Presence of SARS-CoV-2-like coronaviruses in bats from east coast Malaysia.

Most of the public health importance coronaviruses, such as Severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and SARS-CoV-2 are likely originated from bats and spread to humans through intermediate hosts; civet cats, dromedary camel and Malayan pangolin, respectively. SARS-CoV-2-like coronaviruses were detected in Thailand, which is neighbouring with Kelantan in East Coast Malaysia. To date, there is no report on the presence of public health concerns (SARS-CoV, SARS-CoV-2 and MERS-CoV) coronaviruses in bats from Malaysia. This study was aimed to elucidate the presence of these coronaviruses in bat samples from East Coast, Malaysia. A total of hundred seventy oropharyngeal swab samples were collected from three states of East Coast Malaysia. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was conducted based on partial 3' Untranslated region (3'UTR) or ORF10 gene and the products were sequenced. The sequences were compared with all coronavirus sequences from the National Center for Biotechnology Information-GenBank (NCBI-GenBank) using NCBI-Basic Local Alignment Search Tool (NCBI-BLAST) software. A phylogenetic tree was constructed to determine the genetic relationship among the detected coronaviruses with the reference coronaviruses from the NCBI-GenBank. Our results showed that SARSCoV-2-like viruses were present in 3% (5/170) of the bats from East Coast Malaysia that have 98-99% sequence identities and are genetically related to SARS-CoV-2 from humans. This finding indicates the presence of SARS-CoV-2-like viruses in bats from East Coast Malaysia that may become a public health concern in the future.

Presence of SARS-CoV-2-like coronaviruses in bats from east coast Malaysia

@#Most of the public health importance coronaviruses, such as Severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and SARS-CoV-2 are likely originated from bats and spread to humans through intermediate hosts; civet cats, dromedary camel and Malayan pangolin, respectively. SARS-CoV-2-like coronaviruses were detected in Thailand, which is neighbouring with Kelantan in East Coast Malaysia. To date, there is no report on the presence of public health concerns (SARS-CoV, SARS-CoV-2 and MERS-CoV) coronaviruses in bats from Malaysia. This study was aimed to elucidate the presence of these coronaviruses in bat samples from East Coast, Malaysia. A total of hundred seventy oropharyngeal swab samples were collected from three states of East Coast Malaysia. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was conducted based on partial 3’ Untranslated region (3’UTR) or ORF10 gene and the products were sequenced. The sequences were compared with all coronavirus sequences from the National Center for Biotechnology Information-GenBank (NCBI-GenBank) using NCBI-Basic Local Alignment Search Tool (NCBI-BLAST) software. A phylogenetic tree was constructed to determine the genetic relationship among the detected coronaviruses with the reference coronaviruses from the NCBI-GenBank. Our results showed that SARSCoV-2-like viruses were present in 3% (5/170) of the bats from East Coast Malaysia that have 98-99% sequence identities and are genetically related to SARS-CoV-2 from humans. This finding indicates the presence of SARS-CoV-2-like viruses in bats from East Coast Malaysia that may become a public health concern in the future.

First report of equine Setaria digitata (von Linstow 1906) infestation in Malaysia.

The occurrence of Setaria digitata in a horse is reported for the first time in Malaysia. An 8-year-old Thoroughbred cross mare was referred to the University Veterinary Clinic with the primary complaint of corneal opacity and excessive eye discharge. After initial treatment with Terramycin eye ointment, corneal opacity cleared partially to reveal a moving thread-like cylindrical worm in the anterior chamber of the eye. The parasite was successfully removed surgically, and examination under the light microscope revealed that the isolated worm (length = 45 mm) was a 5th stage larva of S. digitata based on morphological criteria. Confirmation of the species of the worm was through molecular methods. The 12S rRNA gene was PCR-amplified, and the purified amplicon was directly sequenced. Phylogenetic analyses revealed that the isolated roundworm showed 100% sequence similarity with that of S. digitata in NCBI GenBank database (Accession no.: KY284626.1). This report is the first confirmed case of equine ocular setariasis by S. digitata in Malaysia. The current study provides evidence that S. digitata is an etiological agent of ocular infection and its presence in Malaysia.

Renal depletion of myo-inositol is associated with its increased degradation in animal models of metabolic disease.

