A systematic review of case-control studies of cytokines in blister fluid and skin tissue of patients with Stevens Johnson syndrome and toxic epidermal necrolysis.

Stevens Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are severe cutaneous adverse reactions characterised by keratinocyte apoptosis, necroptosis and epidermal detachment. Several cytokines and cytotoxic proteins have been shown to be elevated in the blood and skin of SJS/TEN sufferers and biologics such as intravenous immune globulin and tumour necrosis factor (TNF)-alpha inhibitors have demonstrated good therapeutic potential. The exact pathogenic model of SJS/TEN however remains elusive. This systematic review aimed to evaluate the case-control studies of cytokines and cytotoxic proteins in the blister fluid and skin of adults with Stevens Johnson syndrome and/or toxic epidermal necrolysis. This review was registered with INPLASY and conducted in accordance with the PRISMA reporting guidelines. Potential bias was assessed using the NIH criteria. Eleven articles describing results from 96 cases and 170 controls were included. Fas, Fas ligand, Interleukin (IL)-8 and B-cell lymphoma (Bcl)-2 were elevated in SJS/TEN blister fluid and skin tissue, compared with healthy controls. IL-2, IL-6, TNF-alpha, tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), interferon-gamma and matrix metalloproteinase-2 were elevated in SJS/TEN blister fluid compared with fluid sampled from lesional controls. Granulysin, IL-33, TGF-beta-1 and IL-13 were elevated in SJS/TEN skin tissue compared with lesional lichen planus tissue, as was IL-13, IFN-gamma, IL-2 and IL-5, when compared with erythema multiforme tissue. A wide array of cytokines and cytotoxic proteins are present at higher concentrations in the blister fluid and skin tissue of SJS/TEN patients compared with healthy and lesional controls. Our findings suggest that these proteins may be pathogenic, as well as possibly markers for diagnosis, disease severity and course. They may also prove to be useful therapeutic targets. More research is needed.

Dead-infant carrying by chimpanzee mothers in the Budongo Forest.

It has been suggested that non-human primates can respond to deceased conspecifics in ways that suggest they experience psychological states not unlike humans, some of which could indicate they exhibit a notion of death. Here, we report long-term demographic data from two East African chimpanzee groups. During a combined 40-year observation period, we recorded 191 births of which 68 died in infancy, mostly within the first year. We documented the post-mortem behaviour of the mothers and describe nine occasions where Budongo chimpanzee mothers carried infants for 1-3 days after their death, usually until the body started to decompose. We also observed three additional cases of extended carrying lasting for more than 2 weeks, one of which was followed by the unusual extended carrying of an object and another which lasted 3 months. In each case, the corpses mummified. In addition, we report four instances of recurring dead-infant carrying by mothers, three of whom carried the corpse for longer during the second instance. We discuss these observations in view of functional hypotheses of dead-infant carrying in primates and the potential proximate mechanisms involved in this behaviour.

Advancing respiratory-cardiovascular physiology with the working heart-brainstem preparation over 25 years.

Twenty-five years ago, a new physiological preparation called the working heart-brainstem preparation (WHBP) was introduced with the claim it would provide a new platform allowing studies not possible before in cardiovascular, neuroendocrine, autonomic and respiratory research. Herein, we review some of the progress made with the WHBP, some advantages and disadvantages along with potential future applications, and provide photographs and technical drawings of all the customised equipment used for the preparation. Using mice or rats, the WHBP is an in situ experimental model that is perfused via an extracorporeal circuit benefitting from unprecedented surgical access, mechanical stability of the brain for whole cell recording and an uncompromised use of pharmacological agents akin to in vitro approaches. The preparation has revealed novel mechanistic insights into, for example, the generation of distinct respiratory rhythms, the neurogenesis of sympathetic activity, coupling between respiration and the heart and circulation, hypothalamic and spinal control mechanisms, and peripheral and central chemoreceptor mechanisms. Insights have been gleaned into diseases such as hypertension, heart failure and sleep apnoea. Findings from the in situ preparation have been ratified in conscious in vivo animals and when tested have translated to humans. We conclude by discussing potential future applications of the WHBP including two-photon imaging of peripheral and central nervous systems and adoption of pharmacogenetic tools that will improve our understanding of physiological mechanisms and reveal novel mechanisms that may guide new treatment strategies for cardiorespiratory diseases.

