Evaluating Bacillus flexus as bioremediators for ammonia removal in shrimp culture water and wastewater and characterizing microbial communities in shrimp pond sludge.

The accumulation of nitrogen compounds in shrimp farming water and effluent presents a major challenge. Ammonia is a form of nitrogen that limits shrimp growth due to its potential toxicity and effects on shrimp health and water quality. This study is aimed at identifying promising bioremediators from shrimp pond sludge to mitigate ammonia levels in both culture water and wastewater and at determining major bacterial communities in sludge using metagenomic analysis. A sludge sample was collected from a shrimp pond in Selangor, Malaysia, to isolate potential ammonia-removing bacteria. Out of 64 isolated strains, Bacillus flexus SS2 showed the highest growth in synthetic basal media (SBM) containing ammonium sulfate at a concentration of 70 mg/L as the sole nitrogen source. The strain was then incubated in SBM with varying pH levels and showed optimal growth at pH 6.5-7. After 24 h of incubation, B. flexus SS2 reduced the ammonia concentration from an initial concentration of 5 to 0.01 mg/L, indicating a 99.61% reduction rate, which was highest in SBM at pH 7. Moreover, the strain showed ammonia removal ability at concentrations ranging from 5 to 70 mg/L. Metagenomic analysis revealed that Proteobacteria was the most abundant phylum in the sludge, followed by Cyanobacteria, Actinobacteria, Chloraflexi, Firmicutes, and Campilobacterota. Bacillus flexus SS2 belongs to the Bacillota phylum and has the potential to serve as a bioremediator for removing ammonia from shrimp culture water and wastewater.

Assessment of Fasciola and Paramphistomes co-infection in large ruminants through faecal egg counts around Taiping, Malaysia.

Emerging cases of Fasciola and Paramphistomes co-infection have been reported, especially in tropical regions. Thisis due to Fasciola and Paramphistomes sharing biological factors which influence the pattern of transmission, especially in faecal egg shedding due to interaction and competition in the definitive host. Most reports surveyed the occurrence of fasciolosis in ruminants with a lack of observation of faecal egg distribution. Therefore, present study is aimed to assess the distribution of Fasciola and Paramphistomes faecal egg count (fec) in co-infected large ruminants in Larut, Matang, and Selama areas (Taiping). A total of 371 faecal samples were collected at random from 23 ruminant herds. Flukefinder® sedimentation was used to quantify the Fasciola and Paramphistomes eggs. Descriptive analyses were performed to determine the prevalence of co-infections, and Spearman correlation analysis was used to correlate the fec. Overall, the prevalence of Fasciola and Paramphistomes co-infection was 23.7% (n=89/371) in Taiping. Prevalence of paramphistomosis was always higher than fasciolosis in overall and single infection, with 46.9% (n=174/371) and 22.9% (n=85/371) compared to 36.9% (n=137/371) and 12.9% (n=48/371) respectively. Egg per gram (epg) of both parasites were positively skewed with a median of 1.5 epg in fasciolosis and 10.5 epg in paramphistomosis. Spearman correlation analysis of the epg in co-infected bovine was found to have a moderately positive correlation with rs=0.39 (p-value<0.01). The recent study observed a moderate prevalence of Fasciola and Paramphistomes coinfection in a large ruminant population from Taiping, with the prevalence of paramphistomosis being higher than fasciolosis. Hence, this suggests that infection with one of these parasites increases the chance of infection with another. There is a need to integrate fec in parasite surveillance to monitor the trend of parasite transmission. Findings in the present study could tailor control strategies, especially for fasciolosis to limit the economic loss and prevent zoonotic transmission.

Alcohol Dependence Modulates Amygdalar mTORC2 and PKCε Expression in a Rodent Model.

