In vitro cytotoxicity assay of Fucoidan extracted from Turbinaria conoides against cancer cell lines MCF7, A549, and normal cell line L929

Abstract The main aim of the study is to quantify the cytotoxic property of the Fucoidan extracted from the Turbinaria conoides using the MTT assay with the standard fucose. Fucoidan was extracted using the soaked water method and it was determined using the HPLC procedure the obtained Test sample Fucoidan extracted from the Turbinaria conoides and standard fucose was subjected to the cytotoxicity assay against the MCF7 Human breast cancer cell line, A549 lung cancer cell line, and L929 normal mouse fibroblast cell line. From the results it was found that the Test sample showed good IC50 value for MCF7 cell line then A549 with an increasing concentration 24 hours incubation at 37°C The IC50 for MCF7 was 115.21 µg/ml and A549 396.46µg/ml and the Fucoidan extract was checked for its cytotoxicity against the normal mouse fibroblast cell line L929, Fucoidan was found non-lethal to the L929 mouse fibroblast normal cell line. Standard fucose also gave a significant result towards MCF7 and against the L929. This indicates that the Fucoidan extracted from Tubinaria conoides shows better anticancer potential in it. Hence its application can be further extended in the pharmacological fields.

p53 & Cyclin D1 expression in surgically resected clear margins of oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is one of the most well-known malignancies that affect the human population worldwide. The early diagnosis and early intervention of OSCC help improve the survival rate of the patients. The tumour free surgical margins are a positive prognostic factor for recurrence-free survival. The molecular markers can be used to detect the tumour free surgical margins. Aim: The aim of the study is to evaluate the expression of p53 & Cyclin D1 marker in resected surgical apparently clear margins and to correlate the p53 & Cyclin D1 expression with clinicopathological characteristics and patient outcome. Methods: The study population included retrospective cases of OSCC with apparently clear margins (2017-18) n=10 and Clinicopathological variables relevant to survival analysis were recorded. Finally, two margins were selected from each case, a total of 20 margins were included in this study. Paraffin-embedded wax blocks retrieved and tissue sections were made. Expression of cyclin D1 and p 53 was assessed by the immunohistochemical staining procedure Results: Positive expressions Cyclin D1 in 40% of mild dysplasia margins and 60% in clearance adequate margins were present. p53 expression was seen in 16% of mild dysplasia margins and 84% in clearance adequate margins. The expression of p53 and Cyclin D1 molecular markers are noted in the basal & parabasal layer of epithelium. Conclusion: Molecular markers could play a more reliable method for the assessment of dysplasia at the margins

Enzyme-Linked Immunosorbent Assay Detection For Malignancy Using Anti-P53 Antibodies

@#Tumour protein 53 (p53) plays an important role in the instruction of the cell cycle. In a variety of transformed cell lines, tumour protein is expressed in high amounts, and it is believed to contribute to transformation and malignancy. This research aimed to detect the anti-p53 antibodies in sera of patients with various malignant tumours and to evaluate the sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA). A case-control study was conducted on samples from 49 patients with various types of malignant tumours at Sultanah Bahiyah Hospital, Alor Setar, Kedah, Malaysia, and 32 healthy control cases with non‐malignant disease collected from Universiti Sains Malaysia clinic, Penang, Malaysia. The antibodies against p53 protein in the serum samples were analysed using the commercial ELISA kit, Calbiochem® p53- ELISAPLUS. The results showed that the rate of anti-p53 antibodies in patients with various malignant tumours was 13 out of 49 (26.5 %), compared with only 2 out of 32 (6.25%) in healthy controls (p < 0.001). The sensitivity of this kit reached 28.6% and the specificity was 93.8%. In conclusion, these results suggest that the anti-p53 antibodies can be detected in different sera of malignant tumour patients and the ELISA kit is highly specific; nevertheless, its discrimination power is not perfect because of its low sensitivity to determine the anti-p53 antibodies.

