Methadone Maintenance Treatment for Opioid Dependents: a Retrospective Study.

OBJECTIVES: To determine the prevalence of continued illicit drug use among people enrolled in methadone maintenance treatment (MMT), the association between hepatitis C status and methadone dosage, and the predictors for illicit drug abstinence during MMT. METHODS: Clinical records of active opioid dependents who underwent MMT between 1 January 2007 and 31 March 2021 in Hospital Tuanku Fauziah, Perlis, Malaysia were retrospectively reviewed. Data collected included baseline demographics, history of illicit drug use, temporal trend in methadone dosage modulation, and co-use of illicit drugs during the MMT. RESULTS: A total of 87 patients (mean age, 43.9 ± 8.33 years) were included. Their mean duration of involvement in MMT was 7.8 ± 3.69 years. The most commonly used drug was heroin (88.5%), followed by kratom (51.7%). Between 2019 and 2021, 61 (70.1%) patients had ceased abusing opioid, but 51 (58.6%) patients continued using any of the illicit drugs. Methamphetamine and amphetamine co-use was most common (n = 12, 37.5%). Hepatitis C status was not associated with the current methadone dose (U = 539.5, p = 0.186) or the highest dose required (t = -0.291, df = 74, p = 0.772). No predictor for illicit drug abstinence during MMT was identified. Methadone dose positively correlated with frequency of defaulting treatments (r = 0.22, p = 0.042). CONCLUSION: Among our patients, MMT for opioid dependents cannot sufficiently curb illicit drug use, and there is a shift toward stimulants abuse.

Application of a new system for classifying root and canal anatomy in studies involving micro-computed tomography and cone beam computed tomography: Explanation and elaboration.

Adequate knowledge and accurate characterization of root and canal anatomy is an essential prerequisite for successful root canal treatment and endodontic surgery. Over the years, an ever-increasing body of knowledge related to root and canal anatomy of the human dentition has accumulated. To correct deficiencies in existing systems, a new coding system for classifying root and canal morphology, accessory canals and anomalies has been introduced. In recent years, micro-computed tomography (micro-CT) and cone beam computed tomography (CBCT) have been used extensively to study the details of root and canal anatomy in extracted teeth and within clinical settings. This review aims to discuss the application of the new coding system in studies using micro-CT and CBCT, provide a detailed guide for appropriate characterization of root and canal anatomy and to discuss several controversial issues that may appear as potential limitations for proper characterization of roots and canals.

Deep learning architectures analysis for age-related macular degeneration segmentation on optical coherence tomography scans.

BACKGROUND AND OBJECTIVES: Aged people usually are more to be diagnosed with retinal diseases in developed countries. Retinal capillaries leakage into the retina swells and causes an acute vision loss, which is called age-related macular degeneration (AMD). The disease can not be adequately diagnosed solely using fundus images as depth information is not available. The variations in retina volume assist in monitoring ophthalmological abnormalities. Therefore, high-fidelity AMD segmentation in optical coherence tomography (OCT) imaging modality has raised the attention of researchers as well as those of the medical doctors. Many methods across the years encompassing machine learning approaches and convolutional neural networks (CNN) strategies have been proposed for object detection and image segmentation. METHODS: In this paper, we analyze four wide-spread deep learning models designed for the segmentation of three retinal fluids outputting dense predictions in the RETOUCH challenge data. We aim to demonstrate how a patch-based approach could push the performance for each method. Besides, we also evaluate the methods using the OPTIMA challenge dataset for generalizing network performance. The analysis is driven into two sections: the comparison between the four approaches and the significance of patching the images. RESULTS: The performance of networks trained on the RETOUCH dataset is higher than human performance. The analysis further generalized the performance of the best network obtained by fine-tuning it and achieved a mean Dice similarity coefficient (DSC) of 0.85. Out of the three types of fluids, intraretinal fluid (IRF) is more recognized, and the highest DSC value of 0.922 is achieved using Spectralis dataset. Additionally, the highest average DSC score is 0.84, which is achieved by PaDeeplabv3+ model using Cirrus dataset. CONCLUSIONS: The proposed method segments the three fluids in the retina with high DSC value. Fine-tuning the networks trained on the RETOUCH dataset makes the network perform better and faster than training from scratch. Enriching the networks with inputting a variety of shapes by extracting patches helped to segment the fluids better than using a full image.

