PRR16/Largen Induces Epithelial-Mesenchymal Transition through the Interaction with ABI2 Leading to the Activation of ABL1 Kinase

Advanced or metastatic breast cancer affects multiple organs and is a leading cause of cancer-related death. Cancer metastasis is associated with epithelial-mesenchymal metastasis (EMT). However, the specific signals that induce and regulate EMT in carcinoma cells remain unclear. PRR16/Largen is a cell size regulator that is independent of mTOR and Hippo signalling pathways. However, little is known about the role PRR16 plays in the EMT process. We found that the expression of PRR16 was increased in mesenchymal breast cancer cell lines. PRR16 overexpression induced EMT in MCF7 breast cancer cells and enhances migration and invasion. To determine how PRR16 induces EMT, the binding proteins for PRR16 were screened, revealing that PRR16 binds to Abl interactor 2 (ABI2). We then investigated whether ABI2 is involved in EMT. Gene silencing of ABI2 induces EMT, leading to enhanced migration and invasion. ABI2 is a gene that codes for a protein that interacts with ABL proto-oncogene 1 (ABL1) kinase. Therefore, we investigated whether the change in ABI2 expression affected the activation of ABL1 kinase. The knockdown of ABI2 and PRR16 overexpression increased the phosphorylation of Y412 in ABL1 kinase. Our results suggest that PRR16 may be involved in EMT by binding to ABI2 and interfering with its inhibition of ABL1 kinase. This indicates that ABL1 kinase inhibitors may be potential therapeutic agents for the treatment of PRR16-related breast cancer.

Loss of EMP2 Inhibits Melanogenesis of MNT1 Melanoma Cells via Regulation of TRP-2

Melanogenesis is the production of melanin from tyrosine by a series of enzyme-catalyzed reactions, in which tyrosinase and DOPA oxidase play key roles. The melanin content in the skin determines skin pigmentation. Abnormalities in skin pigmentation lead to various skin pigmentation disorders. Recent research has shown that the expression of EMP2 is much lower in melanoma than in normal melanocytes, but its role in melanogenesis has not yet been elucidated. Therefore, we investigated the role of EMP2 in the melanogenesis of MNT1 human melanoma cells. We examined TRP-1, TRP-2, and TYR expression levels during melanogenesis in MNT1 melanoma cells by gene silencing of EMP2. Western blot and RT-PCR results confirmed that the expression levels of TYR and TRP-2 were decreased when EMP2 expression was knocked down by EMP2 siRNA in MNT1 cells, and these changes were reversed when EMP2 was overexpressed. We verified the EMP2 gene was knocked out of the cell line (EMP2 CRISPR/Cas9) by using a CRISPR/Cas9 system and found that the expression levels of TRP-2 and TYR were significantly lower in the EMP2 CRISPR/Cas9 cell lines. Loss of EMP2 also reduced migration and invasion of MNT1 melanoma cells. In addition, the melanosome transfer from the melanocytes to keratinocytes in the EMP2 KO cells cocultured with keratinocytes was reduced compared to the cells in the control coculture group. In conclusion, these results suggest that EMP2 is involved in melanogenesis via the regulation of TRP-2 expression.

Seroprevalence of acute dengue in a Malaysian tertiary medical centre

Objectives: The aims of this study were to determine the seroprevalence of acute dengue in Universiti Kebangsaan Malaysia [UKM] Medical Centre and its correlation with selected haematological and biochemical parameters

Methods: This cross-sectional study was conducted from January to June 2015. A patient was serologically diagnosed with acute dengue if the dengue virus IgG, IgM or NS-1 antigen was reactive

Results: Out of 1,774 patients suspected to have acute dengue, 1,153 were serologically diagnosed with the infection, resulting in a seroprevalence of 64.9%. Dengue-positive patients had a lower mean platelet count [89 x 10[9]/L] compared to the dengue-negative patients [171 x 10[9]/L] [p<0.0001]. The mean total white cell count was also lower in the dengue-positive cases [4.7 x 10[9]/L vs. 7.2 x 10[9]/L; p<0.0001]. The mean haematocrit was higher in patients with acute dengue [42.5% vs. 40.0%; p<0.0001]. Likewise, the serum alanine transaminase level was also higher in patients with acute dengue [108 U/L vs. 54 U/L; p<0.0001]

Conclusions: Dengue is very prevalent in UKM Medical Centre as most patients suspected to have acute dengue had serological evidence of the infection. The platelet count was the single most likely parameter to be abnormal [i.e. low] in patients with acute dengue

Screening for New Delhi metallo-Beta-lactamase-1 in Enterobacteriaceae: is there a role for the modified Hodge test?

