Risk factors associated with soil transmitted helminth (STH) infection in two indigenous communities in Malaysia.

Soil-transmitted helminth (STH) could possibly cause mild to severe health effects such as diarrhea, weakness, intestinal blood loss, and impaired cognitive development and growth. In Malaysia, previous studies depicted a high prevalence rate of STH was due to poor hygiene practice and low efficacies of anthelminthic drugs. This study was conducted to investigate hand hygiene practice and WASH criteria's (Water, sanitation and hygiene) related to STH infection among two indigenous tribes in Peninsular Malaysia. A cross-sectional study was carried out to study the relationship among STH infection compared to water quality, sanitation, and hygiene conditions. A total of 190 individuals from two indigenous villages participated in the study, with ages ranging from 5 to 60 years old. In addition, Pearson's Chisquare (X2) test was utilized to test the relationship among STH with demographic socioeconomic and behavioral factors. The confidence interval (CI) of 95% is used to estimate the precision of the odds ratio (OR). Multivariate logistic regression models were also used to identify the risk factors associated with STH infections. The overall findings indicated a prevalence rate of 72% for STH, and distributed mainly among children aged < 12 years. Furthermore, multivariate analyses using logistic regression revealed chronic health problems, incorrect hand washing, and walking bare footed were associated with STH infection. Overall results indicated high prevalence of STH among the indigenous villagers, which aligns with the published literature and proves to be a problem need to be addressed as neglected disease. Interestingly, there was a significant relationship between the presences of chronic diseases and STH infection, which prompted other questions the awareness needs to be educated and the simple and low-cost intervention on the proper way of hand washing may help to reduce STH infection in these indigenous communities.

Risk factors associated with soil transmitted helminth (STH) infection in two indigenous communities in Malaysia

@#Soil-transmitted helminth (STH) could possibly cause mild to severe health effects such as diarrhea, weakness, intestinal blood loss, and impaired cognitive development and growth. In Malaysia, previous studies depicted a high prevalence rate of STH was due to poor hygiene practice and low efficacies of anthelminthic drugs. This study was conducted to investigate hand hygiene practice and WASH criteria’s (Water, sanitation and hygiene) related to STH infection among two indigenous tribes in Peninsular Malaysia. A cross-sectional study was carried out to study the relationship among STH infection compared to water quality, sanitation, and hygiene conditions. A total of 190 individuals from two indigenous villages participated in the study, with ages ranging from 5 to 60 years old. In addition, Pearson’s Chisquare (X2) test was utilized to test the relationship among STH with demographic socioeconomic and behavioral factors. The confidence interval (CI) of 95% is used to estimate the precision of the odds ratio (OR). Multivariate logistic regression models were also used to identify the risk factors associated with STH infections. The overall findings indicated a prevalence rate of 72% for STH, and distributed mainly among children aged < 12 years. Furthermore, multivariate analyses using logistic regression revealed chronic health problems, incorrect hand washing, and walking bare footed were associated with STH infection. Overall results indicated high prevalence of STH among the indigenous villagers, which aligns with the published literature and proves to be a problem need to be addressed as neglected disease. Interestingly, there was a significant relationship between the presences of chronic diseases and STH infection, which prompted other questions the awareness needs to be educated and the simple and low-cost intervention on the proper way of hand washing may help to reduce STH infection in these indigenous communities.

Kytococcus sedentarius and Micrococcus luteus: highly prevalent in indoor air and potentially deadly to the immunocompromised - should standards be set?

The multifarious types of infections contracted from indoor environments show that buildings can serve as a reservoir for infectious bacteria. This study is an investigation into the type and concentrations of bacteria in the indoor and outdoor environments of an electronic factory, an office and a winery in Malaysia. Trypticase soy agar (TSA) (with ambient air incubation) and TSA supplemented with haemin and NADH (with CO2 enhanced incubation) were used for the isolation of bacteria. The plates were incubated at 37ºC for 3 days. A random selection of bacterial isolates were Gram stained and identified using the BD BBL Crystal Identification Systems. Kytococcus sedentarius and Micrococcus luteus were the predominant bacterial species identified from indoor air. These bacteria were present at relatively high concentrations in indoor air, at times, above 800 colony forming units per cubic meter (CFU/m3) of air. This indicates that both K. sedentarius and M. luteus can survive a wide range of adverse conditions, including chemical contamination and ultraviolet exposure. M. luteus is a known cause of pneumonia in immunocompromised individuals and has also been implicated in skin infections. Recent reports suggest species of kytococci as emerging opportunistic pathogens of the immunocompromised, paediatrics and the elderly. We postulate that opportunistic bacteria, such as the kytococci and the micrococci, may also have a potential role in instigating subclinical, more subtle symptoms of disease in inmmunocompetent individuals.

