Displaying publications 1 - 20 of 65 in total

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  1. Abdul Murad NA, Abdullah N, Kamaruddin MA, Abd Jalal N, Ismail N, Yusof NAM, et al.
    J ASEAN Fed Endocr Soc, 2021;36(2):127-132.
    PMID: 34966195 DOI: 10.15605/jafes.036.02.02
    OBJECTIVE: In this present study, we aim to evaluate the accuracy of the HbA1c relative to fasting plasma glucose (FPG) in the diagnosis of diabetes and pre-diabetes among The Malaysian Cohort (TMC) participants.

    METHODOLOGY: FPG and HbA1c were taken from 40,667 eligible TMC participants that have no previous history of diabetes, aged between 35-70 years and were recruited from 2006 - 2012. Participants were classified as normal, diabetes and pre-diabetes based on the 2006 World Health Organization (WHO) criteria. Statistical analyses were performed using ANOVA and Chi-square test, while Pearson correlation and Cohen's kappa were used to examine the concordance rate between FPG and HbA1c.

    RESULTS: The study samples consisted of 16,224 men and 24,443 women. The prevalence of diabetes among the participants was 5.7% and 7.5% according to the FPG and HbA1c level, respectively. Based on FPG, 10.6% of the participants had pre-diabetes but this increased to 14.2% based on HbA1c (r=0.86; P<0.001). HbA1c had a sensitivity of 58.20 (95% CI: 56.43, 59.96) and a specificity of 98.59 (95% CI: 98.46, 98.70).

    CONCLUSION: A higher prevalence of pre-diabetes and diabetes was observed when using HbA1c as a diagnosis tool, suggesting that it could possibly be more useful for early detection. However, given that HbA1c may also have lower sensitivity and higher false positive rate, several diagnostic criteria should be used to diagnose diabetes accurately.

  2. Abdullah AA, Ramli N, Yaacob NM, Hussain S
    J ASEAN Fed Endocr Soc, 2022;37(2):15-22.
    PMID: 36578895 DOI: 10.15605/jafes.037.02.03
    OBJECTIVE: This study aimed to determine the proportion, clinical characteristics, hormonal status, median time for normalization of serum thyroxine (FT4) and thyroid-stimulating hormone (TSH) and factors affecting time to thyroid function test (TFT) normalization of neonates born to mothers with maternal hyperthyroidism admitted in our institution.

    METHODOLOGY: This was a retrospective cohort study that included 170 newborns admitted to the Neonatal Intensive Care Unit (NICU) of Hospital Universiti Sains Malaysia (HUSM) with a history of maternal hyperthyroidism from January 2013 until December 2018. We analyzed their baseline demographic and clinical characteristics, maternal thyroid status and antibody levels. Finally, we analyzed newborn thyroid function and thyroid antibodies.

    RESULTS: The proportion of neonates born to mothers with maternal hyperthyroidism was 0.8% (170 of 20,198 neonates within the study period). Seven (4.1%) developed overt hyperthyroidism, while four (2.4%) had thyroid storm. The median time for thyroid function test normalization was 30 days (95% CI: 27.1 to 32.8). The median time for TFT normalization was longer among neonates of mothers with positive thyroid antibodies [46.6 days (95% CI, 20.6 to 39.4)] and of mothers who received anti-thyroid treatment [31.7 days (95% CI, 23.5 to 39.9)].

    CONCLUSION: Neonates born to mothers with hyperthyroidism is uncommon. These babies were observed to have a longer time for normalization of thyroid function tests if their mothers had thyroid antibodies or received anti-thyroid treatment.

  3. Abdullah NRA, Akbar RZA
    J ASEAN Fed Endocr Soc, 2017;32(1):50-53.
    PMID: 33442085 DOI: 10.15605/jafes.032.01.09
    Autoimmune thyroiditis in the course of other autoimmune diseases such as systemic lupus erythematosus (SLE) is common because these disorders are attributed to the production of autoantibodies against various autoantigens. Beyond this association, autoimmune thyroiditis can occur before, during or after the development of SLE. In this report, we describe a female who presented with facial puffiness, lethargy and progressive abdominal distension. She was diagnosed with autoimmune thyroiditis followed by the diagnosis of SLE complicated by a massive ascites, a rare form of lupus peritonitis, which is sterile ascites that results from severe serositis. Her presentation was complex and posed a diagnostic challenge and dilemma to the physicians involved in her care.
  4. Anuar Zaini A, Feng Tung Y, Ahmad Bahuri NF, Yazid Jalaludin M
    J ASEAN Fed Endocr Soc, 2020;35(1):62-67.
    PMID: 33790495 DOI: 10.15605/jafes.035.01.11
    Introduction: The causes of congenital hypothyroidism (CHT) are thyroid dysgenesis (TD), dyshormonogenesis (TDH) or transient hypothyroidism (TH).

