephalic veins (BCV) preferentially; unilateral stenting of only one BCV in addition to the SVC was performed based on operator preference or inability to access both sides. Technical success was defined as the ability to stent the SVC and at least one BCV; clinical success was the elimination of SVCS symptoms. Technical success was achieved in 83 (99%) patients, using the unilateral technique in 22 and bilateral stenting in 61 patients. The groups did not differ with regard to age, sex, underlying diseases, or location and extent of venous compromise. Immediate clinical success was achieved in 20 (91%) of 22 patients in the unilateral group and 55 (90%) of 61 patients in the bilateral group. Two patients suffered late occlusion in the unilateral group, while in the bilateral group, 8 patients had early occlusion and 9 had late occlusion. Thus, the total occlusion rate was significantly (p<0.05) lower in the unilateral group. There was 1 other complication (pericardial tamponade) in the bilateral group, for a 28% total complication rate, which was significantly higher (p=0.039) than the 9% in the unilateral group. The 1, 3, 6, and 12-month primary stent patency rates were 90%, 81%, 76%, and 69%, respectively. Patency tended to last longer in the unilateral group, but the difference was not significant (p=0.11). Although bilateral Wallstent placement achieved equal technical and clinical success, it tended to confer shorter-lived patency and caused more complications.


Fibrolamellar hepatocellular carcinoma arising in a background of focal nodular hyperplasia: a report of 2 cases.
Fibrolamellar carcinoma, a rare variant of hepatocellular carcinoma, and focal nodular hyperplasia, a benign lesion, are rare hepatic lesions that are known to occur in young women with noncirrhotic livers. Some have suggested that fibrolamellar carcinoma might be the malignant counterpart of focal nodular hyperplasia. The coexistence of the 2 lesions is very rare. Two cases of fibrolamellar hepatocellular carcinoma arising in a background of focal nodular hyperplasia followed long-term oral contraception, and 1 of the 2 occurred during pregnancy. Distinguishing fibrolamellar carcinoma from focal nodular hyperplasia has important implications for treatment and prognosis. One should be aware of such conditions, especially in patients with a long history of oral contraception.


Changes in expression of genes related to cell proliferation in human mesenchymal stem cells during in vitro culture in comparison with cancer cells.
We investigated the expression levels of several genes related to cell proliferation in human mesenchymal stem cells (hMSCs) during in vitro culture for use in clinical applications. In this study, we focused on the relationship between hMSC proliferation and transforming growth factor beta (TGFbeta) signaling during in vitro culture. The proliferation rate of hMSCs gradually decreased and marked changes in hMSC morphology were not observed in 3 months of in vitro culture. The mRNA expressions of TGFbeta1, TGFbeta2, and TGFbeta receptor type I (TGFbetaRI) in hMSCs increased with the length of cell culture. There had been no change in the TGFbeta3, TGFbetaRII, and TGFbetaRIII mRNA expressions by the 12th passage from the primary culture (at about 3 months). The mRNA expression of Smad3 increased, but those of c-myc and nucleostemin decreased with the length of hMSC in vitro culture. In addition, the expression profiles of the genes that regulate cellular proliferation in hMSCs were significantly different from those of cancer cells. In conclusion, hMSCs derived from bone marrow seldom underwent spontaneous transformation during 1-2 months of in vitro culture for use in clinical applications. In hMSCs as well as in epithelial cells, growth might be controlled by the TGFbeta family signaling.


High-melting lipid mixtures and the origin of detergent-resistant membranes studied with temperature-solubilization diagrams.
The origin of resistance to detergent solubilization in certain membranes, or membrane components, is not clearly understood. We have studied the solubilization by Triton X-100 of binary mixtures composed of egg sphingomyelin (SM) and either ceramide, diacylglycerol, or cholesterol. Solubilization has been assayed in the 4-50°C range, and the results are summarized in a novel, to our knowledge, form of plots, that we have called temperature-solubilization diagrams. Despite using a large detergent excess (lipid/detergent 1:20 mol ratio) and extended solubilization times (24-48 h) certain mixtures were not amenable to Triton X-100 solubilization at one or more temperatures. DSC of all the lipid mixtures, and of all the lipid + detergent mixtures revealed that detergent resistance was associated with the presence of gel domains at the assay temperature. Once the system melted down, solubilization could occur. In general adding high-melting lipids limited the solubilization, whereas the addition of low-melting lipids promoted it. Lipidomic analysis of Madin-Darby canine kidney cell membranes and of the corresponding detergent-resistant fraction indicated a large enrichment of the nonsolubilized components in saturated diacylglycerol and ceramide. SM-cholesterol mixtures were special in that detergent solubilization was accompanied, for certain temperatures and compositions, by an independent phenomenon of reassembly of the partially solubilized lipid bilayers. The temperature at which lysis and reassembly prevailed was ∼25°C, thus for some SM-cholesterol mixtures solubilization occurred both above and below 25°C, but not at that temperature. These observations can be at the origin of the detergent resistance effects observed with cell membranes, and they also mean that cholesterol-containing detergent-resistant membrane remnants cannot correspond to structures existing in the native membrane before detergent addition.


