Initiation of simian virus 40 DNA replication in vitro: aphidicolin causes accumulation of early-replicating intermediates and allows determination of the initial direction of DNA synthesis.
Aphidicolin, a specific inhibitor of DNA polymerase alpha, provided a novel method for distinguishing between initiation of DNA synthesis at the simian virus 40 (SV40) origin of replication (ori) and continuation of replication beyond ori. In the presence of sufficient aphidicolin to inhibit total DNA synthesis by 50%, initiation of DNA replication in SV40 chromosomes or ori-containing plasmids continued in vitro, whereas DNA synthesis in the bulk of SV40 replicative intermediate DNA (RI) that had initiated replication in vivo was rapidly inhibited. This resulted in accumulation of early RI in which most nascent DNA was localized within a 600- to 700-base-pair region centered at ori. Accumulation of early RI was observed only under conditions that permitted initiation of SV40 ori-dependent, T-antigen-dependent DNA replication and only when aphidicolin was added to the in vitro system. Increasing aphidicolin concentrations revealed that DNA synthesis in the ori region was not completely resistant to aphidicolin but simply less sensitive than DNA synthesis at forks that were farther away. Since DNA synthesized in the presence of aphidicolin was concentrated in the 300 base pairs on the early gene side of ori, we conclude that the initial direction of DNA synthesis was the same as that of early mRNA synthesis, consistent with the model proposed by Hay and DePamphilis (Cell 28:767-779, 1982). The data were also consistent with initiation of the first DNA chains in ori by CV-1 cell DNA primase-DNA polymerase alpha. Synthesis of pppA/G(pN)6-8(pdN)21-23 chains on a single-stranded DNA template by a purified preparation of this enzyme was completely resistant to aphidicolin, and further incorporation of deoxynucleotide monophosphates was inhibited. Therefore, in the presence of aphidicolin, this enzyme could initiate RNA-primed DNA synthesis at ori first in the early gene direction and then in the late gene direction, but could not continue DNA synthesis for an extended distance.


Mitochondrial DNA quantification as a tool for embryo viability assessment: retrospective analysis of data from single euploid blastocyst transfers.
Does the amount of mitochondrial DNA (mtDNA) in blastocyst biopsy specimens have the potential to serve as a biomarker of euploid embryo implantation ability, independent of morphology? The results of this study strongly suggest that elevated mtDNA levels, above a previously defined threshold, are strongly associated with blastocyst implantation failure and represent an independent biomarker of embryo viability. Improved methods of embryo selection are highly desirable in order to increase the efficiency of IVF treatment. At present, even the transfer of chromosomally normal embryos of high morphological grade cannot guarantee that a pregnancy will follow. Recently, it has been proposed that the quantity of mtDNA in embryonic cells may be an indicator of developmental potential, with higher levels of mtDNA associated with reduced implantation. However, thus far reported data sets have been relatively small and in some cases have lacked appropriate validation. This large, blinded, retrospective study involved the analysis of relative mtDNA levels in 1505 euploid blastocysts obtained from 490 couples undergoing preimplantation genetic testing for aneuploidy. Implantation outcomes were compared to mtDNA levels in order to determine the capacity of the method to predict viability and to assess the validity of previously established thresholds. DNA from blastocyst biopsy samples was amplified and then subjected to aneuploidy analysis using next generation sequencing or array comparative genomic hybridization. Only those embryos classified as chromosomally normal had their mtDNA levels assessed. This analysis was undertaken retrospectively using quantitative real-time PCR, without knowledge of the outcome of embryo transfer. Predictions of implantation failure, based upon mtDNA levels were subsequently compared to the observed clinical results. All cycles involved the transfer of a single embryo. Of all blastocysts analyzed, 9.2% (139/1505) contained mtDNA levels above a previously established viability threshold and were therefore predicted to have reduced chances of implantation. To the date of analysis, 282 euploid blastocysts had been transferred with an overall implantation rate of 65.6% (185/282). Of the transferred embryos, 249 contained levels of mtDNA in the normal range, 185 of which produced a pregnancy, giving an implantation rate of 74.3% for euploid embryos with 'normal' quantities of mtDNA. However, 33 of the transferred embryos were determined to have elevated mtDNA quantities. None of these led to a pregnancy. Therefore, the negative predictive value of mtDNA assessment in this cohort was 100% (33/33). The difference between the implantation rates for embryos with normal and elevated mtDNA levels was highly significant (P < 0.0001). The mtDNA thresholds, used for classification of embryos, were unaffected by female age or the clinic in which the IVF was undertaken. The probability of an embryo having elevated levels of mtDNA was not influenced by variation in embryo morphology. This study provides strong evidence that mtDNA quantification can serve as a valuable tool to assist the evaluation of blastocyst viability. However, to determine the true extent of any clinical benefits, other types of investigations, such as non-selection studies and randomized controlled trials, will also be necessary. The results of this study suggest that mtDNA quantity can serve as an independent biomarker for the prediction of euploid blastocyst implantation potential. Prospective studies should now be undertaken to confirm these results. Additionally, investigations into the underlying biological cause(s) of elevated mtDNA levels and an enhanced understanding of how they relate to diminished implantation potential would be invaluable. This study was supported by funding provided by Reprogenetics. None of the authors have any competing interests.


Where are kin recognition signals in the human face?
We report two experiments that aimed to determine where in the face the cues that signal kinship fall. In both experiments, participants were shown 30 pairs of photographs of children's faces. Half of the pairs portrayed siblings and half did not. The 220 participants were asked to judge whether each pair of photographs portrayed siblings. We measured the effect on kin recognition performance of masks that covered the upper half or the lower half of the face (Experiment 1) and the eye region or the mouth region (Experiment 2). In Experiment 1, we found that the signal detection estimate of performance d' decreased only 5.3% (ns) when the lower face was masked but by more than 65% when the upper face was masked. We tested whether the combination of kinship information from the two halves of the face can be treated as optimal combination of independent cues and found that it could be. In Experiment 2, we found that masking the eye region led to only a 20% reduction (ns) in performance whereas masking the mouth region led to a nonsignificant increase in performance. We also found that the eye region contains only slightly more information about kinship than the upper half of the face outside of the eye region.


MiR-337-3p Promotes Adipocyte Browning by Inhibiting TWIST1.
The prevalence of metabolic syndrome (MetS) and obesity is an alarming health issue worldwide. Obesity is characterized by an excessive accumulation of white adipose tissue (WAT), and it is associated with diminished brown adipose tissue (BAT) activity. Twist1 acts as a negative feedback regulator of BAT metabolism. Therefore, targeting Twist1 could become a strategy for obesity and metabolic disease. Here, we have identified miR-337-3p as an upstream regulator of Twist1. Increased miR-337-3p expression paralleled decreased expression of TWIST1 in BAT compared to WAT. Overexpression of miR-337-3p in brown pre-adipocytes provoked a reduction in Twist1 expression that was accompanied by increased expression of brown/mitochondrial markers. Luciferase assays confirmed an interaction between the miR-337 seed sequence and Twist1 3'UTR. The inverse relationship between the expression of TWIST1 and miR-337 was finally validated in adipose tissue samples from non-MetS and MetS subjects that demonstrated a dysregulation of the miR-337-Twist1 molecular axis in MetS. The present study demonstrates that adipocyte miR-337-3p suppresses Twist1 repression and enhances the browning of adipocytes.


Pathogenesis of edema formation in the nephrotic syndrome.
The development of edema in the nephrotic syndrome has traditionally been viewed as an underfill mechanism. According to this view, urinary loss of protein results in hypoalbuminemia and decreased plasma oncotic pressure. As a result, plasma water translocates out of the intravascular space and results in a decrease in intravascular volume. In response to the underfilled circulation, effector mechanisms are then activated that signal the kidney to secondarily retain salt and water. While an underfill mechanism may be responsible for edema formation in a minority of patients, recent clinical and experimental findings would suggest that edema formation in most nephrotic patients is the result of primary salt retention. Direct measurements of blood and plasma volume or measurement of neurohumoral markers that indirectly reflect effective circulatory volume are mostly consistent with either euvolemia or a volume expanded state. The ability to maintain plasma volume in the setting of a decreased plasma oncotic pressure is achieved by alterations in transcapillary exchange mechanisms known to occur in the setting of hypoalbuminemia that limit excessive capillary fluid filtration. The intrarenal mechanism responsible for primary sodium retention is not yet known, but may involve tubular resistance to the natriuretic effect of atrial natriuretic peptide.


Effects of extra-corporeal shock waves on penile hemodynamics and histopathology in rats.
To study the effect of extra-corporeal shock wave (ESW) on the penile hemodynamics and histopathology in rats. Adult male Sprague-Dawley rats were divided at random into 3 groups. ESW application was performed with a Siemens Lithostar with the rats under anesthesia lying prone on the balloon probe. Rats in Group I received a total of 1000 shocks at 18 kV and immediately underwent hemodynamic evaluation performed by direct electrostimulation of the cavernous nerve and measurement of intracavernous pressure (ICP). Rats in Group II received 3 times 1000 shocks at 18 kV at weekly intervals and hemodynamic evaluation was performed 1 month after the last ESW application. Group III served as the control. Histopathological examinations of penile tissues were done on Masson's trichrome and hematoxylin and eosin stained sections. Penile hemodynamic evaluation showed a trend toward a diminished mean maximal ICP, duration of erection, ICP during the plateau phase and maximal ICP/ blood pressure ratio in Group I, although there was no significant significance. The mean latency period in Groups I and II was prolonged. Petechial bleeding within tunical layers and small foci of hemorrhage within the corpora cavernosa were observed in Group I. However, histopathological examination failed to reveal any significant differences between the groups in terms of smooth muscle content, tunical thickness, organization of collagen bundles and elastic fiber-lattice framework. ESW has certain damaging effects on the penis.