Evaluation of muscle involvement & degeneration in advanced oral submucous fibrosis using three different stains ­ a retrospective cross-sectional study / Avaliação do envolvimento e degeneração muscular na fibrose submucosa oral avançada usando três colorações diferentes - um estudo transversal retrospectivo

Background: Oral submucous fibrosis (OSMF) is graded according to various histological factors which include the epithelial changes and the connective tissue changes. These features could be identified in routine Hematoxylin and Eosin (H and E) staining in shades of pink. However, the use of special stains provides a contrast to various connective tissue components thereby aiding in better visualization of these connective tissue changes in advanced OSMF cases. Objective:To compare and evaluate muscle involvement and degeneration in advanced oral submucous fibrosis using three different stains namely, H&E, Van Gieson, and Masson's Trichrome. Material and Methods: 10 Formalin-fixed paraffin-embedded tissue sections of advanced OSMFwere stained using 3 different stains namely Hematoxylin and Eosin (H&E), Van-Gieson, and Masson trichrome. The results obtained were tabulated and statistically analyzed using Kruskal Wallis ANOVA. Results: The hyalinization and fibrosis involving the skeletal muscle were better visualized in Masson's Trichrome but were not statistically significant. The muscle degeneration in deeper areas was better visualized in Masson's trichrome when compared to the H&E and Van Gieson. Conclusion: Hematoxylin and eosin stains all the connective tissue components in various shades of pink, use of special stains bestows contrast between different components of connective tissue, thus improvising grading of OSMF. Masson's trichrome stain can be used as a single adjunct to hematoxylin and eosin stain as changes in the superficial and deeper connective tissue could be ascertained (AU)

Contexto: A fibrose submucosa oral (FSO) é classificada de acordo com vários fatores histológicos que incluem alterações epiteliais e do tecido conjuntivo. Essas características podem ser identificadas na coloração de rotina com Hematoxilina e Eosina (H&E) em tons de rosa. No entanto, o uso de colorações especiais fornece um contraste para vários componentes do tecido conjuntivo, auxiliando assim na melhor visualização dessas alterações do tecido conjuntivo em casos avançados de FSO. Objetivo: Comparar e avaliar o envolvimento e degeneração muscular da fibrose submucosa oral avançada usando três colorações diferentes: H&E, Van Gieson e Tricrômio de Masson. Material e Métodos: 10 seções de tecido fixadas em formalina e embebidas em parafina de FSO avançada foram coradas usando essas três colorações. Os resultados obtidos foram tabulados e analisados estatisticamente por meio do teste de Kruskal Wallis. Resultados: A hialinização e fibrose envolvendo o músculo esquelético foram melhor visualizadas no tricromo de Masson, mas não foram estatisticamente significativas. A degeneração muscular em áreas mais profundas foi melhor visualizada no tricrômico de Masson, quando comparado ao H&E e Van Gieson. Conclusão: Hematoxilina e Eosina coloram todos os componentes do tecido conjuntivo em vários tons de rosa. O uso de colorações especiais confere contraste entre os diferentes componentes do tecido conjuntivo, melhorando assim a avaliação da FSO. A coloração com tricrômio de Masson pode ser usada como um único adjunto para a coloração de Hematoxilina e Eosina, pois assim é possível verificar alterações superficiais e profundas no tecido conjuntivo. (AU)

Efficacy of Anhydrous Copper Sulphate as a Solid Dehydrant in Tissue Processing Procedure

