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Abstract Background Despite impressive strides in health, social protection, and education, children continue to experience high rates of child maltreatment in Malaysia. This mixed-methods study assessed the feasibility of a five-session, social learning-based parenting program delivered by government staff in a community setting to reduce violence against children. Methods Parents of children from birth to 17 years were recruited from two communities near Kuala Lumpur to participate in the government-run program called the Naungan Kasih Positive Parenting Program (“Protecting through Love” in Bahasa Melayu). Quantitative data from female caregivers (N = 74) and children ages 10–17 (N = 26) were collected along with qualitative interviews and focus groups with parents, children, and facilitators. The primary outcome was child maltreatment with secondary outcomes including neglect, positive parenting, acceptability of corporal punishment, harsh parenting, positive discipline, and child behavior problems. Multilevel Poisson regression and multilevel linear regression were conducted to compare baseline and post-test outcomes. Qualitative interviews and focus groups examined how participants experienced the program utilizing a thematic analysis approach. Results Quantitative analyses found pre-post reductions in overall child maltreatment, physical abuse, emotional abuse, attitudes supporting corporal punishment, parent sense of inefficacy, and child behavior problems. There were no reported changes on positive and harsh parenting, parental mental health, and marital satisfaction, nor were there any other significant changes reported by children. Qualitative findings suggested that the program had tangible benefits for female caregivers involved in the program, with the benefits extending to their family members. Conclusions This feasibility study is one of the few studies in Southeast Asia that examined the feasibility and initial program impact of a parenting program delivered by government staff to families with children across the developmental spectrum from birth to 17 years. Promising results suggest that the program may reduce child maltreatment across a range of child ages. Findings also indicate areas for program improvement prior to further delivery and testing, including additional training and content on sexual and reproductive health, parenting children with disabilities, and online child protection.

Intro: A national SARS-CoV-2 genome sequencing consortium was established in Aug 2021 to help monitor the emergence of the COVID-19 variants. Tropical Infectious Diseases Research and Education Centre (TIDREC) received samples from the states of Kedah, Kelantan, Perak, and Terengganu between Aug 2021 and March 2022 and samples from Kedah and Perlis from 1 Apr 2022. In the current study, we sequenced and analyzed the SARS-CoV-2 strains received from Kedah between July 2021 to June 2022. Methods: The RNA was extracted and screened for the presence of SARS-CoV-2 RNA using qRT-PCR. Samples with a Ct value less than 32 were selected for sequencing. The genome libraries were prepared using Ion AmpliSeq™ SARSCoV-2 Research Panel and sequenced using the Ion GeneStudio™ S5 Prime System. The genome sequences were assembled, and SARS-CoV-2 lineages were assigned using PANGOLIN COVID-19 Lineage Assigner. Findings: A total of 310 samples were sequenced and analyzed. Results showed that all 310 samples belonged to two groups of Variants of Concern (VOC), Delta and Omicron. Our results revealed that between Aug 2022 and mid-December, the Delta variant was the only circulating VOC in Kedah, with AY.59 as the dominant circulating lineage. The Omicron, BA.1.1, was first detected on 21 Dec 2022 in our collection. The Delta and Omicron variants were co-circulated in Kedah between Jan and Feb 2022. After that, the Omicron BA.2 lineage was reported in Feb 2022, and it replaced the BA.1 and its descendent lineages as the dominant lineage starting in March 2022. The Omicron BA.5 lineage was first detected in samples collected on 26 May 2022, suggesting the emergence of BA.5 in Kedah. Conclusion: We reported the continuous introduction, dispersal, and replacement of SARS-CoV-2 variants in Kedah, Malaysia. Continuous monitoring of the circulating SARS-CoV-2 is vital to detect the emergence of newer SARSCoV-2 variants.