Renal depletion of myo-inositol (MI) is associated with the pathogenesis of diabetic nephropathy in animal models, but the underlying mechanisms involved are unclear. We hypothesized that MI depletion was due to changes in inositol metabolism and therefore examined the expression of genes regulating de novo biosynthesis, reabsorption, and catabolism of MI. We also extended the analyses from diabetes mellitus to animal models of dietary-induced obesity and hypertension. We found that renal MI depletion was pervasive across these three distinct disease states in the relative order: hypertension (-51%)>diabetes mellitus (-35%)>dietary-induced obesity (-19%). In 4-wk diabetic kidneys and in kidneys derived from insulin-resistant and hypertensive rats, MI depletion was correlated with activity of the MI-degrading enzyme myo-inositol oxygenase (MIOX). By contrast, there was decreased MIOX expression in 8-wk diabetic kidneys. Immunohistochemistry localized the MI-degrading pathway comprising MIOX and the glucuronate-xylulose (GX) pathway to the proximal tubules within the renal cortex. These findings indicate that MI depletion could reflect increased catabolism through MIOX and the GX pathway and implicate a common pathological mechanism contributing to renal oxidative stress in metabolic disease.

Childhood adrenocortical carcinoma as a sentinel cancer for detecting families with germline TP53 mutations.

Li-Fraumeni syndrome (LFS) is a highly penetrant, autosomal dominant disorder where affected individuals carry a 50% risk of developing cancer before 30 years of age. It is most commonly associated with mutations in the tumour suppressor gene, TP53. Adrenocortical carcinoma (ACC) is a very rare paediatric cancer, and up to 80% of affected children are found to carry germline TP53 mutations. Hence, we propose using childhood ACC incidence as selection criteria for referral for TP53 mutation testing, independent of familial cancer history. Under the auspices of the Malaysian Society of Paediatric Haematology-Oncology, four eligible children diagnosed with ACC over a 30-month study period were referred for mutation testing. Three had a germline TP53 mutation. Subsequent TP53 testing in relatives showed two inherited mutations and one de novo mutation. These findings strongly support paediatric ACC as a useful sentinel cancer for initiating a germline TP53/LFS detection programme, particularly in countries where the lack of structured oncogenetic practice precludes the identification of families with LFS features.

Development of an algorithm of satellite markers for monitoring chimerism status in post-allogeneic haematopoietic stem cell transplantation patients.

INTRODUCTION: Analysis of variable number tandem repeats (VNTRs) by polymerase chain reaction (PCR) is a common method used to predict engraftment status in post-allogeneic haematopoeitic stem cell transplantation (HSCT) patients. Different populations have different copies of repeated DNA sequence and hence, different percentage of informativeness between patient and donor. METHODS: PCR amplification of four highly polymorphic VNTR markers (YNZ-22, D1S80, D1S111 and ApoB) was conducted on 60 patient-donor pairs. The informativeness of the markers was analysed using 3% agarose gel electrophoresis. RESULTS: We developed an algorithm for identification of informative VNTR markers in 60 post-allogeneic HSCT patients. YNZ-22 was the most informative (72%), followed by D1S80 (63%) and D1S111 (60%), while the least informative was ApoB (47%). The degree of informativeness achieved was 95%, which could discriminate 57 patient-donor pairs, when all four markers were combined. CONCLUSION: Since population genetic studies on VNTR loci are not well established in Southeast Asia, the present study is useful to determine reliable markers during the initial screening step of chimerism analysis. By following this algorithm, we are able to reduce time and cost of finding a suitable VNTR marker in our cohort.

Percutaneous urinary drainage and ureteric stenting in malignant disease.

Patients with malignant ureteric obstruction often have a poor life expectancy, even if relief of urinary obstruction is achieved. Careful discussion between the patient, their family and all health care professionals involved in the case must be undertaken before any intervention. The goal of treatment in the palliative setting may be to offer symptom relief, avoid complications from renal insufficiency or allow further oncological systemic therapy. The obstruction can be relieved by placement of a percutaneous nephrostomy tube, a ureteric stent or, more rarely, due to the palliative nature of the patients, a more complex open surgical procedure. Nephrostomy tubes offer excellent drainage and can be placed under a local anaesthetic, although they do require and a bag, which can reduce a patient's quality of life. Bleeding, sepsis, bowel transgression and pleural complications can all be encountered when inserting a nephrostomy. JJ stents in comparison have a higher failure rate due to extrinsic compression and usually require a general anaesthetic for placement, although significant bleeding and other complications are reduced. Quality of life is still affected by irritative lower urinary tract symptoms. Metallic ureteric stents have recently been developed to try and offer better relief of obstruction than polymeric stents with fewer side-effects and increased intervals between changes. Initial data appear encouraging, although further trials are needed. No blueprint for treatment of malignant ureteric obstruction exists and each case must be decided on an individual basis with a fully informed patient.