The importance of local specialists in science: Where are the local researchers in primatology?

Catherine Hobaiter, John Walter Akankwasa, Geresomu Muhumuza, Moreen Uwimbabazi and Inza Koné discuss the importance of local specialists in science.

Evaluation of biosurfactant production potential of Lysinibacillus fusiformis MK559526 isolated from automobile-mechanic-workshop soil.

BACKGROUND: Biosurfactants are amphipathic biological compounds with surface active potential and are produced by many microorganisms. Biosurfactant production by Lysinibacillus fusiformis MK559526 isolated from automobile-mechanic-shop soil was investigated with a view to assessing its potential for production and potential for optimization. MATERIALS AND METHODS: Effects of carbon and nitrogen sources, pH, temperature and incubation periods on biosurfactant production were evaluated with a view to optimizing the processes. Fourier Transform Infra-Red absorption peaks and Gas chromatography mass spectrometry were used to determine the functional groups of the chemical make-up and the chemical profile of the biosurfactant respectively. RESULTS: Lysinibacillus fusiformis surfactant had emulsification index of 65.15 ± 0.35 %, oil displacement of 2.7 ± 0.26 mm, zone of haemolysis of 7.3 ± 0.16 mm and a positive drop collapse test. Optimized culture conditions for biosurfactant production: temperature, 35 ºC; pH, 7.0; starch solution, 40 g/L and urea, 1.5 g/L showed a reduction in surface tension to 28.46 ± 1.11 mN/m and increased emulsification index to 93.80 ± 0.41 %. Maximum biosurfactant production of 2.92 ± 0.04 g/L was obtained after 72 h. The biosurfactant contained peptides and fatty acids. The predominant fatty acid was 9-Octadecenoic acid (80.80%). CONCLUSIONS: The above results showing high emulsification potential and remarkable reduction in the surface tension are good biosurfactant attributes. Consequently, Lysinibacillus fusiformis MK559526 is a good candidate for biosurfactant production.

Effect of online instruction on volunteers who support people with complex communication needs in active recreation.

This study was conducted to teach and evaluate the effectiveness of an online training called The CAPTURE & Share program. The program was taught using online instruction in an asynchronous manner with six volunteers who facilitate recreational participation for individuals with complex communication needs. The online training focused on teaching volunteers how to support opportunities for individuals to capture photos and videos from recreation and then share them across their social networks. The first aim of the study was to teach the volunteers about implementation and use of the 8-step CAPTURE & Share program while the second aim was to evaluate the programs' overall effectiveness. Two multiple baselines designs across three participants each were used to evaluate the volunteers' written responses to probes delivered during baseline, intervention, maintenance, and generalization phases. Additionally, social validation data were collected. Results indicated that volunteers were not only successful in learning the program, but were also highly satisfied with the online training methods. Implications for using online instruction are discussed for speech-language pathologists, caregivers, individuals with complex communication needs, and recreational professionals.

Association between retinopathy and cardiovascular disease in patients with chronic kidney disease (from the Chronic Renal Insufficiency Cohort [CRIC] Study).

Patients with chronic kidney disease experience co-morbid illnesses, including cardiovascular disease (CVD) and retinopathy. The purpose of the present study was to assess the association between retinopathy and self-reported CVD in a subgroup of the participants in the Chronic Renal Insufficiency Cohort study. For this observational, ancillary investigation, 2,605 Chronic Renal Insufficiency Cohort participants were invited to participate in the present study, and nonmydriatic fundus photographs in both eyes were obtained for 1,936 subjects. The photographs were reviewed in a masked fashion at a central photograph reading center. The presence and severity of retinopathy (diabetic, hypertensive, or other) and vessel diameter caliber were assessed using standard protocols by trained graders who were masked to the information about the study participants. A history of self-reported CVD was obtained using a medical history questionnaire. Kidney function measurements and traditional and nontraditional risk factors for CVD were obtained from the Chronic Renal Insufficiency Cohort study. A greater severity of retinopathy was associated with a greater prevalence of any CVD, and this association persisted after adjustment for the traditional risk factors for CVD. The presence of vascular abnormalities usually associated with hypertension was also associated with increased prevalence of CVD. We found a direct relation between CVD prevalence and mean venular caliber. In conclusion, the presence of retinopathy was associated with CVD, suggesting that retinovascular pathology might indicate macrovascular disease, even after adjustment for renal dysfunction and traditional CVD risk factors. This would make the assessment of retinal morphology a valuable tool in CKD studies of CVD outcomes.