INTRODUCTION: Online interventions have emerged as innovative solutions to address health issues, notably obesity, while minimizing the necessity for in-person interactions during the recent COVID-19 pandemic. There is urgency in designing and developing an appropriate online intervention module within a web-based application. This study aims to describe the design process and usability of the online Canteen Operators, Parents, and Teachers (CoPT Nutri Trail) module for an upcoming school-based obesity intervention. MATERIALS AND METHODS: The design methodology consists of four phases: content development, design, web app development, and usability testing. The web application was designed to personalize usage for teachers, parents, and canteen operators to foster healthy environments for children. Thirteen experts, including teachers, nutritionists, and IT professionals, participated in the usability testing. RESULTS: All experts agreed that the design of the web application met the requirements of target users, requiring slight improvements in graphic design, language, and information elements. The web application tool was found acceptable, as the usability of alpha scores for all interface elements ranged from 68.2% to 86.6%, falling between good and very good scores. The content validation indices indicate good levels of validity (ranging from 0.83 to 1.00). CONCLUSION: The CoPT Nutri Trail web application is ready to be launched as a school-based intervention for obesity. [ABSTRACT FROM AUTHOR]

Objective: hCG is commonly used as an ovulation trigger in IVF. Its usage is associated with OHSS. GnRH agonist is an alternative to hCG and is associated with reduced incidence of OHSS. This study compared the cycle outcomes of GnRH agonists with hCG as an ovulation trigger in IVF cycles., Methods: The medical notes of 209 IVF cycles receiving GnRH agonist and hCG as ovulation trigger over 18 months were reviewed in this retrospective study. The number and quality of mature oocytes, the number and quality of embryos, pregnancy rates, and outcomes were compared using Independent T-test or One-way ANOVA for normal distribution. The Mann-Whitney test or Kruskal-Wallis test was used for not normally distributed. p<0.05 was considered statistically significant., Results: The cycle outcomes of 107 GnRH agonist-trigger and 102 hCG-trigger were compared. The MII oocytes retrieved and 2PN count was significantly higher in the GnRH agonist trigger group (p<0.001). Clinical pregnancy rate and ongoing pregnancy were higher in the GnRH agonist trigger group but were not statistically significant. The GnRH agonist trigger group was associated with low OHSS than the hCG trigger group (n=2(1.9%) and n=12(11.8%) respectively, p=0.004)., Conclusion: GnRH agonist trigger is an option as a final maturation trigger in high-responder women undergoing IVF or ICSI cycles.

The first-line treatment of diabetes ketoacidosis (DKA) involves fluid resuscitation with normal saline infusion to correct hypovolemia. Hyperchloremic metabolic acidosis from aggressive normal saline administration was associated with worse clinical outcomes in managing DKA. Other choices for normal saline include balanced electrolyte solutions (BESs). This study aimed to compare the clinical effects between BESs and normal saline in managing DKA. This study was a systematic review of probing articles published from inception to October 2021 in Cochrane Central Register of Controlled Trials, Medical Literature Analysis and Retrieval System Online, Google Scholar, and Scopus. Eight randomized controlled trials with a total of 595 individuals were included. The data were analyzed at 95% confidence level using random-effects models. For the primary outcomes, there was no difference in the duration of DKA resolution. (Mean difference [MD] -4.73, 95% confidence interval [CI] -2.72-4.92; I 2 = 92%; P = 0.180). However, there was a significantly lower postresuscitation chloride concentration in the BES (MD 2.96 95% CI - 4.86 to - 1.06; I 2 = 59%; P = 0.002). For the secondary outcomes, there was a significant reduction in duration for normalization of bicarbonate in the BES group (MD 3.11 95% CI - 3.98-2.23; I 2 = 5%; P = 0.0004). There were no significant differences between groups in duration for recovery of pH, intensive unit admission, and adverse events (mortality and acute renal failure). Resuscitation with BES was associated with decreased chloride and increased bicarbonate values in DKA patients. It suggests that BES prevents DKA patients from hyperchloremic metabolic acidosis., Competing Interests: None Declared., (Copyright: © 2023 Turkish Journal of Emergency Medicine.)