Multiple alcohol use disorder (AUD)-related behavioral alterations are governed by protein kinase C epsilon (PKCε), particularly in the amygdala. Protein kinase C (PKC) is readily phosphorylated at Ser729 before activation by the mTORC2 protein complex. In keeping with this, the current study was conducted to assess the variations in mTORC2 and PKCε during different ethanol exposure stages. The following groups of rats were employed: control, acute, chronic, ethanol withdrawal (EW), and EW + ethanol (EtOH). Ethanol-containing and non-ethanol-containing modified liquid diets (MLDs) were administered for 27 days. On day 28, either saline or ethanol (2.5 g/kg, 20% v/v) was intraperitoneally administered, followed by bilateral amygdala extraction. PKCε mRNA levels were noticeably increased in the amygdala of the EW + EtOH and EW groups. Following chronic ethanol consumption, the stress-activated map kinase-interacting protein 1 (Sin1) gene expression was markedly decreased. In the EW, EW + EtOH, and chronic ethanol groups, there was a profound increase in the protein expression of mTOR, Sin1, PKCε, and phosphorylated PKCε (Ser729). The PKCε gene and protein expressions showed a statistically significant moderate association, according to a correlation analysis. Our results suggest that an elevated PKCε protein expression in the amygdala during EW and EW + EtOH occurred at the transcriptional level. However, an elevation in the PKCε protein expression, but not its mRNA, after chronic ethanol intake warrants further investigation to fully understand the signaling pathways during different episodes of AUD.

Assessment of Fasciola and Paramphistomes co-infection in large ruminants through faecal egg counts around Taiping, Malaysia

@#Emerging cases of Fasciola and Paramphistomes co-infection have been reported, especially in tropical regions. Thisis due to Fasciola and Paramphistomes sharing biological factors which influence the pattern of transmission, especially in faecal egg shedding due to interaction and competition in the definitive host. Most reports surveyed the occurrence of fasciolosis in ruminants with a lack of observation of faecal egg distribution. Therefore, present study is aimed to assess the distribution of Fasciola and Paramphistomes faecal egg count (fec) in co-infected large ruminants in Larut, Matang, and Selama areas (Taiping). A total of 371 faecal samples were collected at random from 23 ruminant herds. Flukefinder® sedimentation was used to quantify the Fasciola and Paramphistomes eggs. Descriptive analyses were performed to determine the prevalence of co-infections, and Spearman correlation analysis was used to correlate the fec. Overall, the prevalence of Fasciola and Paramphistomes co-infection was 23.7% (n=89/371) in Taiping. Prevalence of paramphistomosis was always higher than fasciolosis in overall and single infection, with 46.9% (n=174/371) and 22.9% (n=85/371) compared to 36.9% (n=137/371) and 12.9% (n=48/371) respectively. Egg per gram (epg) of both parasites were positively skewed with a median of 1.5 epg in fasciolosis and 10.5 epg in paramphistomosis. Spearman correlation analysis of the epg in co-infected bovine was found to have a moderately positive correlation with rs=0.39 (p-value<0.01). The recent study observed a moderate prevalence of Fasciola and Paramphistomes coinfection in a large ruminant population from Taiping, with the prevalence of paramphistomosis being higher than fasciolosis. Hence, this suggests that infection with one of these parasites increases the chance of infection with another. There is a need to integrate fec in parasite surveillance to monitor the trend of parasite transmission. Findings in the present study could tailor control strategies, especially for fasciolosis to limit the economic loss and prevent zoonotic transmission.

Gastrointestinal parasitic infections of buffaloes (Bubalus bubalis) in Sarawak Borneo: Prevalence, risk factors, and farming practices.

The objectives of this study were to investigate the prevalence and associated risk factors for gastrointestinal (GI) parasites in buffaloes from various areas of Sarawak, and to assess current management practices of GI parasites among farmers. Faecal samples were collected from 15 farms and 129 animals, as well as data on farm and animal-based characteristics. A total of 129 faecal samples were examined for GI parasites using a modified McMaster and sedimentation. Association between potential risk factors and the prevalence of GI parasites was investigated using Chi-square statistic. The prevalence of Paramphistomum sp., strongyles, and coccidia were 75.2% (95% CI±7.5), 52.7% (95% CI±8.6) and 48.1% (95% CI±8.6), respectively. Farms which had a grazing area less than 50 acres in size had significantly higher prevalence of strongyles (70.5%, χ2 = 8.34, P = 0.004) and paramphistomes (88.6%, χ2 = 6.46, P = 0.01) relative to farms with a larger grazing area (43.5% and 68.2%, respectively). Prevalence of strongyles was lower in farms that did not implement a cut- and-carry system (45.6%, χ2 = 4.17, P = 0.04) in comparison to those that did (64%). The prevalence of paramphistomes was higher on farms with more than 40 animals (80.6%, χ2 = 3.18, P = 0.05) relative to farms with fewer animals. The majority of farmers surveyed (67.9%) showed awareness of GI parasite infection and reported that they recognized the associated symptoms. Most farmers practised deworming, and ivermectin was the most commonly used anthelminthic (60.4%); only 1.9% of farmers used albendazole. Overall this study revealed a high prevalence of GI parasites in buffalo in Sarawak. Although farmers report they are aware of parasitic diseases, further education is still required. This could include how they can successfully implement on-farm changes to reduce the prevalence of GI parasites in their herds.