A prospective, multicentre, randomized, double-blind, placebo-controlled trial of onabotulinumtoxinA to treat plantarflexor/invertor overactivity after stroke.

OBJECTIVE: To examine the safety and efficacy of onabotulinumtoxinA (Botox) for plantarflexor overactivity following stroke. DESIGN: Double-blind randomized controlled trial, open-label extension phase. SETTING: Neurology rehabilitation facilities. SUBJECTS: Eighty-five subjects with lower limb hypertonia received 200 U (n = 28) or 300 U (n = 28) of onabotulinumtoxinA or saline (n = 29) injection. PRIMARY MEASURES: Plantarflexor Ashworth scores at 12 weeks post injection and adverse events. Secondary measures: self-reported spasm frequency and pain, physician rating of hypertonia severity, gait quality and active dorsiflexion. RESULTS: Differences were not seen between onabotulinumtoxinA groups; hence data were pooled. Incidence of adverse events was not different between groups (P = 0.61). Reduction in hypertonia was not different between groups at 12 weeks (P = 0.53); however for subjects with Ashworth scores of >3 at baseline, 14/31 in the onabotulinumtoxinA group demonstrated a reduction of >1 grade versus 1/17 receiving placebo injection (P = 0.01). Overall, onabotulinumtoxinA-injected subjects demonstrated significantly greater improvement in spasm frequency (22/54 versus 4/29, P = 0.01), pain reduction (8/54 versus 1/29, P = 0.02), active dorsiflexion (8/54 versus 1/29 P = 0.03) and gait quality (17/54 versus 6/29, P = 0.02) than controls. In the open-label phase, a second onabotulinumtoxinA injection was associated with greater hypertonia reduction (P = 0.005) and gait quality (P = 0.002) compared with single injection. CONCLUSIONS: OnabotulinumtoxinA injection for ankle flexor overactivity after stroke was safe and well tolerated but did not alter local spasticity at 12 weeks; it did reduce spasms and improve gait quality. There were no detectable differences between higher and lower doses. A second injection may be associated with greater change.

Mylohyoid discharge of the in situ rat: a probe of pontile respiratory activities in eupnea and gasping.

Our purpose was to characterize respiratory-modulated activity of the mylohyoid nerve. Since its motoneurons are in the trigeminal motor nucleus, mylohyoid discharge could serve as a probe of the role of pontile mechanisms in the generation of respiratory rhythms. Studies were performed in the decerebrate, perfused in situ preparation of the rat. Phrenic discharge was recorded as the index of the respiratory rhythm. In eupnea, the mylohyoid nerve discharged primarily during neural expiration, in the period between phrenic bursts. This expiratory discharge increased greatly in hypoxia and fell in hypercapnia. The hypoxia-induced increase in mylohyoid discharge was due, at least in part, to a direct influence of hypoxia on the brain stem. In ischemia, phrenic discharge increased, and then declined to apnea, which was succeeded by gasping. The mylohyoid nerve discharged tonically during the apneic period, but still declined during each of the phrenic bursts of gasping. This maintenance of a respiratory-modulation of the mylohyoid discharge in gasping supports the concept that a release of medullary mechanisms, rather than a ubiquitous suppression of pontile influences, underlies the neurogenesis of gasping. Results also provide additional support for our conclusion that activity of any single cranial nerve does not provide an accurate index of the type of respiratory rhythm, be it eupnea or gasping.

Noeud vital for breathing in the brainstem: gasping--yes, eupnoea--doubtful.