Field evaluation of a rapid diagnostic test to detect antibodies in human toxocariasis.

Human toxocariasis which is caused mainly by the larvae of Toxocara canis and Toxocara cati, is a worldwide zoonotic disease that can be a potentially serious human infection. The enzyme-linked immunosorbent assay (ELISA) using T. canis excretory-secretory (TES) antigens harvested from T. canis larvae is currently the serological test for confirming toxocariasis. An alternative to producing large amounts of Toxocara TES and improved diagnosis for toxocariasis is through the development of highly specific recombinant antigens such as the T. canis second stage larva excretory-secretory 30 kDa protein (recTES-30). The aim of this study was to evaluate the sensitivity and specificity of a rapid diagnostic kit (RDT, named as iToxocara kit) in comparison to recTES-30 ELISA in Serendah Orang Asli village in Selangor, Malaysia. A total of 133 subjects were included in the study. The overall prevalence rates by ELISA and RDT were 29.3% and 33.1%, respectively, with more positive cases detected in males than females. However, no association was found between toxocariasis and gender or age. The percentage sensitivity, specificity, positive predictive value and negative predictive value of RDT were 85.7%, 90.1%, 80% and 93.2%, respectively. The prevalence for toxocariasis in this population using both ELISA and RDT was 27.1% (36/133) and the K-concordance test suggested good agreement of the two tests with a Cohen's kappa of 0.722, P<0.01. In addition, the followed-up Spearman rank correlation showed a moderately high correlation at R=0.704 and P<0.01. In conclusion, the RDT kit was faster and easier to use than an ELISA and is useful for the laboratory diagnosis of hospitalized cases of toxocariasis.

Involvement of G-quadruplex structures in regulation of human RET gene expression by small molecules in human medullary thyroid carcinoma TT cells.

The RET (rearranged during transfection) proto-oncogene encodes a receptor tyrosine kinase for members of the glial cell line-derived neurotrophic factor family of extracellular signaling molecules. The activating germline point mutations in the RET, which are known to induce oncogenic activation of RET tyrosine kinase, are associated with the development of medullary thyroid carcinoma (MTC) and pathogenesis of multiple endocrine neoplasia type 2 (MEN2). The polypurine/polypyrimidine tract in the proximal promoter region of the human RET gene (-51 to -33 relative to transcription start site) is essential for basal transcriptional activity of this gene. This tract consists of a guanine-rich sequence containing five runs of at least three contiguous guanines separated by one or more bases, conforming to a general motif capable of forming an intramolecular G-quadruplex. Here, we show that specific G-quadruplex structures formed in the RET promoter region act to repress the transcription of this gene, and transcription of this gene can be controlled by ligand-mediated G-quadruplex stabilization. In this study, NSC194598, a derivative of indeno[1,2,3-de]quinazoline, was found to be a novel G-quadruplex interactive agent that interfered with transcriptional activation of mutated RET gene in human medullary thyroid carcinoma TT cells. This compound significantly reduced endogenous RET protein levels and increased apoptosis in these cells. Our results provide further support for the idea that G-quadruplex structures may have a critical role in transcriptional regulation of the RET gene in vivo, providing insight into a novel strategy for transcriptional repression of this gene by small molecules.

A comparative evaluation of dengue diagnostic tests based on single-acute serum samples for laboratory confirmation of acute dengue.

A prospective study was carried out to evaluate the sensitivity of dengue NS1 antigen-capture ELISA in comparison with dengue virus isolation, conventional RT-PCR and real-time RT-PCR for laboratory confirmation of acute dengue based on single-acute serum samples. Four primary healthcare centres were involved to recruit patients with clinical diagnosis of dengue illness. Patient's demographic, epidemiological and clinical information were collected on a standardized data entry form and 5 ml of venous blood was collected upon consent. In the laboratory, six types of laboratory tests were performed on each of the collected acute serum sample. Of the 558 acute serum samples collected from 558 patients with clinical diagnosis of dengue from mid-August 2006 to March 2009, 174 serum samples were tested positive by the dengue NS1 antigen-capture ELISA, 77 by virus isolation, 92 by RT-PCR and 112 by real-time RT-PCR. A total of 190 serum samples were tested positive by either one or a combination of the four methods whereas, only 59 serum samples were tested positive by all four methods. Thus, based on single-acute serum samples, 190 of the 558 patients (34.1%) were laboratory-confirmed acute dengue. The overall test sensitivity was 91.6%, 40.5%, 48.4% and 58.9% for dengue NS1 antigen-capture ELISA, virus isolation, conventional RT-PCR and real-time RT-PCR respectively. Statistically, dengue NS1 antigen-capture ELISA was the most sensitive and virus isolation was the least sensitive test for the laboratory confirmation of acute dengue based on single-acute serum specimens. Real-time RT-PCR was significantly more sensitive than the conventional RT-PCR.