The interactions between plasma membrane depolarization and glutamate receptor activation in the regulation of cytoplasmic free calcium in cultured cerebellar granule cells.
The complex modulation of cytoplasmic free calcium concentration ([Ca2+]c) in primary cultures of cerebellar granule cells in response to glutamate receptor agonists has been the subject of several contradictory reports. We here show that 3 components of the [Ca2+]c response can be distinguished: (1) Ca2+ entry through voltage-dependent Ca2+ channels, following KCl- or receptor-evoked depolarization, (2) Ca2+ entry through NMDA receptor channels, and (3) liberation of internal Ca2+ via a metabolotropic receptor. Depolarization with KCl induced a transient [Ca2+]c response (subject to voltage inactivation) decaying to a sustained plateau (largely inhibited by nifedipine). The NMDA response was potentiated by glycine, totally inhibited by (+)5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate (MK-801), and blocked by Mg2+ in a voltage-sensitive manner. Polarized cells displayed small responses to quisqualate (QA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA). Depolarization enhanced a transient response to QA, but not to AMPA. Trans-1-amino-1,3-cyclopentanedicarboxylic acid (trans-ACPD), a selective agonist for the metabolotropic glutamate receptor, caused a transient elevation of [Ca2+]c, which was blocked by prior exposure to QA but not AMPA. The prolonged [Ca2+]c response to kainate (KA) can be resolved into 2 major components: an indirect NMDA receptor-mediated response due to released glutamate and a nifedipine-sensitive component consistent with depolarization-mediated entry via Ca2+ channels. 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX), QA at greater than 10 microM, and AMPA (but not trans-ACPD) reversed the KA response, consistent with an inactivation of the KA receptor.(ABSTRACT TRUNCATED AT 250 WORDS)


Therapeutic ultrasound as a potential male contraceptive: power, frequency and temperature required to deplete rat testes of meiotic cells and epididymides of sperm determined using a commercially available system.
Studies published in the 1970s by Mostafa S. Fahim and colleagues showed that a short treatment with ultrasound caused the depletion of germ cells and infertility. The goal of the current study was to determine if a commercially available therapeutic ultrasound generator and transducer could be used as the basis for a male contraceptive. Sprague-Dawley rats were anesthetized and their testes were treated with 1 MHz or 3 MHz ultrasound while varying power, duration and temperature of treatment. We found that 3 MHz ultrasound delivered with 2.2 Watt per square cm power for fifteen minutes was necessary to deplete spermatocytes and spermatids from the testis and that this treatment significantly reduced epididymal sperm reserves. 3 MHz ultrasound treatment reduced total epididymal sperm count 10-fold lower than the wet-heat control and decreased motile sperm counts 1,000-fold lower than wet-heat alone. The current treatment regimen provided nominally more energy to the treatment chamber than Fahim's originally reported conditions of 1 MHz ultrasound delivered at 1 Watt per square cm for ten minutes. However, the true spatial average intensity, effective radiating area and power output of the transducers used by Fahim were not reported, making a direct comparison impossible. We found that germ cell depletion was most uniform and effective when we rotated the therapeutic transducer to mitigate non-uniformity of the beam field. The lowest sperm count was achieved when the coupling medium (3% saline) was held at 37 degrees C and two consecutive 15-minute treatments of 3 MHz ultrasound at 2.2 Watt per square cm were separated by 2 days. The non-invasive nature of ultrasound and its efficacy in reducing sperm count make therapeutic ultrasound a promising candidate for a male contraceptive. However, further studies must be conducted to confirm its efficacy in providing a contraceptive effect, to test the result of repeated use, to verify that the contraceptive effect is reversible and to demonstrate that there are no detrimental, long-term effects from using ultrasound as a method of male contraception.


The generation of rhythmic activity in a distributed motor system.
Rhythmic activity that is distributed to the brain and buccal ganglia and which underlies several types of behaviour, can be evoked from isolated nervous systems of Pleurobranchaea californica by tonic nerve stimulation. The experiments presented here were designed to test whether this rhythmic activity is produced by independent neuronal oscillators located in each ganglion or whether the rhythmic activity arises from a single oscillatory locus in the buccal ganglion and is transmitted passively to the brain. By interrupting the conduction of activity in the cerebrobuccal connectives (CBC) between brain and buccal ganglia we show that motor output from the brain depends on sustained, cycle to cycle input from the buccal ganglion and cannot be reset with respect to the buccal activity. The production of rhythmic activity in the brain depends on the generation of rhythmic activity in the buccal ganglia whether the rhythms are activated by stimulation of buccal roots or paracerebral command cells in the brain. Simultaneous intracellular recordings from brain motoneurones and buccal interneurones which project to the brain indicate that these interneurones provide both the drive and the pattern for rhythmic motor output in the brain. Tonic stimulation of the CBC can produce rhythmic activity in isolated brains in which all nerve roots and connectives have been cut. This can be explained by the fact that tonic stimulation of the connectives is transformed into phasic activity by the axons within the connective. We conclude therefore, that rhythmic, coordinated activity in the brain and buccal ganglia of Pleurobranchaea arises from oscillatory circuits that are located only in the buccal ganglia.


Current anti-doping policy: a critical appraisal.
Current anti-doping in competitive sports is advocated for reasons of fair-play and concern for the athlete's health. With the inception of the World Anti Doping Agency (WADA), anti-doping effort has been considerably intensified. Resources invested in anti-doping are rising steeply and increasingly involve public funding. Most of the effort concerns elite athletes with much less impact on amateur sports and the general public. We review this recent development of increasingly severe anti-doping control measures and find them based on questionable ethical grounds. The ethical foundation of the war on doping consists of largely unsubstantiated assumptions about fairness in sports and the concept of a "level playing field". Moreover, it relies on dubious claims about the protection of an athlete's health and the value of the essentialist view that sports achievements reflect natural capacities. In addition, costly antidoping efforts in elite competitive sports concern only a small fraction of the population. From a public health perspective this is problematic since the high prevalence of uncontrolled, medically unsupervised doping practiced in amateur sports and doping-like behaviour in the general population (substance use for performance enhancement outside sport) exposes greater numbers of people to potential harm. In addition, anti-doping has pushed doping and doping-like behaviour underground, thus fostering dangerous practices such as sharing needles for injection. Finally, we argue that the involvement of the medical profession in doping and anti-doping challenges the principles of non-maleficience and of privacy protection. As such, current anti-doping measures potentially introduce problems of greater impact than are solved, and place physicians working with athletes or in anti-doping settings in an ethically difficult position. In response, we argue on behalf of enhancement practices in sports within a framework of medical supervision. Current anti-doping strategy is aimed at eradication of doping in elite sports by means of all-out repression, buttressed by a war-like ideology similar to the public discourse sustaining international efforts against illicit drugs. Rather than striving for eradication of doping in sports, which appears to be an unattainable goal, a more pragmatic approach aimed at controlled use and harm reduction may be a viable alternative to cope with doping and doping-like behaviour.


T-cell therapy for EBV-associated nasopharyngeal carcinoma: preparative lymphodepleting chemotherapy does not improve clinical results.
We and others have demonstrated that adoptive cell therapy with Epstein-Barr virus (EBV)-specific autologous cytotoxic T lymphocytes (CTLs) may control disease progression in patients with EBV-associated nasopharyngeal carcinoma (NPC). With the aim of favoring in vivo T-cell expansion, we optimized our cell therapy approach by administering higher doses of EBV-specific CTLs, following lymphodepleting chemotherapy. Eleven patients with EBV-related NPC in whom conventional treatment failed have been enrolled. Patients received nonmyeloablative lymphodepleting chemotherapy consisting of cyclophosphamide and fludarabine. Two doses of autologous EBV-specific CTLs were subsequently infused, 2 weeks apart. Study end points were feasibility and clinical outcome. All patients enrolled completed the treatment and were assessable for analysis. The median dose of CTLs per infusion was 3.7 × 10(8). Therapy was well tolerated, with no severe adverse events ascribable to either chemotherapy or cell therapy. Disease control (defined as either tumor regression or disease stabilization lasting >4 months) was obtained in 6 of 11 patients, in keeping with previously published results. Our data confirm that EBV-specific CTL therapy is safe and associated with antitumor activity in patients with advanced NPC. The use of lymphodepleting chemotherapy before high-dose CTL infusion did not enhance the clinical benefit observed in our previous series.


Epitheliotropic T-cell lymphoma in a guinea pig.
A 4-year-old female guinea pig was presented with pruritic, erythematous, hypotrichotic and scaling skin lesions, mainly involving the abdomen and the hindlegs. Clinical abnormalities were limited to the skin, and the guinea pig was otherwise healthy. The dermatological diagnostic work-up excluded ectoparasites, dermatophytosis and an endocrinopathy as differential diagnoses for the pruritus and hair loss. An allergic dermatitis was under investigation when the general and skin condition deteriorated. A neoplastic skin disorder was suspected, and skin biopsies revealed changes consistent with a cutaneous epitheliotropic T-cell lymphoma. Pautrier's microabscesses and immunophenotyped CD3(+) CD79(-) lymphocytes were identified. Owing to the deteriorating condition of the animal and the unknown but most likely poor prognosis of this disease, the guinea pig was euthanized. To the authors' knowledge, this is the first case report of cutaneous epitheliotropic T-cell lymphoma in the guinea pig with characteristic histopathological and immunohistochemical features.


Ethnicity and time to institutionalization of dementia patients: a comparison of Latina and Caucasian female family caregivers.
To compare rates of institutionalization of dementia patients cared for by Latina and Caucasian female caregivers and to explore which caregiver and care-recipient characteristics predicted institutionalization. Longitudinal. San Francisco Bay area, California. Two hundred sixty-four female caregivers of dementia patients (154 Caucasian women, 110 Latinas) who participated in an intervention project designed to reduce caregiver stress and may represent a sample that is more stressed and motivated than a general sample of caregivers. Number of days between baseline interview and institutionalization was recorded over an 18-month period. Kaplan-Meier survival analysis with the log rank statistic was used to test for ethnic differences in time to institutionalization. Cox regression analyses were conducted to determine whether care-recipient or caregiver characteristics (e.g., care-recipient age and memory and behavior problems; caregiver depression, years of education, income, and views of the caregiving role) significantly interacted with ethnicity to explain time to institutionalization. Latinas delayed institutionalization significantly longer than their Caucasian counterparts; care-recipient characteristics or caregiver demographics did not explain these results, although Latinas who identified greater benefits or more-positive aspects of the caregiving process at baseline were less likely to institutionalize their loved one than those who reported fewer benefits of caregiving. Less-acculturated Latinas were significantly more likely to identify positive aspects of caregiving than more-acculturated Latinas. Latina dementia caregivers delay institutionalization significantly longer than female Caucasian caregivers. In addition, Latino cultural values and positive views of the caregiving role are important factors that may significantly influence their decision to institutionalize loved ones with dementia.