    Methodology: This is a cross-sectional study looking at data over a period of 16 years (2000-2016). Confirmed cases had thyroid scan at the age of 3-years-old and repeated TFT (after 6 weeks off medications). Relevant data was collected retrospectively.

    Results: Forty (60% female) children with CHT were included in the study. Thirty (75%) children presented with high cord TSH. Nine (23%) presented after 2 weeks of life. Majority were diagnosed with TDH (42.5%) with TD and TH of 40% and 17.5% respectively. Median cord TSH of children with TD was significantly higher compared to TDH and TH (p=0.028 and p=0.001 respectively). L-thyroxine doses were not significantly different between TD, TDH and TH at diagnosis or at 3 years.

    Conclusions: TDH is highly prevalent in our population. TD may present after 2 weeks of life. One in five children treated for CHT had TH. Differentiating TD, TDH and TH before initiating treatment remains a challenge in Malaysia. This study provides clinicians practical information needed to understand the possible aetiologies from a patient's clinical presentation, biochemical markers and treatment regime. Reassessing TH cases may be warranted to prevent unnecessary treatment.

  5. Araneta MR
    J ASEAN Fed Endocr Soc, 2019;34(2):126-133.
    PMID: 33442147 DOI: 10.15605/jafes.034.02.02
    Type 2 diabetes prevalence is rising rapidly in Southeast Asia (SEA) where urbanization and adoption of 'western' behavioral lifestyles are attributed as predominant risk factors. The Southeast Asian diaspora to the United States has resulted in a sizable portion of migrant and US born SEAs, with approximately 4 million Filipino Americans, 2 million Vietnamese-Americans, Cambodians (330,000), and Thai (300,000) as the most populous. Their longer exposure to a western lifestyle and participation in clinical studies with other racial/ethnic groups, provide opportunities to evaluate etiologic factors which might inform trends and intervention opportunities among residents of Southeast Asia. Epidemiologic studies in the US have identified higher T2D prevalence among Filipinos (16.1%) compared to groups perceived to be at highest risk for T2D, namely Latinos (14.0%), Black (13.7%), and Native Americans (13.4%), while SEAs (including Burmese, Cambodian, Indonesian, Laotian, Malaysian, and Thai, 10.5%) and Vietnamese (9.9%) had higher T2D risk compared to Whites (7.7%), despite their absence of general obesity. Asian-Americans, including SEAs, East and South Asians, collectively have higher rates of undiagnosed T2D compared to other racial/ethnic groups in the US. Almost half (44%) of Filipinos with newly diagnosed T2D have isolated post-challenge hyperglycemia and will remain undiagnosed if current screening practices remain limited to measures of glycosylated hemoglobin and fasting plasma glucose. The University of California San Diego Filipino Health Study found excess visceral adipose tissue accumulation, low ratio of muscle to total abdominal mass area, low adiponectin concentration, multiparity (≥ 6 live births), and sleep insufficiency (<7 hours) to be unique T2D risk factors among Filipino-American women, even after adjusting for established T2D risk factors including hypertension and parental history of T2D. Social determinants such as low educational attainment (less than college completion), and sustained social disadvantage during childhood and adulthood were independently associated with T2D risk. Gestational diabetes is a known risk factor for future T2DM among women; Northern California data shows that following Asian Indians, gestational diabetes was highest among Filipina and SEA parturients, who had twice the GDM prevalence as Black, Hispanic, and White women. Identification of novel T2D risk factors among SEAs may guide early diagnosis, inform pathophysiology, and identify unique opportunities for T2D prevention and management.
  6. Aziz H, Hussein Z
    J ASEAN Fed Endocr Soc, 2021;36(1):104-107.
    PMID: 34177097 DOI: 10.15605/jafes.036.01.11
