Potensi antikandida ekstrak madu secara in vitro dan in vivo

ABSTRAK Ning Rintiswati, Novi Eko Winarsih, Rusdy Ghazali Malueka: Potensi antikandida ekstrak madu secara in vitro dan in vivo Latar belakang: Telah diteliti potensi madu sebagai antikandida dalam rangka mendapatkan suatu agen anti jamur alternatif. Usaha ini didasarkan pada kenyataan bahwa anti jamur yang merupakan obat andalan untuk menanggulangi infeksi jamur yang ada tidak memiliki spektrum luas, dan sangat terbatas macamnya. Tujuan: Penelitian ini bertujian untuk mengetahui efek madu murni, ekstrak madu, dan residunya terhadap Candida albicans secara in vitro dan in vivo. Bahan dan cara: Penelitian dilakukan secara in vitro dan in vivo Penelitian ini merupakan penelitian eksperimental laboratorik secara in vitro dan eksperimental acak sederhana bagi uji in vivo. Pada uji in vitro, potensi madu murni dibandingkan dengan ekstrak madu dan residunya, mikroba uji yang digunakan adalah Candida albicans Pengujian dilakukan menggunakan metode dilusi cair untuk kandida. Data yang diperoleh berupa angka KHM (kadar hambat minimal) dan KBM (kadar bunuh minimal), sedangkan pada uji in vivo hewan uji yang digunakan adalah mencit, data yang diperoleh berupa data histopatologi dan kecepatan kesembuhan antara kelompok terapi dan kontrol. Hasil: Penelitian menunjukkan bahwa ekstrak madu memiliki daya hambat dan daya bunuh terhadap Candida albicans secara in vivo dan KHM serta KBM dicapai pada konsentrasi 0,3125 %. Madu murni tidak berefek terhadap Candida albicans, sedangkan residu madu menunjukkan daya hambat dan daya bunuh lemah dengan KHM 25% dan KBM 50%. Simpulan: Ekstrak madu mampu mempercepat dan meningkatkan kesembuhan mencit yang diinfeksi dengan Candida albicans secara in viv

Categorization of 77 dystrophin exons into 5 groups by a decision tree using indexes of splicing regulatory factors as decision markers

<p>Abstract</p> <p>Background</p> <p>Duchenne muscular dystrophy, a fatal muscle-wasting disease, is characterized by dystrophin deficiency caused by mutations in the <it>dystrophin </it>gene. Skipping of a target <it>dystrophin </it>exon during splicing with antisense oligonucleotides is attracting much attention as the most plausible way to express dystrophin in DMD. Antisense oligonucleotides have been designed against splicing regulatory sequences such as splicing enhancer sequences of target exons. Recently, we reported that a chemical kinase inhibitor specifically enhances the skipping of mutated <it>dystrophin </it>exon 31, indicating the existence of exon-specific splicing regulatory systems. However, the basis for such individual regulatory systems is largely unknown. Here, we categorized the <it>dystrophin </it>exons in terms of their splicing regulatory factors.</p> <p>Results</p> <p>Using a computer-based machine learning system, we first constructed a decision tree separating 77 authentic from 14 known cryptic exons using 25 indexes of splicing regulatory factors as decision markers. We evaluated the classification accuracy of a novel cryptic exon (exon 11a) identified in this study. However, the tree mislabeled exon 11a as a true exon. Therefore, we re-constructed the decision tree to separate all 15 cryptic exons. The revised decision tree categorized the 77 authentic exons into five groups. Furthermore, all nine disease-associated novel exons were successfully categorized as exons, validating the decision tree. One group, consisting of 30 exons, was characterized by a high density of exonic splicing enhancer sequences. This suggests that AOs targeting splicing enhancer sequences would efficiently induce skipping of exons belonging to this group.</p> <p>Conclusions</p> <p>The decision tree categorized the 77 authentic exons into five groups. Our classification may help to establish the strategy for exon skipping therapy for Duchenne muscular dystrophy.</p

Mutation spectrum analysis of DMD gene in Indonesian Duchenne and Becker muscular dystrophy patients [version 3; peer review: 2 approved]

Background Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations in the DMD gene. The full mutation spectrum of the DMD gene in Indonesian patients is currently unknown. Mutation-specific therapies are currently being developed, such as exon skipping or stop codon read-through therapy. This study was conducted with the aim of identifying the mutation spectrum of the DMD gene in Indonesia to guide future development and application of feasible therapeutic strategies. Methods This study is a cross sectional study that enrolled 43 male patients with a clinical suspicion of DMD or BMD. Multiplex ligation-dependent probe amplification (MLPA) reaction was performed to screen for the common mutations in the DMD gene. Results Out of 43 subjects, deletions accounted for 69.77% (n=30) cases, while duplications were found in 11.63% (n=5) cases. One novel duplication spanning exons 2 to 62 was identified. Deletion mutations clustered around the distal (66.67%) and proximal (26.67%) hot spot regions of the DMD gene while duplication mutations were observed solely at the proximal region. Two false positive cases of single exon deletion detected through MLPA were attributed to sequence mutations affecting primer ligation sites, confirming the need to validate all single exon deletions when using this screening method. Analysis of available maternal DNA samples showed that the rate of de novo mutations (48.15%) appears higher than expected in this population. Out of 31 patients who were classified as DMD based on clinical and genotype characterizations, 60.47% (n=26) of cases were suitable for exon skipping therapy. Conclusion This is the first comprehensive study showing the feasibility of implementing the MLPA method for routine screening of DMD patients in Indonesia. This is also the first study showing the potential applicability of exon skipping therapy in the majority of DMD cases in the country