Diagnosis by Documentary: Professional Responsibilities in Informal Encounters.
Most work addressing clinical workers' professional responsibilities concerns the norms of conduct within established professional-patient relationships, but such responsibilities may extend beyond the clinical context. We explore health workers' professional responsibilities in such "informal" encounters through the example of a doctor witnessing the misdiagnosis and mistreatment of a serious long-term condition in a television documentary, arguing that neither internalist approaches to professional responsibility (such as virtue ethics or care ethics) nor externalist ones (such as the "social contract" model) provide sufficiently clear guidance in such situations. We propose that a mix of both approaches, emphasizing the noncomplacency and practical wisdom of virtue ethics, but grounding the normative authority of virtue in an external source, is able to engage with the health worker's responsibilities in such situations to the individual, the health care system, and the population at large.


Does deliberation make a difference? Results from a citizens panel study of health goals priority setting.
How to involve the public in setting health and health care priorities is a constant challenge for health system decisions. Policy maker interest in involving the public in increasingly complex and value-laden priority setting processes has led to the use of deliberative public involvement methods designed to promote discussion and debate among participants with the objective of obtaining more informed and consensual views. These methods have not been evaluated rigorously using controlled designs with pre- and post-test measurements. We examined, using a controlled design, the effects of introducing different opportunities for deliberation into a process for obtaining public input into a community health goals priority setting process. Our findings indicate that deliberation does make a difference to participant views. As more deliberation is introduced, participant views may be more amenable to change. Deliberation also offers the potential for views to become more rather than less entrenched. While we are beginning to understand the difference deliberation makes to participant views, we are still at an early stage in understanding the process through which these differences come about and what difference deliberation makes to broader outcomes such as civic competence, civic engagement and health policy decisions.


Interleukin-13 in combination with CD40 ligand potently inhibits apoptosis in human B lymphocytes: upregulation of Bcl-xL and Mcl-1.
Interleukin-13 (IL-13) is a novel T-cell-derived cytokine with IL-4-like effects on many cell types. In human B lymphocytes, IL-13 induces activation, stimulates proliferation in combination with anti-IgM or anti-CD40 antibodies, and directs Ig isotype switching towards IgE and IgG4 isotypes. We show here that IL-13 also regulates human B-cell apoptosis. IL-13 reduced spontaneous apoptosis of peripheral blood B cells in vitro, as shown by measurement of DNA fragmentation using the TUNEL and Nicoletti assays. The inhibition of cell death by IL-13 alone was significant but modest, but was potently enhanced in combination with CD40 ligand (CD40L), a survival stimulus for B cells by itself. Interestingly, IL-13 increased the expression of CD40 on peripheral blood B cells, providing a possible mechanism for the observed synergy. IL-13 alone was a less potent inhibitor of apoptosis than IL-4. Moreover, there was no additive effect of combining IL-4 and IL-13 at supraoptimal concentrations, which is consistent with the notion that the IL-4 and IL-13 binding sites share a common signaling subunit. The combination of IL-13 with CD40L augmented the expression of the Bcl-2 homologues Bcl-xL and Mcl-1, suggesting this as a possible intracellular mechanism of induced survival. By contrast, levels of Bcl-2, and two other Bcl-2 family members, Bax and Bak, remained unaltered. Given the importance of the CD40-CD40L interaction in B-cell responses, these results suggest a significant role of IL-13 in the regulation of B-cell apoptosis.


Spatiotemporally controlled overexpression of cyclin D1 triggers generation of supernumerary cells in the postnatal mouse inner ear.
The retinoblastoma family of pocket proteins (pRBs), composed of Rb1, p107, and p130 are negative regulators of cell-cycle progression. The deletion of any individual pRB in the auditory system triggers hair cells' (HCs) and supporting cells' (SCs) proliferation to different extents. Nevertheless, accessing their combined role in the inner ear through conditional or complete knockout methods is limited by the early mortality of the triple knockout. In quiescent cells, hyperphosphorylation and inactivation of the pRBs are maintained through the activity of the Cyclin-D1-cdk4/6 complex. Cyclin D1 (CycD1) is expressed in the embryonic and neonatal inner ear. In the mature organ of Corti (OC), CycD1 expression is significantly downregulated, paralleling the OC mitotic quiescence. Earlier studies showed that CycD1 overexpression leads to cell-cycle reactivation in cultures of inner ear explants. Here, we characterize a Cre-activated, Doxycycline (Dox)-controlled, conditional CycD1 overexpression model, which when bred to a tetracycline-controlled transcriptional activator and the Atoh1-cre mouse lines, allow for transient CycD1 overexpression and pRBs' downregulation in the inner ear in a reversible fashion. Analyses of postnatal mice's inner ears at various time points revealed the presence of supernumerary cells throughout the length of the cochlea and in the vestibular end-organs. Notably, most supernumerary cells were observed in the inner hair cells' (IHCs) region, expressed myosin VIIa (M7a), and showed no signs of apoptosis at any of the time points analyzed. Auditory and vestibular phenotypes were similar between the different genotypes and treatment groups. The fact that no significant differences were observed in auditory and vestibular function supports the notion that the supernumerary cells detected in the adult mice cochlea and macular end-organs may not impair auditory functions.


Breastfeeding may protect from developing attention-deficit/hyperactivity disorder.
Breastfeeding has a positive influence on physical and mental development. Attention-deficit/hyperactivity disorder (ADHD) is a common neurobehavioral disorder with major social, familial, and academic influences. The present study aimed to evaluate whether ADHD is associated with a shorter duration of breastfeeding. In this retrospective matched study, children 6-12 years old diagnosed at Schneider's Children Medical Center (Petach Tikva, Israel) with ADHD between 2008 and 2009 were compared with two control groups. The first one consisted of healthy (no ADHD) siblings of ADHD children; the second control group consisted of children without ADHD who consulted at the otolaryngology clinic. A constructed questionnaire about demographic, medical, and perinatal findings, feeding history during the first year of life, and a validated adult ADHD screening questionnaire were given to both parents of every child in each group. In children later diagnosed as having ADHD, 43% were breastfed at 3 months of age compared with 69% in the siblings group and 73% in the control non-related group (p=0.002). By 6 months of age 29% of ADHD children were breastfed compared with 50% in the siblings group and 57% in the control non-related group (p=0.011). A stepwise logistic regression that included the variables found to be significant in univariate analysis demonstrated a significant association between ADHD and lack of breastfeeding at 3 months of age, maternal age at birth, male gender, and parental divorce. Children with ADHD were less likely to breastfeed at 3 months and 6 months of age than children in the two control groups. We speculate that breastfeeding may have a protective effect from developing ADHD later in childhood.


Exercise during school hours when added to patient education improves outcome for 2 years in adolescent patellofemoral pain: a cluster randomised trial.
Patellofemoral pain (PFP) is common among adolescents and associated with long-lasting pain and disability. Patient education and exercise therapy are commonly used treatments in primary and secondary care but the effect of these treatments in adolescents is unknown. We aimed to determine the effect of exercise therapy as an add-on therapy to patient education compared with education alone. 121 adolescents from 15-19 years of age were cluster randomised to patient education or patient education combined with exercise therapy. Patient education covered self-management of pain and information on PFP. Exercise therapy consisted of supervised exercises on school premises (3/week for 3 months) and instructions on home-based exercises. Adherence to exercises was assessed as attendance and weekly text messages. Primary outcome measure was self-reported recovery (seven-point Likert scale) at 12 months with additional follow-ups at 3, 6 and 24 months. Adolescents randomised to patient education and exercise therapy were more likely to have recovered at 12 months (OR, 1.73, 95% CI 1.02 to 2.93, number needed to treat (NNT) of 11). Similar results were observed at 3 and 6 months (OR 1.88 and 1.43) while the effect was further increased at 24 months (OR of 2.52, NNT of 5). A higher total number of weekly exercise sessions increased the odds of recovery. In adolescent PFP, the addition of exercise therapy for 3 months was more effective than patient education alone. The effect was apparent at 3 months and increased up to 2 years. Adherence to exercises was important and improved the odds of recovery. clinicaltrials.gov reference: NCT01438762.


The protease Omi cleaves the mitogen-activated protein kinase kinase MEK1 to inhibit microglial activation.
Inflammation in Parkinson's disease is closely associated with disease pathogenesis. Mutations in Omi, which encodes the protease Omi, are linked to neurodegeneration and Parkinson's disease in humans and in mouse models. The severe neurodegeneration and neuroinflammation that occur in mnd2 (motor neuron degeneration 2) mice result from loss of the protease activity of Omi by the point mutation S276C; however, the substrates of Omi that induce neurodegeneration are unknown. We showed that Omi was required for the production of inflammatory molecules by microglia, which are the resident macrophages in the central nervous system. Omi suppressed the activation of the mitogen-activated protein kinases (MAPKs) extracellular signal-regulated kinase 1 and 2 (ERK1/2) by cleaving the upstream kinase MEK1 (mitogen-activated or extracellular signal-regulated protein kinase kinase 1). Knockdown of Omi in microglial cell lines led to activation of ERK1/2 and resulted in degradation of IκBα [α inhibitor of nuclear factor κB (NF-κB)], resulting in NF-κB activation and the expression of genes encoding inflammatory molecules, such as tumor necrosis factor-α and inducible nitric oxide synthase. The production of inflammatory molecules induced by the knockdown of Omi was blocked by the MEK1-specific inhibitor U0126. Furthermore, expression of the protease-deficient S276C Omi mutant in a microglial cell line had no effect on MEK1 cleavage or ERK1/2 activation. In the brains of mnd2 mice, we observed increased transcription of several genes encoding inflammatory molecules, as well as activation of astrocytes and microglia. Therefore, our study demonstrates that Omi is an intrinsic cellular factor that inhibits neuroinflammation.


