Physical health, breastfeeding problems and maternal mood in the early postpartum: a prospective cohort study.

This study aimed to investigate prospectively the contribution of maternal physical health and/or breastfeeding problems to maternal mood (depression, anxiety, fatigue, irritability, confusion, vigor) at 8-weeks postpartum. A prospective study was conducted. Participants were recruited antenatally from a public and a private maternity hospital in Melbourne, Australia. Nulliparous pregnant women (N = 229), ≥ 18 years of age, ≥ 36-week gestation, singleton pregnancy and with sufficient English were eligible. Data were collected by self-report questionnaire (pregnancy, weeks 1-4 postpartum) and telephone interview (week 8 postpartum). A high burden of physical problems was classified as ≥ 3 problems (caesarean/perineal pain; back pain; constipation; haemorrhoids; urinary and bowel incontinence) for ≥ 2 time points. A high burden of breastfeeding problems was having ≥ 2 problems (mastitis; nipple pain; frequent expressing; over- or under-supply of milk) for ≥ 2 time points. Multivariate linear regression was used to investigate the relationship between maternal mood, assessed using Profile of Mood States (8-week postpartum), and a high burden of breastfeeding and/or physical health problems. Forty-six women (20.1%) had a high burden of physical symptoms, 44 (19.2%) a high burden of breastfeeding problems only and 25 women (11.0%) had both. A high burden of breastfeeding problems alone (ß = 10.6, p = 0.01) or with co-morbid physical problems (ß = 15.35, p = 0.002) was significantly associated with poorer maternal mood at 8 weeks. Early, effective postnatal treatment of maternal health and breastfeeding problems could reduce women's risk for poor mental health.

Survey of Reproductive Experiences and Outcomes of Cancer Survivors Who Stored Reproductive Material Before Treatment.

STUDY QUESTION: What are the reproductive experiences and outcomes of people who store reproductive material before cancer treatment? SUMMARY ANSWER: Of respondents who had tried to achieve pregnancy since completing cancer treatment almost all had succeeded, in most cases through natural conception. WHAT IS KNOWN ALREADY: People of reproductive age who are diagnosed with cancer can cryopreserve reproductive material to guard against the adverse effects on fertility of gonadotoxic treatment. Little is known about the reproductive outcomes of people who undergo fertility preservation before cancer treatment. STUDY DESIGN, SIZE, DURATION: Cross-sectional survey. PARTICIPANTS/MATERIALS, SETTING, METHODS: Women and men who had stored reproductive material before cancer treatment at two private and one public fertility clinics up to June 2014 and were at least 18 years old at the time were identified from medical records and invited to complete an anonymous questionnaire about their reproductive experiences. MAIN RESULTS AND THE ROLE OF CHANCE: Of the 870 potential respondents 302 (171 female and 131 male) returned completed questionnaires yielding a response rate of 34.5% (39.5% and 29.7% for female and male respondents, respectively). Current age was similar for women and men (37.2 years) but men had been diagnosed with cancer significantly earlier in life than women (28.2 versus 30.3 years, P = 0.03). Almost two-thirds of respondents wished to have a child or another child in the future, some of whom knew that they were unable to. One in ten respondents was a parent before the cancer diagnosis and around one-third had had a child since diagnosis or was pregnant (or a partner in pregnancy) at the time of the survey. Of those who had tried to conceive since completing cancer treatment (N = 119) 84% (79% of women and 90% of men) had had a child or were pregnant (or a partner in pregnancy). Most of the pregnancies since the diagnosis of cancer occurred after natural conception (58/100, 58%). Of the 22 women (13% of all women) and 35 men (27% of all men) who had used their stored reproductive material four women (18%) and 28 men (80%) had had a child or were pregnant or a partner in pregnancy at the time of completing the survey. The most commonly stated reason for not using the stored material was not being ready to try for a baby. LIMITATIONS, REASON FOR CAUTION: The relatively low response rate, particularly among men, means that participation bias may have influenced the findings. As type of cancer was self-reported and we did not ask questions about respondents' cancer treatments, it is not possible to link reproductive outcomes to type of cancer or cancer treatment. Also, there is no way of comparing the sample with the populations they were drawn from as data on reproductive outcomes of people who store reproductive material before cancer treatment are not collected routinely. This might have led to over- or underestimates of the reproductive experiences and outcomes reported in this paper. WIDER IMPLICATIONS OF THE FINDINGS: The findings add to the limited evidence about the reproductive outcomes of this growing group of people and can be used to inform the advice given to those contemplating fertility preservation in the context of cancer. STUDY FUNDING/COMPETING INTERESTS: The study was funded by the National Health and Medical Research Council (APP1042347). The authors have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: Not applicable.

