171 Indeed, treatment of postsymptomatic, 90-day-old SOD1G93A mice (a model of ALS) with WIN 55,212-2, significantly delayed disease progression. Furthermore, genetic ablation of the FAAH enzyme, which results in raised levels of the endocannabinoid anandamide, prevented the appearance

of disease signs in these mice. Surprisingly, elevation of cannabinoid levels with either WIN 55,212-2 or FAAH ablation had no effect on life span. Ablation of the CB1 receptor, in contrast, had no effect on disease onset in these mice, but significantly extended life span. Together these results show that cannabinoids have significant Inhibitors,research,lifescience,medical neuroprotective effects in this model of ALS, and suggest that these beneficial effects may be mediated by nonCB1 receptor mechanisms.172 THC was also found to delay the progression of disease.173,174 Treatment

with AM1241, a CB2-selective Inhibitors,research,lifescience,medical agonist, was effective at slowing signs of disease progression, when administered after onset of signs in an ALS mouse model. Administration at the onset of tremors delayed motor impairment in treated mice when compared with vehicle controls175; moreover, AM-1241 prolonged survival in these mice.176 In a survey among ALS patients, cannabis was reported to be moderately effective in reducing symptoms of appetite loss, depression, pain, spasticity, and drooling.177 Cannabinoids were also proposed to have a role in Inhibitors,research,lifescience,medical Inhibitors,research,lifescience,medical the treatment of Alzheimer’s disease (AD). THC competitively inhibits acetylcholinesterase (AChE) and prevents AChE-induced amyloid beta-peptide (Abeta) aggregation, the key pathological marker of AD.178 THC treatment also decreased severity of disturbed behavior, and this effect persisted during the placebo period in patients who had received THC.179 Compared with baseline, THC led to a reduction in nocturnal motor activity These findings were corroborated by improvements in the Neuropsychiatrie Inventory total score, as well as in subscores for agitation, aberrant motor, and nighttime behaviors; no side effects were observed.180 Studies on cannabinoid anticonvulsant

activity Inhibitors,research,lifescience,medical began in 1975, when CBD, and four CBD derivatives, (CBD-aldehyde-diacetate, 6-oxo-CBD-diacetate, 6-hydroxy-CBD-tri-acetate and 9-hydroxy-CBD-triacetate) were shown to protect Selleckchem MK 8776 against maximal electroshock convulsions in mice, to potentiate pentobarbital for sleeping-time and to reduce spontaneous motor activity.181 Later additional CBD analogs were shown to be active182-184 CBD was found to be an effective anticonvulsant with specificity more comparable to drugs clinically effective in major, but not in minor seizures. Furthermore, it appears that CBD enhances the anticonvulsant effects of drugs in major seizures and reduces their effects in minor seizures.185,186 Hence, CBD was suggested as a drug for the treatment of children with pharmacoresistant epilepsy.

165 Other fields where chronopharmacology is of demonstrated relevance are those of asthma and cardiovascular disorders.166,167 In the field of psychotropic agents or illegal drugs, aside from research on laboratory animals, and research on alcohol,

little is known on circadian changes in pharmacodynamics. In particular, the extent of changes in the concentration/response curve over the course of the day has not been well evaluated in humans. The pharmacology of alcohol shows circadian changes.168 Several studies during the last 50 years have shown that alcohol is absorbed more rapidly, with higher blood levels when taken during the morning, but that it is also eliminated faster.169 #SCR7 mw keyword# Each addicted person has his or her own daily schedule to start the consumption of alcohol 170 Alcohol given during one day to nonaddicted volunteers does not influence the circadian rhythm, nor the concentration of Cortisol, but it increases that of testosterone171 and suppresses the nocturnal Inhibitors,research,lifescience,medical increase of TSH, and decreases the mean concentration of the later hormone.168 Clinical consequences of chronopharmacology There is little information as to whether giving a psychotropic medication once a day Inhibitors,research,lifescience,medical or as a divided dose shows any benefit in terms of efficacy or side effects. It might be with substances that themselves influence the

physiology of biological clocks that chronopharmacology will find its major application. Inhibitors,research,lifescience,medical Benzodiazepines and other sedatives can influence the phase position of circadian rhythms, and lithium, as well as a few antidepressants, might modify the functioning of the SCN.172 However, lithium and most antidepressants and many benzodiazepines have a half -life of elimination that is longer that 12 hours; thus their effects persist throughout the nicthemere. Melatonin has a short half-life and its timing of administration is quite