Copper(II)-selective chelation improves function and antioxidant defences in cardiovascular tissues of rats as a model of diabetes: comparisons between triethylenetetramine and three less copper-selective transition-metal-targeted treatments.

AIMS/HYPOTHESIS: Treatment with the Cu(II)-selective chelator triethylenetetramine (TETA) improves cardiovascular disease in human patients, and cardiac and vascular/renal disease in rats used as a model of diabetes. Here we tested two hypotheses: first, that TETA elicits greater improvement in organ function than less Cu-selective transition-metal-targeted treatments; second, that the therapeutic actions of TETA are consistent with mediation through suppression of oxidative stress. METHODS: Rats were made diabetic with streptozotocin (55 mg/kg, i. v.) and treated from 8 weeks after disease induction for the following 8 weeks with effective dosages of oral TETA, or one of three less Cu-selective transition-metal-targeted treatments: D-penicillamine, deferiprone or Zn acetate. Treatment effects were measured in ex vivo cardiac and aortic tissues, plasma and urine. RESULTS: Diabetes damaged both cardiac and renal/vascular function by impairing the ability of cardiac output to respond physiologically to rising afterload, and by significantly elevating the urinary albumin/creatinine ratio. Diabetes also lowered total antioxidant potential and heparan sulphate levels in cardiac and arterial tissues, and serum ferroxidase activity, whereas it elevated urinary heparan sulphate excretion. TETA treatment rectified or partially rectified all these defects, whereas the other three experimental treatments were ineffectual. By contrast, none of the four drug treatments lowered diabetes-mediated elevations of plasma glucose or lipid concentrations. CONCLUSIONS/INTERPRETATION: TETA may limit the cardiac and renal/vascular damage inflicted by diabetes through its actions to reinforce antioxidant defence mechanisms, probably acting through selective chelation of 'loosely-bound'/chelatable Cu(II). It may also improve heparan sulphate homeostasis and bolster antioxidant defence by increasing vascular extracellular superoxide dismutase activity. Urinary albumin/creatinine ratio might prove useful for monitoring TETA treatment.

Expression of metastasis-associated molecules in non-small cell lung cancer and their prognostic significance.

Molecules associated with tumor invasion and metastasis have been actively investigated, but their prognostic significance has been variously reported by investigators. We evaluated the expression of matrix metalloproteinase (MMP)-1, MMP-2, MMP-3, MMP-9, MMP-10, tissue inhibitor of MMPs (TIMP)-1, TIMP-2, ß-catenin, E-cadherin and cyclooxygenase-2 (COX-2) in 43 cases of non-small cell lung cancer (NSCLC). Immunohistochemistry of each marker was performed on tissue microarray paraffin blocks, and the results were determined by a semi-quantitative method using an intensity score (0-3) and percentage score (0-3). The expression scores of each marker were correlated with TNM stage and patient survival data. The expression of MMP-3 and COX-2 was significantly increased in higher stage tumors (P<0.001 and P=0.046, respectively), while a correlation with patient survival length was observed for MMP-1 and COX-2 (P=0.034 and 0.019, respectively). All stage I or II cases with increased MMP-1 expression succumbed to NSCLC within 34.1 months. Cases with low expression of both MMP-1 and COX-2 had a significantly longer survival time than cases with high expression of either of the two markers (P=0.002). These results suggest that MMP-1 and COX-2 are plausible candidate survival markers for NSCLC.

Effect of maternal age, birth weight and infant sex on total nucleated cell (TNC) count and volume of umbilical cord blood (UCB) collected.

This study evaluates the effect of maternal age, birth weight and infant sex on two main UCB parameters for use and long-term cryopreservation: TNC and volume. Data from 1000 UCB units were collected and analyzed in this study. The results indicate that TNC is correlated to infant birth weight and sex but not maternal age at delivery. Volume is only correlated to birth weight but not maternal age and infant sex.

Exploring the relationship between caffeine intake and essential tremor.

BACKGROUND: It has been suggested that environmental factors may be associated with essential tremor (ET). This study was carried out to evaluate the association of caffeine intake with ET. METHOD: In a case control study, patients diagnosed with ET and healthy controls underwent a standardized questionnaire interview to evaluate the exposure to coffee and tea intake. A multivariate logistic regression analysis was carried out to evaluate the association of caffeine intake and other environmental factors with risk of ET. RESULTS: 179 subjects including 79 ET patients and 100 controls matched for age, gender and ethnicity were included in the analysis. Univariate analysis revealed that caffeine consumption in ET patients was higher than control group (median and 90th percentile range: 2300 (0, 9000) mg-years versus 1500 (0, 6090) mg-years, p=0.01). However, the multivariate logistic regression analysis demonstrated that caffeine was no longer a significant factor associated with ET (p=0.119). There was no significant correlation between amount of caffeine intake and disease duration (Spearman's r=0.194; p=0.202) or total tremor score (Spearman's r=0.045; p=0.771) in ET patients. CONCLUSION: Caffeine consumption was not associated with risk of ET in our study population. Further studies are needed to investigate the significance of gene-environmental interaction in ET.