For the past 200 years, various regions of the brainstem have been proposed as a 'noeud vital' for breathing-a critical region which, when destroyed, results in an irreversible cessation of breathing and death. Complicating this search for a noeud vital is the extensive network of neurons that comprises the brainstem respiratory control system of pons and medulla. Does a cessation of breathing following ablation of any region reflect the removal of a critical set of neurons whose activity generates the respiratory rhythm or does it reflect the interruption of one component of the neuronal circuit, such that this circuit cannot function, at least temporarily? An additional complication is that in contemporary neuroscience, a number of in vitro preparations have been introduced for the study of the generation of the respiratory rhythms. However, how are the rhythms that these preparations generate related to normal breathing? Are these rhythms similar to those of gasping, which is recruited when normal, eupnoeic breathing fails, or are these rhythms unique to the in vitro preparation and not related to any breathing pattern in vivo?

Location and properties of respiratory neurones with putative intrinsic bursting properties in the rat in situ.

Using the in situ arterially perfused preparations of both neonatal and juvenile rats, we provide the first description of the location, morphology and transmitter content of a population of respiratory neurones that retains a bursting behaviour after ionotropic receptor blockade. All burster neurones exhibited an inspiratory discharge during eupnoeic respiration. These neurones were predominantly glutamatergic, and were located within a region of the ventral respiratory column that encompasses the pre-Bötzinger complex and the more caudally located ventral respiratory group. Bursting behaviour was both voltage and persistent sodium current dependent and could be stimulated by sodium cyanide to activate this persistent sodium current. The population of burster neurones may overlap with that previously described in the neonatal slice in vitro. Based upon the present and previous findings, we hypothesize that this burster discharge may be released when the brain is subject to severe hypoxia or ischaemia, and that this burster discharge could underlie gasping.

Discharge of the hypoglossal nerve cannot distinguish eupnea from gasping, as defined by phrenic discharge, in the in situ mouse.

If normal, eupneic breathing fails, gasping is recruited. Serotonin was proposed as essential for gasping, based on findings using an in vitro mouse preparation. This preparation generates rhythmic activities of the hypoglossal nerve that are considered to be akin to both eupnea and gasping. In previous studies, gasping of in situ rat and mouse preparations continued unabated following blockers of receptors for serotonin. However, hypoglossal activity was not recorded in the mouse, and we hypothesized that its discharge during gasping might be dependent on serotonin. In the in situ mouse preparation, hypoglossal discharge had varying and inconsistent patterns during eupnea, discharging concomitant with the phrenic burst, at varying intervals between phrenic bursts, or was silent in some respiratory cycles. In eupnea, phrenic discharge was incrementing, whereas hypoglossal discharge was decrementing in 15 of 20 preparations. During ischemia-induced gasping, peak phrenic height was reached at 205 +/- 17 ms, compared with 282 +/- 27.9 ms after the start of the eupneic burst (P < 0.002). In contrast, rates of rise of hypoglossal discharge in gasping (peak at 233 +/- 25 ms) and eupnea (peak at 199 +/- 19.2 ms) were the same. The uncoupling of hypoglossal from phrenic discharge in eupnea was exacerbated by methysergide, an antagonist of serotonin receptors. These findings demonstrate that hypoglossal discharge alone cannot distinguish eupnea from gasping nor, in eupnea, can hypoglossal activity be used to differentiate neural inspiration from expiration. These findings have significant negative implications for conclusions drawn from the in vitro medullary slice of mouse.

Genesis of gasping is independent of levels of serotonin in the Pet-1 knockout mouse.