Lactobacillus plantarum DR7 alleviates stress and anxiety in adults: a randomised, double-blind, placebo-controlled study.

Probiotics have been reported to exert beneficial effects along the gut-brain axis. This randomised, double-blind and placebo-controlled human study aimed to evaluate such properties of Lactobacillus plantarum DR7 and its accompanying mechanisms in stressed adults. One hundred and eleven (n=111; DR7 n=56, placebo n=55) stressed adults were recruited based on moderate stress levels using the PSS-10 questionnaire. The consumption of DR7 (1×109 cfu/day) for 12 weeks reduced symptoms of stress (P=0.024), anxiety (P=0.001), and total psychological scores (P=0.022) as early as 8 weeks among stressed adults compared to the placebo group as assessed by the DASS-42 questionnaire. Plasma cortisol level was reduced among DR7 subjects as compared to the placebo, accompanied by reduced plasma pro-inflammatory cytokines, such as interferon-γ and transforming growth factor-α and increased plasma anti-inflammatory cytokines, such as interleukin 10 (P<0.05). DR7 better improved cognitive and memory functions in normal adults (>30 years old), such as basic attention, emotional cognition, and associate learning (P<0.05), as compared to the placebo and young adults (<30 years old). The administration of DR7 enhanced the serotonin pathway, as observed by lowered expressions of plasma dopamine ß-hydroxylase (DBH), tyrosine hydroxylase (TH), indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase accompanied by increased expressions of tryptophan hydroxylase-2 and 5-hydroxytryptamine receptor-6, while stabilising the dopamine pathway as observed via stabilised expressions of TH and DBH over 12 weeks as compared to the placebo (P<0.05). Our results indicated that DR7 fulfil the requirement of a probiotic strain as per recommendation of FAO/WHO and could be applicable as a natural strategy to improve psychological functions, cognitive health and memory in stressed adults.

ß-Cyclodextrin conjugated bifunctional isocyanate linker polymer for enhanced removal of 2,4-dinitrophenol from environmental waters.

In this work, we reported the synthesis, characterization and adsorption study of two ß-cyclodextrin (ßCD) cross-linked polymers using aromatic linker 2,4-toluene diisocyanate (2,4-TDI) and aliphatic linker 1,6-hexamethylene diisocyanate (1,6-HDI) to form insoluble ßCD-TDI and ßCD-HDI. The adsorption of 2,4-dinitrophenol (DNP) on both polymers as an adsorbent was studied in batch adsorption experiments. Both polymers were well characterized using various tools that include Fourier transform infrared spectroscopy, thermogravimetric analysis, Brunauer-Emmett-Teller analysis and scanning electron microscopy, and the results obtained were compared with the native ßCD. The adsorption isotherm of 2,4-DNP onto polymers was studied. It showed that the Freundlich isotherm is a better fit for ßCD-TDI, while the Langmuir isotherm is a better fit for ßCD-HMDI. The pseudo-second-order kinetic model represented the adsorption process for both of the polymers. The thermodynamic study showed that ßCD-TDI polymer was more favourable towards 2,4-DNP when compared with ßCD-HDI polymer. Under optimized conditions, both ßCD polymers were successfully applied on various environmental water samples for the removal of 2,4-DNP. ßCD-TDI polymer showed enhanced sorption capacity and higher removal efficiency (greater than 80%) than ßCD-HDI (greater than 70%) towards 2,4-DNP. The mechanism involved was discussed, and the effects of cross-linkers on ßCD open up new perspectives for the removal of toxic contaminants from a body of water.

Simultaneous use of adzuki beans (Vigna angularis) flour as meat extender and fat replacer in reduced-fat beef meatballs (bebola daging).

Adzuki bean is high in protein and fiber with a potential to be used as meat extender and fat replacer in the meat product. Replacement of both the corn flour and fat with different percentages of adzuki beans flour (ABF) has successfully produced acceptable reduced fat meatballs. Meatballs with 100% (w/w) ABF replacement exhibited highest cooking yield and higher moisture content compared to meatballs without the flour, which indicates its ability to bind water. Increasing the ABF content also increased the hardness and chewiness of the meatballs, whilst decreasing their lightness and yellowness. Replacing the corn flour and fat contents with ABF has obviously decreased the fat and calorie contents of the meatballs, yet their protein and carbohydrate contents remained the same compared to control. The sensory test revealed that meatball samples with 25% (w/w) and 50% (w/w) ABF showed no significant difference compared to control but received highest overall acceptability among the panelists. This indicates that replacement of corn flour and fat with ABF especially at 50% (w/w) in the production of reduced fat meatballs resulted with better physicochemical properties and acceptable sensory compared to original meatballs.