Effect of temperature stress on circulating biogenic amines in bovine.
A sensitive, simple and selective chromatographic method using high performance liquid chromatography was developed to measure circulating levels of histamine (HI), norepinephrine (NE), dopamine (DA) and serotonin (5-HT) as indicators of response to thermal stress in two breeds of cattle. Duplicate exposures for 8 hr to 13 degrees C resulted in significantly elevated plasma NE and DA in longhorns but not in Herefords and suggest a relatively greater sensitivity to the cold (13 degrees C) in the longhorn. Environmental temperatures of 32 and 42 degrees C significantly increased rectal temperatures of both breeds with much higher rectal temperatures in Herefords. The 32 and 42 degrees C effects on circulating NE and DA of Herefords were highly significant but not in the longhorn. However, heat stress significantly elevated HI and 5-HT in the longhorn which may account for their relatively greater heat tolerance based on their lower rectal temperatures.


On the extent and origins of genic novelty in the phylum Nematoda.
The phylum Nematoda is biologically diverse, including parasites of plants and animals as well as free-living taxa. Underpinning this diversity will be commensurate diversity in expressed genes, including gene sets associated specifically with evolution of parasitism. Here we have analyzed the extensive expressed sequence tag data (available for 37 nematode species, most of which are parasites) and define over 120,000 distinct putative genes from which we have derived robust protein translations. Combined with the complete proteomes of Caenorhabditis elegans and Caenorhabditis briggsae, these proteins have been grouped into 65,000 protein families that in turn contain 40,000 distinct protein domains. We have mapped the occurrence of domains and families across the Nematoda and compared the nematode data to that available for other phyla. Gene loss is common, and in particular we identify nearly 5,000 genes that may have been lost from the lineage leading to the model nematode C. elegans. We find a preponderance of novelty, including 56,000 nematode-restricted protein families and 26,000 nematode-restricted domains. Mapping of the latest time-of-origin of these new families and domains across the nematode phylogeny revealed ongoing evolution of novelty. A number of genes from parasitic species had signatures of horizontal transfer from their host organisms, and parasitic species had a greater proportion of novel, secreted proteins than did free-living ones. These classes of genes may underpin parasitic phenotypes, and thus may be targets for development of effective control measures.


2D DIGE proteomic analysis of early post mortem muscle exudate highlights the importance of the stress response for improved water-holding capacity of fresh pork meat.
Variation in water-holding capacity (WHC), which presents a major economic burden to the swine industry, is considered to be underpinned by variation at a molecular and biochemical level. High-resolution 2D DIGE followed by MS analysis and Western blot were used to unravel the proteome of muscle exudate, collected following centrifugation, in the pH 4-7 range. A first 2DE-based protein map of this substrate was produced where 89 spots were successfully characterised. Two phenotypes divergent for WHC plus one intermediate were compared with a view to deciphering the biochemical processes impacting on variation in WHC. Twenty spots were observed to be altered across the phenotypes. Of these, 14 represented sixteen proteins including metabolic enzymes, stress response proteins and structural proteins. Triosephosphate isomerase and transferrin showed a major difference between the two extreme phenotypes, and may have potential as biological markers for WHC prediction. Several members of the HSPs family were highlighted. This proteomic study makes an important contribution towards a more detailed molecular view of the processes behind WHC and will provide a valuable resource for future investigations.