Women's reflections on fertility and motherhood after breast cancer and its treatment.

Breast cancer and its treatment have complex ramifications for women of reproductive age, including reduced fertility. With the aim of increasing understanding of what it means to women to manage fertility and motherhood in the years after a diagnosis of breast cancer, in-depth qualitative interviews were conducted with 10 women aged 26-45 years, living in Victoria, Australia, who had been diagnosed with breast cancer aged 25-41. Transcripts were analysed thematically and interpreted within narrative theory. Six themes linking breast cancer to fertility and motherhood were identified: diagnosis as a pivotal life event, robbed of time and choice, significance of fertility, being a mother, narrative justification, and life after breast cancer treatment. Women without children described a preoccupying sorrow about lost fertility. Women's accounts yielded evidence of narrative meaning-making, including justifying their decisions and actions in relation to survival, treatment and fertility, and coping with adversity by developing consoling plots. Breast cancer, fertility and reproductive health are inter-linked in diverse ways which have immediate and long-term consequences. Even if women are receiving optimum fertility management, it is evident that some women of reproductive age will need continuing post-cancer care to manage and ameliorate ramifications of diminished or lost fertility.

Looking downstream: a review of the literature on physical and psychosocial health outcomes in adolescents and young adults who were conceived by ART.

BACKGROUND: The use of assisted reproductive technology (ART) is now well established in many countries and the first generations of offspring are reaching maturity. We reviewed the published literature to describe the available evidence about health outcomes in ART-conceived young people who were of an adolescent age or older. METHODS: The EMBASE, Medline and PsychINFO databases were searched from January 1998 to October 2010. Key inclusion criteria were that the study sample have a mean age of ≥ 12 years or a mean follow-up period of ≥ 12 years and were conceived by ART. RESULTS: Seven publications reported physical health outcomes and 10 reported psychosocial health outcomes in ART offspring. Compared with control groups, some differences in physiological outcomes in relation to growth and development, chronic illness and risk of cancer have been reported. Overall, psychosocial studies of ART-conceived young people indicate that their cognitive function and psychological and social adjustment are similar to that of comparison groups. CONCLUSIONS: Overall, nine ART-conceived populations of this age group have been studied. Most samples included < 300 participants and methodologies varied between studies. Health information on this age group is therefore limited and the clinical significance of the findings remains unclear. Further research focusing on ART-conceived young adults is needed, particularly in relation to neurological health outcomes where no studies have been reported to date.

Breastfeeding after assisted conception: a prospective cohort study.

AIM: The aims were to investigate the prevalence of breastfeeding after conception with assisted reproductive technology (ART) and identify risk factors for breastfeeding duration <6 weeks and cessation of breastfeeding before the baby is 8 months old. METHODS: A consecutive cohort of women who had conceived at one of two ART centres in Melbourne, Australia, was recruited in early pregnancy. The women completed telephone interviews and postal questionnaires in pregnancy and 3, 8 and 18 months after the birth. RESULTS: Of 239 eligible women, 183 (77%) agreed to take part. Participants were more likely than the general population of childbearing Australian women to initiate breastfeeding (89% vs 83.3%, p=0.05) but by 3 months, a smaller proportion was breastfeeding exclusively (46% vs 57.3%, p=0.004). The proportions not providing any breast milk at 6 weeks and 8 months were 23% and 57%, respectively. More anxiety in late pregnancy and sub-optimal breastfeeding advice predicted breastfeeding duration <6 weeks and breastfeeding cessation before 8 months. CONCLUSION: The predictors of less favourable breastfeeding outcomes after ART identified may be modifiable. Antenatal strategies to reduce anxiety in pregnancy and postnatal strategies to ensure consistent breastfeeding advice may improve breastfeeding outcomes among women who give birth after ART.

Men's experiences of infertility and infertility treatment 5 years after diagnosis of male factor infertility: a retrospective cohort study.