Tissue processing in histotechnique is an important procedure after specimen collection involving three main procedures namely, dehydration using alcohol, clearing using xylene, and infiltration by paraffin wax. Isopropyl alcohol, the widely used dehydrating agent is toxic and when exposed to heated isopropyl alcohol fumes, it leads to numerous health hazards. Anhydrous copper sulphate is less toxic than isopropyl alcohol and requires less amount of exposure to the chemicals. The aim of the study was to investigate the dehydrating potential of anhydrous CuSO4 as an isopropyl alcohol substitute. METHODSA descriptive study of forty paired soft tissue specimens were obtained from the Department of Oral Pathology and Microbiology and were subjected to routine histopathological tissue processing with isopropyl alcohol and anhydrous CuSO4 as dehydrating agents. Histomorphological criteria namely nuclear staining, cytoplasmic staining, artefacts, and background staining were evaluated and the scores were tabulated, and statistical analysis was done using SPSS version 20 by IBM. RESULTSOur study results showed that 40 % of alcohol dehydrated tissue samples had better nuclear staining than tissue samples dehydrated by anhydrous copper sulphate and 15 % of alcohol dehydrated tissue samples had better cytoplasmic staining than tissues dehydrated by Anhydrous CuSO4. Background staining of more than 20 % was seen in anhydrous CuSO4 dehydrated tissue specimens than alcohol dehydrated tissue specimens. Artefacts were seen in similar ratios in both alcohol (80 %) and anhydrous copper sulphate (75 %) dehydrated tissue specimens. CONCLUSIONSAnhydrous CuSO4 has superior dehydrating properties than isopropyl alcohol which lead to over-dehydration of the tissue specimens. Further studies are required to validate the findings.

Peripheral ameloblastoma of the gingiva

Peripheral Ameloblastoma (PA) is a benign odontogenic tumor, arising from the cell rest of Serres, reduced enamel epithelium and basal cells of the surface epithelium. Peripheral ameloblastoma is a rare odontogenic neoplasm occurring commonly in the mandibular gingiva. PA clinically resembles other peripherally occurring lesions like pyogenic granuloma, peripheral ossifying fibroma, peripheral giant cell granuloma, and squamous papilloma. The recurrence rate of PA is 16-19% which demands a straight follow up. We report a case of recurrent peripheral ameloblastoma occurring in a 72-year old male located in the mandibular lingual gingiva in relation to the 44, 45 element's regions. The patient had a primary lesion excised from the same site 6 years before which was diagnosed as ameloblastoma.

Pattern of acute kidney injury and its outcome in a tertiary care centre

Background: Acute kidney injury is a multiplex disease with severe morbidity and mortality. The trends of acute kidney injury vary according to the regions and the population under study. The aim of this study is to evaluate the trends of acute kidney injury and its outcome in a tertiary care hospital.Methods: The study was a prospective observational study conducted at a tertiary care hospital in a metropolitan city. A total of 102 patients of acute kidney injury were selected based on the Kidney Disease Improving Global Outcomes guidelines of acute kidney injury. The main trends of acute kidney injury presentation and its outcome were assessed.Results: Of 102 patients admitted, 42 had a sepsis related diagnosis (42.41%), 17 patients (17.16%) had cardiovascular disease related acute kidney injury and 12 patients (12.12%) had developed acute kidney injury due to drugs and poisons. According to RIFLE (risk of renal failure, injury to kidney, failure and loss of function and end-stage kidney disease) category, 43.96% of patients belonged to the risk category and 30.77% to the injury category. Of 34 patients in failure category, 23 recovered and 11 did not recover. Authors compared the trends of acute kidney injury in patients who recovered and who deteriorated. The mean serum creatinine values were 3.42 mg/dl in patients who didn’t recover from acute kidney injury and 2.05 mg/dl in patients who recovered. In patients of the recovered group, the mean urine output value is 783 ml/day; in deterioration group, 445 ml/day.Conclusions: Most common etiologies of acute kidney injury in this study include sepsis, drugs and poisons, cardiovascular diseases and diarrheal diseases in order of occurrence. High serum creatinine at admission and oliguria were the most common factors that contributed to deterioration in acute kidney injury.