    Giant parathyroid adenoma (GPA) is defined as adenoma larger than 3.5 g. Twenty-one cases of parathyroid mass >3.5 g in patients with primary hyperparathyroidism who underwent parathyroidectomy in Hospital Putrajaya, Malaysia were identified. Most cases presented with nephrolithiasis. Two cases are reported as parathyroid cancer. GPA has significantly higher serum calcium and iPTH levels and can be asymptomatic. Parathyroid carcinoma patients are frequently symptomatic, with large tumors. Differentiating GPA from parathyroid cancer is important as it determines the subsequent surgical intervention.
  7. Azman SSBW, Mohamad MB
    J ASEAN Fed Endocr Soc, 2021;36(2):201-212.
    PMID: 34966206 DOI: 10.15605/jafes.036.02.05
    Acute suppurative thyroiditis is a rare and potentially fatal condition. We present a case of an 18-year-old Malay female who presented with one-week history of painful right sided neck swelling, fever and odynophagia. Neck CT confirms ruptured multiloculated abscess with posterosuperior extension into prevertebral space. Pus and tissue cultured Streptococcus anginosus and Eikenella corrodens with positive TB PCR. She responded well to ampicillin/sulbactam and anti-tuberculosis treatment with no evidence of residual collection from ultrasound.
  8. Bernard DMAA, Yi OX, Veerasamy HLA, Bidin MBL, Keong SC
    J ASEAN Fed Endocr Soc, 2021;36(2):200-204.
    PMID: 34966205 DOI: 10.15605/jafes.036.02.04
    Presentation of Cushing's syndrome during pregnancy is extremely rare. We report a 21-year-old female with Cushing's syndrome diagnosed at 23 weeks of gestation and had recurrent acute pulmonary oedema during the antepartum and postpartum period. She delivered prematurely via emergency caesarean section at 28 weeks of gestation. This case highlights the rare occurrence of recurrent acute pulmonary oedema during pregnancy and consequential premature birth in a patient with adrenal Cushing's. She was diagnosed with adrenal Cushing's during the postpartum period based on unsuppressed serum cortisol after overnight and low-dose dexamethasone suppression test with a suppressed ACTH. CT scan of the adrenal glands revealed a right adrenal cortical adenoma. The risk of complications in infants and mothers who suffer from Cushing's syndrome needs to be handled carefully. The diagnosis of Cushing's syndrome in pregnant women often overlaps and is difficult to establish in early pregnancy.
  9. Borhan MK, Tan FHS, Basry NSA
    J ASEAN Fed Endocr Soc, 2022;37(2):89-94.
    PMID: 36578883 DOI: 10.15605/jafes.037.02.09
    A collision tumor involving metastasis to a pituitary adenoma is rare. We describe a case of a 68-year-old Bidayuh woman with underlying treatment-responsive lung adenocarcinoma, who presented with mass effect, panhypopituitarism and polyuria. Her initial imaging study reported pituitary macroadenoma, and she was treated with hormone replacement therapy. She then underwent transsphenoidal tumor debulking surgery with subsequent histopathological findings of a collision tumor of an adenocarcinoma with metastasis to a non-functioning pituitary adenoma.
  10. Cheah JM, Ng D, Low MY, Foo SH
    J ASEAN Fed Endocr Soc, 2019;34(2):206-209.
    PMID: 33442157 DOI: 10.15605/jafes.034.02.12
    Thyroid crisis is a life-threatening form of thyrotoxicosis characterized by multi-system dysfunction. Therapeutic plasma exchange has been reported to be effective in removing excessive circulating thyroid hormones. We present a 46-year-old female with recently diagnosed Graves' disease associated with thyrotoxic cardiomyopathy admitted for thyroid crisis complicated by propranolol-induced circulatory collapse, acute kidney injury and ischemic hepatitis. The tachyarrhythmia was refractory to conventional therapy. Initiation of TPE resulted in rapid clinical and biochemical stabilization.
  11. Ching WL, Idris AN, Nor NK, Giok LP
    J ASEAN Fed Endocr Soc, 2023;38(2):13-19.
    PMID: 38045679 DOI: 10.15605/jafes.038.02.10
    INTRODUCTION: Type 1 diabetes mellitus (T1DM) is an autoimmune disorder that requires a lifelong treatment regimen which may affect psychosocial development.