Gastrointestinal parasitic infections of buffaloes (Bubalus bubalis) in Sarawak Borneo: Prevalence, risk factors, and farming practices

@#The objectives of this study were to investigate the prevalence and associated risk factors for gastrointestinal (GI) parasites in buffaloes from various areas of Sarawak, and to assess current management practices of GI parasites among farmers. Faecal samples were collected from 15 farms and 129 animals, as well as data on farm and animal-based characteristics. A total of 129 faecal samples were examined for GI parasites using a modified McMaster and sedimentation. Association between potential risk factors and the prevalence of GI parasites was investigated using Chi-square statistic. The prevalence of Paramphistomum sp., strongyles, and coccidia were 75.2% (95% CI±7.5), 52.7% (95% CI±8.6) and 48.1% (95% CI±8.6), respectively. Farms which had a grazing area less than 50 acres in size had significantly higher prevalence of strongyles (70.5%, χ² = 8.34, P = 0.004) and paramphistomes (88.6%, χ² = 6.46, P = 0.01) relative to farms with a larger grazing area (43.5% and 68.2%, respectively). Prevalence of strongyles was lower in farms that did not implement a cut- and-carry system (45.6%, χ² = 4.17, P = 0.04) in comparison to those that did (64%). The prevalence of paramphistomes was higher on farms with more than 40 animals (80.6%, χ² = 3.18, P = 0.05) relative to farms with fewer animals. The majority of farmers surveyed (67.9%) showed awareness of GI parasite infection and reported that they recognized the associated symptoms. Most farmers practised deworming, and ivermectin was the most commonly used anthelminthic (60.4%); only 1.9% of farmers used albendazole. Overall this study revealed a high prevalence of GI parasites in buffalo in Sarawak. Although farmers report they are aware of parasitic diseases, further education is still required. This could include how they can successfully implement on-farm changes to reduce the prevalence of GI parasites in their herds.

Expression of oestrogen-α receptor in papillary thyroid carcinoma and its association with metastasis.

INTRODUCTION: Papillary thyroid carcinoma (PTC) is the ninth most common malignancy among women. Although the disease prognosis is good, less favourable outcomes are predicted in those with higher disease stages and nodal metastasis. Oestrogen- α (ER-α) expression has been associated with aggressive presentation and greater disease progression and has been proposed as a predictor for lymph node metastases. The objective of this study was to evaluate the association between ER expression and clinicopathological features i.e. lymph node metastasis, tumour size, extrathyroidal extension, histological variants of PTC , age groups , ethnic and gender. METHODS: We studied ER-α expression in 84 cases of PTC obtained within an eight-year period (2011-2018) by immunohistochemical technique (IHC). Associations between ER-α expression and clinicopathological features were evaluated using Fisher's exact test. The statistical significance was set at p < 0.05. RESULTS: ER-α was expressed in 13.1% of all the PTC cases examined (n=11/84). There were no associations observed between ER-α expression and lymph node metastasis (p=1.000), tumour size (p=0.970), extrathyroidal extension (p=0.677), variants of PTC (p=1.000), age groups (p=0.188), gender (p=0.725) or race (p=0.920). CONCLUSION: There was no evidence in this study to support the application of ER-α as prediction marker for lymph node metastasis or disease aggressiveness in PTC. Given that the scope of this study was limited to the protein expression of ER- α, we also propose the inclusion of molecular analysis of ESR1 gene expression, as well as inclusion of detailed clinical and radiological findings in future research investigating the role of ER-α in prognostication of PTC.

Diagnostic performance of prostate health index (PHI) in predicting prostate cancer on prostate biopsy.