relevant for its efficacy in SAD treatment.77 The following general clinical and general rules prevail: a stimulating medication should be given Inhibitors,research,lifescience,medical in the morning and not late in the afternoon or in the evening, a medication that is sedative should be given at the time of sleep, and a medication that induces nausea might be better tolerated when given with a meal. Conclusion The time structure of biology is as essential as is its spatial structure, yet the relevance of chronobiology for pathophysiology remains underestimated in internal medicine, neurology, or psychiatry This might be because measuring the rhythms Dipeptidyl peptidase of most biological variables over the long term is complex, because feedback loops and regulations maintain vital phenomena within apparently stable ranges labeled as norms, and finally because our knowledge about the temporal structure of biology remains incomplete. Indeed, the relation between endogenous biological oscillators is far less well established than are mechanisms in other domains of biology, for example endocrine feedback mechanisms.

The randomisation was stratified for lung function (FEV1 > or ≤ 40% predicted), 6-minute walk distance (> or ≤ 50% predicted) (Troosters et al 1999), and the main limiting symptom in the initial endurance cycle test (ie, dyspnoea, leg fatigue, or a combination of both symptoms). Participants

undertook three sessions per week of supervised group training in their allocated exercise mode for eight weeks. Each participant maintained his/her medication regimen during the intervention period. ALK activation An assessor, blinded to group allocation, performed the outcome measures at the end of the intervention period. Participants were included if they had COPD stage I to IV (Global Initiative for COPD classification (GOLD) 2008). Participants were excluded if any of the following criteria applied: acute exacerbation of COPD within the last 4 weeks, significant co-morbidity including malignancy, symptomatic Dasatinib mw cardiovascular disease, or other systemic or musculoskeletal disease that could hinder the exercise training. As well, participants were excluded if they had a body mass index (weight in kg/height in m2) ≥ 35 kg/m2, required supplemental

oxygen during exercise training, or used a walking aid. The study participants underwent pulmonary function testing including spirometry, lung volumes, and carbon monoxide transfer factor, and the six-minute walk test. Pulmonary function tests were performed according to the recommended standards (ATS/ERS Task Force 2005a, 2005b, 2005c) and results were compared with predicted normal values (Quanjer et al 1993). In the walk group, participants trained on a 26-m circular indoor track with the

initial training speed set at 75% of the participant’s peak walking speed, achieved in the incremental shuttle walk test (Hernandez et al 2000). Each participant was given a goal of completing a set number of laps in each five-minute period. All participants used a lap counter to monitor the number of laps walked during the prescribed duration. In the cycle group, participants were trained on an upright cycle ergometer with the initial training intensity set at 60% of the peak work capacity achieved in the incremental cycle test (Maltais et al 1997). The initial training intensities were chosen based on previous studies that reported that these training intensities were tolerated by participants Linifanib (ABT-869) with COPD (Hernandez et al 2000, Maltais et al 1997). The training intensities for both groups were progressed as symptoms permitted so that the dose of training was maximised, with participants in the walk group walking at a faster pace and those in the cycle group cycling at a higher work rate. In the walk group, if walking speed Modulators became limited by stride length, further progress of training intensity was achieved by adding weights in 2 kg increments to a backpack. The duration of training for both groups was 30 minutes in the first week and increased by five minutes every two weeks to a maximum of 45 minutes by Week 6.

Discussion One important step in the management of a patient with a history of seizure(s) is correct diagnosis of the illness. An important and sometimes problematic issue in the management of seizure is the differentiation of epileptic seizures from non-epileptic ones.1 Basically, epilepsy is a clinical diagnosis. Unless one happens to observe a seizure while recording

the EEG, which is a rare event in general clinical practice, the diagnosis relies on the judgment of a physician Inhibitors,research,lifescience,medical or other health care providers. This judgment ultimately rests on the history provided by the patient or others, which may be misleading and results in maltreatment, similar to the scenario, which happened in the case of the present patient. The history of seeing flashing light was interpreted as epileptic aura. The triggering factors such as pain and inspecting blood were overlooked,