Effects of combination of phenazine-1-carboxylic acid from Streptomyces kebangsaanensis with amphotericin B on clinical Fusarium solani species

Aim@#A novel endophyte, Streptomyces kebangsaanensis was isolated from the stem of a Malaysian ethnomedicinal plant, Portulaca oleracea in 2013. Studies on S. kebangsaanensis crude extract showed that it had antifungal activities and further work led to isolation of a novel compound, phenazine-1-carboxylic acid (PCA). This study investigated the combinatorial effect of PCA isolated from S. kebangsaanensis with amphotericin B on the growth of four clinical Fusarium solani isolates. @*Methodology and results@#Disk diffusion assay showed that the crude extract of S. kebangsaaneesis inhibited growth of all four F. solani isolates. Whereas, the compound PCA from this extract inhibited two of the tested F. solani isolates, UZ541/12, and UZ667/13 at minimum inhibitory concentration of 18.00 µg/mL Combinations of this compound with amphotericin B, reduced the minimum inhibitory concentration of amphotericin B for these two isolates from 8 to 0.13 µg/mL and 4 to 0.03 µg/mL respectively. Analysis of fractional inhibitory concentration index showed that a borderline synergism is present between the compound and amphotericin B. @*Conclusion, significance and impact of the study@#These results indicate PCA may be useful in improving actions of available drugs against antimicrobial resistant microorganisms.

Phytochemicals with antifungal properties: Cure from nature

Aims@#The exploration of natural products with innovative uses is dynamic and expanding rapidly. Medicinal plants have fascinated many researchers that subsequently lead to research publications highlighting plant extracts with wide range of secondary metabolites such as flavonoids, alkaloids, glycosides, quinones, terpenoids, tannins and saponins that exhibit antimicrobial activities and disease control. The concentration of these bioactive compounds in each plant species varies based on the pathosystem and environmental conditions. This study aims to uncover the various types of phytochemicals with antifungal properties.@*Methodology and results@#Seven categories of plant-based antifungal compounds were reviewed, which are terpenoids, saponins, phenolic compounds, coumarins, alkaloids, essential oils and peptides, with examples and structures of some available compounds. The mechanism of action of each category of phytochemical was discussed. Also, the impact of some compounds was explained and elaborated. @*Conclusion, significance and impact of study@#It is of a great importance to explore natural plant fighters against fungal infection. Those active plant components do not only have antifungal properties, but they also help in the healing process and some even exhibit anticancer activities. The development and knowledge of antifungal activities from plant extracts have the potential for applications in antifungal therapy. Since the exact description of how antifungal compounds function in the human body is still unclear more studies are required to unveil phytochemicals’ properties and to elucidate their effects on living cells.

Construction of an Escherichia coli expression vector for the non-structural (NS)-1 protein of avian influenza virus H5N1 (Pembangunan vektor pengekspresan Escherichia coli untuk protein non-struktural (NS)-1 virus influenza unggas H5N1) / Jurnal Sains Kesihatan Malaysia

@#In the search for universal vaccine candidates for the prevention of avian influenza, the non-structural (NS)-1 protein of avian influenza virus (AIV) H5N1 has shown promising potential for its ability to effectively stimulate the host immunity. This study was aimed to produce a bacterial expression plasmid using pRSET B vector to harbour the NS1 gene of AIV H5N1 (A/Chicken/Malaysia/5858/2004 (H5N1)) for protein expression in Escherichia coli (E. coli). The NS1 gene (687 bp) was initially amplified by polymerase chain reaction (PCR) and then cloned into a pGEM-T Easy TA vector. The NS1 gene was released from pGEM-T-NS1 using EcoRI and XhoI restriction enzymes (RE). The pRSET B vector was also linearized using the same RE. The digested NS1 gene and linearized pRSET B were ligated using T4 DNA ligase to form the expression plasmid, pRSET B-NS1. The NS1 gene sequence in pRSET B-NS1 was confirmed by DNA sequencing. To prepare recombinant bacterial cells for protein expression in the future, pRSET B-NS1 was transformed into E. coli strain BL21 (DE3) by heat-shock. Colonies bearing the recombinant plasmid were screened using PCR. The DNA sequencing analysis revealed that the NS1 gene sequence was 97% homologous to that of AIV H5N1 A/Chicken/Malaysia/5858/2004 (H5N1). These results indicated that the NS1 gene of influenza A/Chicken/Malaysia/5858/2004 (H5N1) was successfully amplified and cloned into a pRSET B vector. Bacterial colonies carrying pRSET B-NS1 can be used for the synthesis of NS1-based influenza vaccine in the future and thereby aid in the prevention of avian influenza.