Fabricating and imaging carbon-fiber immobilized enzyme ultramicroelectrodes with scanning electrochemical microscopy.
The scanning electrochemical microscope (SECM) is used to image the activity of enzymes immobilized on the surfaces of disk-shaped carbon-fiber electrodes. SECM was used to map the concentration of enzymatically produced hydroquinone or hydrogen peroxide at the surface of a 33-microm diameter disk-shaped carbon-fiber electrode modified by an immobilized glucose-oxidase layer. Sub-monolayer coverage of the enzyme at the electrode surface could be detected with micrometer resolution. The SECM was also employed as a surface modification tool to produce microscopic regions of enzyme activity by using a variety of methods. One method is a gold-masking process in which microscopic gold patterns act as mask for producing patterns of chemical modification. The gold masks allow operation in both a positive or negative process for patterning enzyme activity. A second method uses the direct mode of the SECM to produce covalently attached amine groups on the carbon surface. The amine groups are anchors for attachment of glucose oxidase by use of a biotin/avidin process. The effect of non-uniform enzyme activity was investigated by using the SECM tip to temporarily damage an immobilized enzyme surface. SECM imaging can observe the spatial extent and time-course of the enzyme recovery process.


Elucidating the mechanism of miRNA-214 in the regulation of gingival carcinoma.
The aim of the present study was to evaluate the expression levels of microRNA (miRNA)-214 in tumor tissue, blood and saliva of patients with gingival carcinoma, and to investigate the mechanisms underlying the infiltration and invasion of gingival carcinoma. Between January 2013 and March 2015, blood and saliva samples, gingival carcinoma tumor specimens and peritumoral tumor tissues were harvested from 56 patients with gingival carcinoma. Blood and saliva samples were also harvested from 33 control patients without gingival carcinoma. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to detect miRNA-214 and protein tyrosine phosphatase gene (PTEN) mRNA levels. Western blotting and ELISA were performed to detect PTEN protein levels. The results of RT-qPCR demonstrated that the expression of PTEN mRNA in tumor tissues, blood and saliva of patients with gingival carcinoma were significantly decreased compared with that of the control group (P<0.05). These findings were consistent were consistent with the results of PTEN protein expression detected via western blotting and ELISA in these samples (P<0.05). Conversely, the expression levels of miRNA-214 in these samples were significantly increased (P<0.05) in patients with gingival carcinoma compared with the control group. The decreased expression of PTEN may be associated with the expression of miRNA-214. miRNA-214 may regulate infiltration and invasion of gingival carcinoma via PTEN. These results suggest that miRNA-214 may be used as a marker of gingival carcinoma.


A new polymorph of 2,6-bis-(trifluoro-meth-yl)benzoic acid.
The asymmetric unit of a second polymorph of the title compound, C(9)H(4)F(6)O(2), contains five independent mol-ecules, which form hydrogen-bonded O-H⋯O dimers about inversion centers. The most significant structural difference between this structure and that of the first polymorph [Tobin & Masuda (2009 ▶). Acta Cryst. E65, o1217] is the hydrogen-bonded, dimeric orientation of the carb-oxy-lic acid functionalities.


3-Nitropropionic acid increases frataxin expression in human lymphoblasts and in transgenic rat PC12 cells.
Friedreich ataxia (FRDA) is the most common recessive ataxia caused by reduced expression of frataxin, a nuclear encoded mitochondrial protein. In this study we examined the effects of 3-nitropropionic acid (3-NP) on frataxin expression in FRDA patient and control lymphoblasts and in rat pheochromocytoma cell line (PC12) overexpressing human frataxin. Our studies showed an up-regulation of frataxin expression in both FRDA and control lymphoblasts following exposure to 3-NP. In addition, in transgenic frataxin overexpressing cells 3-NP caused an increase of frataxin protein.


Inter-group conflict in health care: UK students' experiences of bullying and the need for organisational solutions.
This paper addresses bullying of United Kingdom (UK) nursing students whilst on work placement as a specific issue of inter-group difficulty that currently affects nurses and students working in the UK National Health Service. The authors begin by discussing the concept of bullying and sharing the types of bullying reported in two recent studies involving UK nursing students. Both studies illustrate the effects that negative workplace experiences can have on new entrants to the profession. After reviewing various individual solutions which have been recommended for reducing bullying, they suggest that the most effective solution is for health care organisations offering placement training to become much more proactive in creating a culture that will not tolerate bullying behaviour by staff at any level. The literature suggests bullying is a phenomenon affecting workplaces in many countries. Thus the issues described in this article, and the solutions offered have relevance to a variety of health care settings.