BACKGROUND: The aim of this study was to describe the perceptions of infertile men regarding the impact of infertility on their intimate relationships, their experience of treatment and their sources of information and support. METHODS: A cross-sectional survey of a consecutive cohort of men diagnosed 5 years earlier as infertile at Melbourne IVF and the Royal Women's Hospital Reproductive Services, Melbourne was conducted. Study-specific questions assessed the impact of male factor infertility on the intimate relationships, their perceived quality of infertility-related health care and their preferred sources of infertility-related information and personal support and the effectiveness of these. RESULTS: The response rate was 41% (112/276). Male factor infertility was reported to have had a negative impact on the intimate partner relationship by 25% of men, and 32% reported a negative effect on their sexual satisfaction. Satisfaction with medical care and clinic information was high and not influenced by the outcome of the treatment. Clinic-provided information and discussion with clinic staff were the most strongly preferred sources of information, and the partner and clinic staff were the most valued sources of personal support. Very few men found support groups useful and less than half confided in friends. CONCLUSIONS: The findings suggest that for a significant subgroup of men, male factor infertility affects their intimate relationship negatively. Wider sources of social support are not used by infertile men as they rely predominantly on clinic-provided information and support. This indicates that psychologically informed supportive clinical care is particularly important for men diagnosed as infertile.

Early post-partum adjustment and admission to parenting services in Victoria, Australia after assisted conception.

BACKGROUND: Higher rates of admission to residential early parenting services (REPSs) after assisted conception compared with spontaneous conception have been reported. The aim of this study was to characterize early post-partum psychological functioning and the rate of, and risks factors for, admission to REPSs in women conceiving with assisted reproductive technology (ART) in Australia. METHODS: A consecutive cohort of women who had conceived through ART was recruited systematically in early pregnancy. At 3 months post-partum, participants completed postal questionnaires which included a new measure of the degree of difficulty involved in conceiving, the Burden of Infertility and Treatment (BIT) scale. RESULTS: Of 166 women who participated, 8% had already been admitted to a REPS within 3 months, which is a higher rate compared with other women in the first 12 months (5%). Compared with community samples of new mothers, there was no difference in rate of depression. A higher proportion reported dysregulated infant behaviours (P < 0.0001) and a smaller proportion was breast feeding exclusively (P < 0.0001). Greater difficulty conceiving (higher BIT score) was associated with lower maternal confidence. CONCLUSIONS: Clinical care of the increasing group of women who conceive with ART should include explicit assessment of early post-partum psychological functioning and early intervention if difficulties in managing infant behaviour are reported.

Psychological and social aspects of pregnancy, childbirth and early parenting after assisted conception: a systematic review.

BACKGROUND: It is known that infertility affects emotional well-being, satisfaction with life and self-esteem and that failed assisted reproductive technology (ART) treatment is associated with diminished life satisfaction, reduced self-confidence and substantial psychological distress. Investigations of whether these persist when treatment results in a pregnancy and live birth have been undertaken. METHODS: A systematic search for English-language research articles on psychological and social aspects of pregnancy, childbirth and the first post-partum year after ART conception. RESULTS: Of 466 retrieved papers, 46 met inclusion criteria. These reported data from 28 studies. There is consistent evidence that marital satisfaction, emotional well-being and self-regard in pregnancy, attachment to the fetus and parent-infant relationship in ART groups are similar to comparison groups. Anxiety about the survival of the fetus and early parenting difficulties appear to be higher and post-natal self-confidence lower. Evidence about adjustment to pregnancy and parenthood and the experience of childbirth is inconclusive and reports of parental perceptions of infant temperament and behaviour are contradictory. Between-study methodological differences may explain the lack of consistency in findings of the influence of infertility and ART on some aspects of the transition to parenthood. CONCLUSIONS: Overall, this body of evidence is best described as emergent. It is possible that in pregnancy after ART, parenthood might be idealized and this might then hinder adjustment and the development of a confident parental identity.

Women's experiences of childbirth and post-natal healthcare after assisted conception.

BACKGROUND: The proportion of women who give birth after assisted reproduction technology (ART) treatment is increasing. To date, little is known about their experience of childbirth. One of the aims of this study was to investigate the experience of childbirth and the post-natal healthcare after ART. METHODS: A prospective, longitudinal study of a systematically recruited consecutive cohort of women who had conceived with ART in Melbourne, Australia, in 2001 was investigated using telephone interviews and self-report questionnaires. The experience of birth was explored 3 months post-partum. RESULTS: One hundred and sixty-six women who had conceived through ART participated. Compared with other Australian women, participants were more likely to have a Caesarean birth (51% versus 25%, P < 0.0001). Women who had a Caesarean birth were less likely to report having had an active say about the management of the birth (P < 0.01) and to have held their baby at birth (P < 0.0001) and more likely to report disappointment with the birth event (P < 0.0001), severe post-natal pain (P = 0.02) and needing a lot of help or advice with infant feeding (P = 0.001) than those who had a vaginal birth. CONCLUSIONS: After ART, there are highly elevated rates of operative birth which appear to influence early post-natal adjustment.