Association between Increased Matrix Metalloproteinase-9 (MMP-9) Levels with Hyperglycaemia Incidence in Acute Ischemic Stroke Patients

BACKGROUND: Hyperglycemia is common in acute stroke patients. Hyperglycemia can induce the production of reactive oxygen species, causing increased activity of matrix metalloproteinase-9 (MMP-9). AIM: This study aimed to determine an association between the increased levels of MMP-9 and the incidence of hyperglycemia in acute ischemic stroke patients. METHODS: This is a case-control study. Acute ischemic stroke patients admitted to the Stroke Unit of a reference hospital in Yogyakarta, Indonesia was divided into the hyperglycemic and non-hyperglycemic group. Demographic and clinical characteristics of each subject were recorded, and blood levels of MMP-9 were measured. Seventy-one patients were recruited, 40 subjects in the hyperglycemic group and 31 subjects in the non-hyperglycemic group. RESULTS: The median levels of blood MMP-9 level in the hyperglycemic and non-hyperglycemic group were 974.37 and 748.48 ng/mL, respectively, and the difference was statistically not significant (95% CI, 191.24-2849.53; p = 0.07). When the calculated cut-off point of 600.99 ng/mL was used, the proportion of patients with higher MMP-9 levels was significantly more in the hyperglycemic group compared with the ones in the non-hyperglycemic group (82.5% and 54.8%, respectively; OR = 3.88; p = 0.011). CONCLUSION: We concluded that the proportion of patients with MMP-9 level &gt;600.99 ng/mL was significantly higher in acute ischemic stroke patients with hyperglycemia.ng/mL was significantly higher in acute ischemic stroke patients with hyperglycemia

The Association between Pesticide Exposure and Neurological Signs and Symptoms in Farmers in Magelang District, Central Java, Indonesia

BACKGROUND: Excessive use of pesticides is known to cause neurotoxicity. Chronic effects of pesticide poisoning include neuropathy and tremors. AIM: This study aimed to determine the association between pesticide exposure and the occurrence of neurological signs and symptoms, especially neuropathy and tremor, in farmers. METHODS: This was a cross-sectional study. The study location was Seloprojo Village, Ngablak District, Magelang Regency, Central Java Province. Farmers as subjects were recruited to determine neuropathy using Diabetic Neuropathy Symptom (DNS) and Diabetic Neuropathy Examination (DNE) scoring. Tremor events were measured with Tremor Rating Scale (TRS). Cholinesterase levels were examined using venous blood samples to determine the level of pesticide poisoning. RESULTS: Of the 120 farmers studied, 68.3% experienced pesticide poisoning with cholinesterase levels below normal values. Weakness of the upper limb was found in 10 subjects (8.33%), while weakness of the lower limbs was found in 6 subjects (5%). There were 59.2% farmers who met the neuropathy criteria from the DNS score and those who met the neuropathic criteria from the DNE score were 6.7%. Tremor symptoms were found in 71.7% of the farmers. There was no significant association between cholinesterase levels and DNS score (p = 0.737), but there were significantly lower levels of cholinesterase (p = 0.046) in the neuropathy group measured with DNE score. There was no significant association between cholinesterase levels and TRS (p = 0.204). CONCLUSION: Cholinesterase levels were significantly associated with neuropathy incidence measured with DNE criteria but statistically not related to tremors in farmers exposed to pesticides

Blood Cholinesterase Level is Associated with Cognitive Function in Indonesian School-age Children Exposed to Pesticides

BACKGROUND: Pesticides are known as depressors of acetylcholinesterase (AChE) activity, resulting in the nervous system toxicity. The previous studies have described associations between AChE, a stable marker of pesticide poisoning, and cognitive performance in children. AIM: This study aimed to identify the association between blood AChE level and cognitive function in children exposed to pesticides in the Magelang Regency, Indonesia. METHODS: A cross-sectional study involving school-age children with a history of pesticide exposure in Ngablak, Magelang Regency, Central Java, Indonesia, was conducted. Blood AChE level was evaluated, and the Modified Mini–Mental State Examination for Children (MMMSEC) was used to analyze the cognitive function of the children. RESULTS: In total, 56 subjects aged between 9 and 11 years were included in this study. Median blood AChE level was 9.64 kIU/L, and 24 subjects (42.9%) had low blood AChE levels. Median MMMSEC score was 33. Eleven subjects (19.6%) were found to have abnormal cognitive function. Bivariate analysis showed that blood AChE level was associated with MMMSEC score (r = 0.343, p = 0.010). Multiple linear regression showed that blood AChE level had a positive association with cognitive function in children, assessed using the MMMSEC score (β = 0.360; p = 0.006). Further analysis showed that the attention and orientation (memory function) domains of the MMMSEC were significantly associated with blood AChE level (β: 0.371 and 0.297, respectively, p &lt; 0.05). CONCLUSIONS: Blood AChE level, a stable marker of pesticide poisoning, was positively associated with cognitive function in children, as assessed using the MMMSEC score. In particular, the orientation and attention domains of the MMMSEC were associated with blood AChE level