Determining structure of highly flexible protein with multiple conformations can be challenging. This paper aims to combine molecular dynamics (MD) and small angle X-ray diffraction (SAX) techniques as a solution to overcome issues related to protein conformation in hardly crystallized protein. Based on prior studies, a cold-active lipase AMS8 was simulated in solvents showing stability in its N-terminal and high flexibility in its C-terminal. However, MD in its own algorithm could not explain the basis of macromolecule conformational transitions or changes related to protein through folding. Hence, by combining SAXS with MD, it is possible to understand the structure of flexible AMS8 lipase in natural space. Based on the findings, SAXS ab-initio model of AMS8 lipase was identified as a monomeric protein in which the optimized model of cold-active lipase AMS8 derived from SAXS data was found to be aligned with AMS8 homology model under series of MD timeframe., Competing Interests: Declaration of competing interest The authors declare no competing financial interests., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Introduction: biodegradable stents of various designs are reportedly used in pancreato-biliary conditions with promising results. Their major advantage is the avoidance of repeat endoscopic procedure for stent removal, thereby reducing overall costs and endoscopic retrograde cholangiopancreatography (ERCP) associated adverse events. The aim of the study was to evaluate the feasibility and safety of a new biodegradable stent in patients with pancreato-biliary diseases., Methods: a prospective multicenter pilot study was performed. All consecutive patients ≥ 18 years old who underwent biliary or pancreatic stenting using the new biodegradable Archimedes stent were included in the study. There were three biodegradation profiles. Technical and clinical success and feasibility and safety were assessed during a pre-established follow-up schedule., Results: fifty-three patients (mean age: 48.54 ± 19.29, 66 % male) with biliary (n = 29, 54.7 %) or pancreatic (n = 24, 45.3 %) indications were included. The distribution of stents used according to degradation properties were as follows: fast (n = 11, 20.8 %), medium (n = 16, 30.2 %) and slow (n = 26, 49.1 %). The technical and clinical success were 100 % and 77.8 %, respectively. Thirty-five patients were followed for a median of 26 weeks (range: 4-56, 66 %). There were nine procedure-related adverse events (17 %), all mild, including one uneventful stent-related event (external migration)., Conclusion: the biodegradable Archimedes stent placement is feasible and safe in pancreato-biliary diseases.

Background: Schools serve as a focal point in which to engage students and school communities, including teachers, parents and other healthcare providers (e.g. nutritionists), in implementing obesity interventions. Bringing them together is beneficial to ensuring healthy eating among schoolchildren and to creating healthy school environments. However, there are barriers and success factors related to the implementation of such an intervention. Therefore, this study aimed to explore the barriers and success factors to school-based obesity interventions to improve obesity implementation., Methods: An exploratory case study with multiple units of analysis was conducted, including interviews, document analyses and a survey. The interview sessions involved six participants, including teachers and nutritionists. The survey was conducted among 121 participants involving 30 teachers, 78 parents and 13 school canteen operators. Pattern-matching techniques were employed to formulate the results., Results: The barriers consisted of the school system, parental awareness, staff availability, and children's compliance. The success factors included the teachers' role, the nutritional approach used, compliance with the healthy school canteen guidelines, and strong collaboration with the Parent-Teacher Association (PTA) and other agencies., Conclusion: It is necessary to digitise an obesity intervention module with some improvements, including strengthening the broader school community's role and integrating nutrition education with technology to improve implementation., Competing Interests: Conflict of Interest None., (© Penerbit Universiti Sains Malaysia, 2022.)

Introduction: Data on the prevalence of nontraumatic headaches with red flag symptoms in Asia are sparse. Therefore, the objectives of this study were to determine the final diagnosis and prevalence of abnormal CT scans in patients who presented to the emergency department (ED) with red flag symptoms., Materials and Methods: This was a retrospective study based on the Radiology Department database of patients who presented to the ED with complaints of nontraumatic headache, had red flag symptoms, and underwent cranial CT scans. The inclusion criteria were adult patients presenting through the ED with nontraumatic headache who underwent cranial CT scans. Multivariate analysis was performed based on logistic regression to analyse the significance of the predictive value for abnormal CT scans., Results: A total of 216 patients underwent cranial CT scans in the ED. More than half of the cases (53.7%) were male. A total of 146 patients (67.6%) had no obvious abnormalities in the CT scans, while 41 patients (19.0%) had cranial infarction, 9 patients (4.2%) had intracranial bleeding, and 20 patients (9.3%) had brain tumours. The most common diagnosis was primary headache syndrome, followed by cerebral vascular accident (CVA). Multivariate analysis showed that three factors were associated with abnormal CT scans: age, systolic blood pressure (SBP), and mean arterial pressure (MAP). New onset of headache at the age of 50 years or older (Odds Ration, OR 3.21, 95% Confidence Interval, CI 1.15, 8.94), SBP (OR 4.82. 95%CI 2.29, 10.40) and MAP (OR 6.21, 95%CI 2.71, 14.70) were significant., Conclusion: The prevalence of abnormal CT scan findings in nontraumatic headache patients with red flag symptoms was 32%. Primary headache syndrome is the most common diagnosis. An age greater than 50 years old during the onset of headache, SBP greater than 180mmHg and MAP greater than 120mmHg were associated with a higher risk of abnormal cranial CT scans.