INTRODUCTION: Prostate health index (PHI) has been shown to have better diagnostic accuracy in predicting prostate cancer (PCa) in men with total prostate-specific antigen (PSA) levels between 4-10ng/ml. However, little is known of its value in men with elevated PSA beyond this range. This study aimed to evaluate the diagnostic performance of PHI in Malaysian men with elevated PSA values ≤ 20ng/ml. MATERIALS AND METHODS: From March 2015 to August 2016, all men consecutively undergoing transrectal ultrasound (TRUS)-guided prostate biopsy with total PSA values ≤ 20ng/ ml were recruited. Blood samples were taken immediately before undergoing prostate biopsy. The performance of total PSA, %fPSA, %p2PSA and PHI in determining the presence of PCa on prostate biopsy were compared. RESULTS: PCa was diagnosed in 25 of 84 patients (29.7%). %p2PSA and PHI values were significantly higher (p<0.05) in patients with PCa than those without PCa. The areas under the receiver operating characteristic curves for total PSA, %fPSA, %p2PSA and PHI were 0.558, 0.560, 0.734 and 0.746, respectively. At 90% sensitivity, the specificity of PHI (42.4%) was five times better than total PSA (8.5%) and two times better than %fPSA (20.3%). By utilising PHI cut-off >22.52, 27 of 84 (32.1%) patients could have avoided undergoing biopsy. CONCLUSION: Findings of our study support the potential clinical effectiveness of PHI in predicting PCa in a wider concentration range of total PSA up to 20ng/ml.

Asthma control and asthma treatment adherence in primary care: results from the prospective, multicentre, non-interventional, observational cohort ASCOPE study in Malaysia.

INTRODUCTION: As the first point of contact for those presenting with asthma symptoms, primary healthcare plays a crucial role in asthma management. This is a nationwide study of assessment of asthma symptom control and adherence to asthma medication among outpatients in public health clinics in Malaysia. METHODS: This is a prospective, observational multicentre study (ASCOPE; NCT03804632). Data on asthma control, assessment of control symptoms, and adherence to treatment were collected from medical records and interviews of patients. The level of asthma control was assessed using the Global Initiative for Asthma (GINA) Assessment of Symptom Control. Adherence of patient to medication for asthma was assessed through interview of patients using four questions adapted from the Malaysian Medication Adherence Scale. RESULTS: Among the 1011 patients recruited, 416 (41%) had well controlled asthma, 388 (38%) were partly controlled, and 207 (21%) had uncontrolled asthma. Majority (81%) had mild asthma and all patients were on asthma medication. Most patients did not have spirometry data (97%) but underwent peak flow rate measurements (98%). Poor adherence occurred at all levels of asthma control but was worst among those with uncontrolled asthma. This was statistically significant across all four questions on adherence (p<0.05). For example, more patients with uncontrolled asthma forgot doses (56%) or stopped treatment (39%) than those with well-controlled asthma (44% and 27%respectively). CONCLUSIONS: Among Malaysian primary care patients with asthma, less than 50% had well-controlled asthma, and low adherence to treatment was common. More effort is needed to improve asthma control among patients in Malaysia, including those with mild asthma.

Identification of the amino acids in the Major Histocompatibility Complex class II region of Scottish Blackface sheep that are associated with resistance to nematode infection.

Lambs with the Major Histocompatibility Complex DRB1*1101 allele have been shown to produce fewer nematode eggs following natural and deliberate infection. These sheep also possess fewer adult Teladorsagia circumcincta than sheep with alternative alleles at the DRB1 locus. However, it is unclear if this allele is responsible for the reduced egg counts or merely acts as a marker for a linked gene. This study defined the MHC haplotypes in a population of naturally infected Scottish Blackface sheep by PCR amplification and sequencing, and examined the associations between MHC haplotypes and faecal egg counts by generalised linear mixed modelling. The DRB1*1101 allele occurred predominately on one haplotype and a comparison of haplotypes indicated that the causal mutation or mutations occurred in or around this locus. Additional comparisons with another resistant haplotype indicated that mutations in or around the DQB2*GU191460 allele were also responsible for resistance to nematode infections. Further analyses identified six amino acid substitutions in the antigen binding site of DRB1*1101 that were significantly associated with reductions in the numbers of adult T. circumcincta.

Dosimetric response of fabricated Ge-doped optical fibres in computed tomography RQT beam quality x-ray beams.

