Hematopoietic expression of HOXB4 is regulated in normal and leukemic stem cells through transcriptional activation of the HOXB4 promoter by upstream stimulating factor (USF)-1 and USF-2.

The homeobox genes encode a family of transcription factors that regulate development and postnatal tissue homeostasis. Since HOXB4 plays a key role in regulating the balance between hematopoietic stem cell renewal and differentiation, we studied the molecular regulation of HOXB4 expression in human hematopoietic stem cells. HOXB4 expression in K562 cells is regulated at the level of transcription, and transient transfection defines primary HOXB4 regulatory sequences within a 99-bp 5' promoter. Culture of highly purified human CD34(+) bone marrow cells in thrombopoietin/Flt-3 ligand/stem cell factor induced HOXB4 3-10-fold, whereas culture in granulocyte/macrophage colony-stimulating factor, only increased HOXB4/luciferase expression 20-50%. Mutations within the HOXB4 promoter identified a potential E box binding site (HOX response element [HXRE]-2) as the most critical regulatory sequence, and yeast one hybrid assays evaluating bone marrow and K562 libraries for HXRE-2 interaction identified upstream stimulating factor (USF)-2 and micropthalmia transcription factor (MITF). Electrophoretic mobility shift assay with K562 extracts confirmed that these proteins, along with USF-1, bind to the HOXB4 promoter in vitro. Cotransfection assays in both K562 and CD34(+) cells showed that USF-1 and USF-2, but not MITF, induce the HOXB4 promoter in response to signals stimulating stem cell self-renewal, through activation of the mitogen-activated protein kinase pathway. Thus hematopoietic expression of the human HOXB4 gene is regulated by the binding of USF-1 and USF-2, and this process may be favored by cytokines promoting stem cell self-renewal versus differentiation.

Emergence of Vibrio parahaemolyticus as an important cause of diarrhea in Malaysia.

Vibrio parahaemolyticus was isolated from 47 of 2,699 stools examined primarily for the exclusion of cholera. All strains grew well in alkaline peptone water containing 0.5% NaCl and in Monsur's medium. Serotyping showed them to be of various types. Adults were mainly affected. The importance of looking for V. parahaemolyticus in clinical specimens from cases of diarrhea is emphasized.

Animal salmonellosis in peninsular Malaysia. II. Annual and zoological distribution of Salmonella serotypes over the 10-year period 1966-1975.

A total of 860 Salmonella isolations were made in Peninsular Malaysia from 15 animal species (domestic and wild), eggs, molluscs, flies, and animal feed. The isolations were distributed among 31 serotypes in eight groups. The most common serotype isolated was Salmonella pullorum, followed by S. choleraesuis and S. infantis. S. typhimurium had the widest zoological distribution. The importance of controlling animal salmonellosis is emphasized.

Febrile illness in Malaysia--an analysis of 1,629 hospitalized patients.

We studied 1,629 febrile patients from a rural area of Malaysia, and made a laboratory diagnosis in 1,025 (62.9%) cases. Scrub typhus was the most frequent diagnosis (19.3% of all illnesses) followed by typhoid and paratyphoid (7.4%); flavivirus infection (7.0%); leptospirosis (6.8%); and malaria (6.2%). The hospital mortality was very low (0.5% of all febrile patients). The high prevalence of scrub typhus in oil palm laborers (46.8% of all febrile illnesses in that group) was confirmed. In rural Malaysia, therapy with chloramphenicol or a tetracycline would be appropriate for undiagnosed patients in whom malaria has been excluded. Failure to respond to tetracycline within 48 hours would usually suggest a diagnosis of typhoid, and indicate the need for a change in therapy.

Significance of circumsporozoite-specific antibody in the natural transmission of Plasmodium falciparum, Plasmodium vivax, and Plasmodium malariae in an aboriginal (Orang Asli) population of central peninsula Malaysia.

Two hundred and seventy-five Orang Asli volunteers living in nine villages in the Pos Legap Valley of Perak State, peninsular Malaysia, participated in a prospective study designed to characterize the epidemiological, parasitological, and entomological characteristics of Plasmodium falciparum, P. vivax, and P. malariae malaria transmission. Prevalence rates for the three plasmodial species at initiation of the study ranged from 56% in the 0-4-year-old age group to 0% in individuals over the age of 40. Entomological surveys were conducted, enabling us to determine mosquito salivary gland-positive rates and entomological inoculation rates of 1.2 infectious mosquito bites per person per month for P. falciparum, 2.4 for P. vivax, and 0.3 for P. malariae. Cumulative incidence rates over the 16 weeks of the study, following radical cure of all volunteers, were 22.5% for P. falciparum, 12.7% for P. vivax, and 1.5% for P. malariae. The median baseline antibody titer against the immunodominant repetitive B cell epitope of P. falciparum or P. vivax circumsporozoite protein was significantly higher for volunteers who did not become parasitemic. Volunteers were selected for further study if they had evidence of being challenged with P. falciparum sporozoites during the study, based on a two-fold or greater increase in antibody titer against the immunodominant repetitive B cell epitope of the circumsporozoite protein. Resistance to infection was seen in six of 10 individuals who had high (greater than 25 OD units) baseline ELISA titers, compared with only three of 24 individuals who had low baseline ELISA titers (chi 2 P less than 0.02). A similar analysis for P. vivax did not show a significant correlation.