and motor phenomena, loss of consciousness and post-ictal confusion led to the misdiagnosis Inhibitors,research,lifescience,medical of the illness. She had been diagnosed Inhibitors,research,lifescience,medical as having epilepsy for 16 years, however, normal EEGs and AED-unresponsiveness had been ignored. As others have mentioned in similar case reports, no one, even physicians who witness the events, is immune to making a false clinical diagnosis of epilepsy.3 Syncope is a condition that is most commonly misdiagnosed as epilepsy.4 The precise mechanism Inhibitors,research,lifescience,medical of vasovagal syncope is not fully understood. It is proposed that the see more failure of sympathetic efferent vasoconstrictor traffic occurs episodically and in response to a triggering agent such as fear, anger or pain.5 There is evidence for the involvement of both neural and chemical pathways.6 Presyncopal symptoms include lightheadedness, generalized muscle weakness, tinnitus Inhibitors,research,lifescience,medical and visual blurring, but up to a third of patients will have little or no prodrome. Often, unlike epileptic seizures, tonic-clonic convulsion, other motor phenomena, and

post-ictal confusion are uncommon features, but may occur.7 In the present case, injection (psychogenically)-induced asystole led to hypoxia, which in turn caused a typical tonic-clonic convulsion. Conclusion Idoxuridine The possibility of vasovagal syncope should always be taken into consideration when evaluating patients with medically-refractory or unusual pattern of seizures. In such circumstances, simultaneous video-EEG/ECG monitoring may help achieve the correct diagnosis, particularly if the physician applies a triggering agent(s) after obtaining the patient’s consent. Conflict of Interest: None declared
Background: Brillantaisia lamium is an erect branched herb, which grows to a height of 1.50 m in moist tropical areas, both in full sun and partial shade. In , the aerial part of this plant is used in the treatment of various microbial infections such as skin diseases and infections of urinary tract.

As far as we know, the only data about gene therapy on man concern one case of Mucopolysaccharidosis II (18), three patients affected by Mucopolysaccharidosis I (19) and some patients suffering from Gaucher disease (20). Considering the researches carried out so far, we think that many problems have still to be PI3K inhibitor solved before proving unequivocally effectiveness and safety of this treatment in man: a patient’s optimal age for undergoing gene therapy, the possible development of immunologic reactions,

the capability to modulate both levels and duration of enzyme activity, the need of finding a specific ablative regimen for BMT Inhibitors,research,lifescience,medical approach. Conclusions As above reported, therapeutic approaches toward finding treatment options fit to face the underlying causes are many: so far positive results, unanimously Inhibitors,research,lifescience,medical accepted by the international scientific community, have been obtained only for few lysosomal disorders. However, LSDs, though quite rare diseases, are getting more and more investments from private enterprises interested in orphan drug production. Inhibitors,research,lifescience,medical The above mentioned fact lets us hope that, in a near future, the natural development of more and more diseases will be influenced and thus modified.
McArdle disease or Glycogenosis type V is an autosomal recessive metabolic disorder caused by a deficiency of the muscle isoform of glycogen phosphorylase (myophosphorylase,

PYGM), the specific skeletal muscle enzyme that initiates glycogen breakdown. Since the first clinical description by Brian McArdle in 1951, several patients have been identified worldwide and significant advances have been made in the study of molecular genetics of the disease. Inhibitors,research,lifescience,medical Molecular heterogeneity has been demonstrated by the identification to date of more than 65 mutations in the PYGM gene. In this paper, we will present an update on the mutations Inhibitors,research,lifescience,medical reported to date in the PYGM gene. Keywords: McArdle disease, Glycogenosis type V, PYGM gene Clinical data McArdle disease or Glycogen Storage Disease

type V (GSD-V; MIM # 232600) is the most common muscle glycogenosis caused by the deficiency muscle glycogen phosphorylase (myophosphorylase, EC 2.4.1.1) activity. GSD-V is clinically characterized by exercise intolerance with premature fatigue, cramps, myalgia, moderate to high levels of serum creatine kinase (CK) at rest, and by episodic myoglobinuria (1). All McArdle patients experience Thymidine kinase the so-called ‘second-wind’ phenomenon, characterized by the ability to resume exercise with less difficulty, after following a short period of rest at the first appearance of fatigue (2). A subset of patients develops fixed weakness and wasting with aging, indicating phenotypic variability). The diagnosis is based on the clinical phenotype and DNA analysis is suggested as the first choice in the diagnostic approach.

