Mammalian reverse genetics without crossing reveals Nr3a as a short-sleeper gene

The identification of molecular networks at the system level in mammals is accelerated by next-generation mammalian genetics without crossing, which requires both the efficient production of whole-body biallelic knockout (KO) mice in a single generation and high-performance phenotype analyses. Here, we show that the triple targeting of a single gene using the CRISPR/Cas9 system achieves almost perfect KO efficiency (96%–100%). In addition, we developed a respiration-based fully automated noninvasive sleep phenotyping system, the Snappy Sleep Stager (SSS), for high-performance (95.3% accuracy) sleep/wake staging. Using the triple-target CRISPR and SSS in tandem, we reliably obtained sleep/wake phenotypes, even in double-KO mice. By using this system to comprehensively analyze all of the N-methyl-D-aspartate (NMDA) receptor family members, we found Nr3a as a short-sleeper gene, which is verified by an independent set of triple-target CRISPR. These results demonstrate the application of mammalian reverse genetics without crossing to organism-level systems biology in sleep research.

Mass transfer effects on the generalised vortex flow over a stationary surface with or without magnetic field

The effect of injection and suction on the generalised vortex flow of a steady laminar incompressible fluid over a stationary infinite disc with or without magnetic field under boundary-layer approximations has been studied. The coupled nonlinear ordinary differential equations governing the self-similar flow have been numerically solved using the finite-difference scheme. The results indicate that the injection produces a deeper inflow layer and de-stabilises the motion while suction or magnetic field suppresses the inflow layer and produces stability. The effect of decreasingn, the parameter characterising the nature of vortex flow, is similar to that of increasing the injection rate.

Self-reported bruxism : Associated factors among media personnel with or without irregular shift work