Prevalence of optimal cardiovascular screening activities and associated factors among apparently healthy school teachers in Kota Bharu, Kelantan.

OBJECTIVES: This study aimed to determine the prevalence of optimal cardiovascular disease (CVD) screening activities and the associated factors among apparently healthy school teachers. METHODS: A cross-sectional analytical observational study was conducted among 380 secondary school teachers in Kelantan, Malaysia. A self-administered questionnaire addressing sociodemographic data and factors influencing CVD screening activities was administered. Descriptive analysis, simple and multiple logistic regression analyses were performed. RESULTS: A total of 348 teachers responded to the questionnaire, with a response rate of 91.6%. The prevalence of optimal CVD screening activities was 29.3% (95% CI: 24.52, 34.08). Age, knowledge of CVD screening, family history of CVD and availability of health facilities were significantly linked to CVD screening. CONCLUSION: The prevalence of optimal screening activities was low. A great majority of the factors contributing to optimal screening were modifiable. Health care providers should widely implement global health-oriented rather than disease-orientated assessment in their daily practice.

Understanding the Perception of Islamic Medicine Among the Malaysian Muslim Community.

This study was conducted to identify and describe the patients' perceptions of Islamic medicine based on gender, age, marital, educational level and working status among the Malaysian Muslim population. A nationwide interviewer-administered questionnaire survey was conducted in 2013. An open-ended questionnaire pertaining to Islamic medicine was used to increase the probability of capturing maximum data. This survey implemented a multistage design, stratified by state, proportionate to the size of the state population and was representative of the Malaysian population. Post-survey classification of results was performed accordingly. Complex data analysis was carried out using SPSS 16.0. The discussion was identified and categorised into various sections. The paper concludes that Islamic medicine has a major influence in the Malaysian Muslim community compared to other alternatives. Further, its potential for growth and importance especially for treating spiritual ailments cannot be denied. The respondents indicated that two factors motivate Islamic medicine in Malaysia: (1) the Muslim community opts for alternative healing because of their dissatisfaction with conventional methods; (2) Islamic medicine focuses only on healing spiritual-related problems. The average perception of respondents is that the function of Islamic medicine in healing physical diseases is undervalued and that it is not suitable to replace the functions of modern health institutions.

Detection of α-thalassaemia in neonates on cord blood and dried blood spot samples by capillary electrophoresis.

INTRODUCTION: Haemoglobin Bart's (Hb Bart's) level is associated with α-thalassaemia traits in neonates, enabling early diagnosis of α-thalassaemia. The study aimed to detect and quantify the Hb Bart's using Cord Blood (CB) and CE Neonat Fast Hb (NF) progammes on fresh and dried blood spot (DBS) specimen respectively by capillary electrophoresis (CE). METHODS: Capillarys Hemoglobin (E) Kit (for CB) and Capillarys Neonat Hb Kit (for NF) were used to detect and quantify Hb Bart's by CE in fresh cord blood and dried blood spot (DBS) specimens respectively. High performance liquid chromatography (HPLC) using the ß-Thal Short Programme was also performed concurrently with CE analysis. Confirmation was obtained by multiplex ARMS Gap PCR. RESULTS: This study was performed on 600 neonates. 32/600 (5.3%) samples showed presence of Hb Bart's peak using the NF programme while 33/600 (5.5%) were positive with CB programme and HPLC methods. The range of Hb Bart's using NF programme and CB programme were (0.5-4.1%) and (0.5-7.1%), respectively. Molecular analysis confirmed all positive samples possessed α-thalassaemia genetic mutations, with 23/33 cases being αα/--SEA, four -α3.7/-α3.7, two αα/-α3.7 and three αα/ααCS. Fifty Hb Bart's negative samples were randomly tested for α-genotypes, three were also found to be positive for α-globin gene mutations. Thus, resulting in sensitivity of 91.7% and 88.9% and specificity of 100% for the Capillarys Cord Blood programme and Capillarys Neonat Fast programme respectively. CONCLUSION: Both CE programmes using fresh or dried cord blood were useful as a screening tool for α-thalassaemia in newborns. All methods show the same specificity (100%) with variable, but acceptable sensitivities in the detection of Hb Bart.