Effect of photoperiod (PP) and photosynthetic photon flux intensity (PPFI) on nutrients consumption, growth and lipid profile of unusual microalga Picochlorum maculatum (PSDK01) in shrimp culture effluent

Microalgae have been known for broad spectrum of ecological and industrial applications. However, continuous mass culturing of microalgae is still a challenging task. In this context, studies on optimization of culturing conditions like photoperiod and light intensity on nutrients consumption are not uncommon. In the present study, we tried to evaluate the effect of different photoperiod (PP) and photosynthetic photon flux intensity (PPFI) on the nutrient consumption and algal growth of Picochlorum maculatum in shrimp culture effluent. The experiment lasted for 15 days at five different PP cycles (24:0, 18:06, 14:10, 10:14 and 6:18 h light:dark) and four PPFI (50, 100, 150 and 200 µmol m-2 s-1) at 28ºC temperature. Every five days the samples were analyzed and results showed that the PP of 18 h light and 6 h dark and 150 µmol m-2 s-1 of PPFI favouring the growth of P. maculatum with a maximum nutrients consumption (PO43+ 91%, NO3- 68%, NO2- 64%, NH3+ 77%), maximum growth (0.876±0.001 abs) and biomass production (9.93±0.19 g L−1) with highest yield of 9.2% total lipid. Among the photoperiod and photosynthetic photon flux intensity tested, 18:6 h light:dark and 150 µmol m-2 s-1 showed the highest nutrients removal and algal growth. This study strongly suggests that P. maculatum PSDK01 was a potential candidate for further commercial development in larger scale

Correlation between bizygomatic and maxillary central incisor width for gender identification / Correlação entre a distância bizigomática e a espessura de incisivo central maxilar para identificação de gênero

Objective: The aim of the present study is to analyze and evaluate the applicability of bizygomatic and maxillary central incisor width in identifying the sex of an individual for anthropological studies. Material and Methods: The study was conducted on 100 individuals in a private dental institution. The width of the central incisor was measured by requesting the subject to bite onto a sheet of modelling wax. The bizygomatic width was calculated with the help of a divider by taking the most prominent area of the zygomatic arch as the reference point bilaterally. Berry's formula was used to calculate the width of the maxillary central incisor from the bizygomatic width. Berry's Formula "Width of the maxillary central incisor = Bizygomatic width / 16." The data obtained was tabulated and analyzed statistically. Results: The results in our study indicated that the widths of both maxillary central incisors and the bizygomatic width were found to be higher in males when compared to females with a positive strong correlation. Conclusion: The Berry's index can be used for identifying the gender and can also be used for facial reconstruction. (AU)

Objetivo: O objetivo do presente estudo é analisar e avaliar a aplicabilidade da distância bizigomática e espessura de incisivos centrais maxilares na identificação do sexo de um indivíduo para estudos antropológicos. Material e métodos: O estudo foi conduzido com 100 indivíduos de uma instituição odontológica privada. A espessura do incisivo central foi medida pedindo ao sujeito que mordesse em uma folha de cera. A espessura bizigomática foi calculada com o auxílio de uma régua pegando a área mais proeminente do arco zigomático como ponto de referência bilateralmente. A fórmula de Berry foi usada para calcular a espessura do incisivo central maxilar da espessura zigomática. Fórmula de Berry: "Espessura do incisivo central maxial = Espessura bizigomática / 16". Os dados obtidos foram tabulados e analisados estatisticamente. Resultados: Os resultados em nosso estudo indicaram que as espessuras de ambos os incisivos centrais maxilares como as espessuras bizigomáticas foram maiores no sexo masculino do que no sexo feminino, com uma correlação positiva forte. Conclusão: O índice de Berry pode ser usado para identificação de gênero e também pode ser usado para reconstrução facial. (AU)