Novel germanium (Ge)-doped silica glass fibres tailor-made in Malaysia are fast gaining recognition as potential media for thermoluminescence (TL) dosimetry, with active research ongoing into exploitation of their various beneficial characteristics. Investigation is made herein of the capability of these media for use in diagnostic imaging dosimetry, specifically at the radiation dose levels typically obtained in conduct of Computed Tomography (CT). As a first step within such efforts, there is need to investigate the performance of the fibres using tightly defined spectra, use being made of a Philips constant potential industrial x-ray facility, Model MG165, located at the Malaysian Nuclear Agency Secondary Standards Dosimetry Lab (SSDL). Standard radiation beam qualities (termed RQT) have been established for CT, in accord with IEC 61267: 2003 and IAEA Technical Reports Series No. 457: 2007. A calibrated ionisation chamber has also been utilised, forming a component part of the SSDL equipment. The fabricated fibres used in this study are 2.3 mol% flat fibre (FF) of dimensions 643 × 356 µm2 and 2.3 mol% cylindrical fibre (CF) of 481 µm diameter, while the commercial fibre used is 4 mol% with core diameter of 50 µm. The dopant concentrations are nominal preform values. The fibres have been irradiated to doses of 20, 30 and 40 milligray (mGy) for each of the beam qualities RQT 8, RQT 9 and RQT 10. For x-rays generated at constant potential values from 100 to 150 kV, a discernible energy-dependent response is seen, comparisons being made with that of lithium fluoride (LiF) thermoluminescence dosimeters (TLD-100). TL yield versus dose has also been investigated for x-ray doses from 2 to 40 mGy, all exhibiting linearity. Compared to TLD-100, greater sensitivity is observed for the fibres.

Gleason scores in prostate needle biopsy and prostatectomy specimens in prostatic adenocarcinoma: A correlation study.

INTRODUCTION: Gleason scoring (GS) categorised prostatic adenocarcinoma into five prognostic grade groups (PGGs); associated with different prognosis and treatment. This study aims to correlate between Gleason scores of needle biopsies with the corresponding total prostatectomy specimens, and to assess the relationship between the percentage of Gleason 4 tumour pattern (GP4) within Gleason score 7 (GS7) needle biopsy groups with the pathological staging. MATERIALS AND METHODS: Seventy-eight specimens of needle prostate biopsy and its subsequent radical prostatectomy were retrospectively studied. The GSs of the needle biopsy were compared with the corresponding prostatectomy specimens. The percentage of GP4 in GS7 needle biopsy groups was calculated and correlated with the pathological staging. RESULTS: More than half (60%) of GS 6 needle biopsy cases (PGG 1) were upgraded in the prostatectomy specimen, while the majority (80%) of the GS7 needle biopsy groups (PGG 2 and 3) remain unchanged. Cohen's Kappa shows fair agreement in the Gleason scoring between needle biopsies and prostatectomy specimens, K = 0.324 (95% CI, 6.94 to 7.29), p <0.0005 and in the percentage of GP4 in GS7 needle biopsy groups and their corresponding radical prostatectomy specimens, K = 0.399 (95% CI 34.2 - 49.2), p<0.0005. A significant relationship was seen between the percentage of GP4 in GS7 needle biopsy with the pT and pN stage of its radical prostatectomy (p = 0.008 and p=0.001 respectively). CONCLUSION: A higher percentage of GP4 in GS7 tumour is associated with worse tumour behaviour, therefore it is crucial for clinicians to realise this in deciding the optimal treatment.

The diagnostic utility of cytokeratin 19 in differentiating malignant from benign thyroid lesions.

INTRODUCTION: Thyroid carcinoma is classically diagnosed based on certain histological criteria. In some cases, definitive diagnoses may be challenging when morphological features are equivocal. This study evaluated the usefulness of Cytokeratin 19 (CK 19) as an immunohistochemical marker to differentiate the different histological types of malignant thyroid neoplasms, particularly papillary thyroid carcinoma (PTC) from benign thyroid lesions. MATERIALS AND METHODS: We collected 54 malignant and 65 benign thyroid lesions diagnosed by histology in Universiti Kebangsaan Malaysia Medical Centre between January 2010 and December 2015. All cases were immunohistochemically stained with CK 19 and evaluated by 3 independent observers. The immunostaining patterns were scored based on the intensity and proportion of staining and finally graded as negative, weak positive, moderate positive or strong positive. In addition, the immunostaining scores of the malignant cases were correlated with their TNM pathological tumour stages. RESULTS: Cytokeratin 19 staining expression was higher in malignant than benign thyroid lesions (p < 0.001) which was most prominent among classical PTC. The four PTC cases that showed negative or weak staining were all follicular variant of PTC. Benign conditions were mostly negative or showed weak positivity. There was no correlation between CK 19 expression and TNM primary tumour stage (pT). CONCLUSION: Cytokeratin 19 is a useful marker in differentiating malignant from benign thyroid conditions particularly the classical PTC, provided its interpretation is by correlation with morphology and takes into consideration the intensity and proportion of positive staining.