    OBJECTIVE: To identify behavioural and emotional problems in children and adolescents with T1DM.

    METHODOLOGY: A cross-sectional study using the Child Behaviour Check List (CBCL) was conducted among all T1DM patients receiving treatment at the Paediatric Endocrine Unit, Hospital Tunku Azizah Kuala Lumpur, Malaysia.

    RESULTS: Forty T1DM patients were included. The mean age of the participants was 12.4 years (SD = 2.69), with 52.5% males, and 75% Malay. The average duration of illness was 4.8 years, 9 were pre-pubertal, while mean HbA1c was 9.4%. Thirty-five percent of the respondents had parent-reported internalizing problems and 17.5% had parent-reported externalizing problems. Those >12 years old had more internalizing problems (p = 0.004) compared to those ≤12 years old. The differences were in the anxious/depressed syndrome subscale (p = 0.001) and withdrawn/depressed syndrome subscale (p = 0.015). There were no statistically significant differences in the 3 main global scores by gender, glycaemic control, duration of illness and pubertal status by univariate analysis.

    CONCLUSION: T1DM patients >12 years old were at higher risk of developing psychosocial difficulties. This highlighted the benefit of screening of behavioural and emotional issues in children and adolescents with T1DM.

  12. Dharan SS, Kamaruddin NA
    J ASEAN Fed Endocr Soc, 2017;32(2):169-172.
    PMID: 33442102 DOI: 10.15605/jafes.032.02.12
    Acromegaly is a rare disease with an annual incidence of 3 to 4 cases in a million.1 Diagnosis is often delayed due to the slow progression of the disease. Persistent elevation of growth hormone (GH) in acromegaly causes a reduction in life expectancy by 10 years. Aside from multiple cardiovascular, respiratory and metabolic co-morbidities, it has also been proven to cause an increased incidence of cancer. The main treatment of acromegaly is surgical excision of the functioning pituitary adenoma. Multiple comorbidities, including obstructive sleep apnea (OSA), left ventricular hypertrophy (LVH) and soft tissue swelling, make surgery complicated, if not impossible. Medical therapy to reduce comorbidities may be indicated in certain situations. Somatostatin receptor ligands (SRL) are able to reduce, and possibly normalize, IGF-1 levels.2 Reduction of insulin-like growth factor-1 (IGF-1), the main mediator of GH, is able to resolve headache, sweating, fatigue and soft tissue swelling, and also reduce ventricular hypertrophy. This case report illustrates the successful use of the SRL octreotide LAR in treating acromegaly. It also confirms the observation from several case series that thyroid cancer is the most common malignancy in acromegaly.
  13. Guan GK, Rajoo S, Dusa NM, Yahya NHN, Bidin MBL
    J ASEAN Fed Endocr Soc, 2021;36(2):220-222.
    PMID: 34966210 DOI: 10.15605/jafes.036.02.09
    Somatostatin analogue is useful in carcinoid crisis for symptom control. Optimal dosing of somatostatin analogues for carcinoid symptoms is not known. This case highlighted management issues using combination short-acting octreotide infusion with long-acting lanreotide during carcinoid crisis. The patient had left lung neuroendocrine tumour that metastasized to his liver and bone, post left lobectomy. Due to extensive metastasis to the liver causing recurrent carcinoid crisis, he required shorter interval long-acting lanreotide with continuous infusion of short-acting octreotide, which led to transient diabetes insipidus. Symptoms resolved with discontinuation of treatment. Somatostatin analogues, especially in combination, may inhibit the posterior pituitary resulting in diabetes insipidus. Prompt withdrawal of short-acting somatostatin analogue and initiation of desmopressin can reverse the complication. It is important to recognize this complication with combination of octreotide and lanreotide injections to avoid serious complications.
  14. Helmi MAM, Hussain S
    J ASEAN Fed Endocr Soc, 2020;35(1):125-128.
    PMID: 33442181 DOI: 10.15605/jafes.035.01.22
    Developmental delay, Epilepsy and Neonatal Diabetes (DEND) syndrome is the most severe form of Permanent Neonatal Diabetes with KCNJ11 gene mutation which accounts for most of the cases. We report the first DEND syndrome in Malaysia with heterozygous missense mutation Q52R at KCNJ11 (Kir6.2) gene with delayed presentation beyond 6 months of age and failure to transition to glibenclamide. This report signifies the phenotypical variability among patients with the same genetic mutation and the different response to treatment.
  15. Hussain S, Elnajeh M
    J ASEAN Fed Endocr Soc, 2020;35(2):176-180.
    PMID: 33442189 DOI: 10.15605/jafes.035.02.05
    OBJECTIVE: We aim to study the prevalence and risk factors of hypovitaminosis D among healthy adolescents in Kota Bharu, Kelantan based on the most recent Paediatric Consensus guideline.