Tutkimuksen kohderyhmänä oli mediatyöntekijöitä, joiden toimenkuva on viime vuosina muuttunut yhä kuormittavammaksi epäsäännöllisen vuorotyön sekä jatkuvien teknisten, organisatoristen ja taloudellisten tekijöiden ristipaineessa. Väitöskirjatutkimus on osa laajempaa tutkimushanketta, joka suunniteltiin selvittämään epäsäännöllisen vuorotyön mahdollisia haittoja. Tutkimusta tukivat taloudellisesti Työsuojelurahasto ja Suomen Hammaslääkäriseura Apollonia sekä resurssipanostuksin Hammaslääketieteen laitos (HY), Työterveyslaitos ja Yleisradio Oy. Bruksismi on tahdosta riippumatonta hampaiden narskuttelua tai yhteenpuristamista. Hampaiden narskuttelu on rytmistä jaksoittain toistuvaa puremalihasten toimintaa, joka esiintyy nukkuessa -tavallisimmin kevyen unen ja havahtumisjaksojen yhteydessä. Valveilla ollessa bruksismi on terveillä ihmisillä lähinnä hampaiden yhteenpuristamista. Yleisen käsityksen mukaan toistuvaa unibruksismia esiintyy noin 10 %:lla ja valveilla tapahtuvaa hampaiden yhteenpuristamista noin 20 %:lla. Aiemmin bruksismi kuului kansainvälisen unihäiriöluokituksen (ICSD 1997) mukaan unen erityishäiriöihin, mutta tuorein luokitus (ICSD 2005) listaa sen unen liikehäiriöihin. Väitöstutkimuksen yleisenä tavoitteena oli kartoittaa koetun bruksismin ja uni- valvehäiriöiden yhteyttä. Tutkimus oli poikittainen vertailututkimus epäsäännöllistä vuorotyötä ja säännöllisiä päivävuoroja tekevien välillä. Mielenkiinto kohdistui myös bruksismin ja kasvojen alueen kivun mahdolliseen yhteyteen. Lisäksi tutkimuksessa selvitettiin joidenkin tunnetusti unen laatua huonontavien psykososiaalisten, neurologisten ja fysiologisten tekijöiden yhteyttä koettuun bruksismiin. Tutkimuksen kohderyhmän muodosti 750 Yleisradion epäsäännöllistä vuorotyötä tekevää työntekijää. Vertailuryhmänä käytettiin samansuuruista satunnaistetusti valittua kaltaistettua Yleisradion työntekijäjoukkoa, joka tekee samankaltaista työtä, mutta säännöllisenä päivätyönä. Kohderyhmälle lähetettiin kyselylomakkeet, jotka kartoittivat koetun bruksismin lisäksi mm. tutkittavien taustatiedot, yleisen terveydentilan, yleisiä koettuja stressioireita ja tuntemuksia, kipuoireita, sekä unen laatua. Lisäksi esitettiin jaksamista ja työympäristöä koskevia kysymyksiä. Kyselyyn vastasi kaikkiaan 874 henkilöä. Kokonaisvastausprosentti oli 58,3 % (53,7 % miehiä). Epäsäännöllistä vuorotyötä tekevien vastausprosentti oli 82,3 % ja säännöllistä päivätyötä tekevien ryhmässä 34,3 %. Työtehtävät sisälsivät ohjelmien toimitus- ja tuottamistyötä, teknistä tuotanto- ja tukityötä, sekä esimies- ja hallintotyötä. Miesten keski-ikä vuorotyöryhmässä oli 45,0 (± 10,6) vuotta ja naisten keski-ikä 42,6 (± 10,7) vuotta, vastaavat luvut päivätyötä tekeville olivat 47,4 (± 9,7) ja 45,5 (± 10,1) vuotta. Vuorotyötä tekevistä oli miehiä 56,6 %, päivätyöryhmässä miehien osuus oli 46,7 %. Usein koettua bruksismia havaittiin koko tutkimusjoukossa 10,6 %:lla. Bruksismin esiintyvyydessä ei ollut merkitsevää eroa epäsäännöllistä vuorotyötä ja päivätyötä tekevien välillä. Kun bruksismia ja stressiä arvioitiin suhteessa tyytyväisyyteen nykyiseen työaikamuotoon, molemmat olivat merkitsevästi vallitsevimpia niillä, jotka halusivat vaihtaa nykyistä työaikamuotoaan. Epäsäännöllistä vuorotyötä tekevät lisäksi ilmoittivat kokevansa enemmän stressiä kuin päivätyötä tekevät sekä olivat tyytymättömämpiä työaikamuotoonsa. Tutkittavista henkilöistä katkonaista unta esiintyi 43,6 %:lla sekä 36,2 % koki unensa virkistämättömäksi. Kasvokipua esiintyi 19,6 %:lla. Usein toistuva bruksaus sekä tyytymättömyys työaikamuotoon olivat erittäin merkitsevästi yhteydessä unihäiriöiden sekä riittämättömän unen oireiden kanssa. Bruksismi ja katkonainen uni osoittautuivat myös kasvokivun taustatekijöiksi. Tutkimus osoitti, että koetulla bruksismilla oli merkitsevä yhteys unihäiriöihin, kasvokipuun, koettuun stressiin ja ahdistuneisuuteen, nuorempaan ikään, runsaampiin hammaslääkäri- ja lääkärikäynteihin sekä siihen että oli tyytymätön työaikamuotoonsa (itse työaikamuoto ei ollut merkitsevä tekijä). Tutkimuksen yhtenä johtopäätöksenä todettiin, että koettu bruksismi voi terveillä työikäisillä henkilöillä olla osa stressaavaa tilannetta ja siihen liittyvää käyttäytymistä. Tämän tiedostaminen terveydenhuollossa voisi olla hyödyllistä.

Drug Analysis without Primary Reference Standards : Application of LC-TOFMS and LC-CLND to Biofluids and Seized Material