Radiologi diagnostik

viii, 216 hal; 21 c

Effectiveness of Gotu Kola Extract 750 mg and 1000 mg Compared with Folic Acid 3 mg in Improving Vascular Cognitive Impairment after Stroke

This study aimed to determine the effectiveness of gotu kola (Centella asiatica) in improving cognitive function in patients with vascular cognitive impairment (VCI). This study uses a quasi-experimental design. Subjects in this study were patients with poststroke cognitive impairment who were treated at two hospitals in Yogyakarta, Indonesia. The number of subjects was 48: 17 subjects were treated with 1000 mg/day of gotu kola extract, 17 subjects treated with 750 mg/day of gotu kola extract, and 14 subjects treated with 3 mg/day of folic acid for 6 weeks. A Montreal Cognitive Assessment-Indonesian version (MoCA-Ina) was conducted at the beginning of treatment and after 6 weeks of therapy. It was found that all trials effectively improved poststroke VCI based on MoCA-Ina scores over the course of the study. There is no significant difference in ΔMoCA-Ina (score at the 6th week of treatment − score at the beginning) mean score among the three groups, indicating that gotu kola is as effective as folic acid in improving poststroke VCI. Gotu kola was shown to be more effective than folic acid in improving memory domain. This study suggested that gotu kola extract is effective in improving cognitive function after stroke

Prevalence of Stroke and Associated Risk Factors in Sleman District of Yogyakarta Special Region, Indonesia

Background. Stroke remains one of the most common noncommunicable diseases among Indonesian populations. This study aimed to identify the prevalence of stroke and its associated risk factors in the Sleman District of Yogyakarta Special Region, Indonesia. Method. This study was a secondary analysis of community-based data collected by the Sleman Health and Demographic Surveillance System (HDSS) in 2016. Basic demographic and socioeconomic data were collected. Additional questions about history of stroke and other chronic diseases were interviewed as a self-reported diagnosis. History of hormonal contraceptives use and dietary patterns were also collected. We examined the association between the prevalence of stroke and risk factors, namely, age, gender, self-reported history of chronic diseases, hormonal contraceptives use, and high-risk dietary patterns. Results. The survey included 4,996 households composed of 20,465 individuals. Data regarding stroke incidents were available from 13,605 subjects aged ≥20 years old. Among them, a total of 4,884 subjects also have data regarding stroke risk factors. The overall prevalence of stroke in Sleman District was 1.4% (0.5% men and 0.90% women). The prevalence increased with additional decades of age (p<0.001). In a multivariable model, increasing age, self-reported history of hypertension (OR=8.37, 95%CI: 4.76 to 14.69), and self-reported history of diabetes mellitus (OR=2.87, 95%CI: 1.54 to 5.35) were significantly associated with stroke. Conclusions. A community-based survey in Indonesia showed a high prevalence of stroke which was associated with increasing age, hypertension, and diabetes mellitus. These findings suggest that preventive actions against the aforementioned modifiable risk factors should be prioritized

Phosphorothioate Modification of Chimeric 2'-O-Methyl RNA/Ethylene-Bridged Nucleic Acid Oligonucleotides Increases Dystrophin Exon 45 Skipping Capability and Reduces Cytotoxicity

Backgrounds, Antisense oligonucleotide (AO)-mediated exon skipping is the most promising way to express internally deleted dystrophin in Duchenne muscular dystrophy (DMD), by correcting the reading frame of dystrophin mRNA. An antisense chimeric oligonucleotide consisting of 2'-O-methyl RNA and ethylene-bridged nucleic acid (ENA), targeting exon 45 of the dystrophin gene, AO85, has been shown to induce exon 45 skipping efficiently. Since phosphorothioate (PS)-modification of AO85 has never been explored, we produced a PS-modified AO85 (AO88) and examined its exon skipping capability and cytotoxicity. Methods, Exon 45 skipping activity was examined in primary muscle cells established from a DMD patient carrying a deletion of dystrophin exon 44. Cytotoxicity was assessed by MTT assay. Results, AO88 induced dystrophin exon 45 skipping from 50 nM. More than 90% of products lacked exon 45 at 400 nM. AO88 showed significantly higher exon skipping activity than AO85. The EC50 of AO88 was 94.8 nM, while EC50 of AO85 was 66.7 nM. Cytotoxicity was lower for AO88 than for AO85. Conclusion, the PS-modified RNA/ENA chimera displayed stronger exon skipping activity and lower cytotoxicity than the phosphodiester-RNA/ENA chimera. AO88 has better potential for clinical use than AO85