A comparative evaluation of dengue diagnostic tests based on single acute serum samples for laboratory confi rmation of acute dengue

A prospective study was carried out to evaluate the sensitivity of dengue NS1 antigen-capture ELISA in comparison with dengue virus isolation, conventional RT-PCR and real-time RT-PCR for laboratory confi rmation of acute dengue based on single-acute serum samples. Four primary healthcare centres were involved to recruit patients with clinical diagnosis of dengue illness. Patient’s demographic, epidemiological and clinical information were collected on a standardized data entry form and 5 ml of venous blood was collected upon consent. In the laboratory, six types of laboratory tests were performed on each of the collected acute serum sample. Of the 558 acute serum samples collected from 558 patients with clinical diagnosis of dengue from mid-August 2006 to March 2009, 174 serum samples were tested positive by the dengue NS1 antigen-capture ELISA, 77 by virus isolation, 92 by RT-PCR and 112 by real-time RT-PCR. A total of 190 serum samples were tested positive by either one or a combination of the four methods whereas, only 59 serum samples were tested positive by all four methods. Thus, based on singleacute serum samples, 190 of the 558 patients (34.1%) were laboratory-confi rmed acute dengue. The overall test sensitivity was 91.6%, 40.5%, 48.4% and 58.9% for dengue NS1 antigen-capture ELISA, virus isolation, conventional RT-PCR and real-time RT-PCR respectively. Statistically, dengue NS1 antigen-capture ELISA was the most sensitive and virus isolation was the least sensitive test for the laboratory confi rmation of acute dengue based on single-acute serum specimens. Real-time RT-PCR was signifi cantly more sensitive than the conventional RT-PCR.

A comparative study on the performance of two commercial anti-dengue IgM assay kits.

The performance of a commercial rapid immunochromatographic dengue IgG/IgM assay device was evaluated against an in-place dengue IgM-capture ELISA in the National Public Health laboratory. Of the 239 serum samples from patients with clinical diagnosis of acute dengue illness, 140 and 99 samples were tested positive and negative respectively for anti-dengue IgM by the in-placed ELISA. Comparatively, 72 and 76 samples were tested positive and negative respectively, and 91 samples gave equivocal results by the rapid dengue test device. The rapid immunochromatographic assay device gave a relative sensitivity of 49.3% and a relative specificity of 62.6%. Though the rapid immunochromatographic assay device has the advantages of rapid testing which simultaneously detects both IgG and IgM and can also be performed with whole blood, serum or plasma, the user has to exercise extreme caution with the interpretation of the test result.

Evaluating the sensitivity of a commercial dengue NS1 antigen-capture ELISA for early diagnosis of acute dengue virus infection.

INTRODUCTION: The aim of this report is to establish an accurate diagnosis of acute dengue virus infection early, in order to provide timely information for the management of patients and early public health control of dengue outbreak. METHODS: 224 serum samples from patients with a clinical diagnosis of acute dengue infection, which were subsequently confirmed by laboratory tests, were used to evaluate the performance of a commercially-available dengue NS1 antigen-capture ELISA kit. RESULTS: The dengue NS1 antigen-capture ELISA gave an overall sensitivity rate of 93.3 percent (209/224). The sensitivity rate was significantly higher in acute primary dengue (97.4 percent) than in acute secondary dengue (68.8 percent). In comparison, the virus isolation gave an overall positive isolation rate of 64.7 percent, with a positive rate of 70.8 percent and 28.1 percent, for acute primary dengue and acute secondary dengue, respectively. Molecular detection of dengue RNA by RT-PCR gave an overall positive detection rate of 63.4 percent, with a positive rate of 62.5 percent and 68.8 percent, for acute primary dengue and acute secondary dengue, respectively. Of the 224 acute serum samples from patients with laboratory-confirmed acute dengue infection, dengue IgM was detected in 88 specimens, comprising 68 acute primary dengue specimens and 20 acute secondary dengue specimens. NS1 antigen-capture ELISA kit gave an overall sensitivity rate of 88.6 percent in the presence of anti-dengue IgM and 96.3 percent in the absence of anti-dengue IgM. CONCLUSION: Of the 224 acute serum samples, the sample ages of 166 acute serum samples are known. The positive detection rate of dengue NS1 antigen-capture ELISA, on the whole, was higher than the other three established diagnostic test methods for laboratory diagnosis of acute dengue infection.