Drug Analysis without Primary Reference Standards: Application of LC-TOFMS and LC-CLND to Biofluids and Seized Material Primary reference standards for new drugs, metabolites, designer drugs or rare substances may not be obtainable within a reasonable period of time or their availability may also be hindered by extensive administrative requirements. Standards are usually costly and may have a limited shelf life. Finally, many compounds are not available commercially and sometimes not at all. A new approach within forensic and clinical drug analysis involves substance identification based on accurate mass measurement by liquid chromatography coupled with time-of-flight mass spectrometry (LC-TOFMS) and quantification by LC coupled with chemiluminescence nitrogen detection (LC-CLND) possessing equimolar response to nitrogen. Formula-based identification relies on the fact that the accurate mass of an ion from a chemical compound corresponds to the elemental composition of that compound. Single-calibrant nitrogen based quantification is feasible with a nitrogen-specific detector since approximately 90% of drugs contain nitrogen. A method was developed for toxicological drug screening in 1 ml urine samples by LC-TOFMS. A large target database of exact monoisotopic masses was constructed, representing the elemental formulae of reference drugs and their metabolites. Identification was based on matching the sample component s measured parameters with those in the database, including accurate mass and retention time, if available. In addition, an algorithm for isotopic pattern match (SigmaFit) was applied. Differences in ion abundance in urine extracts did not affect the mass accuracy or the SigmaFit values. For routine screening practice, a mass tolerance of 10 ppm and a SigmaFit tolerance of 0.03 were established. Seized street drug samples were analysed instantly by LC-TOFMS and LC-CLND, using a dilute and shoot approach. In the quantitative analysis of amphetamine, heroin and cocaine findings, the mean relative difference between the results of LC-CLND and the reference methods was only 11%. In blood specimens, liquid-liquid extraction recoveries for basic lipophilic drugs were first established and the validity of the generic extraction recovery-corrected single-calibrant LC-CLND was then verified with proficiency test samples. The mean accuracy was 24% and 17% for plasma and whole blood samples, respectively, all results falling within the confidence range of the reference concentrations. Further, metabolic ratios for the opioid drug tramadol were determined in a pharmacogenetic study setting. Extraction recovery estimation, based on model compounds with similar physicochemical characteristics, produced clinically feasible results without reference standards.

Gene-based pleiotropy across migraine with aura and migraine without aura patient groups

Introduction: It is unclear whether patients diagnosed according to International Classification of Headache Disorders criteria for migraine with aura (MA) and migraine without aura (MO) experience distinct disorders or whether their migraine subtypes are genetically related. Aim: Using a novel gene-based (statistical) approach, we aimed to identify individual genes and pathways associated both with MA and MO. Methods: Gene-based tests were performed using genome-wide association summary statistic results from the most recent International Headache Genetics Consortium study comparing 4505 MA cases with 34,813 controls and 4038 MO cases with 40,294 controls. After accounting for non-independence of gene-based test results, we examined the significance of the proportion of shared genes associated with MA and MO. Results: We found a significant overlap in genes associated with MA and MO. Of the total 1514 genes with a nominally significant gene-based p value (pgene-based ≤ 0.05) in the MA subgroup, 107 also produced pgene-based ≤ 0.05 in the MO subgroup. The proportion of overlapping genes is almost double the empirically derived null expectation, producing significant evidence of gene-based overlap (pleiotropy) (pbinomial-test = 1.5 × 10–4). Combining results across MA and MO, six genes produced genome-wide significant gene-based p values. Four of these genes (TRPM8, UFL1, FHL5 and LRP1) were located in close proximity to previously reported genome-wide significant SNPs for migraine, while two genes, TARBP2 and NPFF separated by just 259 bp on chromosome 12q13.13, represent a novel risk locus. The genes overlapping in both migraine types were enriched for functions related to inflammation, the cardiovascular system and connective tissue. Conclusions: Our results provide novel insight into the likely genes and biological mechanisms that underlie both MA and MO, and when combined with previous data, highlight the neuropeptide FF-amide peptide encoding gene (NPFF) as a novel candidate risk gene for both types of migraine.

Genome-wide association analysis identifies susceptibility loci for migraine without aura

Migraine without aura is the most common form of migraine, characterized by recurrent disabling headache and associated autonomic symptoms. To identify common genetic variants associated with this migraine type, we analyzed genome-wide association data of 2,326 clinic-based German and Dutch individuals with migraine without aura and 4,580 population-matched controls. We selected SNPs from 12 loci with 2 or more SNPs associated with P values of <1 x 10(-5) for replication testing in 2,508 individuals with migraine without aura and 2,652 controls. SNPs at two of these loci showed convincing replication: at 1q22 (in MEF2D; replication P = 4.9 x 10(-4); combined P = 7.06 x 10(-11)) and at 3p24 (near TGFBR2; replication P = 1.0 x 10(-4); combined P = 1.17 x 10(-9)). In addition, SNPs at the PHACTR1 and ASTN2 loci showed suggestive evidence of replication (P = 0.01; combined P = 3.20 x 10(-8) and P = 0.02; combined P = 3.86 x 10(-8), respectively). We also replicated associations at two previously reported migraine loci in or near TRPM8 and LRP1. This study identifies the first susceptibility loci for migraine without aura, thereby expanding our knowledge of this debilitating neurological disorder.