Objective: The New Delhi metallo-Beta-lactamase-1 [NDM-1] enzyme is a plasmid-encoded enzyme that inactivates carbapenem antibiotics. This study aims to ascertain if the modified Hodge test [MHT] has a role in screening for NDM-1 in Enterobacteriaceae with reduced carbapenem susceptibility

Methods: Over a period of one year, all Enterobacteriaceae isolates from all clinical specimens with reduced susceptibility to at least one carbapenem were subjected to MHT and conventional polymerase chain reaction [PCR] detection of the NDM-1 gene

Results: A total of 13,098 Enterobacteriaceae isolates were screened and 63 [0.48%] had reduced susceptibility to at least one carbapenem. Out of the 63 isolates, 45 [71.4%] were MHT-positive. The NDM-1 gene was detected in 18 of the 63 isolates [28.6%]. All 18 PCR-positive isolates were also MHT-positive. Thus, the sensitivity and specificity of the MHT in detecting NDM-1 in Enterobacteriaceae with reduced carbapenem susceptibility are 100% and 40%, respectively

Conclusion: The MHT is a useful test to screen for the presence of NDM-1 in Enterobacteriaceae with reduced carbapenem susceptibility. However, due to its rather low specificity, all MHT-positive isolates should be subjected to alternative tests [e.g. PCR] for confirmation, especially if other types of carbapenemases [e.g. KPC] are prevalent

Paecilomyces lilacinus fungaemia in an AIDS patient:the importance of mycological diagnosis

Fungaemia due to Paecilomyces lilacinus is generally not considered in AIDS patients because this condition is not categorised as an AIDS-indicator illness. We report a case of a 25-year-old lady who presented to our hospital with Guillain-Barre Syndrome, with the subsequent development of refractory fungaemia, multi-organ failure and disseminated intravascular coagulopathy. Amphotericin B was given as empirical antifungal therapy. HIV screening was reactive and Paecilomyces lilacinus was isolated from her blood. The fungaemia did not resolve after one week of amphotericin B treatment. The addition of itraconazole was also unsuccessful in clearing the fungaemia. Accurate mycological diagnosis is important in the care of AIDS patients with fungaemia because of the risk of treatment failure with empirical therapy

In-house developed light cycler real time polymerase chain reaction for rapid detection of respiratory syncytial virus at University Kebangsaan Medical Centre, Malaysia

Respiratory syncytial virus [RSV] is a major cause of respiratory illness in young children worldwide. Its detection by viral culture and immunofluorescent assay is time consuming and has low sensitivity. The objective of the study was to develop In House Light Cycler Real Time Polymerase Chain Reaction for Rapid Detection of Respiratory Syncytial Virus. In house developed Light Cycler Real-Time reverse transcriptase polymerase chain reaction [LC-RT-PCR] assay was standardized using designed primers for the rapid identification of clinical samples of RSV. A total of 130 samples [throat swab and nasopharyngeal aspirate] obtained from University Kebangsaan Malaysia Medical Centre, were efficiently tested and of which 54 [42%] were RSV positive. This study indicated that this newly developed in house rRT-PCR assay proved to be an excellent method for the rapid detection of RSV.

Dengue in Malaysia: an epidemiological perspective study

Dengue is a mosquito-transmitted infection and is an important health problem in Asia for its fatality. The objective of the study was to determine some epidemiological parameters relating to age, gender, community, and prevalence nature, serology and disease severity. One hundred forty nine dengue suspected sera samples were obtained from suspected patients presenting with dengue symptoms. The samples collected were analyzed by serological detection of Immunoglobulin-M [IgM], Immunoglobulin G [IgG], virus culture in cell- line C36/36 and Reverse Transcriptase Polymerase Chain Reaction [RT-PCR]. The studies showed that out of 149 sera samples, 48 [32.2%] were detected dengue virus by cell culture and RT-PCR and 108 [72.5%] positive by serology. Of the 108 cases, 40 [26.8%] were positive to IgM, 61 [41%] were positive to both IgG and IgM, and 7[4.7%] cases were positive to IgG only. Prevalence of dengue was higher in age group 20-29 years and infection was more in male and in Malay population. Epidemic intensity was the highest in the month of June. Dengue is highly endemic in Malaysia and age group 20-29 was vulnerable to infection, male infected more than female and infection was higher in Malay community. Effective preventive and control measures may be strengthened to reduce the infections

Molecular characterization of two Bangladeshi infectious bursal disease virus isolates using the hypervariable sequence of VP2 as a genetic marker

Two Bangladeshi infectious bursal disease virus (IBDV) isolates collected in 2007, termed GB1 and GB3, were subjected to comparative sequencing and phylogenetic analyses. Sequence analysis of a 474-bp hypervariable region in the VP2 gene revealed that among four major amino acid substitutions observed in the strains, two were unique to GB1 and GB3 (Ser217Leu and Ala270Thr) while one substitution was only found in GB1 (Asn299Ser). Among IBDVs from Bangladesh including GB1 and GB3, the rate of identity and homology was around 97~99%. The amino acid sequences of GB1 and GB3 differ from those of previous Bangladeshi IBDV isolates and contain amino acid substitutions Pro222Ala and Asn299Ser (in GB3 only). Phylogenetic analysis revealed that GB1 and GB3 are grouped with other very virulent IBDVs of European and American origin in contrast to two previously isolated Bangladeshi IBDV strains (GenBank accession Nos. AF362776 and AF260317), which belong to the Asian group. It was concluded that GB1 and GB3 belong to a very virulent group of IBDVs. However, amino acid sequences of GB1 and GB3 differ from those of the other Bangladeshi IBDVs by one or two amino acids encoded in the hypervariable region of the VP2 gene.