Alternatives to Traditional Pedicle Screws for Posterior Fixation of the Degenerative Lumbar Spine.
Traditional pedicle screws are currently the gold standard to achieve stable 3-column fixation of the degenerative lumbar spine. However, there are cases in which pedicle screw fixation may not be ideal. Due to their starting point lateral to the pars interarticularis, pedicle screws require a relatively wide dissection along with a medialized trajectory directed toward the centrally located neural elements and prevertebral vasculature. In addition, low bone mineral density remains a major risk factor for pedicle screw loosening, pullout, and pseudarthrosis. The purpose of this article is to review the indications, advantages, disadvantages, and complications associated with posterior fixation techniques of the degenerative lumbar spine beyond the traditional pedicle screws. Comprehensive literature searches of the PubMed, Scopus, and Web of Science databases were performed for 5 methods of posterior spinal fixation, including (1) cortical bone trajectory (CBT) screws, (2) transfacet screws, (3) translaminar screws, (4) spinous process plates, and (5) fusion mass screws and hooks. Articles that had been published between January 1, 1990, and January 1, 2020, were considered. Non-English-language articles and studies involving fixation of the cervical or thoracic spine were excluded from our review. After reviewing over 1,700 articles pertaining to CBT and non-pedicular fixation techniques, a total of 284 articles met our inclusion criteria. CBT and transfacet screws require less-extensive exposure and paraspinal muscle dissection compared with traditional pedicle screws and may therefore reduce blood loss, postoperative pain, and length of hospital stay. In addition, several methods of non-pedicular fixation such as translaminar and fusion mass screws have trajectories that are directed away from or posterior to the spinal canal, potentially decreasing the risk of neurologic injury. CBT, transfacet, and fusion mass screws can also be used as salvage techniques when traditional pedicle screw constructs fail. CBT and non-pedicular fixation may be preferred in certain lumbar degenerative cases, particularly among patients with osteoporosis. Limitations of non-pedicular techniques include their reliance on intact posterior elements and the lack of 3-column fixation of the spine. As a result, transfacet and translaminar screws are infrequently used as the primary method of fixation. CBT, transfacet, and translaminar screws are effective in augmenting interbody fixation and have been shown to significantly improve fusion rates and clinical outcomes compared with stand-alone anterior lumbar interbody fusion. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Role of Nogo-A in neuronal survival in the reperfused ischemic brain.
Nogo-A is an oligodendroglial neurite outgrowth inhibitor, the deactivation of which enhances brain plasticity and functional recovery in animal models of stroke. Nogo-A's role in the reperfused brain tissue was still unknown. By using Nogo-A(-/-) mice and mice in which Nogo-A was blocked with a neutralizing antibody (11C7) that was infused into the lateral ventricle or striatum, we show that Nogo-A inhibition goes along with decreased neuronal survival and more protracted neurologic recovery, when deactivation is constitutive or induced 24 h before, but not after focal cerebral ischemia. We show that in the presence of Nogo-A, RhoA is activated and Rac1 and RhoB are deactivated, maintaining stress kinases p38/MAPK, SAPK/JNK1/2 and phosphatase-and-tensin homolog (PTEN) activities low. Nogo-A blockade leads to RhoA deactivation, thus overactivating Rac1 and RhoB, the former of which activates p38/MAPK and SAPK/JNK1/2 via direct interaction. RhoA and its effector Rho-associated coiled-coil protein kinase2 deactivation in turn stimulates PTEN, thus inhibiting Akt and ERK1/2, and initiating p53-dependent cell death. Our data suggest a novel role of Nogo-A in promoting neuronal survival by controlling Rac1/RhoA balance. Clinical trials should be aware of injurious effects of axonal growth-promoting therapies. Thus, Nogo-A antibodies should not be used in the very acute stroke phase.


Establishment of human fetal hepatocyte organoids and CRISPR-Cas9-based gene knockin and knockout in organoid cultures from human liver.
The liver is composed of two epithelial cell types: hepatocytes and liver ductal cells. Culture conditions for expansion of human liver ductal cells in vitro as organoids were previously described in a protocol; however, primary human hepatocytes remained hard to expand, until recently. In this protocol, we provide full details of how we overcame this limitation, establishing culture conditions that facilitate long-term expansion of human fetal hepatocytes as organoids. In addition, we describe how to generate (multi) gene knockouts using CRISPR-Cas9 in both human fetal hepatocyte and adult liver ductal organoid systems. Using a CRISPR-Cas9 and homology-independent organoid transgenesis (CRISPR-HOT) approach, efficient gene knockin can be achieved in these systems. These gene knockin and knockout approaches, and their multiplexing, should be useful for a variety of applications, such as disease modeling, investigating gene functions and studying processes, such as cellular differentiation and cell division. The protocol to establish human fetal hepatocyte organoid cultures takes ~1-2 months. The protocols to genome engineer human liver ductal organoids and human fetal hepatocyte organoids take 2-3 months.


Hepatoprotective activity of Sonchus asper against carbon tetrachloride-induced injuries in male rats: a randomized controlled trial.
Sonchus asper (SAME) is used as a folk medicine in hepatic disorders. In this study, the hepatoprotective effects of the methanol extract of SAME was evaluated against carbon tetrachloride (CCl4)-induced liver injuries in rats. To evaluate the hepatoprotective effects of SAME, 36 male Sprague-Dawley rats were equally divided into 6 groups. Rats of Group I (control) were given free access to approved feed and water. Rats of Group II were injected intraperitoneally with CCl4 (3 ml/kg) as a 30% solution in olive oil (v/v) twice a week for 4 weeks. Animals of Groups III (100 mg/kg) and IV (200 mg/kg) received SAME, whereas those of Group V were given silymarin via gavage (100 mg/kg) after 48 h of CCl4 treatment. Group VI received SAME (200 mg/kg) twice a week for 4 weeks without CCl4 treatment. Various parameters, such as the serum enzyme levels, serum biochemical marker levels, antioxidant enzyme activities, and liver histopathology were used to estimate the hepatoprotective efficacy of SAME. The administration of SAME and silymarin significantly lowered the CCl4-induced serum levels of hepatic marker enzymes (aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase), cholesterol, low-density lipoprotein, and triglycerides while elevating high-density lipoprotein levels. The hepatic contents of glutathione and activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase, and glutathione reductase were reduced. The levels of thiobarbituric acid-reactive substances that were increased by CCl4 were brought back to control levels by the administration of SAME and silymarin. Liver histopathology showed that SAME reduced the incidence of hepatic lesions induced by CCl4 in rats. SAME may protect the liver against CCl4-induced oxidative damage in rats.


Generalized uncertainty relations and coherent and squeezed states.
Characteristic uncertainty relations and their related squeezed states are briefly reviewed and compared in accordance with the generalizations of three equivalent definitions of canonical coherent states. The standard SU(1, 1) coherent states are shown to be the unique states that minimize the Schrödinger uncertainty relation for every pair of the three generators and the Robertson relation for the three generators. The characteristic uncertainty inequalities are naturally extended to the case of several states. It is shown that these inequalities can be written in the equivalent complementary form.


A New Framework for the Assessment of Cerebral Hemodynamics Regulation in Neonates Using NIRS.
We present a new framework for the assessment of cerebral hemodynamics regulation (CHR) in neonates using near-infrared spectroscopy (NIRS). In premature infants, NIRS measurements have been used as surrogate variables for cerebral blood flow (CBF) in the assessment of cerebral autoregulation (CA). However, NIRS measurements only reflect changes in CBF under constant changes in arterial oxygen saturation (SaO2). This condition is unlikely to be met at the bedside in the NICU. Additionally, CA is just one of the different highly coupled mechanisms that regulate brain hemodynamics. Traditional methods for the assessment of CA do not take into account the multivariate nature of CHR, producing inconclusive results. In this study we propose a newly developed multivariate methodology for the assessment of CHR. This method is able to effectively decouple the influences of SaO2 from the NIRS measurements, and at the same time, produces scores indicating the strength of the coupling between the systemic variables and NIRS recordings. We explore the use of this method, and its derived scores, for the monitoring of CHR using data from premature infants who developed a grade III-IV intra-ventricular hemorrhage during the first 3 days of life.


Quality assurance questionnaire for professionals fails to improve the quality of informed consent.
The informed consent process for research warrants improvement but approaches designed to enhance informed consent need testing in the context of actual clinical research. Test the cumulative effect of a retrospective quality assurance questionnaire intended to enhance awareness in the person obtaining informed consent on the quality of the informed consent in clinical trials. In the Veterans Affairs Cooperative Study 'Enhancing the Quality of Informed Consent- Self Monitoring', 30 study sites are randomly assigned from five clinical trials to either use a new quality assurance questionnaire after each informed consent encounter or the standard process of obtaining informed consent. The quality of informed consent is evaluated using independent telephone interviews of 836 subjects who had given consent to participate in the clinical trials and the authors' study. The main outcome measures are two previously validated scores derived from an independent telephone interview, measuring the overall quality of consent as well as the degree of 'therapeutic misapprehension'. Patients and assessors are blind to the study arm assignment. Subjects report complete (93%) or some (6%) satisfaction with the consent process of the 'parent' clinical trial, and 91% recognize no consequences to non-participation. Concerning the 'primary purpose' of the parent trial, 67% indicate understanding of the research purpose, 41% that the research is to benefit others, while 14% think the research is directed to their own benefit; 60% report no risk to participation and 65% report at least one expects direct benefit. Interviewers assess 77% of subjects as showing full appreciation of the 'voluntariness' of participation. The quality assurance questionnaire do not provide an appreciable effect on the quality of informed consent. Using mixed model methods to account for the group randomization, near zero, non-significant effects have been found for the overall assessment score (-0.034 on a 0-10 point scale, standard error 0.099, P = 0.73) and for the score measuring 'therapeutic misconception' (-0.005 on a 0-5 point scale, standard error 0.137, P = 0.97). Permutation methods yield similar results. Confidence intervals are narrow enough to exclude any clinically important effect. The intervention may work in a more homogeneous patient population, or one that is not sampled. The outcome measurement relies on a short, anonymous, telephone interview (to minimize burden and eliminate bias), but a longer, face-to-face interview may be more sensitive to differences. A 'checklist' tied directly to the outcome measures may show an effect. Despite prior beliefs, a standardized quality assurance tool do not enhance informed consent in actual clinical trials. Future research is needed to rigorously evaluate proposed methods to enhance informed consent prior to widespread introduction.