8-11 These are summarized in Table

I, together with some of the other findings that are of current interest, but that have not been substantiated to the same extent.12,13 Because the focus of this paper is on postmortem studies, the imaging literature is not discussed in detail. However, it is noteworthy that it is the incontrovertible in vivo imaging evidence that there is a pathology of schizophrenia to be found which has driven the ongoing neuropathological studies. Table I. Macroscopic brain changes in schizophrenia. The imaging data have also allowed other important conclusions to be drawn, which inform and bolster postmortem research (Table II). 14-18 In particular, since the structural Inhibitors,research,lifescience,medical changes are present at the time of disease onset and are, by and large, not progressive thereafter (although there may well be exceptions to this rule17,19), it is reasonable to assume that the Inhibitors,research,lifescience,medical corresponding histological abnormalities share this property, even though it is in practice impossible to prove this in postmortem studies. Inhibitors,research,lifescience,medical Instead, the latter can now focus on the microscopic and molecular aspects of the pathology, which remain out of reach of any imaging modality. Table II. Characteristics of structural imaging findings in schizophrenia. Histological and molecular pathology of schizophrenia Contemporary histological studies have addressed two main

areas: first, to clarify the frequency and nature of neurodegenerative abnormalities in schizophrenia; and, second, to investigate the cellular organization (cytoarchitccture) of the brain. A summary of the most established and the most often cited findings Inhibitors,research,lifescience,medical is given in Table III. Table III. Histological findings in schizophrenia. Gliosis

and neurodegeneration The two most robust and important findings concerning the neuropathology of schizophrenia are Inhibitors,research,lifescience,medical both negative: there is no excess of gliosis, or of Alzheimer’s disease or other neurodegenerative pathology. The issue of gliosis (reactive astrocytosis) has been extensively investigated since a report, that gliosis was common in schizophrenia, especially in the diencephalon around the third ventricle.20 As gliosis is a sign of past inflammation, this invoked scenarios for schizophrenia involving infective, Proteasome inhibitor ischemic, autoimmune, or neurodegenerative processes. However, over a dozen subsequent investigations CYTH4 using more quantitative methods have not replicated this observation, and the consensus is now that gliosis is not a feature of schizophrenia.21 The issue is complicated by the excess of nonspecific and focal abnormalities, including gliosis, seen in brain scries of schizophrenia; however, this is likely to be an epiphenomenon not an intrinsic finding21,22 and, importantly, decreased brain size is still seen after omission of all such brains.

The monoaminooxidase inhibitor phenelzine was shown to be as effective as clomipramine in a double-blind trial in OCD patients,80 while in another one it was no better than placebo.81 A double-blind study with St John’s wort (hypericum perforatum) failed to support efficacy for OCD.82 Trazodone, a 5-HT2 receptor antagonist and SRI, had shown Inhibitors,research,lifescience,medical symptomatic improvements in case series in clomipramine-resistant OCD patients83 and in augmentation of SSRIs.84 However, a double-blind study indicated that trazodone in monotherapy lacks substantial antiobsessive effects.85 For selective serotonin-norepinephrine reuptake inhibitors venlaf

axine and duloxetine, reliable placebo-controlled trials are still absent. In a double-blind comparison of venlafaxine and paroxetine in primary OCD patients no significant differences with regard to response or responder rates were shown.86 In a single -blind study, venlafaxine was as efficacious as clomipramine in the acute treatment of OCD.87 In an open retrospective investigation in treatment-resistant OCD beneficial Inhibitors,research,lifescience,medical effects Inhibitors,research,lifescience,medical of venlafaxine were demonstrated.88 According to case series and reports switching from SSRI to duloxetine in treatment-resistant OCD patients may be helpful.89,90