Evaluation of a commercial dengue NS1 antigen-capture ELISA for laboratory diagnosis of acute dengue virus infection.

A commercial dengue NS1 antigen-capture ELISA was evaluated to demonstrate its potential application for early laboratory diagnosis of acute dengue virus infection. Dengue virus NS1 antigen was detected in 199 of 213 acute serum samples from patients with laboratory confirmation of acute dengue virus infection but none of the 354 healthy blood donors' serum specimens. The dengue NS1 antigen-capture ELISA gave an overall sensitivity of 93.4% (199/213) and a specificity of 100% (354/354). The sensitivity was significantly higher in acute primary dengue (97.3%) than in acute secondary dengue (70.0%). The positive predictive value of the dengue NS1 antigen-capture ELISA was 100% and negative predictive value was 97.3%. Comparatively, virus isolation gave an overall positive isolation rate of 68.1% with a positive isolation rate of 73.9 and 31.0% for acute primary dengue and acute secondary dengue, respectively. Molecular detection of dengue RNA by RT-PCR gave an overall positive detection rate of 66.7% with a detection rate of 65.2 and 75.9% for acute primary dengue and acute secondary dengue, respectively. The results indicate that the commercial dengue NS1 antigen-capture ELISA may be superior to virus isolation and RT-PCR for the laboratory diagnosis of acute dengue infection based on a single serum sample.

Genetic diversity of enterovirus 71 isolated from cases of hand, foot and mouth disease in the 1997, 2000 and 2005 outbreaks, Peninsular Malaysia.

All known field isolates of enterovirus 71 (EV71) can be divided into three distinct genogroups (A, B, C) and 10 subgenogroups (A, B1-5, C1-4) based on VP1 gene sequences. We examined VP1 gene sequences of 10, 12 and 11 EV71 strains isolated in peninsular Malaysia during the outbreaks of hand, foot and mouth disease in 1997, 2000 and 2005 respectively. Four EV71 strains isolated in the hand, foot and mouth disease outbreak of 2006 in Sarawak (Malaysian Borneo) were included to describe their genetic relationship. Four subgenogroups (C1, C2, B3 and B4) of EV71 co-circulated and caused the outbreak of hand, foot and mouth disease in peninsular Malaysia in 1997. Two subgenogroups (C1 and B4) were noted to cause the outbreak in 2000. In the 2005 outbreak, besides EV71 strains of subgenogroup C1, EV71 strains belonged to subgenogroup B5 were isolated but formed a cluster which was distinct from EV71 strains of the subgenogroup B5 isolated in 2003. The four EV71 strains isolated from clinical specimens of patients with hand, foot and mouth disease in the Sarawak outbreak in early 2006 also belonged to subgenogroup B5. Phylogenetic analysis of the VP1 gene sequences showed that the four Sarawak EV71 isolates belonged to the same cluster as the EV71 strains that were isolated in peninsular Malaysia as early as May 2005. The data suggested that the EV71 strains causing the outbreak in Sarawak could have originated from peninsular Malaysia.

Chikungunya virus of Central/East African genotype detected in Malaysia.

Since its isolation in Tanzania in 1953, chikungunya virus has caused periodic outbreaks in both tropical Africa and Asia. In the last decade, the virus has shown not only increased activity but has expanded its geographical locations, thus classical delineation of various genotypes of chikungunya virus to specific geographic locales no longer holds true. Rapid mass movement of people and the constant presence of the right vectors in this region could have contributed to the change in virus ecology. This paper documents the first detection of chikungunya virus of Central/East genotype in Malaysia from a patient who was most likely infected with the virus during her visit to India. Without good Aedes vector measures, only time will tell whether this genotype rather than the existing endemic genotype will subsequently cause the next chikungunya outbreak in Malaysia.

Outbreak of chikungunya due to virus of Central/East African genotype in Malaysia.