The burden of vancomycin-resistant enterococcal infections in US hospitals, 2003 to 2004.
Despite significant concern in the health care community regarding vancomycin-resistant enterococci (VRE), there are no estimates of the total number of VRE infections that occur each year in US hospitals. Using data from a national survey of hospital discharges and a national antimicrobial resistance surveillance system, we estimated the annual number of US hospitalization with VRE bloodstream, urinary tract, and wound or intra-abdominal infections. Because of the inexact nature of hospital discharge diagnosis coding, we made both a conservative and liberal estimate of hospitalization with VRE infection by using a variety of data sources. For the years 2003 and 2004, we conservatively estimated that there were 20777 and 20931 VRE infections, respectively; for those same years, the liberal estimates were 78330 and 85586, respectively. Because there are such a large number of hospital discharges for which an infection is coded without an organism code, it is likely that the conservative estimate is an underestimate of the true burden. These estimates highlight the importance of controlling VRE and the need to develop improved methods for tracking the burden of such infections.


Stroke in systemic lupus erythematosus.
We investigated the clinical and pathologic characteristics of stroke in 234 patients with systemic lupus erythematosus. Thirteen patients (5.6%) developed cerebrovascular disease. Cerebral infarction was noted in eight, cerebral hemorrhage in two, and subarachnoid hemorrhage in three. In seven (54%) of these 13 patients, stroke occurred less than or equal to 5 years after systemic lupus erythematosus was diagnosed. Among the predisposing risk factors for stroke, hypertension was the most important. Lupus anticoagulant was detected in three (38%) and anticardiolipin antibody in three (43% of seven investigated) of the patients with infarction. Evaluation of the clinical manifestations and autoantibodies indicated that renal involvement and high titers of anti-deoxyribonucleic acid antibody were more frequent in the stroke group than in the non-stroke group. Autopsy studies on six of the patients with stroke revealed small infarcts and hemorrhages in all, but in no case was true angiitis observed. Libman-Sacks endocarditis was found in two of the three patients with infarction. In conclusion, the important contributory factor to the development of stroke in patients with systemic lupus erythematosus is considered to be hypertension mediated by immunologic abnormalities. Antiphospholipid antibodies and Libman-Sacks endocarditis are closely associated with occlusive cerebrovascular disease.


Impact of graphyne on structural and dynamical properties of calmodulin.
Carbon-based nanomaterials such as graphyne, graphene, and carbon nanotubes have attracted considerable attention for their applications, but questions remain regarding their biosafety through potential adverse interactions with important biomolecules. Here, we investigate the binding of a graphyne nanosheet and a graphyne quantum dot to calmodulin (CaM), a dynamic Ca2+ binding protein. Simulation results show that both the graphyne nanosheet and graphyne quantum dot are able to bind both Ca2+-bound and Ca2+-free CaM with similar binding affinities. From our simulations, graphyne quantum dots appear particularly worrisome for potential toxicity as they bind to and occlude the hydrophobic binding pocket of both Ca2+-bound and Ca2+-free CaM. In both the nanosheet and quantum dot systems, the binding interactions between graphyne and CaM are markedly hydrophobic and heavily suppress the Ca2+-free CaM dynamics. Our results reiterate that graphitic nanomaterials including graphyne may have negative impacts on both the structure and dynamics of key proteins such as CaM involved in calcium signal transduction.


THE POSTMORTEM RIGOR OF THE MAMMALIAN HEART AND THE INFLUENCE OF AN ANTEMORTEM STIMULATION OF THE PNEUMOGASTRIC NERVES UPON ITS DEVELOPMENT.
IN NORMAL ANIMALS (DOGS, CATS AND RABBITS) WHICH WERE KILLED BY EXSANGUINATION, THE BEGINNING OF RIGOR IN PRACTICALLY ALL CASES WAS SEPARATED FROM RESPIRATORY DEATH BY TWO WELL CHARACTERIZED PERIODS: (1) a period of pulsation during which each ventricle still exhibited some spontaneous contractions, and (2) a period of relaxation during which the ventricles, while showing neither spontaneous contractions nor rigor, are more relaxed than in normal diastole, and during which the ventricles gradually lose their irritability. (The cardiac rigor or tonic contraction which was observed by recent investigators to set in immediately after death is an artificial phenomenon produced by filling the heart with saline and connecting it with a manometer.) In the right ventricle both periods are longer than in the left ventricle, i. e., rigor sets in later. Moreover the development of rigor from onset to maximum is also longer in the right ventricle than in the left, although here the difference is less striking. The stopping of spontaneous beating, the disappearance of irritability, and the development of rigor, manifest in both ventricles a topographical progress from the base toward the apex, i. e., the stoppage of beating, the disappearance of irritability and the setting in of rigor occur first at the uppermost part of the ventricle and last at the lowest point of the ventricle. It often occurred that rigor was already present in the basal part of a ventricle while the apex was still beating. It is probable also that the loss of vital activity and irritability and the development of rigor progress topographically from the endocardial to the epicardial surfaces. Prolonged etherization retards the onset, but hastens the development of rigor; atropinization hastens both onset and development. Repeated prolonged stoppages of the heart caused by antemortem (and postmortem) stimulations of the pneumogastric nerves hasten the onset as well as the development of rigor of the heart. All three periods are affected by the inhibitory influence of the stimulation, the period of relaxation, however, seems to be the one which is shortened most. The most probable interpretation of this phenomenon is the assumption that it is caused by an asphyxiation of the cardiac tissues. The stimulations of the pneumogastric nerves seem to hasten also the onset of the general rigor-probably this, too, is the result of some premature asphyxiation of the skeletal muscles.


Mediator regulates non-coding RNA transcription at fission yeast centromeres.
In fission yeast, centromeric heterochromatin is necessary for the fidelity of chromosome segregation. Propagation of heterochromatin in dividing cells requires RNA interference (RNAi) and transcription of centromeric repeats by RNA polymerase II during the S phase of the cell cycle. We found that the Med8-Med18-Med20 submodule of the Mediator complex is required for the transcriptional regulation of native centromeric dh and dg repeats and for the silencing of reporter genes inserted in centromeric heterochromatin. Mutations in the Med8-Med18-Med20 submodule did not alter Mediator occupancy at centromeres; however, they led to an increased recruitment of RNA polymerase II to centromeres and reduced levels of centromeric H3K9 methylation accounting for the centromeric desilencing. Further, we observed that Med18 and Med20 were required for efficient processing of dh transcripts into siRNA. Consistent with defects in centromeric heterochromatin, cells lacking Med18 or Med20 displayed elevated rates of mitotic chromosome loss. Our data demonstrate a role for the Med8-Med18-Med20 Mediator submodule in the regulation of non-coding RNA transcription at Schizosaccharomyces pombe centromeres. In wild-type cells this submodule limits RNA polymerase II access to the heterochromatic DNA of the centromeres. Additionally, the submodule may act as an assembly platform for the RNAi machinery or regulate the activity of the RNAi pathway. Consequently, Med8-Med18-Med20 is required for silencing of centromeres and proper mitotic chromosome segregation.


The expression profile and function of Satb2 in zebrafish embryonic development.
The present study shows the expression profile and function of the homeobox gene, satb2 during zebrafish embryonic development. Satb2 was ubiquitously expressed from the 1 cell stage to the 10-somite stage in zebrafish embryos. Satb2 showed stage-specific expression profiles such as in the pronephric duct at 24 hpf, the branchial arches at 36 hpf, and the ganglion cell layer of the retina and fins at 48 hpf. Additionally, satb2 knockdown embryos were arrested at 50-60% epiboly, and transplantation experiments with satb2 knockdown cells showed migration defects. Interestingly, satb2 knockdown cells also exhibited down-regulation of dynamin II and VAMP4, which are involved in exocytosis and endocytosis, respectively. Furthermore, satb2 knockdown cells have a disorganized actin distribution and an underdeveloped external yolk syncytial layer, both of which are involved in epiboly. These results suggest that satb2 has a functional role in epiboly. This role may potentially be the regulation of endo-exocytic vesicle transport-dependent cell migration and/or the regulation of the development of the yolk syncytial layer.


Association Between Body Mass Index and Outcomes in Venoarterial Extracorporeal Membrane Oxygenation.
The association between obesity, or elevated body mass index (BMI), and outcomes in patients receiving venoarterial extracorporeal membrane oxygenation (VA-ECMO) has not been well established. Recent studies in patients receiving venovenous ECMO did not detect an association between obesity and increased mortality. The purpose of this retrospective observational study is to evaluate the association between BMI and survival in patients receiving VA-ECMO for cardiogenic shock. All patients >18 years of age supported on VA-ECMO for refractory cardiogenic shock in a single academic center between 2009 and 2019 were included. ECMO outcomes, including successful ECMO decannulation and 30-day survival, were analyzed after stratification according to BMI. Multivariable and univariate logistic regression were used to assess the association between BMI and VA-ECMO outcomes. Of the total patients (n = 355) cannulated for VA-ECMO, 61.7% of the patients survived to ECMO recovery/decannulation, 45.5% of the patients survived to 30 days after ECMO decannulation, and 38.9% of the patients survived to hospital discharge with no statistically significant differences among the BMI groups. Multivariable logistic regression did not reveal any associations between obesity as defined by BMI and survival to ECMO decannulation (odds ratio [OR] 1.07 per 5 unit increase in BMI, 95% confidence interval [CI], 0.86-1.33; P = .57), 30-day survival (OR = 0.91, 95% CI, 0.73-1.14; P = .41) or survival to hospital discharge (OR = 0.95, 95% CI, 0.75-1.20; P = .66). Despite potential challenges to cannulation and maintaining adequate flow during ECMO, this single centered, retrospective observational study did not detect association between BMI and survival to ECMO decannulation, 30-day survival, or survival to hospital discharge for patients requiring VA-ECMO for refractory cardiogenic shock. These data suggest that obesity alone should not exclude candidacy for VA-ECMO.The primary outcome in this retrospective study was survival of the ECMO therapy (survival to ECMO decannulation), defined as surviving >24 hours after decannulation without a withdrawal of care. Secondary outcomes included survival at 30 days and survival to hospital discharge.