Boer goats appear to lack a functional IgA and eosinophil response against natural nematode infection.

Gastrointestinal nematode infection is one of the major diseases affecting small ruminants. Although some breeds of goats are quite resistant, many breeds of goats are relatively susceptible. This study used a combined parasitological, immunological, bioinformatic and statistical approach to examine the role of goat IgA and eosinophils in protection against Teladorsagia circumcincta. Molecular modelling suggested that the transmembrane domain of the high affinity IgA receptor was dysfunctional in goats. Statistical analyses failed to find any association in naturally infected goats between high IgA or eosinophil responses and low faecal egg counts. Together these results indicate that IgA and eosinophil responses against T. circumcincta are less effective in goats than sheep.

CD56 expression in benign and malignant thyroid lesions.

INTRODUCTION: Thyroid cancer is the most common endocrine malignancy with more than 95% originating from follicular epithelial cells. Diagnostic dilemma may arise in occasional cases such as when an encapsulated nodule with a follicular growth pattern exhibits clear nuclei with grooves making it difficult to distinguish a follicular adenoma from encapsulated follicular variant papillary thyroid carcinoma. This study aimed to evaluate the diagnostic utility of an immunohistochemical marker, CD56, to distinguish between benign and malignant thyroid lesions. MATERIALS AND METHODS: We retrospectively studied CD56 expression in 54 benign and 54 malignant thyroid lesions using archival formalin fixed paraffin-embedded tissue blocks for the study period from January 2010 to December 2015, diagnosed in a tertiary hospital. RESULTS: CD56 was expressed in 52/54 (96.3%) of benign specimens and only 24/54 (44.4%) of malignant ones. The malignant specimens comprised 31 (57.4%) papillary thyroid carcinomas (PTC), 11 (20.3%) follicular carcinomas (FC), seven (13%) medullary thyroid carcinomas (MC), one (1.9%) poorly differentiated carcinoma (PC) and four (7.4%) anaplastic carcinomas (AC). CD56 was not expressed in 28/31 (90.3%) of the PTCs, 1/11 (9.1%) FCs, 1/4 (25%) of ACs while all MCs and the PD were positive. The benign group comprised nodular hyperplasias (29/54), lymphocytic thyroiditis (10/54), follicular adenomas (FA) (14/54) and one hyalinising trabecular tumour. CD56 was expressed in all the benign cases except one FA and one nodular hyperplasia. Thirteen of the 14 FAs were CD56 positive. The difference in expression between benign and malignant tumours was statistically significant as the p value was <0.01. CONCLUSION: CD56 is a potentially good immunohistochemical marker for differentiating papillary thyroid carcinoma from other benign follicular lesions of the thyroid especially in differentiating follicular variant PTC from FA in equivocal cases.

Unexplained ascites, a sign for neuroendocrine carcinoma.

Neuroendocrine neoplasm is an epithelial neoplasm with predominant neuroendocrine differentiation that can arise from many organs in the body. We reported a rare case of gastric neuroendocrine carcinoma which accounts for less than 1% of all gastric tumours that is associated with poor prognosis. The recognition of this rare tumour in early stage is challenging and high suspicious into it might bring to early detection and so forth might improve the prognostication.

Proteomics Analysis of Ovarian Cancer Cell Lines and Tissues Reveals Drug Resistance-associated Proteins.