RNA-seq Analysis of δ9-Tetrahydrocannabinol-treated T Cells Reveals Altered Gene Expression Profiles That Regulate Immune Response and Cell Proliferation.
Marijuana has drawn significant public attention and concern both for its medicinal and recreational use. Δ9-Tetrahydrocannabinol (THC), which is the main bioactive component in marijuana, has also been shown to possess potent anti-inflammatory properties by virtue of its ability to activate cannabinoid receptor-2 (CB-2) expressed on immune cells. In this study, we used RNA-seq to quantify the transcriptomes and transcript variants that are differentially regulated by THC in super antigen-activated lymph node cells and CD4(+) T cells. We found that the expressions of many transcripts were altered by THC in both total lymph node cells and CD4(+) T cells. Furthermore, the abundance of many miRNA precursors and long non-coding RNAs was dramatically altered in THC-treated mice. For example, the expression of miR-17/92 cluster and miR-374b/421 cluster was down-regulated by THC. On the other hand miR-146a, which has been shown to induce apoptosis, was up-regulated by THC. Long non-coding RNAs that are expressed from the opposite strand of CD27 and Appbp2 were induced by THC. In addition, THC treatment also caused alternative promoter usage and splicing. The functions of those altered transcripts were mainly related to immune response and cell proliferation.


Effect of Nanodiamond and Nanoplatinum Liquid, DPV576, on Human Primary Keratinocytes.
Nanofabrics are now being used in a wide range of products that come into direct contact with skin, including carpet, clothing, and medical fabrics. In the current study, we examined the effect of a dispersed aqueous mixture of nanodiamond (ND) and nanoplatinum (NP) (DPV576) on human primary keratinocytes with respect to transient receptor potential vanilloid (TRPV) channel expression, secretion of cytokines and production of nerve growth factor (NGF). Keratinocytes were treated with DPV576 at concentrations of 1:10 and 1:100 dilutions for 24 hours in vitro, and their activation of was determined by production of cytokines TNF-α, IL-1β, and prostaglandin (PGE2), and by production of NGF. Inhibitor experiments were carried out by incubating keratinocytes with the TRPV4-selective antagonist HC-067047. TRPV receptor expression (TRPV1, TRPV3 and TRPV4) on keratinocytes as well as changes in Ca2+ potential were examined by flow cytometry. DPV576 treatment of keratinocytes resulted in the following effects: (1) stimulation of keratinocytes as indicated by the significant secretion of cytokines IL-1β, TNF-α, and PGE2, an effect noted only at higher concentration (1:10); (2) significant decrease in the expression of TRPV4 on keratinocytes with a spike in the calcium flux, but no change in the expression of TRPV1 and TRPV3; (3) induction of cytokine secretion independent of TRPV4, as the addition of TRPV4 inhibitor had no significant effect on the cytokine production from keratinocytes; (4) induction of NGF secretion by keratinocytes. These results demonstrate that DPV576 activates keratinocytes via multiple signaling pathways which may reduce stress associated with inflammation, pain, and circadian rhythms. ND/NP-coated fabrics that target the modulation of local inflammation, pain, and circadian rhythms could potentially be of benefit to humans.


Congenital toxoplasmosis in twins: a case report.
We report a case of congenital toxoplasmosis in a twin pregnancy at 21 weeks of gestation. The complete antenatal investigation included ultrasound examination, cordocentesis and amniocentesis. The ultrasonographic evaluation showed a distinct morphological aspect confirmed by the fetal biological profile. The time lag between placental and fetal infection, or the immunological fetal responses could explain the varying stages of the fetal disease. This is another case which early demonstrates that there may exist a distinct clinical pattern in bichorial pregnancy with varying biological fetal responses. It underlines the importance of cordocentesis in the case of maternal toxoplasma seroconversion, enabling us to gather a maximum number of biological data for complete appreciation of the fetal status. The information obtained permits a precise adaptation of the medical decision.