Syringomyelia in children with closed spinal dysraphism: long-term outcomes after surgical intervention.
The nature of the relationship between spinal cord syrinx and tethered cord is not well known. It is unclear if surgical cord untethering results in resolution or improvement of an associated syrinx. The objective of this study was to report the response of spinal cord syrinx to surgical cord untethering. The authors retrospectively reviewed all patients with a syrinx and tethered cord who presented to a single institution over an 11-year interval. Patients with open neural tube defects were excluded. Thirty-one patients were identified, 25 of whom had both clinical and imaging follow-up after surgery. Patients were grouped according to etiology of the tethered cord. Clinical outcomes and syrinx characteristics were recorded. Of the 25 patients with tethered cord, 68% (n = 17) were male. The average age at presentation was 2.5 years (0-10.1 years) and age at surgery was 3.7 years (range 1 day to 17 years). Etiologies of tethered cord were lipomyelomeningocele (n = 8), thickened/fatty filum (n = 7), intradural lipoma (n = 5), myelocystocele (n = 2), meningocele (n = 2), and diastematomyelia (n = 1). Twenty-three of the patients underwent primary untethering, whereas 2 patients had received untethering previously at another institution. The average syrinx length and width prior to surgery were 4.81 vertebral levels (SD 4.35) and 5.19 mm (SD 2.55 mm), respectively. Conus level ranged from L1 to S3. Patients were followed for an average of 8.4 years (1.35-15.85 years). Overall there was no significant change in syrinx length or width postoperatively; the average syrinx length increased by 0.86 vertebral levels (SD 4.36) and width decreased by 0.72 mm (SD 2.94 mm). Seven of 25 patients had improvement in at least one presenting symptom, including scoliosis, weakness, bowel/bladder dysfunction, and pain. Eight patients had stable presenting symptoms. Six patients were asymptomatic and 5 patients had new or worsening symptoms, which included scoliosis, pain, or sensory changes. Although some syrinxes improved after surgery for tethered cord, radiological improvement was not consistent and did not appear to be associated with change in clinical symptoms. The decision to surgically untether a cord should be focused on the clinical symptoms and not the presence of a syrinx alone. Further studies are needed to confirm this finding.


Antigenic diversity thresholds and the development of AIDS.
Longitudinal studies of patients infected with HIV-1 reveal a long and variable incubation period between infection and the development of AIDS. Data from a small number of infected patients show temporal changes in the number of genetically distinct strains of the virus throughout the incubation period, with a slow but steady rise in diversity during the progression to disease. A mathematical model of the dynamic interaction between viral diversity and the human immune system suggests the existence of an antigen diversity threshold, below which the immune system is able to regulate viral population growth but above which the virus population induces the collapse of the CD4+ lymphocyte population. The model suggests that antigenic diversity is the cause, not a consequence, of immunodeficiency disease. The model is compared with available data, and is used to assess how the timing of the application of chemotherapy or immunotherapy influences the rate of progress to disease.


Stromal Senescence By Prolonged CDK4/6 Inhibition Potentiates Tumor Growth.
Senescent cells within the tumor microenvironment (TME) adopt a proinflammatory, senescence-associated secretory phenotype (SASP) that promotes cancer initiation, progression, and therapeutic resistance. Here, exposure to palbociclib (PD-0332991), a CDK4/6 inhibitor, induces senescence and a robust SASP in normal fibroblasts. Senescence caused by prolonged CDK4/6 inhibition is DNA damage-independent and associated with Mdm2 downregulation, whereas the SASP elicited by these cells is largely reliant upon NF-κB activation. Based upon these observations, it was hypothesized that the exposure of nontransformed stromal cells to PD-0332991 would promote tumor growth. Ongoing clinical trials of CDK4/6 inhibitors in melanoma prompted a validation of this hypothesis using a suite of genetically defined melanoma cells (i.e., Ras mutant, Braf mutant, and Ras/Braf wild-type). When cultured in the presence of CDK4/6i-induced senescent fibroblasts, melanoma cell lines exhibited genotype-dependent proliferative responses. However, in vivo, PD-0332991-treated fibroblasts enhanced the growth of all melanoma lines tested and promoted the recruitment of Gr-1-positive immune cells. These data indicate that prolonged CDK4/6 inhibitor treatment causes normal fibroblasts to enter senescence and adopt a robust SASP. Such senescent cells suppress the antitumor immune response and promote melanoma growth in immunocompetent, in vivo models.Implications: The ability of prolonged CDK4/6 inhibitor treatment to induce cellular senescence and a robust SASP in primary cells may hinder therapeutic efficacy and promote long-term, gerontogenic consequences that should be considered in clinical trials aiming to treat melanoma and other cancer types. Mol Cancer Res; 15(3); 237-49. ©2016 AACR.


Fusiform Cervical Mass in a 6-Year Old Boy; Do not Forget the Thymic Cyst.
Cervical thymic cyst is a rare clinical entity, with approximately one hundred cases reported in the literature so far. The purpose of this case report is to highlight some certain features, along with an extensive research of the relevant literature. A 6-year-old boy was admitted to the Otorhinolaryngology Department due to the presence of a left-sided, painless, latero-cervical swelling, first observed by his parents 2 weeks ago. Physical examination revealed a painless, well-delineated mass, with no signs of inflammation. No enlarged cervical nodes were present. The mass extended from the mandibular angle, under the sternocleidomastoid muscle, in proximity with the ipsilateral neurovascular bundle. Ultrasound transverse gray-scale panoramic view detected a wellcircumscribed lesion, with fine echogenic foci, appearing in close proximity with the upper pole of the left thyroid lobe and the ipsilateral common carotid artery. Elective surgical intervention with complete mass excision was performed. Histopathological examination confirmed the diagnosis of a cervical thymic cyst. Cervical thymic remnants represent a group of neck masses that pediatricians and pediatric surgeons should consider in differential diagnosis of both cystic and solid neck masses. Most cystic cervical thymic masses are found in the lower third of the anterior neck, extending deep to the sternocleidomastoid muscle, featuring close anatomic relationship with the composites of the ipsilateral carotid sheath. Elective surgery is kept as the treatment of choice, after ruling out the possibility of subject immunologic disturbance.


Pressure-volume analysis in athyroid patients off and on thyroxine supplementation: a pilot study.
Thyroid hormone importantly affects the cardiovascular system. However, evaluation of stroke volume (SV) and its determinants is confounded by variations in volume status that occur along different thyroid states. This study applied the pressure-volume (PV) framework to obtain relatively load-independent estimates of cardiac function in hypothyroidism as compared to euthyroidism. Ten athyroid patients were assessed echocardiographically after 4 weeks in deep hypothyroid state, and again after supplementation with oral Levothyroxine (LT4) for 3 months. Thyroid hormone levels were assessed and noninvasive pressure-volume (PV) analysis based on dedicated repeated echocardiograms was performed. Changes were assessed using paired tests. Results are presented as medians and interquartile ranges. Hypothyroidism was associated with reduced stroke volume (SV: 67.6 ± 17 vs. 75.7 ± 20.6 mL, P = 0.024), preload (end-diastolic volume, EDV: 122.6 ± 32.5 vs. 135.7 ± 33.6 mL, P = 0.004), and contractility (end-systolic elastance, Ees : 1.7 ± 0.33 vs. 2.58 ± 1.33 mmHg/mL, P = 0.01). Afterload was constant (effective arterial elastance, Ea : 1.66 ± 0.32 vs. 1.79 ± 0.52 mmHg/mL, P = 0.43) and the total energy spent was lower (PVA∙HR: 86.7 ± 28 vs. 110.9 ± 32.1 J, P = 0.04). Hemodynamic manifestations of frank hypothyroidism in humans are characterized by reduced preload and contractility, and unchanged total afterload. LT4 therapy increased work efficiency and heart rate, but not the net energy expenditure. Noninvasive PV analysis may be useful to follow-up different thyroid states.


Active testing search for point cloud matching.
We present a general approach for solving the point-cloud matching problem for the case of mildly nonlinear transformations. Our method quickly finds a coarse approximation of the solution by exploring a reduced set of partial matches using an approach to which we refer to as Active Testing Search (ATS). We apply the method to registration of graph structures by branching point matching. It is based solely on the geometric position of the points, no additional information is used nor the knowledge of an initial alignment. In the second stage, we use dynamic programming to refine the solution. We tested our algorithm on angiography, retinal fundus, and neuronal data gathered using electron and light microscopy. We show that our method solves cases not solved by most approaches, and is faster than the remaining ones.


Natural Mineral-Based Solid Oxide Fuel Cell with Heterogeneous Nanocomposite Derived from Hematite and Rare-Earth Minerals.
Solid oxide fuel cells (SOFCs) have attracted much attention worldwide because of their potential for providing clean and reliable electric power. However, their commercialization is subject to the high operating temperatures and costs. To make SOFCs more competitive, here we report a novel and attractive nanocomposite hematite-LaCePrOx (hematite-LCP) synthesized from low-cost natural hematite and LaCePr-carbonate mineral as an electrolyte candidate. This heterogeneous composite exhibits a conductivity as high as 0.116 S cm(-1) at 600 °C with an activation energy of 0.50 eV at 400-600 °C. For the first time, a fuel cell using such a natural mineral-based composite demonstrates a maximum power density of 625 mW cm(-2) at 600 °C and notable power output of 386 mW cm(-2) at 450 °C. The extraordinary ionic conductivity and device performances are primarily attributed to the heterophasic interfacial conduction effect of the hematite-LCP composite. These superior properties, along with the merits of ultralow cost, abundant storage, and eco-friendliness, make the new composite a highly promising material for commercial SOFCs.