Chikungunya is an acute febrile illness caused by an alphavirus which is transmitted by infective Aedes mosquitoes. Two previous outbreaks of chikungunya in Malaysia were due to chikungunya virus of Asian genotype. The present outbreak involved two adjoining areas in the suburb of Ipoh city within the Kinta district of Perak, a state in the northern part of Peninsular Malaysia. Thirty seven residents in the main outbreak area and two patients in the secondary area were laboratory confirmed to be infected with the virus. The index case was a 44-year Indian man who visited Paramakudi, Tamil Naidu, India on 21st November 2006 and returned home on 30th of November 2006, and subsequently developed high fever and joint pain on the 3rd of December 2006. A number of chikungunya virus isolates were isolated from both patients and Aedes albopictus mosquitoes in the affected areas. Molecular study showed that the chikungunya virus causing the Kinta outbreak was of the Central/East African genotype which occurred for the first time in Malaysia.

Isolation of monoclonal antibodies-escape variant of dengue virus serotype 1.

INTRODUCTION: During an outbreak from December 2004 to March 2005, 138 isolates of dengue virus were prospectively obtained from acute-phase serum samples of 1,067 patients with the provisional clinical diagnosis of acute dengue illness admitted to the adult wards of Hospital Tengku Ampuan Rahimah, Klang, Malaysia. Of the 138 dengue virus isolates, 87, 11, 24 and 3 were typed as dengue serotypes 1, 2, 3 and 4, respectively, by a commercial dengue virus typing kit using monoclonal antibodies (Mab). 13 dengue virus isolates could not be assigned to any specific serotype by serotyping Mab and molecular typing using dengue-type specific molecular typing primer pairs. We report the associated clinical features and limited molecular genetics of this Mab-escape dengue virus variant. METHODS: Limited molecular characterisation of the Mab-escape dengue virus variants with respect to a few concurrently isolated dengue serotype 1 virus was performed by reverse transcriptase polymerase chain reaction (RT-PCR), followed by nucleic acid sequencing of the 500-bp dengue virus partial genomic capsid-PreM fragment. RESULTS: The aligned nucleic acid sequence of RT-PCR products showed that these Mab-escape variants were of identical nucleic acid sequence, and shared the highest sequence homology (99 percent) with dengue virus serotype 1 (GeneBank accession No. AB178040) isolated from a Japanese patient in 2004. Though these Mab-escape dengue virus variants were noted to replicate to a 2-log higher titre than the current circulating dengue virus serotype 1, there was no significant difference between these variants and the currently circulating dengue virus serotype 1 with respect to disease severity (dengue fever versus dengue haemorrhagic fever) and clinical features. CONCLUSION: There was no significant difference in the proportion of patients developing dengue haemorrhagic fever following acute infection by Mab-escape dengue virus 1 variant in comparison with infection by the conventional dengue virus 1. Similarly, there was no significant difference in the pattern of clinical presentations following acute infection by the two different strains of virus.

Outbreak of influenza amongst residential school students in Malaysia.

In the months of July and August 2003, an outbreak of acute respiratory illness caused by influenza A virus occurred among students in seven residential schools situated in the northern part (Perak) of Peninsular Malaysia. Out of 4989 students, aged 13 to 18 years (mean = 15.9), 1419 (28%) were effected by influenza-like illness. All patients were treated as outpatients except for 36 students who required admission for high fever, severe coughing and shortness of breath. Abnormal chest X-ray findings were noted for those that required inpatient management. Influenza A virus was isolated from 37 sputum specimens, 20 throat swabs and three nasal swab specimens from a total of 278 clinical samples obtained from 180 patients. Isolates from each of the outbreaks were sent to WHO Collaborating Centre for Reference and Research on Influenza, Melbourne, Australia for antigenic and genetic analysis. One school outbreak was due to influenza A (H1N1), A/New Caledonia/20/99-like virus while the other six school outbreaks were due to influenza A (H3N2) viruses which were A/Fujian/411/2002-like).

Re-emergence of Chikungunya virus in Malaysia.

An outbreak of Chikugunya (CHIK) fever occurred among the fishing community in Bagan Pancor, Perak. The outbreak was laboratory confirmed within 48 hours after the receipt of the specimens. Fifty-three patients' serum samples were submitted for laboratory investigation and 47 (88.7%) were confirmed to be positive for CHIK infection by RT-PCR, and/or virus isolation, and/or in-house immunoflourescent test. RT-PCR and virus isolation were the tests of choice for patients with illness of four days or less and detection of CHIK specific IgM for those with more than four days of fever. The nucleic acid sequence based on the 354- and 294-bp of the nsP1 and E1 genes of the CHIK virus detected from pools of adults Aedes aegypti mosquitoes were identical to those CHIKV virus isolated from humans in the same locality. Phylogenetic analysis of the CHIK virus based on the 257 nts partial E1 gene indicates that Bagan Panchor's strain was closely related to the first CHIK virus isolated during the outbreak in Klang in 1998.