For the selective noradrenaline reuptake inhibitor reboxetine, successful augmentation of citalopram was reported in a single case.91 For augmentation of SSRIs with pindolol, a 5-HT1A and (3-adrenergic antagonist, a double-blind placebo-controlled trial found significant improvement of OCD symptoms in treatment resistant patients,92 while an open trial only showed such effects after supplemental addition of tryptophan.93 After double-blind primary addition of pindolol versus placebo to fluvoxamine, Inhibitors,research,lifescience,medical the latency of antiobsessional response to the SSRI was not shortened.94 A double -blind study of adjuvant buspirone, a 5-HT1A partial agonist, in OCD patients,

who had shown to some extent an effect of clomipramine, did not yield significant further clinical improvement.95 For lithium two double-blind augmentation studies have been published that Inhibitors,research,lifescience,medical do not support its usefulness in OCD. In fluvoxamine -refractory patients, a small though statistically significant reduction of OCD symptoms was reported, but the authors Phosphoprotein phosphatase doubted the clinical meaningfulness of these findings.96 A crossover study with adjuvant lithium or thyroid hormone in clomipraminetreated patients showed no significant change of OCD symptoms after either treatment.97 Benzodiazepine and opioid receptor ligands have been tested in OCD. A double-blind combination study of clonazepam with sertraline did not reveal significant effects during 12 weeks of treatment.98 While in a double-blind crossover study clonazepam in monotherapy produced a significant www.selleckchem.com/products/ulixertinib-bvd-523-vrt752271.html decrement in OCD symptoms during the first 3 weeks of treatment,99 it was found to be without effect in a 10-week double-blind placebo-controlled trial.

Indeed, it remains surprising that amitriptyline is still commonly prescribed in the USA, apparently in preference to nortriptyline or desipramine, and to trazodone and several newer alternatives. Nortriptyline has been the most frequently directly studied TCA in elderly patients, involving 300 or more

patients in 22 clinical trials.2,4 It is the only antidepressant to have been directly and extensively studied in very elderly patients (>80 year olds).2 Results with nortriptyline suggest that the range of plasma concentrations needed for a therapeutic this website effect is the same in both younger and older patients. However, despite treating patients at plasma levels within a presumed Inhibitors,research,lifescience,medical therapeutic “window” (between 50 and 150 ng/mL), significant residual depressive symptoms have been noted in one half of patients in the clinical trials, and specific dosage recommendations cannot be derived from these studies.5 Clinical practice suggests that effective daily doses in the elderly range from 50 mg to 100

Inhibitors,research,lifescience,medical mg, but this should be taken as a guide at best. There is considerable evidence that clinical response to antidepressant drug therapy depends not only on adequate dose and – in the case of TCAs – blood levels of medication, but adequate length of treatment as well. There is a general Inhibitors,research,lifescience,medical consensus that significant response often occurs later in elderly patients than in younger patients, often after 6 to 12 weeks of therapy. Medication compliance with TCAs by elderly patients is especially important and difficult to achieve. It has been estimated that 70% of patients fail to take 25% to 50% of their medication.6 Lack of adherence to instructions results in wide fluctuations in plasma levels, which has been shown to be predictive of poor Inhibitors,research,lifescience,medical Inhibitors,research,lifescience,medical outcome. Thus, the measurement of plasma blood levels in elderly patients is even more important than in younger patients, both to verify compliance and to confirm that therapeutic drug concentrations have been reached while remaining below toxic levels. Antidepressant

treatment in the 1990s Many treatment recommendations emanate from the 1991 NIH Consensus Development Conference7,8 and from the 1993 AHCPR guidelines.9 At that time, the SSRI fluoxetine had been available for only a few years, and sertraline and paroxetine had not yet been released. Many clinicians favored these medications ADAMTS5 because of the decreased likelihood of anticholinergic and cardiovascular side effects. Two other SSRIs have been introduced in the USA since then, fluvoxamine in 1996, and citalopram, at the end of 1998. (Fluvoxamine is indicated only for obsessive compulsive disorders in the US, although it is indicated for depression in other parts of the world.) In addition, three non-SSRIs, all with complex neurotransmitter actions, have recently been marketed, nefazodone and venlafaxine, as well as a noradrenergic medication, mirtazapine.