Campylobacter jejuni type VI secretion system: roles in adaptation to deoxycholic acid, host cell adherence, invasion, and in vivo colonization.
The recently identified type VI secretion system (T6SS) of proteobacteria has been shown to promote pathogenicity, competitive advantage over competing microorganisms, and adaptation to environmental perturbation. By detailed phenotypic characterization of loss-of-function mutants, in silico, in vitro and in vivo analyses, we provide evidence that the enteric pathogen, Campylobacter jejuni, possesses a functional T6SS and that the secretion system exerts pleiotropic effects on two crucial processes--survival in a bile salt, deoxycholic acid (DCA), and host cell adherence and invasion. The expression of T6SS during initial exposure to the upper range of physiological levels of DCA (0.075%-0.2%) was detrimental to C. jejuni proliferation, whereas down-regulation or inactivation of T6SS enabled C. jejuni to resist this effect. The C. jejuni multidrug efflux transporter gene, cmeA, was significantly up-regulated during the initial exposure to DCA in the wild type C. jejuni relative to the T6SS-deficient strains, suggesting that inhibition of proliferation is the consequence of T6SS-mediated DCA influx. A sequential modulation of the efflux transporter activity and the T6SS represents, in part, an adaptive mechanism for C. jejuni to overcome this inhibitory effect, thereby ensuring its survival. C. jejuni T6SS plays important roles in host cell adhesion and invasion as T6SS inactivation resulted in a reduction of adherence to and invasion of in vitro cell lines, while over-expression of a hemolysin co-regulated protein, which encodes a secreted T6SS component, greatly enhanced these processes. When inoculated into B6.129P2-IL-10(tm1Cgn) mice, the T6SS-deficient C. jejuni strains did not effectively establish persistent colonization, indicating that T6SS contributes to colonization in vivo. Taken together, our data demonstrate the importance of bacterial T6SS in host cell adhesion, invasion, colonization and, for the first time to our knowledge, adaptation to DCA, providing new insights into the role of T6SS in C. jejuni pathogenesis.


Development of a risk stratification and prevention index for stratified care in chronic low back pain. Focus: yellow flags (MiSpEx network).
Chronic low back pain (LBP) is a major cause of disability; early diagnosis and stratification of care remain challenges. This article describes the development of a screening tool for the 1-year prognosis of patients with high chronic LBP risk (risk stratification index) and for treatment allocation according to treatment-modifiable yellow flag indicators (risk prevention indices, RPI-S). Screening tools were derived from a multicentre longitudinal study (n = 1071, age >18, intermittent LBP). The greatest prognostic predictors of 4 flag domains ("pain," "distress," "social-environment," "medical care-environment") were determined using least absolute shrinkage and selection operator regression analysis. Internal validity and prognosis error were evaluated after 1-year follow-up. Receiver operating characteristic curves for discrimination (area under the curve) and cutoff values were determined. The risk stratification index identified persons with increased risk of chronic LBP and accurately estimated expected pain intensity and disability on the Pain Grade Questionnaire (0-100 points) up to 1 year later with an average prognosis error of 15 points. In addition, 3-risk classes were discerned with an accuracy of area under the curve = 0.74 (95% confidence interval 0.63-0.85). The RPI-S also distinguished persons with potentially modifiable prognostic indicators from 4 flag domains and stratified allocation to biopsychosocial treatments accordingly. The screening tools, developed in compliance with the PROGRESS and TRIPOD statements, revealed good validation and prognostic strength. These tools improve on existing screening tools because of their utility for secondary preventions, incorporation of exercise effect modifiers, exact pain estimations, and personalized allocation to multimodal treatments.


Charitable Food Systems' Capacity to Address Food Insecurity: An Australian Capital City Audit.
Australian efforts to address food insecurity are delivered by a charitable food system (CFS) which fails to meet demand. The scope and nature of the CFS is unknown. This study audits the organisational capacity of the CFS within the 10.9 square kilometres of inner-city Perth, Western Australia. A desktop analysis of services and 12 face-to-face interviews with representatives from CFS organisations was conducted. All CFS organisations were not-for⁻profit and guided by humanitarian or faith-based values. The CFS comprised three indirect services (IS) sourcing, banking and/or distributing food to 15 direct services (DS) providing food to recipients. DS offered 30 different food services at 34 locations feeding over 5670 people/week via 16 models including mobile and seated meals, food parcels, supermarket vouchers, and food pantries. Volunteer to paid staff ratios were 33:1 (DS) and 19:1 (IS). System-wide, food was mainly donated and most funding was philanthropic. Only three organisations received government funds. No organisation had a nutrition policy. The organisational capacity of the CFS was precarious due to unreliable, insufficient and inappropriate financial, human and food resources and structures. System-wide reforms are needed to ensure adequate and appropriate food relief for Australians experiencing food insecurity.


Treatments for people living with schizophrenia in Sub-Saharan Africa: an adapted realist review.
To identify the treatments and interventions available and their impact on people living with schizophrenia in Sub-Saharan Africa. Help-seeking behaviour and the choice of treatment are largely influenced by socio-cultural factors and beliefs about the causes of mental illness. This review addresses the gap in knowledge regarding the treatment options available to people living with schizophrenia in Sub-Saharan Africa. Adapted realist literature review. Electronic databases searched in June 2016 included PubMed, EMBASE, PsycINFO, ProQuest and CINAHL. The adapted realist review approach used to synthesize the published research involved identifying the review aim, searching and selecting relevant studies, extracting, iteratively analysing and synthesizing relevant data and reporting results. Forty studies from eight countries were reviewed. Most people were treated by both faith/traditional healers and modern psychiatry. Common treatments included antipsychotics, electroconvulsive therapy and psychosocial interventions. Few treatment options were available outside major centres, there was poor adherence to medication and families reported a high level of burden associated with caring for a relative. Major limitations of this review were the lack of studies, variable quality and low level of evidence available from most countries from Sub-Saharan Africa and lack of generalizability. People living with schizophrenia in Sub-Saharan Africa were treated by faith, traditional healers and modern psychiatry, if at all. Further research is needed to better understand the local situation and the implications for caring for people from this region. Mental health services in Sub-Saharan Africa are limited by fiscal shortages, lack of mental health services and qualified mental health professionals. This review provides evidence to inform nursing and healthcare policy, including recruiting and training mental health professionals and ensuring access to evidence-based, person-centred and culturally relevant mental health services within the primary care context.


Plasmid-mediated high-level resistance to aminoglycosides in Enterobacteriaceae due to 16S rRNA methylation.
A self-transferable plasmid of ca. 80 kb, pIP1204, conferred multiple-antibiotic resistance to Klebsiella pneumoniae BM4536, which was isolated from a urinary tract infection. Resistance to beta-lactams was due to the bla(TEM1) and bla(CTX-M) genes, resistance to trimethroprim was due to the dhfrXII gene, resistance to sulfonamides was due to the sul1 gene, resistance to streptomycin-spectinomycin was due to the ant3"9 gene, and resistance to nearly all remaining aminoglycosides was due to the aac3-II gene and a new gene designated armA (aminoglycoside resistance methylase). The cloning of armA into a plasmid in Escherichia coli conferred to the new host high-level resistance to 4,6-disubstituted deoxystreptamines and fortimicin. The deduced sequence of ArmA displayed from 37 to 47% similarity to those of 16S rRNA m(7)G methyltransferases from various actinomycetes, which confer resistance to aminoglycoside-producing strains. However, the low guanine-plus-cytosine content of armA (30%) does not favor an actinomycete origin for the gene. It therefore appears that posttranscriptional modification of 16S rRNA can confer high-level broad-range resistance to aminoglycosides in gram-negative human pathogens.


The location of descending fibres to sympathetic neurons supplying the eye and sudomotor neurons supplying the head and neck.
Evidence is given of the location in man of the fibres going to the sympathetic neurons of the lateral horn that supply the intrinsic and extrinsic muscles of the eye and the sweat glands of the head and neck. For the region of the pons and medulla, the evidence is abstracted from the literature. For the cervical spinal cord, the evidence is from our cases of anterolateral cordotomy. In the medulla, thrombosis of the artery of the fossette latérale destroys the fibres; this locates the fibres in the posterolateral retro-olivary area. But not all fibres to the sudomotor neurons lie there: some run elsewhere, though they probably remain ipsilateral. In the cervical cord, the fibres supplying the sympathetic neurons of the intrinsic and extrinsic muscles of the eye run near the posterior angle of the anterior horn. Most of the fibres supplying the sudomotor neurons lie in the same region, though some lie outside this area but on the same side of the cord.


The effects of electronic health record implementation on medical student educators.
Electronic heath records (EHRs) are being rapidly adopted. Little attention has been paid to the effects of EHR implementation on teaching. The goal was to determine the effect of EHR implementation on medical student educators and to characterize features that might influence their responses. A survey of all clinical faculty at Oregon Health & Science University was taken. The authors inquired about the effect of EHR implementation on enthusiasm for teaching and characterized factors that might contribute to responses. Nearly half of the faculty reported decreased enthusiasm for teaching following EHR implementation (48.2%). Most (65.1%) reported that the EHR distracted from teaching, and few saw its advantages. Most reported that EHR implementation led them to teach less (62.3%); the most enthusiastic teachers were most affected. EHR implementation in an academic medical center has substantial effects on educators. Further work is needed to determine how to train medical educators and modify the EHR to best support teaching.


Catalases of Aspergillus fumigatus.
Upon infection of a host, the pathogenic fungus Aspergillus fumigatus is attacked by the reactive oxygen species produced by phagocytic cells. Detoxification of hydrogen peroxide by catalases was proposed as a way to overcome this host response. A. fumigatus produces three active catalases; one is produced by conidia, and two are produced by mycelia. The mycelial catalase Cat1p was studied previously. Here we characterized the two other catalases, their genes, and the phenotypes of gene-disrupted mutants. CatAp, a spore-specific monofunctional catalase, is resistant to heat, metal ions, and detergent. This enzyme is a dimeric protein with 84.5-kDa subunits. The 749-amino-acid polypeptide exhibits high levels of similarity to the Aspergillus nidulans CatA catalase and to bacterial catalase HPII of Escherichia coli. In spite of increased sensitivity to H(2)O(2), killing of DeltacatA conidia by alveolar macrophages and virulence in animals were similar to the killing of conidia by alveolar macrophages and virulence in animals observed for the wild type. In contrast to the Cat1p and CatAp catalases, the mycelial Cat2p enzyme is a bifunctional catalase-peroxidase and is sensitive to heat, metal ions, and detergent. This enzyme, an 82-kDa monomer, is homologous to catalase-peroxidases of several fungi and bacteria. Surprisingly, mycelium of the double Deltacat1Deltacat2 mutant with no catalase activity exhibited only slightly increased sensitivity to H(2)O(2) and was as sensitive to killing by polymorphonuclear neutrophils as mycelium of the wild-type strain. However, this mutant exhibited delayed infection in the rat model of aspergillosis compared to infection by the wild-type strain. These results indicate that conidial catalase is not a virulence factor and that mycelial catalases transiently protect the fungus from the host.