BACKGROUND: Carboplatin and paclitaxel form the cornerstone of chemotherapy for epithelial ovarian cancer, however, drug resistance to these agents continues to present challenges. Despite extensive research, the mechanisms underlying this resistance remain unclear. MATERIALS AND METHODS: A 2D-gel proteomics method was used to analyze protein expression levels of three human ovarian cancer cell lines and five biopsy samples. Representative proteins identified were validated via western immunoblotting. Ingenuity pathway analysis revealed metabolomic pathway changes. RESULTS: A total of 189 proteins were identified with restricted criteria. Combined treatment targeting the proteasome-ubiquitin pathway resulted in re-sensitisation of drug-resistant cells. In addition, examination of five surgical biopsies of ovarian tissues revealed α-enolase (ENOA), elongation factor Tu, mitochondrial (EFTU), glyceraldehyde-3-phosphate dehydrogenase (G3P), stress-70 protein, mitochondrial (GRP75), apolipoprotein A-1 (APOA1), peroxiredoxin (PRDX2) and annexin A (ANXA) as candidate biomarkers of drug-resistant disease. CONCLUSION: Proteomics combined with pathway analysis provided information for an effective combined treatment approach overcoming drug resistance. Analysis of cell lines and tissues revealed potential prognostic biomarkers for ovarian cancer.

The genetic architecture of the MHC class II region in British Texel sheep.

Understanding the structure of the major histocompatibility complex, especially the number and frequency of alleles, loci and haplotypes, is crucial for efficient investigation of the way in which the MHC influences susceptibility to disease. Nematode infection is one of the most important diseases suffered by sheep, and the class II region has been repeatedly associated with differences in susceptibility and resistance to infection. Texel sheep are widely used in many different countries and are relatively resistant to infection. This study determined the number and frequency of MHC class II genes in a small flock of Texel sheep. There were 18 alleles at DRB1, 9 alleles at DQA1, 13 alleles at DQB1, 8 alleles at DQA2 and 16 alleles at DQB2. Several haplotypes had no detectable gene products at DQA1, DQB1 or DQB2, and these were defined as null alleles. Despite the large numbers of alleles, there were only 21 distinct haplotypes in the population. The relatively small number of observed haplotypes will simplify finding disease associations because common haplotypes provide more statistical power but complicate the discrimination of causative mutations from linked marker loci.

H3K9me3 facilitates hypoxia-induced p53-dependent apoptosis through repression of APAK.

Regions of hypoxia occur in most solid tumors, and they are associated with a poor prognostic outcome. Despite the absence of detectable DNA damage, severe hypoxia (<0.1% O2) induces a DNA damage response, including the activation of p53 and subsequent induction of p53-dependent apoptosis. Factors affecting hypoxia-induced p53-dependent apoptosis are unclear. Here we asked whether H3K9me3, through mediating gene repression, could regulate hypoxia-induced p53-dependent apoptosis. Under hypoxic conditions, increases in H3K9me3 occur in an oxygen-dependent but HIF-1-independent manner. We demonstrate that under hypoxic conditions, which induce p53 activity, the negative regulator of p53, APAK, is repressed by increases in H3K9me3 along the APAK loci. APAK repression in hypoxia is mediated by the methyltransferase SETDB1 but not Suv39h1 or G9a. Interestingly, increasing hypoxia-induced H3K9me3 through pharmacological inhibition of JMJD2 family members leads to an increase in apoptosis and decreased clonogenic survival and again correlates with APAK expression. The relevance of understanding the mechanisms of APAK expression regulation to human disease was suggested by analysis of patients with colorectal cancer, which demonstrates that high APAK expression correlates with poor prognosis. Together, these data demonstrate the functional importance of H3K9me3 in hypoxia, and they provide a novel mechanistic link between H3K9me3, p53 and apoptosis in physiologically relevant conditions of hypoxia.

Anti N-Methyl-D-Aspartate Receptor Encephalitis with Ovarian Teratoma: A Dilemma in Diagnosis

We report a rare case of altered mental status in a young patient with immature ovarian teratoma. A 22-year-old woman presented with seizures, hallucination, amnesia and orofacial dyskinesia. Examination and investigation revealed an ovarian massand asalphing-oophorectomy was performed. The histopathological examination result showed an immature teratoma grade 2 with thepresence of immature primitive glial tissue. Her CSF N-Methyl-DAspartic acid receptor (Anti-NMDAR) antibodytest was positive. N-Methyl-D-Aspartic acid receptor antibody associated limbic encephalitis is an autoimmune antibody-mediated neuropsychiatric disorder. Resection of the tumour and immunotherapy resulted in full recovery.