    METHODOLOGY: Ten public schools were selected from Kota Bharu, Kelantan. We analysed their demography (age, gender, ethnicity, income), measured their anthropometry (height, weight, BMI) and finally analysed their vitamin D and intactParathyroid hormone levels.

    RESULTS: The prevalence of hypovitaminosis D was 16.9% among healthy teenagers with mean age of 15.9±1.39 years. Multivariate analysis showed female gender (adjusted OR, 95% CI): 23.7 (5.64, 100.3) and Chinese 0.24 (0.07, 0.84) were the significant predictors for hypovitaminosis D.

    CONCLUSION: The prevalence of healthy adolescents with hypovitaminosis D in Kota Bharu, Kelantan was 16.9% using the most recent cut off value of 30 nmol/L from the global consensus 2016. Female and Malay were the significant risk factors associated with hypovitaminosis D. Higher cut off value would result in overestimation of prevalence rate of hypovitaminosis D.

  16. Hussain S, Men KK, Majid NA
    J ASEAN Fed Endocr Soc, 2017;32(2):132-138.
    PMID: 33442096 DOI: 10.15605/jafes.032.02.06
    Objectives: We aim to compare the clinical and biochemical profile of metabolic syndrome between obese children below and above 10 years attending Paediatric clinic Hospital Universiti Sains Malaysia (HUSM) from 2006 to 2015. This is to determine if age, particularly the transition to puberty, modifies the prevalence of components of metabolic syndrome in obese children.

    Methodology: The medical records of 84 obese children under 18 years of age seen at Paediatric clinic HUSM from 2006 to 2015 were reviewed. Demographic (age, gender, ethnicity), anthropometric (weight and height), clinical [body mass index (BMI), systolic blood pressure (SBP) and diastolic blood pressure (DBP)] and biochemical [serum total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), fasting plasma glucose (FPG)] parameters were recorded, analyzed and compared.

    Results: Majority of subjects in both age groups were boys, with 68.2% <10 years old. Mean age was 9.69 years (±3.36). The clinical and biochemical parameters of metabolic syndrome were similar between those <10 years old and >10 years, with the exception of BMI, waist circumference, SBP and TG level. Multivariate regression analysis showed that the parameters of metabolic syndrome significantly associated with age ≥10 years were systolic hypertension (adjusted OR 7.17, 95% CI, 1.48 to 34.8) and BMI >30 kg/m2 (adjusted OR 3.02, 95% CI, 1.16 to 7.86).

    Conclusion: There were similar clinical and biochemical parameters of metabolic syndrome in both age groups. The proportions of children with metabolic syndrome were similar regardless of age group. The overall prevalence rate of metabolic syndrome was 27.3%. In view of the alarming presence of components of metabolic syndrome even in children less than 10 years of age, efforts aimed at the prevention of childhood obesity in the community should be intensified.