Streptomyces sanglieri which colonised and enhanced the growth of Elaeis guineensis Jacq. seedlings was antagonistic to Ganoderma boninense in in vitro studies.

Actinomycete strain AUM 00500 was 99.5 % similar to Streptomyces sanglieri NBRC 100784(T) and was evaluated for antagonistic activity towards Ganoderma boninense, the causative fungus of basal stem rot of oil palm. The strain showed strong antifungal activity towards G. boninense in in vitro and SEM analysis showed various modes of inhibition of the fungus. Ethyl acetate extracts of single culture and inhibition zone of cross-plug culture by HPLC indicated that strain AUM 00500 produced two different antibiotics of the glutarimide group namely cycloheximide and actiphenol. In greenhouse trials, oil palm seed treated with spores of S. sanglieri strain AUM 00500 at 10(9) cfu/ml showed significant (P < 0.05) increase in oil palm seedlings growth when compared to the control. Streptomyces sanglieri strain AUM 00500 successfully colonised the epidermal surface of the roots of treated oil palm seedlings and it was recovered from root fragments plated on starch casein agar.

Medication adherence in schizophrenia: a comparison between outpatients and relapse cases.

OBJECTIVE: Adherence to medication is essential for maximising the outcomes of patients with schizophrenia as the consequences of poor adherence are devastating. The study aimed to compare medication adherence between patients with relapse schizophrenia and those attending psychiatric follow-up clinics, and to determine the factors affecting adherence. METHODS: This was a cross-sectional study involving 70 patients with schizophrenia who were divided equally into 2 groups. Medication adherence was assessed with the Medication Adherence Rating Scale. Appropriate instruments were used to measure insight, social support, and psychopathology. Various socio-demographic and clinical variables were explored to find associations with medication adherence. RESULTS: Medication adherence among patients with schizophrenia was poor; 51% of the patients did not adhere to a medication regimen. Adherence was better in outpatients with schizophrenia (61%) than in relapse cases (39%), although the difference was not statistically significant (t = 1.70; p = 0.09). Besides, relapse patients had significant higher number of admission (X(2) = 22.95; p < 0.05) and severe psychopathology (t = -29.96; p < 0.05), while perceived social support was significantly better in outpatients with schizophrenia (t = 2.90; p < 0.05). Frequency of admission (adjusted b = -0.55; 95% confidence interval [CI], -0.99 to -0.10; p < 0.05) and psychopathology (adjusted b = -0.12; 95% CI, -0.24 to -0.01; p < 0.05) were also significantly associated with medication adherence. CONCLUSION: Medication adherence among both groups of patients with schizophrenia was poor. If adherence is addressed appropriately, the number of admissions and severity of psychopathology could be improved.

Gender differences in coping skills of parents with hearing-impaired children.

OBJECTIVE: To assess differences between the coping strategies of the mothers and fathers with hearing-impaired children. METHODOLOGY: A cross-sectional study was conducted looking at parents of children with moderate to profound hearing impairment. The parents with more than one child with a hearing impairment, the parents of children with additional disabilities and syndromes, single parents, and parents with their own hearing impairments were excluded from the study. A Brief COPE Scale questionnaire translated into Malay was used to assess the coping strategies. The questionnaire includes 28 items and was rated using a four-point Likert scale. Independent t-testing was used to compare the coping strategies of mothers and fathers. Simple linear regression was used to determine the association between age and coping strategies. RESULTS: There were 72 participants. The number of mothers and fathers was equal. Religion, active coping and acceptance were the highest three scores in the domains, while substance use and behavioural disengagement were least used in both groups. The domains of religion, seeking emotional support and seeking instrumental support scored significantly higher in mothers than in fathers (p < 0.05). There was a significant difference between the two groups in terms of problem-focused strategies (p = 0.016) but not in terms of emotional-focused strategies (p = 0.134). There were significant negative linear correlations between age and seeking emotional support, humour and instrumental support ( p < 0.05). CONCLUSION: There were gender differences in the coping strategies among parents with hearing-impaired children. These are important factors that should be considered when counselling and establishing support groups for the parents of these children.

Interleukin-27 exhibited anti-inflammatory activity during Plasmodium berghei infection in mice.