Prevalence, nature, severity and correlates of postpartum depressive symptoms in Vietnam.

OBJECTIVE: To examine depressive symptomatology in women after childbirth in Ho Chi Minh City, Vietnam. DESIGN: A cross sectional survey. SETTING: Hung Vuong Obstetrics and Gynaecology Hospital and the Maternal, Child Health and Family Planning Center of Ho Chi Minh City, Vietnam. POPULATION: Mothers of infants aged +/- six weeks attending well-baby clinics. METHOD: Participants were recruited consecutively in the postnatal wards and invited to take part in the study at the first clinic visit. Individual structured interviews about health and social circumstances, including the Edinburgh Postnatal Depression Scale (EPDS) were administered during clinic visits. The interview schedule was translated into Vietnamese, back translated for verification and piloted. Interviewers were specifically trained members of staff of the two centres. MAIN OUTCOME MEASURES: EPDS scores and responses to structured questions about specific and non-specific symptoms. RESULTS: Of 506 women who participated, 166 (33%) had EPDS scores in the clinical range of > 12 and 99 (19%) acknowledged suicidal ideation. In a forward stepwise logistic regression analysis, 77% of cases with EPDS scores > 12 were correctly classified in a model which included unwelcome pregnancy, lack of a permanent job, < 30 days complete rest after childbirth, an unsettled baby, not being given special foods, avoiding proscribed foods and being unable to confide in their husbands. CONCLUSION: Depressive symptomatology is more prevalent among parturient women in Ho Chi Minh City, Vietnam than reported rates in developed countries and is at present unrecognised.

Sampling considerations for garden symphylans (Order: Cephalostigmata) in western Oregon.

Sampling recommendations were developed for a potato bait sampling method used to estimate garden symphylan (Scutigerella immaculata Newport) densities in western Oregon. Sample size requirements were developed using Taylor's power law to describe the relationship between sample means and variances. Developed sampling recommendations performed well at sample sizes of 30 and greater, when validated by resampling a cohort of 40 independent data sets. Sample size requirements for the bait sampling method were 1.5 times greater than the requirements for the soil sampling method over densities from 1 to 20 S. immaculata per sample unit. As S. immaculata densities increased from April to May, sample size requirements decreased by 36% for fixed precision levels. For sampling in April, decreasing the damage threshold from 20, to 10 and five S. immaculata per sample unit, required a 1.6 and 2.5 times greater sample size requirement, respectively, for a fixed precision level (c) appropriate for pest management (c = 0.25). The bait sampling method provides an efficient reliable alternative to the standard soil sampling method used to monitor garden symphylan populations.

Rapid analysis of a plasmid by hydrophobic-interaction chromatography with a non-porous resin.

A HPLC technique has been developed, based on hydrophobic-interaction Chromatography with a non-porous packing (TSKgel Butyl-NPR, Tosoh Biosep LLC), that allows separation of the open circular (nicked) and supercoiled forms of five DNA plasmids, ranging in size from 4 to 30 kilo base pairs (kb). The identity of the bands was determined through light scattering and gel electrophoresis. Several buffers, gradients, flow-rates and temperatures were evaluated in determining the optimum operating conditions for the separation. For all plasmids a reversed ammonium sulfate in phosphate buffer (pH 7.1) gradient was established. The chromatographic resolution between the supercoiled and nicked peaks was found to be a function of flow-rate and temperature. The resolution and the elution order did not vary with plasmid size, with the open-circular form always being eluted before the supercoiled form. Hydrophobic-interaction chromatography is a useful alternative to ion exchange or size exclusion for the chromatography of large plasmids, up to 30 kb.

Nature, severity and correlates of psychological distress in women admitted to a private mother-baby unit.