Brain age is a neuroimaging-based biomarker of aging. This study examined whether the difference between brain age and chronological age (brain-PAD) is associated with cognitive function at baseline and longitudinally. Participants were relatively healthy, predominantly white community-dwelling older adults (n = 531, aged ≥70 years), with high educational attainment (61% ≥12 years) and socioeconomic status (59% ≥75th percentile). Brain age was estimated from T1-weighted magnetic resonance images using an algorithm by Cole et al., 2018. After controlling for age, gender, education, depression and body mass index, brain-PAD was negatively associated with psychomotor speed (Symbol Digit Modalities Test) at baseline (Bonferroni p < 0.006), but was not associated with baseline verbal fluency (Controlled Oral Word Association Test), delayed recall (Hopkins Learning Test Revised), or general cognitive status (Mini-Mental State Examination). Baseline brain-PAD was not associated with 3-year change in cognition (Bonferroni p > 0.006). These findings indicate that even in relatively healthy older people, accelerated brain aging is associated with worse psychomotor speed, but future longitudinal research into changes in brain-PAD is needed., (Copyright © 2021. Published by Elsevier Inc.)

In this study, fouling mechanism and modelling analysis of synthetic lignocellulose biomass and agricultural palm oil effluent was studied using polyethersulfone (PES) ultrafiltration (UF) 10 kDa membrane. The impact of process variables (transmembrane pressure (TMP), pH and concentration of feed solution) on lignocellulosic flux was analysed using pore blocking model. The feasible approaches on utilising deep learning artificial neural network (ANN) to predict smaller flux datasets are studied. Among the input variables, pH of lignin feed solution has significant control towards flux and lignin rejection coefficient for both lignin and lignocellulosic solution. Alteration in the structure of lignin at different pH conditions contributed in the improvement of lignin rejection coefficient to 0.98 at the feed pH of 9. A maximum steady state flux of 52.03 L/m 2 h was observed at the lower lignin concentration (0.25 g/L), TMP of 200 kPa and feed pH of 3. At high TMP and concentration, lignin rejection decreased due to enhancement of feed concentration on membrane surface. The mechanistic model exhibited that cake layer phenomena was dominant in both lignin and lignocellulosic solution. The proposed ANN model showed good correlation (R 2 -1.00) with experimental non-linear flux dynamic data of both lignin and synthetic lignocellulosic solution. In ANN analysis, activation function, algorithm and neuron effect have significant effect in design of accurate model for prediction of small flux datasets. Aerobically-treated palm oil mill filtration analysis also showed that cake layer phenomenon was dominant. A water recovery of 82 % was achieved even at low TMP under short durations., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Dual-layer hollow fiber (DLHF) nanocomposite membrane prepared by co-extrusion technique allows a uniform distribution of nanoparticles within the membrane outer layer to enhance the membrane performance. The effects of spinning parameters especially the air gap on the physico-chemical properties of ZrO 2 -TiO 2 nanoparticles incorporated PVDF DLHF membranes for oily wastewater treatment have been investigated in this study. The zeta potential of the nanoparticles was measured to be around -16.5 mV. FESEM-EDX verified the uniform distribution of Ti, Zr, and O elements throughout the nanoparticle sample and the TEM images showed an average nanoparticles grain size of ~12 nm. Meanwhile, the size distribution intensity was around 716 nm. A lower air gap was found to suppress the macrovoid growth which resulted in the formation of thin outer layer incorporated with nanoparticles. The improvement in the separation performance of PVDF DLHF membranes embedded with ZrO 2 -TiO 2 nanoparticles by about 5.7% in comparison to the neat membrane disclosed that the incorporation of ZrO 2 -TiO 2 nanoparticles make them potentially useful for oily wastewater treatment., Competing Interests: The authors declare no conflict of interest.