Interleukin-27 (IL-27) has a pleiotropic role either as a pro-inflammatory or anti-inflammatory cytokine in inflammatory related diseases. The role and involvement of IL-27 during malaria was investigated and the effects of modulating its release on the production of major inflammatory cytokines and the histopathological consequences in major affected organs during the infection were evaluated. Results showed that IL-27 concentration was significantly elevated throughout the infection but no positive correlation with the parasitaemia development observed. Augmentation of IL-27 significantly elevated the release of anti-inflammatory cytokine, IL-10 whereas antagonising and neutralising IL-27 produced the opposite. A significant elevation of pro-inflammatory cytokines (IFN-γ and IL-6) was also observed, both during augmentation and inhibition of IL-27. Thus, it is suggested that IL-27 exerts an anti-inflammatory activity in the Th1 type response by signalling the production of IL-10 during malaria. Histopathological examination showed sequestration of PRBC in the microvasculature of major organs in malarial mice. Other significant histopathological changes include hyperplasia and hypertrophy of the Kupffer cells in the liver, hyaline membrane formation in lung tissue, enlargement of the white and red pulp followed by the disappearance of germinal centre of the spleen, and tubular vacuolation of the kidney tissues. In conclusion, it is suggested that IL-27 may possibly acts as an anti-inflammatory cytokine during the infection. Modulation of its release produced a positive impact on inflammatory cytokine production during the infection, suggesting its potential in malaria immunotherapy, in which the host may benefit from its inhibition.

Random nanowires of nickel doped TiO2 with high surface area and electron mobility for high efficiency dye-sensitized solar cells.

Mesoporous TiO(2) with a large specific surface area (~150 m(2) g(-1)) is the most successful material in dye-sensitized solar cells so far; however, its inferior charge mobility is a major efficiency limiter. This paper demonstrates that random nanowires of Ni-doped TiO(2) (Ni:TiO(2)) have a dramatic influence on the particulate and charge transport properties. Nanowires (dia ~60 nm) of Ni:TiO(2) with a specific surface area of ~80 m(2) g(-1) were developed by an electrospinning technique. The band gap of the Ni:TiO(2) shifted to the visible region upon doping of 5 at% Ni atoms. The Mott-Schottky analysis shows that the flat band potential of Ni:TiO(2) shifts to a more negative value than the undoped samples. The electrochemical impedance spectroscopic measurements showed that the Ni:TiO(2) offer lower charge transport resistance, higher charge recombination resistance, and enhanced electron lifetime compared to the undoped samples. The dye-sensitized solar cells fabricated using the Ni:TiO(2) nanowires showed an enhanced photoconversion efficiency and short-circuit current density compared to the undoped analogue. The transient photocurrent measurements showed that the Ni:TiO(2) has improved charge mobility compared with TiO(2) and is several orders of magnitude higher compared to the P25 particles.

Interleukin-27 exhibited anti-inflammatory activity during Plasmodium berghei infection in mice

Interleukin-27 (IL-27) has a pleiotropic role either as a pro-inflammatory or anti-inflammatory cytokine in inflammatory related diseases. The role and involvement of IL-27 during malaria was investigated and the effects of modulating its release on the production of major inflammatory cytokines and the histopathological consequences in major affected organs during the infection were evaluated. Results showed that IL-27 concentration was significantly elevated throughout the infection but no positive correlation with the parasitaemia development observed. Augmentation of IL-27 significantly elevated the release of anti-inflammatory cytokine, IL-10 whereas antagonising and neutralising IL-27 produced the opposite. A significant elevation of pro-inflammatory cytokines (IFN-γ and IL-6) was also observed, both during augmentation and inhibition of IL-27. Thus, it is suggested that IL-27 exerts an anti-inflammatory activity in the Th1 type response by signalling the production of IL-10 during malaria. Histopathological examination showed sequestration of PRBC in the microvasculature of major organs in malarial mice. Other significant histopathological changes include hyperplasia and hypertrophy of the Kupffer cells in the liver, hyaline membrane formation in lung tissue, enlargement of the white and red pulp followed by the disappearance of germinal centre of the spleen, and tubular vacuolation of the kidney tissues. In conclusion, it is suggested that IL-27 may possibly acts as an anti-inflammatory cytokine during the infection. Modulation of its release produced a positive impact on inflammatory cytokine production during the infection, suggesting its potential in malaria immunotherapy, in which the host may benefit from its inhibition.