OBJECTIVE: To characterize psychological distress of women admitted with their infants to a private hospital mother-baby unit. METHODS: A cross-sectional self-report survey including standardized psychometric questionnaires of consecutive admissions to Masada Private Hospital Mother-Baby Unit, Melbourne between 1 June and 30 November 1997. Scores on the Profile of Mood States, Edinburgh Postnatal Depression Scale, Intimate Bonds Measure, Parental Bonding Instrument, Social Support Questionnaire, Vulnerable Personality Scale self-report instruments and responses to a structured self-report questionnaire. Of the women admitted, 109 out of 146 participated in the survey. RESULTS: Clinically significant fatigue was almost universal in this cohort. Psychological distress clustered into three distinct groups: (i) probable depression; (ii) fatigue and distress; and (iii) fatigue only. It may be more accurate to conceptualise postpartum psychological distress either as a continuum, or as a wide range of distinct states, rather than as a dichotomous condition in which individuals are categorized as being or not being cases. In this cohort, the severity of distress was associated most consistently with the quality of a woman's relationship with her partner and with infants who were difficult to settle. CONCLUSIONS: It appears that some cases of psychological distress in women following childbirth may be inaccurately conceptualised as depression. Clinically significant fatigue and moderate distress are distinguishable from depression, and warrant specifically tailored assessment strategies and treatments. Following the birth of a baby, the partner of a mother has a strong influence on her emotional health and should be included in any assessment and intervention.

Operative intervention in delivery is associated with compromised early mother-infant interaction.

OBJECTIVE: To investigate the effects of mode and place of delivery on first mother-infant contact and maternal emotional wellbeing. DESIGN: A prospective, longitudinal study. PARTICIPANTS: A sociodemographically representative sample of 203 consecutive primiparous women was recruited, of whom 81% (n = 164) were followed up. SETTING: Four metropolitan hospitals, including one accredited baby friendly hospital, in Melbourne, Australia, 1997. METHODS: During the postnatal hospital stay women were interviewed and medical records were inspected. PARTICIPANTS: completed two self-report psychometric measures: the Edinburgh Postnatal Depression Scale (EPDS) and the Profile of Moods States (POMS). Follow up at eight months postpartum comprised completion of a postal questionnaire and repeat administration of the standardised self-report psychometric mood questionnaires. RESULTS: The First Contact Index provides a measure of the first contact between a mother and her baby after the birth. Two-way analysis of variance revealed significant differences in First Contact Index between three modes of delivery groups: spontaneous, instrumentally-assisted, and caesarean section (P < 0.001) and four hospital of delivery groups (P < 0.001), but there was no significant interaction effect (P = 0.55). Significant negative correlations existed between the First Contact Index and both the POMS and EPDS scores at two days postpartum (POMS, r = -0.339, 95% Cl -0.46 to -0.22; EPDS, r = -0.279, 95% CI -0.41 to -0.16), even within delivery groups, and these were still present at eight months postpartum (POMS, r = -0.298, 95% CI -0.44 to -0.14, EPDS, r = -0.206, 95% CI -0.36 to -0.03). CONCLUSION: Instrumental and surgical delivery exert a negative impact on first postnatal contact between a mother and her baby which has persistent adverse maternal emotional correlates. Hospital practices associated with obstetric intervention in delivery also varied and are amenable to change, enabling optimal first mother-baby interaction regardless of mode of delivery.

Grapes, galls, and geography: the distribution of nuclear and mitochondrial DNA variation across host-plant species and regions in a specialist herbivore.

Studies of patterns of molecular variation in natural populations can provide important insights into a number of evolutionary problems. Among these, the question of whether geographic factors are more important than ecological factors in promoting population differentiation and ultimately speciation has been an important and contentious area in evolutionary biology. Systems involving herbivorous insects have played a leading role in this discussion. This study examined the distribution of molecular variation in a highly specialized gall-forming insect, grape phylloxera (Daktulosphaira vitifoliae Fitch), that is found on both sympatric and allopatric host-plant species of the genus Vitis. In addition, the relationship of insects in the introduced range in the United States to ancestral populations in the native range was examined. Evidence for differentiation along host-plant lines from both nuclear (RAPD) and mitochondrial (COI) DNA was confounded with the effect of geography. Differentiation was found where hosts were allopatric or parapatric, but no evidence was found for such differentiation on two hosts, V. vulpina and V. aestivalis, that are broadly sympatric. The question of population differentiation onto these sympatric hosts can be considered to be resolved--it has not occurred in spite of a long history of association. Evidence was equivocal, but suggestive of a period of divergence in allopatry prior to reestablishment of contact, for insects associated with another host plant species, V. cinerea, found in both sympatric and parapatric populations. A low level of diversity and placement of samples collected from the grape species V. riparia at the tip of a phylogenetic tree supports the hypothesis that this host has been recently colonized from populations from the Mississippi Valley. A polyphyletic origin for biotype B grape phylloxera was supported: Although most samples collected from vineyards in the introduced range in California had similar haplotypes, they were closely related to natives on V. vulpina from the Atlantic Coast-Piedmont region. All samples collected from vineyards in Oregon and Washington were closely related to natives on V. riparia in the northern United States.