Pseudomonas fluorescens AMS8 lipase lid 1 structure is rigid and holds unclear roles due to the absence of solvent-interactions. Lid 1 region was stabilized by 17 hydrogen bond linkages and displayed lower mean hydrophobicity (0.596) compared to MIS38 lipase. Mutating lid 1 residues, Thr-52 and Gly-55 to aromatic hydrophobic-polar tyrosine would churned more side-chain interactions between lid 1 and water or toluene. This study revealed that T52Y leads G55Y and its recombinant towards achieving higher solvent-accessible surface area and longer half-life at 25 to 37 °C in 0.5% (v/v) toluene. T52Y also exhibited better substrate affinity with long-chain carbon substrate in aqueous media. The affinity for pNP palmitate, laurate and caprylate increased in 0.5% (v/v) toluene in recombinant AMS8, but the affinity in similar substrates was substantially declined in lid 1 mutated lipases. Regarding enzyme efficiency, the recombinant AMS8 lipase displayed highest value of k cat /K m in 0.5% (v/v) toluene, mainly with p NPC. In both hydrolysis reactions with 0% and 0.5% (v/v) toluene, the enzyme efficiency of G55Y was found higher than T52Y for p NPL and p NPP. At 0.5% (v/v) toluene, both mutants showed reductions in activation energy and enthalpy values as temperature increased from 25 to 35 °C, displaying better catalytic functions. Only T52Y exhibited increase in entropy values at 0.5% (v/v) toluene indicating structure stability. As a conclusion, Thr-52 and Gly-55 are important residues for lid 1 stability as their existence helps to retain the geometrical structure of alpha-helix and connecting hinge.

Background: The pharmacy internship program is designed to provide comprehensive training for interns to perform various roles in pharmacy practice. In Malaysia, the current internship program has been in place since 2004 in public health facilities, but has never been extensively evaluated., Aim: To assess the level of job satisfaction and perception of Provisionally Registered Pharmacists (PRP) towards the internship training provided., Methods: A cross-sectional nationwide survey was conducted amongst all PRPs undergoing training between December 2015 and February 2016 to assess their level of job satisfaction and perception towards the training requirements stipulated by the Pharmacy Board of Malaysia., Results: A total of 733 PRPs participated in the survey and the response rate was 95.8%. On the basis of the response received, 92.5% (n = 678) felt that the one year training period was 'just nice', 30.5% (n = 223) felt that the logbooks were 'too complicated', and 33.2% (n = 241) said the targets set by the Pharmacy Board of Malaysia were 'too high'. On a scale of 1 to 5, respondents rated 3.32 (SD 0.54) for mean score of job satisfaction. Significant factors influencing mean score of job satisfaction were: perceived fairness at the work place; perceived self-competence after one year of training; preceptors' competence; and pharmacy as a chosen career pathway., Conclusion: Intern pharmacists were fairly satisfied with the internship training in government facilities in the Ministry of Health, Malaysia. A review of the internship logbooks and the practicality of targets set by the Pharmacy Board warrant further improvement., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The alkaline cold-active lipase from Pseudomonas fluorescens AMS8 undergoes major structural changes when reacted with hydrophobic organic solvents. In toluene, the AMS8 lipase catalytic region is exposed by the moving hydrophobic lid 2 (Glu-148 to Gly-167). Solvent-accessible surface area analysis revealed that Leu-208, which is located next to the nucleophilic Ser-207 has a focal function in influencing substrate accessibility and flexibility of the catalytic pocket. Based on molecular dynamic simulations, it was found that Leu-208 strongly facilitates the lid 2 opening via its side-chain. The K M and K cat /K M of L208A mutant were substrate dependent as it preferred a smaller-chain ester (pNP-caprylate) as compared to medium (pNP-laurate) or long-chain (pNP-palmitate) esters. In esterification of ethyl hexanoate, L208A promotes a higher ester conversion rate at 20 °C but not at 30 °C, as a 27% decline was observed. Interestingly, the wild-type (WT) lipase's conversion rate was found to increase with a higher temperature. WT lipase AMS8 esterification was higher in toluene as compared to L208A. Hence, the results showed that Leu-208 of AMS8 lipase plays an important role in steering a broad range of substrates into its active site region by regulating the flexibility of this region. Leu-208 is therefore predicted to be crucial for its role in interfacial activation and catalysis in toluene., Competing Interests: The authors have declared no conflict of interest.