7Å (MSE = 0.013), that is, a total polypeptide/enzyme thickness of 105 ± 3.7Å. Figure 2 The dry nanofilm thickness of 40 ± 3.4Å nm increased considerably when bovine trypsin adsorbed to the film surface, whereas it remained less affected when exposed to the V8 enzyme, at ambient temperature. The bacterial protease V8 behaved differently. The thickness of the nanofilm did not change much on exposure to the solution of V8 enzyme, as can also be Inhibitors,research,lifescience,medical seen from Figure 2. A four-layer model was used for the V8 enzyme’s adsorption to the film, since

the layer on top of the polypeptide film was very thin. The best fit received a polypeptide/enzyme thickness of 42 ± 2.9Å. The immediate interpretation of this result is that the V8 enzyme did not interact with the polypeptide film, as both enzyme and the polypeptide surfaces were negatively charged. www.selleckchem.com/products/azd6738.html However, Craig et al. showed that this enzyme catalyzed degradation

of the LbL film provided that it was terminated by the anionic PLGA. The reason for this is that the V8 peptidase is known to be reactive in catalyzing Inhibitors,research,lifescience,medical cleavage of Glu-X bonds, Inhibitors,research,lifescience,medical that is, peptide bonds involving a glutamate residue [24, 25]. The LbL film studied in this work had PLGA as the terminating layer. However, the temperatures differed. Whereas the QCM-D measurements were performed at 32°C, which was intended to mimic the temperature of a typical wound, the ellipsometry experiments were performed at ambient temperature. Thus, the temperature is vital and no or little enzymatic degradation occurred at room temperature. This is a practically important piece of information because it indicates Inhibitors,research,lifescience,medical that the wound dressing with the antimicrobial agents covered by the polypeptide lid remains intact until it is contacted by the exudate from a chronic wound at the approximate skin temperature of 32°C. However, the V8 protease may not be entirely inactive also at ambient temperature. The surface Inhibitors,research,lifescience,medical that has been exposed to the V8 peptidase solution seems to be slightly rougher immediately after the treatment (±6Å) than after

one or two days, when all measuring points ended up at the same value (±1Å). This induced roughness of the surface may indicate enzymatic cleavage of the top layer of the film, that is, predominantly of PLGA. 4. Conclusion The (PLL/PLGA)3 nanofilm was measured with ellipsometry to study the thickness in its dry state. When ADAMTS5 comparing with the film’s wet and dry thicknesses, it is clear that about 60% of the wet film consists of water. This result is in accordance with previously reported values from similar systems despite the fact that in the present investigation the polypeptides were adsorbed directly to a tailored gold surface imitating nonwoven and not to a surface treated with a primer such as PEI, which is the normal procedure. This indicates that the character of the film without primer is similar to that with primer.

The risks of mortality and re-hospitalisation are difficult to Wortmannin predict with precision in the population of people with heart failure. Most tests aimed at determining factors that could be used as predictors of morbidity and mortality in this group of patients are complicated and expensive, which prevent them from being cost effective. A marked reduction in the capacity to undertake

physical activity is one of the principal symptoms of heart failure. Therefore, potential associations have been investigated between various methods of assessing physical exercise capacity and prognosis (Sarullo et al 2010, Poggio et al 2010). Many predictor variables from formal cardiopulmonary exercise testing have been proposed, including peak oxygen consumption as a percentage of the predicted value, the chronotropic index, and ventilatory efficiency (Poggio et al 2010). When multiple predictors are available, conflicting predictions can make interpretation difficult (Poggi et al 2010). The 6-minute walk test is a simple and inexpensive method of indirectly assessing physical capacity that is widely available and commonly used (Bellet et al 2011, Rostagno et al 2008,

Faggiano et al 2004). Most previous studies have What is already known on this topic: selleck chemical The 6-minute walk test is a simple and inexpensive method of indirectly assessing exercise tolerance. The distance covered by hospitalised patients during the test is predictive of the 1-year risk of cardiovascular death. What this study adds: Among men with chronic heart failure, the 1- and 3-year mortality risk are greater among those who cover less than 468 m on the 6-minute walk test. The specific research questions for this study were: 1. Are there relationships

between the distance covered during the 6-minute walk test and the clinical characteristics of men with stable heart failure? This was a prospective, longitudinal, Libraries observational study in which the predictive ability of the 6-minute why walk test distance was assessed in men with stable heart failure. Participants were followed up for a minimum of three years. The clinical outcomes assessed were mortality and hospitalisation for cardiovascular reasons. Participants were recruited from the Heart Failure Outpatient Clinic of the Center for Heart Disease in Wroclaw, Poland. Male clinic attendees with stable systolic heart failure were approached consecutively and informed about what participation in the study would entail. Those who expressed interest in participation underwent a cardiac evaluation and this was used to assess whether they met the eligibility criteria.