Self-tuning control of a class of multivariable non-linear systems

Self-tuning is applied to the minimum variance control of non-linear multivariable systems which can be characterized by a ' multivariable Hammerstein model '. It is also shown that such systems are not amenable to self-tuning control if control costing is to be included in the performance criterion.

Migraine with aura and migraine without aura are not distinct entities: further evidence from a large Dutch population study

It is often debated whether migraine with aura (MA) and migraine without aura (MO) are etiologically distinct disorders. A previous study using latent class analysis (LCA) in Australian twins showed no evidence for separate subtypes of MO and MA. The aim of the present study was to replicate these results in a population of Dutch twins and their parents, siblings and partners (N = 10,144). Latent class analysis of International Headache Society (IHS)-based migraine symptoms resulted in the identification of 4 classes: a class of unaffected subjects (class 0), a mild form of nonmigrainous headache (class 1), a moderately severe type of migraine (class 2), typically without neurological symptoms or aura (8% reporting aura symptoms), and a severe type of migraine (class 3), typically with neurological symptoms, and aura symptoms in approximately half of the cases. Given the overlap of neurological symptoms and nonmutual exclusivity of aura symptoms, these results do not support the MO and MA subtypes as being etiologically distinct. The heritability in female twins of migraine based on LCA classification was estimated at .50 (95% confidence intervals [CI] .27 - .59), similar to IHS-based migraine diagnosis (h2 = .49, 95% CI .19-.57). However, using a dichotomous classification (affected-unaffected) decreased heritability for the IHS-based classification (h2 = .33, 95% CI .00-.60), but not the LCA-based classification (h2 = .51, 95% CI .23-.61). Importantly, use of the LCA-based classification increased the number of subjects classified as affected. The heritability of the screening question was similar to more detailed LCA and IHS classifications, suggesting that the screening procedure is an important determining factor in genetic studies of migraine.

Latent class and genetic analysis does not support migraine with aura and migraine without aura as separate entities

Latent class and genetic analyses were used to identify subgroups of migraine sufferers in a community sample of 6,265 Australian twins (55% female) aged 25-36 who had completed an interview based on International Headache Society (IHS) criteria. Consistent with prevalence rates from other population-based studies, 703 (20%) female and 250 (9%) male twins satisfied the IHS criteria for migraine without aura (MO), and of these, 432 (13%) female and 166 (6%) male twins satisfied the criteria for migraine with aura (MA) as indicated by visual symptoms. Latent class analysis (LCA) of IHS symptoms identified three major symptomatic classes, representing 1) a mild form of recurrent nonmigrainous headache, 2) a moderately severe form of migraine, typically without visual aura symptoms (although 40% of individuals in this class were positive for aura), and 3) a severe form of migraine typically with visual aura symptoms (although 24% of individuals were negative for aura). Using the LCA classification, many more individuals were considered affected to some degree than when using IHS criteria (35% vs. 13%). Furthermore, genetic model fitting indicated a greater genetic contribution to migraine using the LCA classification (heritability, h(2)=0.40; 95% CI, 0.29-0.46) compared with the IHS classification (h(2)=0.36; 95% CI, 0.22-0.42). Exploratory latent class modeling, fitting up to 10 classes, did not identify classes corresponding to either the IHS MO or MA classification. Our data indicate the existence of a continuum of severity, with MA more severe but not etiologically distinct from MO. In searching for predisposing genes, we should therefore expect to find some genes that may underlie all major recurrent headache subtypes, with modifying genetic or environmental factors that may lead to differential expression of the liability for migraine.

Corneal confocal microscopy detects neuropathy in patients with type 1 diabetes without retinopathy or microalbuminuria