Eupnea is normal breathing. If eupnea fails, as in severe hypoxia or ischemia, gasping is recruited. Gasping can serve as a powerful mechanism for autoresuscitation. A failure of autoresuscitation has been proposed as a basis of the sudden infant death syndrome. In an in vitro preparation, endogenous serotonin is reported to be essential for expression of gasping. Using an in situ preparation of the Pet-1 knockout mouse, we evaluated such a critical role for serotonin. In this mouse, the number of serotonergic neurons is reduced by 85-90% compared with animals without this homozygous genetic defect. Despite this reduction in the number of serotonergic neurons, phrenic discharge in eupnea and gasping of Pet-1 knockout mice was not different from that of wild-type mice. Indeed, gasping continued unabated, even after administration of methysergide, a blocker of many types of receptors for serotonin, to Pet-1 knockout mice. We conclude that serotonin is not critical for expression of gasping. The proposal for such a critical role, on the basis of observations in the in vitro slice preparation, may reflect the minimal functional neuronal tissue and neurotransmitters in this preparation, such that the role of any remaining neurotransmitters is magnified. Also, rhythmic activity of the in vitro slice preparation has been characterized as eupnea or gasping solely on the basis of activity of the hypoglossal nerve or massed neuronal activities of the ventrolateral medulla. The accuracy of this method of classification has not been established.

Dynamics of 2-N-tert-butylaminoxyladenosine incorporated into oligodeoxynucleotides.

We synthesized oligodeoxynucleotide (ODN, 3) containing 2-N-tert-butylaminoxyladenosine (1) and studied EPR spectra of 1 and its duplexes with varied sequences. Duplex formation resulted in a broadening of spectrum and a significant decrease in peak-to-peak height and peak height ratio values. The h(+)/ho values in EPR spectra well correlated with stability of duplex which indicated ODN containing 1 has the potential to monitor DNA structure.

Eupnea of in situ rats persists following blockers of in vitro pacemaker burster activities.

Two groups of intrinsically bursting neurons, linked to respiration, have been identified using in vitro medullary slice preparations. One group is dependent upon a calcium-activated nonspecific cationic current that is blocked by flufanemic acid. This group is hypothesized as essential for eupnea, but not gasping. The second group is dependent upon conductance through persistent sodium channels that is blocked by riluzole. This group is proposed to underlie both eupnea and gasping. In the decerebrate in situ preparation of the juvenile rat, flufanemic acid caused an increase in frequency and a decrease in peak level of the phrenic and vagus nerve activities in both eupnea and gasping. Similar changes in eupnea followed the simultaneous blockades by flufanemic acid and riluzole. However, gasping was eliminated. These results do not support the hypothesis that conductances through either persistent sodium channels or calcium-activated nonspecific cationic channels are essential for the neurogenesis of eupnea. However, gasping does depend upon a conductance through persistent sodium channels.

Maintenance of gasping and restoration of eupnea after hypoxia is impaired following blockers of alpha1-adrenergic receptors and serotonin 5-HT2 receptors.

In severe hypoxia or ischemia, normal eupneic breathing fails and is replaced by gasping. Gasping serves as part of a process of autoresuscitation by which eupnea is reestablished. Medullary neurons, having a burster, pacemaker discharge, underlie gasping. Conductance through persistent sodium channels is essential for the burster discharge. This conductance is modulated by norepinephrine, acting on alpha 1-adrenergic receptors, and serotonin, acting on 5-HT2 receptors. We hypothesized that blockers of 5-HT2 receptors and alpha 1-adrenergic receptors would alter autoresuscitation. The in situ perfused preparation of the juvenile rat was used. Integrated phrenic discharge was switched from an incrementing pattern, akin to eupnea, to the decrementing pattern comparable to gasping in hypoxic hypercapnia. With a restoration of hyperoxic normocapnia, rhythmic, incrementing phrenic discharge returned within 10 s in most preparations. Following addition of blockers of alpha 1-adrenergic receptors (WB-4101, 0.0625-0.500 microM) and/or blockers of 5-HT2 (ketanserin, 1.25-10 microM) or multiple 5-HT receptors (methysergide, 3.0-10 microM) to the perfusate, incrementing phrenic discharge continued. Fictive gasping was still induced, although it ceased after significantly fewer decrementing bursts than in preparations than received no blockers. Moreover, the time for recovery of rhythmic activity was significantly prolonged. This prolongation was in excess of 100 s in all preparations that received both WB-4101 (above 0.125 microM) and methysergide (above 2.5 microM). We conclude that activation of adrenergic and 5-HT2 receptors is important to sustain gasping and to restore rhythmic respiratory activity after hypoxia-induced depression.