We report a fatal case of post-partum streptococcal toxic shock syndrome in a patient who was previously healthy and had presented to the emergency department with an extensive blistering ecchymotic lesions over her right buttock and thigh associated with severe pain. The pregnancy had been uncomplicated, and the mode of delivery had been spontaneous vaginal delivery with an episiotomy. She was found to have septicemic shock requiring high inotropic support. Subsequently, she was treated for necrotizing fasciitis, complicated by septicemic shock and multiple organ failures. A consensus was reached for extensive wound debridement to remove the source of infection; however, this approach was abandoned due to the patient's hemodynamic instability and the extremely high risks of surgery. Both the high vaginal swab and blister fluid culture revealed Group A beta hemolytic streptococcus infection. Intravenous carbapenem in combination with clindamycin was given. Other strategies attempted for streptococcal toxic removal included continuous veno-venous hemofiltration and administration of intravenous immunoglobulin. Unfortunately, the patient's condition worsened, and she succumbed to death on day 7 of hospitalization.

Lipolytic enzymes with cold adaptation are gaining increasing interest due to their biotechnological prospective. Previously, a cold adapted family I.3 lipase (AMS8 lipase) was isolated from an Antarctic Pseudomonas. AMS8 lipase was largely expressed in insoluble form. The refolded His-tagged recombinant AMS8 lipase was purified with 23.0% total recovery and purification factor of 9.7. The purified AMS8 lipase migrated as a single band with a molecular weight approximately 65kDa via electrophoresis. AMS8 lipase was highly active at 30°C at pH 10. The half-life of AMS8 lipase was reported at 4 and 2h under the incubation of 30 and 40°C, respectively. The lipase was stable over a broad range of pH. It showed enhancement effect in its relative activity under the presence of Li + , Na + , K + , Rb + and Cs + after 30min treatment. Heavy metal ions such as Cu 2+ , Fe 3+ and Zn 2+ inhibited AMS8 activity. This cold adapted alkalophilic AMS lipase was also active in various organic solvent of different polarity. These unique properties of this biological macromolecule will provide considerable potential for many biotechnological applications and organic synthesis at low temperature., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The utilization of cold active lipases in organic solvents proves an excellent approach for chiral synthesis and modification of fats and oil due to the inherent flexibility of lipases under low water conditions. In order to verify whether this lipase can function as a valuable synthetic catalyst, the mechanism concerning activation of the lid and interacting solvent residues in the presence of organic solvent must be well understood. A new alkaline cold-adapted lipase, AMS8, from Pseudomonas fluorescens was studied for its structural adaptation and flexibility prior to its exposure to non-polar, polar aprotic and protic solvents. Solvents such as ethanol, toluene, DMSO and 2-propanol showed to have good interactions with active sites. Asparagine (Asn) and tyrosine (Tyr) were key residues attracted to solvents because they could form hydrogen bonds. Unlike in other solvents, Phe-18, Tyr-236 and Tyr-318 were predicted to have aromatic-aromatic side-chain interactions with toluene. Non-polar solvent also was found to possess highest energy binding compared to polar solvents. Due to this circumstance, the interaction of toluene and AMS8 lipase was primarily based on hydrophobicity and molecular recognition. The molecular dynamic simulation showed that lid 2 (residues 148-167) was very flexible in toluene and Ca(2+). As a result, lid 2 moves away from the catalytic areas, leaving an opening for better substrate accessibility which promotes protein activation. Only a single lid (lid 2) showed the movement following interactions with toluene, although AMS8 lipase displayed double lids. The secondary conformation of AMS8 lipase that was affected by toluene observed a reduction of helical strands and increased coil structure. Overall, this work shows that cold active lipase, AMS8 exhibits distinguish interfacial activation and stability in the presence of polar and non-polar solvents., (Copyright © 2016. Published by Elsevier Inc.)