Drug resistance and susceptibility of mycobacterium tuberculosis identified at university Kebangsaan Malaysia medical centre

The prime objective of the study was to evaluate drug resistance and susceptibility of Mycobacterium tuberculosis isolated at University Kebangsaan Malaysia Medical Centre. A total number of 726 specimens in the form of sputum and bronchial lavage obtained from patients suspected to tuberculosis were analysed for confirmatory identification and antibiotic susceptibility testing. The bacteria was identified initially by culture and staining and finally by BDProbeTecTM ET Mycobacteria kits [2008]. All Mycobacterium tuberculosis isolates were subjected to antibiotic susceptibility against streptomycin, isoniazid, rifampicin and ethambutol using BACTECTM MGITTM 960 system. Out of 726 specimens 16 [32.65%] were identified as mono drug resistance 7 [14.29%] as poly drug resistance, 4 [8.16%] as multi-drug resistance [MDR-TB]. Identified TB bacteria were analysed in the light of sources of samples where brief history of the patients' age, gender and community noted. Bacteriologic testing with antibiotic sensitivity, standardized treatment with supervision and patient support, provision and management of the drugs used in treatment are necessary for effective treatment of tuberculosis

Velocity and elasticity of pregnancy wastage and caesarian deliveries in bangladesh

The aim in this paper is to investigate the effect of age factor of mother as a cause of pregnancy wastage and delivery types. Using the information from 2967 mothers of Rajshahi District within the reproductive span [15-49 years], we have found that the proportion of pregnancy wastage to live birth in two edges of reproductive span is tremendously dodgy whereas in other age groups between 20 and 35 these are comparatively benign. Further, 6717 births were accounted to investigate the flow of caesarian deliveries over the ages. We found 359 caesarian deliveries against 6358 natural [vaginal] deliveries. Some statistical tools were used and the velocity and elasticity curves were drawn to analyze the risk of pregnancy wastage and caesarian deliveries. Our result shows that the risk of caesarian delivery increases with the increased age and this risk moves faster than the age

Antibiotic susceptibility and R-olasmid mediated drug resistance in staphylococcus aureus

A total of twenty-eight Staphylococcus aureus strains were isolated from skin lesion samples. Results of antibiotic susceptibility test showed that these twenty-eight Staphylococcus aureus strains were resistant to ampicillin [72%], amoxycillin [72%], penicillin [72%], cotrimoxazole [15%], cloxacillin [50%] tetracycline [11%], cephradine [22%], cephalexine [7%] and nalidixic acid [18%]. To understand whether this drug resistance phenomenon was plasmid mediated or not, plasmids were isolated from a chosen strain of Staphylococcus aureus [S2] and a 23 KB plasmid was obtained. This 23 KB plasmid was then transferred to an antibiotic sensitive E. coli LE 392 and after which the sensitive E. coli LE 392 strain developed drug resistance. Plasmid analysis of the transformed E. coli LE 392 revealed that it contains a 23 KB plasmid corresponding to that of the donor Staphylococcus aureus strain which may harbor the gene[s] encoding multidrug resistance in the donor Staphylococcus aureus

Effect of hilsa (Tenualosa ilisha) fish in hypercholesterolemic subjects.

Tenualosa ilisha is a popular, tasty fish found in the rivers of Bangladesh round the year. The fish is rich in oil which provides mainly 16:0 and 18:1 fatty acids but also a little of w-3 polyunsaturated fatty acids (w-3 PUFAs). The present study was undertaken to evaluate the effect of eating hilsa fish in hypercholesterolemic subjects. The fish was indeed found to have hypocholesterolemic effect in subjects who had hypercholesterolemia. After 10 months of eating 100g hilsa fish per day, serum total cholesterol level fell from 285.1 to 244.6 mg/dl (14.2% decrease) in the hypercholesterolemic subjects. But the difference was not statistically significant (p < 0.05). The fall in total cholesterol was exclusively due to fall in LDL-cholesterol. Serum triglyceride, serum HDL-cholesterol increased in the experimental subjects by 12.5%. On the other hand, serum total cholesterol, HDL-cholesterol, LDL-cholesterol and triglyceride levels were not changed in control subjects. Both in control and experimental subjects there were no changes in body weight and blood pressure during the study period. The results indicate that hilsa fish, although it is fatty and contains cholesterol, but it may reduce blood cholesterol level in hypercholesterolemic subjects.