Deciding on success criteria for predictability of pharmacokinetic parameters from in vitro studies: an analysis based on in vivo observations.
Prediction accuracy of pharmacokinetic parameters is often assessed using prediction fold error, i.e., being within 2-, 3-, or n-fold of observed values. However, published studies disagree on which fold error represents an accurate prediction. In addition, "observed data" from only one clinical study are often used as the gold standard for in vitro to in vivo extrapolation (IVIVE) studies, despite data being subject to significant interstudy variability and subjective selection from various available reports. The current study involved analysis of published systemic clearance (CL) and volume of distribution at steady state (Vss) values taken from over 200 clinical studies. These parameters were obtained for 17 different drugs after intravenous administration. Data were analyzed with emphasis on the appropriateness to use a parameter value from one particular clinical study to judge the performance of IVIVE and the ability of CL and Vss values obtained from one clinical study to "predict" the same values obtained in a different clinical study using the n-fold criteria for prediction accuracy. The twofold criteria method was of interest because it is widely used in IVIVE predictions. The analysis shows that in some cases the twofold criteria method is an unreasonable expectation when the observed data are obtained from studies with small sample size. A more reasonable approach would allow prediction criteria to include clinical study information such as sample size and the variance of the parameter of interest. A method is proposed that allows the "success" criteria to be linked to the measure of variation in the observed value.


Cardiotocography and Clinical Risk Factors in Early Term Labor: A Retrospective Cohort Study Using Computerized Analysis With Oxford System.
The role of cardiotocography (CTG) in fetal risk assessment around the beginning of term labor is controversial. We used routinely collected clinical data in a large tertiary hospital to investigate whether infants with "severe compromise" at birth exhibited fetal heart rate abnormalities in their first-hour CTGs and/or other clinical risks, recorded as per routine care. Retrospective data from 27,927 parturitions (single UK tertiary site, 2001-2010) were analyzed. Cases were included if the pregnancy was singleton, ≥36 weeks' gestation, cephalic presentation, and if they had routine intrapartum CTG as per clinical care. Cases with congenital abnormalities, planned cesarean section (CS), or CS for reasons other than "presumed fetal compromise" were excluded. We analyzed first-hour intrapartum CTG recordings, using intrapartum Oxford System (OxSys) computer-based algorithms. To reflect the effect of routine clinical care, the data was stratified into three exclusive groups: infants delivered by CS for "presumed fetal compromise" within 2 h of starting the CTG (Emergency CS, n = 113); between 2 and 5 h of starting the CTG (Urgent CS, n = 203); and the rest of deliveries (Others, n = 27,611). First-hour CTG and clinical characteristics were compared between the groups, sub-divided to those with and without severe compromise: a composite outcome of stillbirth, neonatal death, neonatal seizures, encephalopathy, resuscitation followed by ≥48 h in neonatal intensive care unit. Two-sample t-test, X2 test, and Fisher's exact test were used for analysis. Compared to babies without severe compromise, those with compromise had significantly higher proportion of cases with baseline fetal heart rate ≥150 bpm; non-reactive trace; reduced long-term and short-term variability; decelerative capacity; and no accelerations in the first-hour CTG across all groups. Prolonged decelerations(≥3 min) were also more common. Thick meconium and small for gestational age were consistently more common in compromised infants across all groups. There was more often thick meconium, maternal fever ≥38 C, sentinel events, and other clinical risk factors in the Emergency CS and Urgent CS compared to the Others group. A proportion of infants born with severe compromise had significantly different first-hour CTG features and clinical risk factors.


Evaluation of a treatment protocol in dogs with intraspinal spirocercosis.
To evaluate the efficiency and safety of a doramectin-based treatment protocol in dogs affected by intraspinal spirocercosis (Spirocerca lupi). Client-owned dogs that were admitted to a veterinary hospital during 2021 to 2022 with acute onset of neurological signs and diagnosed with intraspinal spirocercosis. All dogs underwent complete neurological evaluation, CSF analysis, PCR confirmation of CNS S lupi infection, and follow-up evaluation of at least 6 months. Upon diagnosis, dogs were treated with doramectin at a dose of 400 μg/kg, SC, q 24 h for 3 consecutive days, followed by the same dose once a week for 6 weeks. Prednisone was administered at a dose of 1 mg/kg, PO, q 24 h and tapered every 3 days. Antimicrobial clindamycin was administered at a dose of 12.5 mg/kg, PO, q 12 h for 7 days to reduce the risk of secondary spinal cord infection. Short- and long-term outcomes (1 week to 56 months) were recorded. 8 dogs fulfilled the inclusion criteria, 7 of which presented with neurological deficits and 1 with cervical pain. Initiation of treatment was associated with stopping the deterioration in 7 of 8 dogs. Seven dogs improved and 6 recovered ambulation. One dog was euthanized due to lack of improvement. Six of the recovered dogs were still ataxic on the last follow-up examination at 6 to 56 months. No adverse effects of the drug were noted. Frequent administration of doramectin was found to be safe and effective in preventing neurological deterioration in dogs with intraspinal spirocercosis.


Slotted multimode-interference devices.
We describe a novel modification of multimode-interference devices that has broad applicability. The modification involves introducing a slot (or slots), of a specific width and effective refractive index and at a specific position, that runs the longitudinal length of the multimode-interference region. Introducing N slots reduces the self-image length by a factor of N + 1. Varying the effective refractive index or width of the slot(s) creates a switch. The slot modification can be accomplished in a variety of ways, actually increases bandwidth, and has good error tolerances.


Elastic measurements of layered nanocomposite materials by Brillouin spectroscopy
Surface Brillouin spectroscopy makes it possible to measure surface elastic wave propagation parameters at frequencies up to 20 GHz or more. This enables us to measure the elastic properties of surface layers only a small fraction of a micrometre thick. The wavelength and incident angle of the light determine the wavenumber of surface elastic waves (SAW) that scatter the light inelastically, and their frequency can be found by measuring the change in wavelength of the scattered light. By analysing the elastic wave modes present in the surface, the elastic properties can be deduced. We have used this technique to measure the elastic properties of layered nanocomposite materials, which are widely used in the packaging industry. 12 microns polymer films (PET) were coated with glass oxide layers of thickness as little as 25 nm, to give transparent nanocomposite structures with excellent gas barrier properties. In order to understand and model the behaviour of these films under deformation, it is necessary to determine the elastic properties of the different layers. Evaluation of the elastic properties presents several challenges. First, the oxide layers are much thinner than the wavelengths of the surface phonons in surface Brillouin spectroscopy (and hence the depth probed), which usually lie in the range 250-500 nm. The anisotropic elastic properties of the PET substrate must therefore be measured accurately, and this can be done using bulk Brillouin spectroscopy. Second, a thin layer of metal (usually 10-20 nm) must be deposited on the glass surface so that the surface phonons scatter the light effectively. The elastic properties of the glass layer can then be deduced from surface Brillouin spectroscopy measurements, by simulating the surface wave modes of the metal/glass/polymer composite, and adjusting the parameters to give the best fit. In this way it is possible to observe how the properties of the glass vary as a function of thickness, and in turn to understand how to improve systematically the properties under deformation.


African refugee and immigrant health needs: report from a community-based house meeting project.
As in other communities in the United States, information is lacking about the health needs of Africans refugees and immigrants living in Portland, Oregon. In 2008, the African Partnership for Health coalition (APH) was formed to carry out research, advocacy and education to improve the health and well-being of Africans in Oregon. This was APH's initial project. The purposes of this study were to gather data about the perceived health needs and barriers to health care Africans encounter, and lay the foundation for a program of action to guide APH's future work. Community-based participatory research (CBPR) methods were used to collect data on how to improve the health of the African community in the Portland area and define an agenda for future projects. Popular education principles guided the engagement and training of African community members, who conducted nine house meetings with 56 Africans from 14 countries. The results were analyzed by African community members and researchers and prioritized at a community meeting. Three themes emerged: The stressfulness of life in America, the challenges of gaining access to health care, and the pervasive feelings of disrespect and lack of understanding of Africans' health needs, culture, and life experiences by health providers and staff members. Using CBPR methods, we identified and prioritized the needs of the African community. This information provides a framework for future work of the African Partnership for Health and other service and advocacy groups.


Crystal structure of CC3 (TIP30): implications for its role as a tumor suppressor.
CC3 (TIP30) is a protein with pro-apoptotic and anti-metastatic properties. The tumor suppressor effect of CC3 has been suggested to result from inhibition of nuclear transport by binding to importin betas or by regulating transcription through interaction in a complex with co-activator independent of AF-2 function (CIA) and the c-myc gene. Previous biochemical studies indicated that CC3 has protein kinase activity, and a structural similarity to cAMP-dependent protein kinase catalytic subunit was proposed. By contrast, bioinformatics studies suggested a relationship of CC3 to the short chain dehydrogenase reductase family. To clarify details of the CC3 structural family and ligand binding properties, we have determined the crystal structure of CC3 at 1.7-A resolution. CC3 has a short chain dehydrogenase reductase fold and binding specificity for NADPH, yet it is unlikely to be normally enzymatically active because it is monomeric. These structural results, in conjunction with data from earlier mutagenesis work on the nucleotide binding motif, suggest that NADPH binding is important for the biological activity of CC3, including interaction with importins and with the CIA/c-myc system. CC3 provides an example of the adaptation of a metabolic enzyme fold to include a regulatory role, as also seen in the case of the NADH-binding co-repressor CtBP.


Regional variation in smoking among African Americans.
The impact of geographic region and metropolitan residence on smoking prevalence among African Americans has not been adequately examined. This study analyzed 5 years of data from the National Health Interview Survey (1990-1994) on current smoking and regional variation among 16,738 African Americans. Results. Respondents in the West had the lowest unadjusted smoking prevalence rates and Midwest residents had the highest. Current smoking was lower among African Americans living in non-central cities than in central cities even after adjusting for several sociodemographic covariates. Multivariate logistic regression analysis revealed that black women in the South were significantly less likely to be smokers compared with any other gender/region group. These findings suggest the significance of gender and regional factors such as the social history of migration, social stress and racism, exposure to tobacco advertisement, variations in cultural influences, community structures, and coping strategies in under standing African American smoking behavior.