Background. Not all yeast alcohol dehydrogenase 2 (ADH2) are repressed by glucose, as reported in Saccharomyces cerevisiae. Pichia stipitis ADH2 is regulated by oxygen instead of glucose, whereas Kluyveromyces marxianus ADH2 is regulated by neither glucose nor ethanol. For this reason, ADH2 regulation of yeasts may be species dependent, leading to a different type of expression and fermentation efficiency. Lachancea fermentati is a highly efficient ethanol producer, fast-growing cells and adapted to fermentation-related stresses such as ethanol and organic acid, but the metabolic information regarding the regulation of glucose and ethanol production is still lacking. Methods. Our investigation started with the stimulation of ADH2 activity from S. cerevisiae and L. fermentati by glucose and ethanol induction in a glucose-repressed medium. The study also embarked on the retrospective analysis of ADH2 genomic and protein level through direct sequencing and sites identification. Based on the sequence generated, we demonstrated ADH2 gene expression highlighting the conserved NAD(P)-binding domain in the context of glucose fermentation and ethanol production. Results. An increase of ADH2 activity was observed in starved L. fermentati (LfeADH2) and S. cerevisiae (SceADH2) in response to 2% (w/v) glucose induction. These suggest that in the presence of glucose, ADH2 activity was activated instead of being repressed. An induction of 0.5% (v/v) ethanol also increased LfeADH2 activity, promoting ethanol resistance, whereas accumulating acetic acid at a later stage of fermentation stimulated ADH2 activity and enhanced glucose consumption rates. The lack in upper stream activating sequence (UAS) and TATA elements hindered the possibility of Adr1 binding to LfeADH2. Transcription factors such as SP1 and RAP1 observed in LfeADH2 sequence have been implicated in the regulation of many genes including ADH2. In glucose fermentation, L. fermentati exhibited a bell-shaped ADH2 expression, showing the highest expression when glucose was depleted and ethanol-acetic acid was increased. Meanwhile, S. cerevisiae showed a constitutive ADH2 expression throughout the fermentation process. Discussion. ADH2 expression in L. fermentati may be subjected to changes in the presence of non-fermentative carbon source. The nucleotide sequence showed that ADH2 transcription could be influenced by other transcription genes of glycolysis oriented due to the lack of specific activation sites for Adr1. Our study suggests that if Adr1 is not capable of promoting LfeADH2 activation, the transcription can be controlled by Rap1 and Sp1 due to their inherent roles. Therefore in future, it is interesting to observe ADH2 gene being highly regulated by these potential transcription factors and functioned as a promoter for yeast under high volume of ethanol and organic acids.

The objective of this study was to investigate the modulatory effect of Chlorella vulgaris on cultured fibroblast cells derived from young and old aged individuals focusing on DNA damage, telomere length and telomerase activity. Dose-response test of the algal extract on cells in both age groups revealed that optimum viability was observed at a concentration of 50 microg/ml. Results obtained showed that Chlorella vulgaris exhibited protective effects against H(2)O(2)-induced oxidative stress as shown by the reduction in damaged DNA caused by H(2)O(2) treatment (p<0.05) in Chlorella vulgaris pre- and post-treated groups (p<0.05). Pre-treatment of Chlorella vulgaris resulted in a significant decrease in DNA damage suggesting a bioprotective effect against free radical attacks. A decline in DNA damage was observed in post-treated cells which proves Chlorella vulgaris to present bioremediative properties. In cells induced with oxidative stress, telomere length decreased significantly coupled with a concomitant decline of telomerase activity (p<0.05). However, these reductions were prevented with prior and post treatment of Chlorella vulgaris. Therefore, we concluded that Chlorella vulgaris exhibited bioprotective effects especially in cells obtained from young donor but were more bioremediative for cells obtained from old donor as indicated by DNA damage, telomere shortening and reduction in telomerase activity.