Knowledge about the genetics of essential tremor in patients and their relatives.

In a face-to-face questionnaire-based survey involving a total of 111 essential tremor (ET) patients and their relatives, we demonstrated a general lack of genetic insight amongst our study subjects. There was no significant correlation between genetic knowledge of ET and age, level of education, disease duration or severity. A proactive effort is needed to encourage physicians managing ET patients to incorporate genetic information and education into their practice.

The effect of image size on screen-positive rates for nuchal translucency screening.

It has been shown previously that the absolute measurement of nuchal translucency (NT) thickness, one of the most effective screening tests for fetal Down syndrome, significantly decreases with increasing image size. We undertook a prospective study to assess whether this effect materially alters the NT-derived risk estimation for fetal Down syndrome. In 350 women, NT was measured at both 100 and 200% image sizes. The median NT measurement at 200% was 8% smaller than at 100%. The screen-positive rates at 100 and 200% image sizes were 5.1% (18/350) and 2.3% (8/350), respectively (P= 0.048). Altering the image magnification settings is likely to decrease the sensitivity of NT-derived Down syndrome risk estimation. Further study is required to test the effect of our finding on the sensitivity of NT screening and to assess whether adjustments in gain settings mitigate the effect.

Management of renal-vein perforation during a challenging percutaneous nephrolithotomy.

A 54-year-old man with complex urinary anatomy as the result of previous surgery sustained a renal-vein injury during percutaneous nephrolithotomy for a staghorn calculus with a blood loss of 1.5 L. He was managed with antibiotics, bed rest, heparin, and a 28F nephrostomy catheter, which was withdrawn gradually as the tract sealed. This case highlights the importance of early diagnosis of this complication and the possibility of conservative management.

Pelvi-calyceal height, a predictor of success when treating lower pole stones with extracorporeal shockwave lithotripsy.

Extra corporeal shockwave lithotripsy (ESWL) is the treatment of choice for the majority of renal stones, however, it has the lowest success rate in complete clearance of stones located in the lower pole. We assess whether pelvi-calyceal height is a useful measurement in predicting successful stone clearance from the lower pole. A total of 105 patients with a solitary lower pole calculus of less than 20 mm treated with ESWL were reviewed. Stone size, location and pelvi-calyceal height were measured by intravenous urogram. Success was defined as complete stone clearance. Fifty-four patients (51.4%) had successful treatments, with the remaining 51 (48.6%) having incomplete stone clearance (including two patients in whom treatment had no effect). There was a statistically significant difference (P<0.0001) in pelvi-calyceal height between the two groups. Mean pelvi-calyceal height in patients with complete stone clearance was 15.1 mm (SD=3.9) compared with 22.9 mm (SD=5.2) for those with incomplete clearance. Pelvi-calyceal height is a useful predictor of success when treating lower pole renal stones with ESWL.

Dystrophic ureteral calcification associated with Churg-Strauss vasculitis.

Churg-Strauss syndrome is an uncommon systemic vasculitis affecting small blood vessels. Renal involvement is typical; however, calcinosis of the urinary tract has not previously been described. Dystrophic calcification in the urinary tract is rare, although it is occasionally associated with schistosomiasis, tuberculosis, and polyarteritis nodosa. We report the case of a 19-year-old man with newly diagnosed Churg-Strauss syndrome who presented to us with dystrophic calcification in both ureters causing bilateral obstruction.

1,25-Dihydroxyvitamin D and three low-calcemic analogs decrease UV-induced DNA damage via the rapid response pathway.