Subgingival dysbiosis in smoker and non‑smoker patients with chronic periodontitis.
Periodontitis is one of the most common oral inflammatory diseases, and results in connective tissue degradation and gradual tooth loss. It manifests with formation of periodontal pockets, in which anaerobic and Gram‑negative bacteria proliferate rapidly. Consequently, alteration of the subgingival microbiota is considered the primary etiologic agent of periodontitis. Previous studies have reported that smokers are at increased risk of periodontal disease, in both prevalence and severity, indicating that smoking is a risk factor for the onset and progression of the pathology. In the present study, 16S rRNA sequencing was employed to assess the subgingival microbiota in 6 smoker patients with chronic periodontitis, 6 non‑smoker patients with chronic periodontitis and 8 healthy controls. The results demonstrated significant alterations in the microbial structure of periodontitis patients. High relative abundance of Parvimonans, Desulfubulbus, Paludibacter, Haemophilus, and Sphaerochaeta genera characterized subgingival microbiota of periodontitis patients, both smokers and non‑smokers. Due to the high precision and sensitivity of the 16S rRNA sequencing method, analysis for low‑abundant genera (including Pedobacter, Granulicatella, Paracoccus, Atopobium, Bifidobacterium, Coprococcus, Oridobacteriu, Peptococcus, Oscillospira and Akkermansia) was feasible, and revealed novel phylotypes associated with periodontitis. Of note, a major microbial community alteration was evident in smoker patients, suggesting an association between smoking and severity of subgingival dysbiosis. The present study confirmed that chronic periodontitis is a polymicrobial disease where changes in the equilibrium of subgingival microbiota contribute to severity of pathology.


HLA-TNF haplotype heterogeneity in Greek SLE patients.
To examine TNF microsatellite allele frequencies in SLE patients in the Greek population, where disease susceptibility is less associated with HLA-DR3 haplotypes. A cohort of 46 Greek SLE patients were investigated. Allele frequencies for the TNF microsatellite markers a, b, c and d were determined using a fluorescence based DNA fragment sizing technique. HLA class II typing was performed using a molecular based technique. Associations between SLE and DRB1*1501, *1601 and *0701 were observed and DRB1*0301 was only marginally increased in patients. Linkage disequilibrium was found between DRB1*1501 and TNF a11 and also for DR3 and TNF a2, b3, d2. Stratification of patients suggested that DRB*1501 and TNF a11 frequencies were higher in SLE patients with renal disease and TNF a2 and b 3 frequencies in those without, although these differences did not reach statistical significance. SLE in this Greek population appears to be associated with a number of HLA-DRB1 alleles. The development of renal complications in these patients may be related to the TNF polymorphism encoded on these HLA haplotypes.


Oxytocin and callous-unemotional traits: towards a social-cognitive approach to forensic analysis.
Callous-unemotional (CU) traits in children and adolescents are an important risk factor for forensic behaviors in youth and in adulthood. There is a strong biologic basis in CU traits, offering the possibility to design biological interventions to address this risk factor. This paper advances the hypothesis that the administration of exogenous oxytocin is a biological intervention worthy of further investigation to address CU traits and moderate future risk factors for forensic behaviors. The evidence for this assertion rests in the empiric evidence that social cognitive deficits play a major role in child and adolescent CU traits and psychopathy. Simultaneously, emerging evidence suggests that youth with CU traits have defined neuroendocrinological correlates within the oxytocin system. A review of the normal neurohormonal physiology of this system is presented to provide a groundwork for the delineation of these correlates. That deficits in this system are found in alternate psychopathologies that present with deficits in social cognition serves as the basis for the hypothesis that these deficits may play a key role in the development and manifestation of CU traits. Their correction through psychopharmacologic and alternate means may provide a future route to addressing a central area of adolescent forensic psychiatry.


Modified Fisher method for unilateral cleft lip-report of cases.
Rehabilitation of normal function and form is essential in cleft lip repair. In 2005, Dr. David M. Fisher introduced an innovative method, named "an anatomical subunit approximation technique" in unilateral cleft lip repair. According to this method, circumferential incision along the columella on cleft side of the medial flap is continued to the planned top of the Cupid's bow in straight manner, which runs parallel to the unaffected philtral ridge. Usually, small inlet incision is needed to lengthen the medial flap. On lateral flap, small triangle just above the cutaneous roll is used to prevent unesthetic shortening of upper lip. This allows better continuity of the Cupid's bow and ideal distribution of tension. As a modification to original method, orbicularis oris muscle overlapping suture is applied to make the elevated philtral ridge. Concomitant primary rhinoplasty also results in good esthetic outcome with symmetric nostrils and correction of alar web. As satisfactory results were obtained in three incomplete and one complete unilateral cleft lip patients, indicating Fisher's method can be useful in cleft lip surgery with functional and esthetic outcome. Clinically applied Fisher's method in unilateral cleft lip patients proved the effectiveness in improving the esthetic results with good symmetry. This method also applied with primary rhinoplasty.