Congenital rubella syndrome with positive serology and virus isolation.

An effective live attenuated rubella vaccine was available since 1969 and congenital rubella syndrome can be prevented with appropriate vaccination. We report a baby with congenital rubella syndrome born in Klang valley to indicate that the Universal Rubella Vaccination Programme adopted by the Ministry of Health Malaysia since 2002 has yet to achieve its effect of eliminating transmission of rubella and preventing congenital rubella infection in the community. To our knowledge, the virus isolate represents the first successful isolation of rubella virus in this country and will serve as the reference strain for future comparison in molecular epidemiological tracking of rubella virus activity this country.

Rubella outbreak amongst residential students in a military vocational school of Malaysia.

An outbreak of rubella occurred amongst 303 newly recruited residential Form IV students in a military vocational training school in Malaysia. Of the 303 Form IV students, 77 gave a history of acute illness. Rubella specific IgM was detected in the sera of 46.5% (141/303) whereas rubella specific IgG was detected in 100% of all Form IV students. Sixty five students with no clinical history of acute illness during the outbreak period had detectable rubella IgM in their sera and rubella specific IgM was detected in the sera of all symptomatic students except one. Maculopapular rash was the commonest presenting clinical feature among students with acute rubella infection in this outbreak (97.4%) followed by fever (88.2%). The duration of rash ranged from one to nine days with a mean of 4.6 days. Of the 65 students that had both fever and rash, 56 (85.2%) students had maculopapular skin eruption on the same day as the date of onset of fever, six (9.2%) developed the rash a day after the onset of fever and three (4.6%) had the rash after two days of fever. The duration of fever ranged from one to eight days with a mean of 3.5 days. The duration of conjunctivitis ranged from one to four days with a mean of 2.3 days, and all those who developed conjunctivitis had mild eye-discharge without photophobia. The duration of arthralgia ranged from one to three days with a mean of 2.1 days. The commonest type of joints affected was knee joints (66.7%, 12/18), followed by elbow and shoulder joints (27.8%, 5/18) and wrist joints (5.6%, 1/18). A good clinical history of the temporal relationship between the occurrence of rash and fever during the outbreak could easily differentiate rubella illness from that of measles.

Effects of cortisol on cardiac myocytes and on expression of cardiac genes in fetal sheep.

In 17 fetal sheep aged 129 days, the effects of large-dose infusions of cortisol (72.1 mg/day for 2-3 days) on proliferation, binucleation, and hypertrophy of cardiac myocytes, cardiac expression of angiotensinogen, angiotensin receptor subtypes 1 and 2, Glut-1, glucocorticoid and mineralocorticoid receptors, proteins of the MAPK pathways and calcineurin were studied. Cortisol levels were 8.7 +/- 2.3 nM (SE) in 8 control and 1,028 +/- 189 nM in 9 treated fetuses (P < 0.001). Cortisol had no effect on myocyte binucleation. Left ventricular free wall (LVFW) uni- and binucleated myocytes were larger in cortisol-treated fetuses (P < 0.001, P < 0.05). Cortisol-treated fetuses had higher right ventricular free wall (RVFW) and LVFW angiotensinogen (Aogen) mRNA levels (treated: 2.30 +/- 0.37, n = 8 and 2.05 +/- 0.45, n = 7 vs. control: 0.94 +/- 0.12, n = 8 and 0.67 +/- 0.09, n = 7, P < 0.02). Levels of the glucose transporter Glut-1 mRNA were lower in the LVFW of treated fetuses (0.83 +/- 0.23 vs. 1.47 +/- 0.30 in control, P < 0.05, n = 7, 8). The higher the cortisol level, the greater the Aogen mRNA level (RVFW, r = 0.61, P < 0.01, n = 16; LVFW, r = 0.83, P < 0.0003, n = 14). There were no other changes in mRNA levels nor in levels of extracellular kinase, JNK, p38, their phosphorylated forms, and calcineurin. Thus high levels of cortisol such as occur after birth do not affect fetal cardiac myocyte binucleation or number but are associated with higher levels of ventricular Aogen mRNA, lower levels of Glut-1 mRNA, and hypertrophy of LVFW myocytes. These effects could impact on postnatal cardiac development.