Objective Corneal innervation is increasingly used as a surrogate marker of human diabetic peripheral neuropathy (DPN) however its temporal relationship with the other microvascular complications of diabetes is not fully established. In this cross-sectional, observational study we aimed to assess whether neuropathy occurred in patients with type 1 diabetes, without retinopathy or microalbuminuria. Materials and Methods All participants underwent detailed assessment of peripheral neuropathy [neuropathy disability score (NDS), vibration perception threshold (VPT), peroneal motor nerve conduction velocity (PMNCV), sural sensory nerve conduction velocity (SSNCV) and in vivo corneal confocal microscopy (IVCCM)], retinopathy (digital fundus photography) and albuminuria status [albumin: creatinine ratio (ACR)]. Results 53 patients with Type 1 diabetes with (n=37) and without retinopathy (n=16) were compared to control subjects (n=27). SSNCV, corneal nerve fibre (CNFD) and branch (CNBD) density and length (CNFL) were reduced significantly (p<0.001) in diabetic patients without retinopathy compared to control subjects. Furthermore, CNFD, CNBD and CNFL were also significantly (p<0.001) reduced in diabetic patients without microalbuminuria (n=39), compared to control subjects. Greater neuropathic severity was associated with established retinopathy and microalbuminuria. Conclusions IVCCM detects early small fibre damage in the absence of retinopathy or microalbuminuria in patients with Type 1 diabetes.

Sleep and self-regulation from birth to 7 years: A retrospective study of children with and without ADHD at 8-9 years

Objective To examine mean level differences, and longitudinal and reciprocal relations among behavioral sleep problems, emotional dysregulation, and attentional regulation across early childhood for children with and without ADHD at 8-9 years. Method This study used data from Growing Up in Australia: The Longitudinal Study of Australian Children (LSAC) – Infant Cohort (n = 4109 analyzed). Children with and without ADHD were identified at age 8-9 years via parent-report of ADHD diagnosis and the 5-item Inattention-Hyperactivity subscale from the Strengths and Difficulties Questionnaire. Maternal report of child sleep problems and self-regulation was collected at 0-1, 2-3, 4-5 and 6-7 years of age. ANOVA was used to compare mean level differences in sleep problems, emotional and attentional regulation by ADHD group. Longitudinal structural equation modeling examined the relations among sleep and self-regulation across time in children with and without ADHD. Results Children with ADHD had persistently elevated levels of sleep problems (from infancy) and emotional and attentional dysregulation compared to controls (from 2-3 years of age). Sleep problems, emotional dysregulation, and attentional regulation were stable over time for both groups. Sleep problems were associated with greater emotional dysregulation two years later from 2-3 years of age for both groups, which in turn was associated with poorer attentional regulation. There was no direct relationship between sleep problems and later attentional regulation. Conclusion Sleep problems in children with and without ADHD are associated with emotional dysregulation, which in turn contributes to poorer attentional functioning. This study highlights the importance of assessing and managing sleep problems in young children.

A comparison of welfare outcomes for weaner and mature Bos indicus bulls surgically or tension band castrated with or without analgesia: 2. Responses related to stress, health and productivity

Tension banding castration of cattle is gaining favour because it is relatively simple to perform and is promoted by retailers of the banders as a humane castration method. Two experiments were conducted, under tropical conditions using Bos indicus bulls comparing tension banding (Band) and surgical (Surgical) castration of weaner (7–10 months old) and mature (22–25 months old) bulls with and without pain management (NSAID (ketoprofen) or saline injected intramuscularly immediately prior to castration). Welfare outcomes were assessed using a range of measures; this paper reports on some physiological, morbidity and productivity-related responses to augment the behavioural responses reported in an accompanying paper. Blood samples were taken on the day of castration (day 0) at the time of restraint (0 min) and 30 min (weaners) or 40 min (mature bulls), 2 h, and 7 h; and days 1, 2, 3, 7, 14, 21 and 28 post-castration. Plasmas from day 0 were assayed for cortisol, creatine kinase, total protein and packed cell volume. Plasmas from the other samples were assayed for cortisol and haptoglobin (plus the 0 min sample). Liveweights were recorded approximately weekly to 6 weeks and at 2 and 3 months post-castration. Castration sites were checked at these same times to 2 months post-castration to score the extent of healing and presence of sepsis. Cortisol concentrations (mean ± s.e. nmol/L) were significantly (P < 0.05) higher in the Band (67 ± 4.5) compared with Surgical weaners (42 ± 4.5) at 2 h post-castration, but at 24 h post-castration were greater in the Surgical (43 ± 3.2) compared with the Band weaners (30 ± 3.2). The main effect of ketoprofen was on the cortisol concentrations of the mature Surgical bulls; concentrations were significantly reduced at 40 min (47 ± 7.2 vs. 71 ± 7.2 nmol/L for saline) and 2 h post-castration (24 ± 7.2, vs. 87 ± 7.2 nmol/L for saline). Ketoprofen, however, had no effect on the Band mature bulls, with their cortisol concentrations averaging 54 ± 5.1 nmol/L at 40 min and 92 ± 5.1 nmol/L at 2 h. Cortisol concentrations were also significantly elevated in the Band (83 ± 3.0 nmol/L) compared with Surgical mature bulls (57 ± 3.0 nmol/L) at weeks 2–4 post-castration. The timing of this elevation coincided with significantly elevated haptoglobin concentrations (mg/mL) in the Band bulls (2.97 ± 0.102 for mature bulls and 1.71 ± 0.025 for weaners, vs. 2.10 ± 0.102 and 1.45 ± 0.025 respectively for the Surgical treatment) and evidence of slow wound healing and sepsis in both the weaner (0.81 ± 0.089 not healed at week 4 for Band, 0.13 ± 0.078 for Surgical) and mature bulls (0.81 ± 0.090 at week 4 for Band, 0.38 ± 0.104 for Surgical). Overall, liveweight gains of both age groups were not affected by castration method. The findings of acute pain, chronic inflammation and possibly chronic pain in the mature bulls at least, together with poor wound healing in the Band bulls support behavioural findings reported in the accompanying paper and demonstrate that tension banding produces inferior welfare outcomes for weaner and mature bulls compared with surgical castration.