Knockdown of transglutaminase-2 prevents early age-induced vascular changes in mice

Abstract Purpose: To determine whether the absence of transglutaminase 2 enzyme (TG2) in TG2 knockout mice (TG2-/-) protect them against early age-related functional and histological arterial changes. Methods: Pulse wave velocity (PWV) was measured using non-invasive Doppler and mean arterial pressure (MAP) was measured in awake mice using tail-cuff system. Thoracic aortas were excised for evaluation of endothelial dependent vasodilation (EDV) by wire myography, as well as histological analyses. Results: PWV and MAP were similar in TG2-/-mice to age-matched wild type (WT) control mice. Old WT mice exhibited a markedly attenuated EDV as compared to young WT animals. The TG2-/-young and old mice had enhanced EDV responses (p<0.01) as compared to WT mice. There was a significant increase in TG2 crosslinks by IHC in WT old group compared to Young, with no stain in the TG2-/-animals. Optical microscopy examination of Old WT mice aorta showed thinning and fragmentation of elastic laminae. Young WT mice, old and young TG2-/-mice presented regularly arranged and parallel elastic laminae of the tunica media. Conclusion: The genetic suppression of TG2 delays the age-induced endothelial dysfunction and histological modifications.

Halophilic benthic diatom Amphora coffeaeformis—A potent biomarker for lipid and biomedical application

Biomarkers offer a powerful means to study life and its interaction with the environment. The evolutionary record of eukaryotic phytoplankton has been studied through fossils and molecular biomarkers, such as lipids and nucleic acids. Algal taxonomic groups containing carotenoids serve as age diagnostic biomarkers associated to photosynthetic pigments or fatty acids. Diatoms are known to thrive even in hypersaline conditions which are restricted to higher salinity. Results of this study have revealed the presence of a specific highly branched isoprenoid (HBI) compound, squalene (0.004 g) in the halophilic benthic diatom Amphora coffeaeformis which is considered to be the marker for hyper salinity. It also corresponds to the biomarker of oxidation processes.

Nitric oxide inhibitory and anti-Bacillus activity of phenolic compounds and plant extracts from Mesua species

ABSTRACT Species from the genus Mesua, Calophyllaceae, are rich source for phenolic compounds such as coumarin xanthone, and benzophenone derivatives. An investigation on the potential biologically active phenolic compounds 1-5 and crude extracts from the stem bark of Mesua hexapetala (Hook. f.) P.S. Ashton and Mesua beccariana (Baill.) Kosterm. for nitric oxide inhibitory activity on RAW 264.7 macrophage as well as anti-Bacillus activity on selected Bacillus were carried out. Hexapetarin (1), which we reported as a new compound isolated from M. hexapetala showed very good nitric oxide inhibitory activity with an IC50 value of 30.79 ± 2.68 µM. This compound also gave very significant activities towards Bacillus subtilis ATCC 6633, Bacillus cereus ATCC 33019, Bacillus megaterium ATCC 14581 and Bacillus pumilus ATCC 14884 in disc diffusion and minimum inhibitory concentrations assay. Moreover, 1,3,7-trihydroxy-2,4-di (3-methyl-2-butenyl)xanthone (2) isolated from M. hexapetala showed very significant nitric oxide inhibitory activity with an IC50 value of 12.41 ± 0.89 µM and does not exhibit anti-Bacillus activity on four types of Bacillus. Meanwhile, compounds 3-5 were inactive in the nitric oxide activity test and anti-Bacillus assay.