Coexistent lyme disease and parvovirus infection in a child.

Infectious diseases commonly cause illnesses that mimic rheumatic diseases. Both Lyme disease and Parvovirus B19 infections produce arthritis, rashes, and a systemic illness that may be thought to represent a chronic rheumatic disease. In the case presented, a child with both infections simultaneously exhibited arthralgias, aseptic meningitis, and a facial rash. The features of Lyme disease and Parvovirus B19 infection that may mimic systemic lupus erythematosus include a facial rash, often in a malar distribution, hematologic abnormalities, arthritis, neurologic disorders, and autoantibody positivity. Given the proper season and geographical location, one must consider the possibility of co-infection with these two organisms, especially in those with atypical rheumatic complaints.

First Report of a Cucumber mosaic virus-Associated Satellite RNA in Lobelia erinus.

Lobelia (Lobelia erinus L.) is a common herbaceous annual used in flower beds and hanging baskets. The plant blooms from early to late summer. In the summer of 2000, Lobelia plants expressing virus-like symptoms were collected from a greenhouse-based production site in Ohio. Affected plants expressed a mild leaf mosaic and stunting. Viral-associated dsRNA was isolated from 7 g of symptomatic leaf tissue (1). Four dsRNAs were observed at 3.9, 3.0, 2.25, and 1.05 kb indicating the presence of a Cucumovirus. A fifth dsRNA at 0.75 kb also was observed, consistent with the presence of a satellite RNA. Enzyme-linked immunosorbent assay (ELISA) analysis (Agdia, Inc., Elkhart, IN) of symptomatic Lobelia tissue confirmed the presence of Cucumber mosaic virus (CMV). A (S)CARNA-5 (-) cDNA clone (American Type Culture Collection #45124) was labeled with digoxygenin (DIG) as per the manufacturer's instructions (Genius II DIG-DNA Labeling Kit, Boehringer Mannheim) and used as a diagnostic probe to detect this satellite RNA. Northern hybridization confirmed the identity of the satellite RNA (2). This is the first report of any satellite RNA associated with a virus infection in Lobelia and the first report of CMV in this host in Ohio. References: (1) J. R. Fisher and S. G. P. Nameth. HortScience 35:230-234, 2000. (2) R. A. Valverde et.al. Plant Dis. 74:255-258, 1990.

Purification of myristoylated and nonmyristoylated neuronal calcium sensor-1 using single-step hydrophobic interaction chromatography.

Neuronal calcium sensors (NCSs) belong to a family of Ca(2+)-binding proteins, which serve important functions in neurotransmission, and are highly conserved from yeast to humans. Overexpression of the neuronal calcium sensor-1, called frequenin in the fruit fly and in frog, increases the release of neurotransmitters. Studying the functional role of frequenin in mammals and understanding its structural dynamics is critically dependent on the availability of active purified protein. Neuronal calcium sensors like other members of the family share common structural features: they contain four EF-hands as potential binding sites for Ca(2+) and an N-terminal consensus sequence for myristoylation. Previously, recoverin, distantly related to NCSs, has been expressed and purified from Escherichia coli, involving a combination of different chromatographic steps. NCS-1 has earlier been purified adopting a two-step procedure used for recoverin purification. We have overexpressed NCS-1 from rat in its myristoylated and nonmyristoylated form in E. coli and purified it from crude lysates using a single-step hydrophobic interaction chromatography. The purified protein was identified by Western blotting and mass spectrometry and assayed for its ability to bind Ca(2+) using a Ca(2+) shift assay, terbium fluorescence, and Stains-all binding. The present protocol provides a rapid, more efficient and simplified, single-step method for purifying NCS-1 for structural and functional studies. This method can also be applied to purify related proteins of the superfamily.