1,25-Dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] is anti-apoptotic in human keratinocytes, melanocytes and fibroblasts after ultraviolet (UV)-exposure. To date, there is no published data on the effects of 1,25(OH)(2)D(3) or its analogs on DNA damage in irradiated skin cells. In these skin cells, 24h pre-treatment with 1,25(OH)(2)D(3) dose-dependently (10(-12) to 10(-8)M) decreased CPD damage by up to 60%. This photoprotective effect was also seen if the 1,25(OH)(2)D(3) was added immediately after irradiation and was mimicked by QW-1624F2-2 (QW), a low-calcemic 1beta-hydroxymethyl-3-epi-16-ene-24,24-difluoro-26,27-bis homo hybrid analog. The well-studied low calcemic, rapid acting agonist analogs 1alpha,25(OH)(2)lumisterol(3) (JN) and 1alpha,25(OH)(2)-7-dehydrocholesterol (JM) also protected skin cells from UV-induced cell loss and CPD damage to an extent comparable with that of 1,25(OH)(2)D(3). In contrast, the rapid response antagonist analog 1beta,25(OH)(2)D(3) (HL) completely abolished the photoprotective effects (reduced cell loss and reduced CPD damage) produced by treatment with 1,25(OH)(2)D(3), JN, JM and QW. Evidence for involvement of the nitric oxide pathway in the protection from CPD damage by 1,25(OH)(2)D(3) was obtained. These data provide further evidence for a role of the vitamin D pathway in the intrinsic skin defenses against UV damage. The data also support the hypothesis that the photoprotective effects of 1,25(OH)(2)D(3) are mediated via the rapid response pathway(s).

Ultrasound prediction of risk of spontaneous miscarriage in live embryos from assisted conceptions.

OBJECTIVE: An accurate method to predict subsequent miscarriage in live embryos has not yet been established. This pilot study aimed to determine the most discriminatory ultrasound-based model for predicting spontaneous miscarriage after embryonic life was first detected in assisted conceptions. A method for estimating individual risk of miscarriage was developed. METHODS: This was a prospective cross-sectional survey of 322 live singleton embryos in women from an assisted reproductive technology program. Mean sac diameter (MSD), crown-rump length (CRL), embryonic heart rate (EHR), maternal age and gestational age at the first transvaginal scan detecting embryonic life (between 42 and 62 days) were observed. These variables were included in a multivariate model for predicting spontaneous miscarriage occurring prior to 20 weeks. MSD, CRL and MSD minus CRL were assessed in univariate logistic regression analyses. The global diagnostic accuracy of each model was compared directly using receiver-operating characteristics (ROC) curves. RESULTS: The multivariate model demonstrated the best ROC curve for predicting miscarriage (ROC area 0.87; 95% CI, 0.80-0.95). The separate univariate analyses had less diagnostic accuracy. In particular, MSD - CRL had a significantly smaller ROC area (0.65) than did the multivariate model (P < 0.01). CONCLUSIONS: The most discriminatory test for predicting spontaneous miscarriage in live embryos was a multivariate model, which allows estimation of individual risk levels.

Epidemiology of paediatric renal stone disease in the UK.

BACKGROUND: The previous epidemiological study of paediatric nephrolithiasis in Britain was conducted more than 30 years ago. AIMS: To examine the presenting features, predisposing factors, and treatment strategies used in paediatric stones presenting to a British centre over the past five years. METHODS: A total of 121 children presented with a urinary tract renal stone, to one adult and one paediatric centre, over a five year period (1997-2001). All children were reviewed in a dedicated stone clinic and had a full infective and metabolic stone investigative work up. Treatment was assessed by retrospective hospital note review. RESULTS: A metabolic abnormality was found in 44% of children, 30% were classified as infective, and 26% idiopathic. Bilateral stones on presentation occurred in 26% of the metabolic group compared to 12% in the infective/idiopathic group (odds ratio 2.7, 95% CI 1.03 to 7.02). Coexisting urinary tract infection was common (49%) in the metabolic group. Surgically, minimally invasive techniques (lithotripsy, percutaneous nephrolithotomy, and endoscopy) were used in 68% of patients. CONCLUSIONS: There has been a shift in the epidemiology of paediatric renal stone disease in the UK over the past 30 years. Underlying metabolic causes are now the most common but can be masked by coexisting urinary tract infection. Treatment has progressed, especially surgically, with sophisticated minimally invasive techniques now employed. All children with renal stones should have a metabolic screen.

Mid-trimester fetal subdural hemorrhage: prenatal diagnosis.

A routine antenatal ultrasound examination at 20 weeks' gestation revealed a space-occupying lesion in the fetal right cerebral hemisphere. The borders of the mass were indistinct and there was no midline shift. A small collection of echogenic bowel was identified at the right iliac fossa. In an attempt to explain both findings a provisional diagnosis of a fetal blood dyscrasia was made. Fetal cranial magnetic resonance imaging 24 h later confirmed the diagnosis of a subdural hemorrhage. Subsequent fetal blood sampling confirmed severe fetal thrombocytopenia. To our knowledge this is the first report of the antenatal diagnosis of spontaneous mid-trimester fetal subdural hemorrhage.