Molecular characterisation for clonality and transmission dynamics of an outbreak of Klebsiella pneumoniae amongst neonates in a tertiary care centre in South India

Purpose: Sepsis is a significant cause of morbidity and mortality amongst neonates. Klebsiella pneumoniae is a common cause of nosocomial outbreaks causing bacteraemia and having potential of acquiring plasmids enhancing antimicrobial resistance. In the present study, we investigate K. pneumoniae outbreak causing bacteraemia amongst neonates over a span of 2 months. Isolates were characterised for antimicrobial resistance, virulence, molecular typing for clonality and plasmid typing for transmission dynamics, and patient outcome was investigated. Methods: Thirteen isolates of K. pneumoniae were obtained during October–November 2016. Antimicrobial susceptibility testing was performed, and multiplex polymerase chain reaction (PCR) for ?-lactamases and PCR for ompK35 and ompK36 were performed. To study hypervirulence, string test and PCR for rmpA and rmpA2 were performed. Multilocus sequence typing and Inc plasmid typing were carried out to study transmission dynamics. Results: Amongst 13 isolates, all isolates harboured blaSHVand blaTEM; 12 isolates carried blaCTX-M-1. ompK35 was present in all, but ompK36 was absent in 12 isolates. Ten isolates belonged to ST48, 6 amongst which contained IncFII (K) plasmid. One isolate each belonged to ST29, ST111 and ST2647 (novel clone). None of the isolates was hypervirulent. Conclusion: Extended-spectrum ?-lactamase K. pneumoniae is commonly seen in Indian hospitals and main mechanisms being production of SHV, TEM and CTX-M enzymes as seen in the present study. Outer membrane porins contribute significantly to antimicrobial resistance. Emergence of new clones such as ST2647 implies continuous evolution of the organism and also potential for rapid genetic recombination leading to multidrug resistance. Outbreaks amongst neonates lead to fatal outcome, and stringent hospital infection control is necessary.

Endophytic fungi from Maliau Basin, Sabah: Identification of strains and antimicrobial compounds

Aims@#Endophytic fungi are microorganisms that live asymptomatically within plant tissues, producing a wide range of metabolites, including compounds potentially useful for drug development. We investigated endophytic fungi from Maliau Basin, Sabah to identify strains producing bioactive compounds, notably with antimicrobial activity. @*Methodology and results@#A total of 23 plants were sampled yielding 345 endophytic fungal isolates. Of these, 44 isolates were screened for antimicrobial activity against nine species of bacteria and fungi, revealing 14 endophytes producing bioactive metabolites. Crude fungal extracts were obtained from broth cultures of endophytic isolates with promising activity while the fungal strains were identified using molecular methods. The crude extract of endophyte MB4 WA10, isolated from Callophyllum sp. (bintangor) showed IC50 of 2.6 mg/mL against S. aureus and 0.6 mg/mL against B. subtilis while the extract of MB22 WA16, an isolate identified as Valsaceae sp., was also active against S. aureus with an IC50 of 1.37 mg/mL. Another isolate, namely MB5 L4 (WA), was identified as a Phomopsis sp. and its extract was the most active against S. aureus with an IC50 of 1 mg/mL. The HPLC fraction of this fungal extract with the highest inhibition (92.37%) of S. aureus was purified for compound isolation and identification. A polyketide compound, 2,3-dihydro-2- hydroxy-2,4-dimethyl-5-trans-propenylfuran-3-one (C9H12O3), with molecular weight of 168.192 was identified based on mass spectral and NMR data analysis. This previously identified compound is known to have other antimicrobial properties. @*Conclusion, significance and impact of study@#Rainforests in Malaysia, especially Maliau Basin, harbour many species of fungal endophytes, producing useful bioactive compounds that may be explored for further potential uses, including antimicrobial activity.