  17. Hussein Z, Bidin ML, Alias A, Thiagarajan M, Latif KA, Ratnasingam J, et al.
    J ASEAN Fed Endocr Soc, 2019;34(1):8-14.
    PMID: 33442131 DOI: 10.15605/jafes.034.01.03
    In Malaysia, acromegaly is under-recognised with only 10-15% of the expected number of cases from prevalence estimates, having been diagnosed and managed in established endocrine centres with access to multidisciplinary care. This is mainly due to lack of awareness and standardised approach in diagnosing this disease resulting in delay in diagnosis and management with suboptimal treatment outcomes. This first Malaysian consensus statement on the diagnosis and management of acromegaly addresses these issues and is based on current best practices and latest available evidence so as to reduce the disease burden on acromegaly patients managed in the Malaysian healthcare system.
  18. Im Teoh JH, Mustafa N, Wahab N
    J ASEAN Fed Endocr Soc, 2023;38(1):125-130.
    PMID: 37252417 DOI: 10.15605/jafes.038.01.19
    Autoimmunity associated with severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been well-described as the mechanism of development of thyroid dysfunction following Coronavirus Disease 19 (COVID-19) infection and SARS-CoV-2 vaccination. However, the occurrence of thyroid eye disease (TED) after SARS-CoV-2 vaccination is scarcely described. The postulated mechanisms include immune reactivation, molecular mimicry and the autoimmune/inflammatory syndrome induced by adjuvants (ASIA). We report a case of new-onset TED after receiving the SARS-CoV-2 vaccine.
  19. Jasul G, Paz-Pacheco E, Jimeno C, Suastika K, Hussein Z, Mustafa N, et al.
    J ASEAN Fed Endocr Soc, 2020;35(1):5-13.
    PMID: 33790494 DOI: 10.15605/jafes.035.01.10
    OBJECTIVES: The COVID-19 pandemic has made a major impact on hospital services globally, including the care of persons with diabetes and endocrine disorders. The aim of this study is to describe the epidemiology of COVID-19 in the ASEAN Federation of Endocrine Societies (AFES) member countries; to describe challenges, changes and opportunities in caring for patients with endocrine diseases, as well as in fellowship training programs, and endocrinerelated research in the AFES countries.

    METHODOLOGY: The AFES ASEAN Survey Of Needs in Endocrinology (AFES A.S.-O.N.E.) was an open-ended questionnaire that was sent to the presidents and representatives of the AFES member countries by email. Responses from Societies were collated and synthesized to obtain perspectives on the emergent issues in endocrinology in the Southeast Asian region during this pandemic.

    RESULTS: The burden of COVID-19 cases varied widely across the AFES member countries, with the least number of cases in Vietnam and Myanmar, and the greatest number of cases in either the most populous countries (Indonesia and the Philippines), or a country with the highest capability for testing (Singapore). The case fatality rate was also the highest for Indonesia and the Philippines at around 6%, and lowest for Vietnam at no fatalities. The percentage with diabetes among patients with COVID-19 ranged from 5% in Indonesia to 20% in Singapore, approximating the reported percentages in China and the United States. The major challenges in managing patients with endocrine diseases involved inaccessibility of health care providers, clinics and hospitals due to the implementation of lockdowns, community quarantines or movement control among the member countries. This led to disruptions in the continuity of care, testing and monitoring, and for some, provision of both preventive care and active management including surgery for thyroid cancer or pituitary and adrenal tumors, and radioactive iodine therapy. Major disruptions in the endocrine fellowship training programs were also noted across the region, so that some countries have had to freeze hiring of new trainees or to revise both program requirements and approaches to training due to the closure of outpatient endocrine clinics. The same observations are seen for endocrine-related researches, as most research papers have focused on the pandemic. Finally, the report ends by describing innovative approaches to fill in the gap in training and in improving patient access to endocrine services by Telemedicine.

    CONCLUSION: The burden of COVID-19 cases and its case fatality rate varies across the AFES member countries but its impact is almost uniform: it has disrupted the provision of care for patients with endocrine diseases, and has also disrupted endocrine fellowship training and endocrine-related research across the region. Telemedicine and innovations in training have been operationalized across the AFES countries in an attempt to cope with the disruptions from COVID-19, but its over-all impact on the practice of endocrinology across the region will only become apparent once we conquer this pandemic.

  20. Jiang CY, Shen BS, Yun TZ, Singarayar C, Hui FS
    J ASEAN Fed Endocr Soc, 2021;36(2):213-215.
    PMID: 34966208 DOI: 10.15605/jafes.036.02.10
    Levothyroxine remains the standard therapy for patients with hypothyroidism worldwide. Levothyroxine allergy is rarely seen and alternative therapies are less efficacious and scarcely available. The use of liothyronine (LT3) monotherapy is less favoured due to its short half-life and unpredictable pharmacological profile. We report a 59-year-old male with a hypersensitivity reaction to levothyroxine who was successfully desensitized with oral levothyroxine within a day using a 14-step protocol.
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