A comparison of welfare outcomes for weaner and mature Bos indicus bulls surgically or tension band castrated with or without analgesia: 1. Behavioural responses

Tension-band castration of cattle is gaining favour because it is relatively simple to perform and is promoted by retailers of the devices as a humane castration method. Furthermore, retailers encourage delaying castration to exploit the superior growth rates of bulls compared with steers. Two experiments were conducted, under tropical conditions, comparing tension banding and surgical castration of weaner (7–10 months old) and mature (22–25 months old) Bos indicus bulls with and without pain management (ketoprofen or saline injected intramuscularly immediately prior to castration). Welfare outcomes were assessed using a wide range of measures; this paper reports on the behavioural responses of the bulls and an accompanying paper reports on other measures. Behavioural data were collected at intervals by direct observation and continuously via data loggers on the hind leg of the bulls to 4 weeks post-castration. Tension-banded bulls performed less movement in the crush/chute than the surgically castrated bulls during the procedures (weaner: 2.63 vs. 5.69, P < 0.001; mature: 1.00 vs. 5.94; P < 0.001 for tension-band and surgical castration, respectively), indicating that tension banding was less painful then surgical castration during conduct. To 1.5 h post-castration, tension-banded bulls performed significantly (all P < 0.05) more active behavioural responses indicative of pain compared with surgical castrates, e.g., percentage time walking forwards (weaner: 15.0% vs. 8.1%; mature: 22.3% vs. 15.1%), walking backwards (weaner: 4.3% vs. 1.4%; mature: 2.4% vs. 0.5%), numbers of tail movements (weaner: 21.9 vs. 1.4; mature: 51.5 vs. 39.4) and leg movements (weaner: 12.9 vs. 0.9; mature: 8.5 vs. 1.5), respectively. In contrast, surgically castrated bulls performed more immobile behaviours compared with tension-banded bulls (e.g., standing in mature bulls was 56.6% vs. 34.4%, respectively, P = 0.002). Ketoprofen administration appeared effective in moderating pain-related behaviours in the mature bulls from 1.5 to 3 h, e.g., reducing abnormal standing (0.0% vs. 7.7%, P = 0.009) and increasing feeding (12.7% vs. 0.0%, P = 0.048) in NSAID- and saline-treated bulls, respectively. There were few behavioural differences subsequent to 24 h post-castration, but some limited evidence of chronic pain (3–4 weeks post-castration) with both methods. Interpretation, however, was difficult from behaviours alone. Thus, tension banding is less painful than surgical castration during conduct of the procedures and pain-related behavioural responses differ with castration method (active restlessness in response to tension banding and minimisation of movement in response to surgical castration). Ketoprofen administered immediately prior to castration was somewhat effective in reducing pain, particularly in the mature bulls.

Self-tuning minimum-variance control of nonlinear systems of the Hammerstein model

Self-tuning is applied to the control of nonlinear systems represented by the Hammerstein model wherein the nonlinearity is any odd-order polynomial. But control costing is not feasible in general. Initial relay control is employed to contain the deviations.