Obstructive sleep apnea among express bus drivers in Malaysia: important indicators for screening.

OBJECTIVES: Obstructive sleep apnea (OSA) has been identified as one of the significant risk factors for motor vehicle crashes (MVCs). In the interest of public safety, this study was conducted to determine the prevalence of OSA and its associated factors among express bus drivers in Malaysia. Identifying factors or conditions related with OSA is very important because they can be used as indicators to subject a person to a confirmatory diagnosis using polysomnography testing. METHODS: Two hundred eighty-nine randomly selected express bus drivers from 5 express bus companies participated in the study. Information on demography, medical history, clinical symptoms, and signs of OSA were collected by a designated medical officer and the diagnosis of OSA was done based on the Apnea Hypopnoea Index (AHI) from polysomnography testing. RESULTS: Based on AHI, 128 (44.3%) subjects were diagnosed as having OSA with 83 (28.7%), 26 (9.0%), and 26 (6.6%) classified as mild, moderate, and severe OSA, respectively. Multiple logistic regression analysis results showed that age (odds ratio [OR] = 1.05, 95% confidence interval [CI] 1.01-1.09), snoring (OR = 3.54, 95% CI 1.91-6.57), body mass index (BMI; OR = 1.17, 95% CI 1.09-1.25), hypertension (OR = 1.87, 95% CI 1.02-3.40), and neck circumference (OR = 1.31, 95% CI 1.18-1.46) were significantly associated with OSA status. CONCLUSIONS: The results supported the need for identifying the risk group for OSA among express bus drivers and the need to diagnose them early for an early intervention.

Bacteriostatic effect of Piper betle and Psidium guajava extracts on dental plaque bacteria.

In this study, the bacteriostatic effect of Piper betle and Psidium guajava extracts on selected early dental plaque bacteria was investigated based on changes in the doubling time (g) and specific growth rates (micro). Streptococcus sanguinis, Streptococcus mitis and Actinomyces sp. were cultured in Brain Heart Infusion (BHI) in the presence and absence of the extracts. The growth of bacteria was monitored periodically every 15 min over a period of 9 h to allow for a complete growth cycle. Growth profiles of the bacteria in the presence of the extracts were compared to those in the absence and deviation in the g and micro were determined and analyzed. It was found that the g and mu were affected by both extracts. At 4 mg mL(-1) of P. betle the g-values for S. sanguinis and S. mitis were increased by 12.0- and 10.4-fold, respectively (p < 0.05). At similar concentration P. guajava increased the g-value by 1.8- and 2.6 -fold, respectively (p < 0.05). The effect on Actinomyces sp. was observed at a much lower magnitude. It appears that P. betle and P. guajava extracts have bacteriostatic effect on the plaque bacteria by creating a stressed environment that had suppressed the growth and propagation of the cells. Within the context of the dental plaque, this would ensure the attainment of thin and healthy plaque. Thus, decoctions of these plants would be suitable if used in the control of dental plaque.

Ultrastructural studies of developing goat oocytes in vitro.

The structure and distribution of organelles within developing goat oocytes at various stages of incubation were studied. In oocytes with 5 or more layers of cumulus cells, at 0 h of incubation, the zona pellucida had developed although zonation was not evident. Lipid bodies were present but no mitochondria were observed. At 20 h, the zona pellucida had differentiated into thicker and thinner regions. Clusters of membrane-bound electron-transparent bodies were present in the perivitelline space. The mitochondria were fully developed, distributed evenly and usually in close proximity with dilated endoplasmic reticula. Cortical granules were distributed at the periphery. At 40 h of incubation, a number of mitochondria was hooded. In oocytes of 2 to 4 layers of cumulus cells at 0 h, the zona pellucida was penetrated by cumulus cell processes, and the mitochondria were not well developed. However, in 20-h incubated oocytes, fully developed mitochondria, many of which were hooded, could be observed. Clusters of membrane-bound electron-transparent bodies were also observed, while cortical granules were at the periphery. In cumulus-free oocytes, zonation within the zona pellucida was indistinct. Very few vesicles and lipid bodies were observed. At 20 h, mitochondria were sparsely distributed and were not well developed and lacked cristae. At 40 h, the zona pellucida was less compact, and the membrane-bound electron-transparent bodies were less numerous compared with those of the other groups. Endoplasmic reticula were not dilated, and cortical granules were few and had no definite pattern of distribution.