Determination of Ergosterol as a Potential Biomarker in Pathogenic Medically Important Fungal Isolates / Jurnal Sains Kesihatan Malaysia

Ergosterol, a component of fungal cell membrane, has been frequently detected as an indicator of fungal presence and massin environmental samples like soil. However, its detection in major pathogenic fungal species has not been investigated.In this study, the ergosterol contents of ten pathogenic fungal species were determined. Liquid chromatography was usedfor the detection and quantification of ergosterol extracted from fungal broth cultures. Results showed that ergosteroleluted as a single, well resolved peak in the chromatogram profiles of all tested fungi. Based upon relative amounts ofergosterol produced per fungal mycelial dry weight, three groups of fungal pathogens were identified, namely low ergosterol(Aspergillus niger, Candida albicans and Cryptococcus neoformans at 4.62, 6.29 and 7.08 μg/mg, respectively), mediumergosterol (Fusarium solani, Aspergillus fumigatus, Mucor sp., Penicillium sp., Cryptococcus gattii and Rhizopus sp.at 9.40, 10.79, 10.82, 11.38, 12.60 and 13.40 μg/mg, respectively), and high ergosterol (Candida tropicalis at 22.84 μg/mg), producers. Ergosterol was not detectable in bacterial samples, which were included as controls. This first report onergosterol detection in major pathogenic fungal species indicates that ergosterol may be used as a biomarker to diagnoseinvasive fungal infections in clinical sampl

Pedigree Analysis and Audiological Investigations of Otosclerosis: An Extended Family Based Study

BACKGROUND AND OBJECTIVES: To analyse the audiometric profile and the pedigree of a large family with otosclerosis to understand the inheritance pattern and its implication in clinical management of the disease. SUBJECTS AND METHODS: Pedigree analysis was performed on the basis of family history and audiometric tests. Pure tone audiometry, tympanometry, and acoustic reflexes were evaluated for the family members. Audiometric analysis was also carried out for the individuals who have already underwent corrective surgery at the time of study. RESULTS: Out of 112 family members, 17 were affected individuals, and 11 of them were surgically confirmed. Hearing loss (HL) started unilaterally and progressed to bilateral form. Otosclerosis was presented in early 20’s in the first and second generations but it was delayed to mid-late 30’s in the fourth generation. An affected female was diagnosed with otosclerosis during her pregnancy. Though the disease was familial, a mother of four affected offspring in this family did not develop otosclerosis until she died at the age of 84. CONCLUSIONS: The five-generation family, which was analysed in the present study, exhibited autosomal dominant inheritance of otosclerosis with reduced penetrance. Bilateral HL and pregnancy-aggravated otosclerosis were observed in this family. It was found for the first time that the age of onset of the disease delayed in the successive generations. The current study indicated the importance of detailed pedigree analysis for better clinical management of otosclerosis.

Commercial Herbal Slimming Products: Evaluation of Heavy Metals and Microorganism Contamination at Different Batch Production / Jurnal Sains Kesihatan Malaysia

Herbal-based slimming products which are consumed orally may be contaminated with heavy metals as well as microorganisms. This study aimed to evaluate the safety level of these slimming products by determining heavy metals and microbial contamination in different batch production. Six different brands of herbal-based slimming products (A, B, C, G, H and I) with three different batch productions (1, 2 and 3) were investigated (n =18). Five heavy metals Arsenic, Cadmium, Chromium, Copper and Zinc were determined using an Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). The presence of microorganisms was determined by total aerobic count and the bacteria were identified. The samples’ moisture content was determined by calculating the percentage of water loss after drying process. All batches of samples A and B had high content of zinc, over the permissible level of 5ppm while, 6 samples contained Chromium above the permissible level (1.5 ppm). All 3 batches of sample A presented with the highest total daily intake of heavy metals. Bacteria were present in all the samples tested with the highest numbers in samples G, H and A followed by B, I and C. The highest number of fungi was found in product A while product I was free from fungal contamination. Aspergillus spp. was the predominant fungus present in the samples. There was a weak correlation between moisture content and bacteria (r = 0.087) and fungal (r = 0.253) presence in the samples. As some herbal slimming products contain heavy metals as well as microorganisms, consumers need to be more vigilant and discerning when selecting products to be consumed.