Relationship of some biochemical characteristics of Staphylococcus aureus to enterotoxin production.

A total of 151 enterotoxigenic and 98 non-enterotoxigenic strains of Staphylococcus aureus were examined for production of coagulase, phosphatase, deoxyribonuclease, haemolytic activity, glucose and mannitol fermentation, typical growth on Baird-Parker medium, pigment production and growth under anaerobic conditions. Enterotoxigenic strains showed a higher tendency to demonstrate haemolytic activity, but none of the other biochemical properties could be correlated with the ability of the staphylococci to produce enterotoxins. It appears that other ancillary tests must be developed to be useful in the differentiation of enterotoxigenic and non-enterotoxigenic staphylococci.

Bacterial pathogens isolated from cockroaches trapped from paediatric wards in peninsular Malaysia.

A survey was conducted in 4 paediatric wards in Malaysia to determine the distribution of various species of cockroaches and to examine their gut contents for bacteria. Cockroaches were trapped from food dispensing areas (kitchens), store rooms, cupboards and open wards. 104 cockroaches were caught, consisting of Periplaneta americana (67.3%), Blattella germanica (26%), P. brunnea (4.8%), and Supella longipalpa (1.9%). Bacteria were isolated from all cockroaches except 3 P. americana. Many bacterial species were identified, including the pathogenic and potentially pathogenic species Shigella boydii, S. dysenteriae, Salmonella typhimurium, Klebseilla oxytoca, K. ozaena and Serratia marcescens.

Effective antigen cross-presentation by prostate cancer patients' dendritic cells: implications for prostate cancer immunotherapy.

Despite the potency with which dendritic cells (DCs) are able to utilize the exogenous MHC I antigen cross-presentation pathway to cross-present antigen for the activation of killer T cells in model systems, concern about defects in immune function in cancer patients has led to uncertainty regarding whether immune cells derived from patients can effectively be used to generate tumor vaccines. We have undertaken a careful analysis of the potency of using DCs obtained from prostate cancer patients to cross-present antigen derived from human prostate tumor cells for the activation of antigen-specific T cells. Such DCs can be matured ex vivo into functionally active cells and are capable of cross-presenting influenza antigen derived from internalized apoptotic prostate tumor cells. Importantly, we demonstrate effective stimulation of both CD4+ and CD8+ T cells, as evident by production of IFN-gamma, and the ability of CD8+ T cells to differentiate into effector CTLs. These results, defining conditions in which prostate cancer patient DCs can efficiently utilize the cross-presentation pathway and in which apoptotic tumor can serve as a source of antigen for DCs to activate T cells, demonstrate that this system warrants clinical study as a potential immunotherapy.

Antimicrobial resistance pattern of bacteria isolated from patients seen by private practitioners in the Klang Valley.

Data on bacterial resistance in patients seen by general practitioners are usually not readily available. The objective of this paper is to present the antimicrobial resistance pattern of bacteria isolated from patients seen by private practitioners in the Klang Valley. A total of 18 clinics participated in this study. From mid August 1991 to end of June 1993, 2,823 specimens were received. Throat swabs and urine specimens constituted 56% of all the specimens. A large proportion of the specimens (55%) yielded no growth or just normal flora. The common bacteria encountered were Staphylococcus aureus (18.4%), Escherichia coli (16.2%), Klebsiella spp (13.7%) and Neisseria gonorrhoeae (9.3%). The S. aureus strains were mainly isolated from wound, pus and ear swabs. Not one out of the 218 strains tested was resistant to methicillin. In vitro susceptibility tests showed that 91% were resistant to penicillin while 23% were resistant to tetracycline and 13% to erythromycin. Eighty-two percent of the E. coli were isolated from urine. It was also the most common isolate from urine. Fifty percent of these strains were resistant to ampicillin, 33% to cotrimoxazole, 17% to cephalothin, 21% to ampicillin-sulbactam, 18% to amoxycillin-clavulanic acid while only 2.3% were resistant to nalidixic acid and nitrofurantoin and none to cefuroxime. Generally the gram negative bacilli encountered in general practice are less resistant to the third generation cephalosporins and aminoglycosides when compared to the hospital strains.

Hydrogen sulphide production as an aid to the identification of Vibrio parahaemolyticus.

In this study 18 strains of Vibrio parahaemolyticus from food and 8 from humans were tested for hydrogen sulphide production on various modifications of Russel's Triple Sugar slopes and on TSI. All strains showed a characteristic surface browning on RTS with Andrade's indicator. This was not seen when RTS with phenol red as indicator or TSI were used. Appearance of this phenomenon allows unknown strains to be suspected as being Vibrio parahaemolyticus.

Heavy metal and disinfectant resistance in clinical isolates of gram-negative rods.

Ninety-five clinical strains of Gram-negative bacteria were examined for resistance to mercury, silver and disinfectants. 41% of the strains possessed resistance to mercury, 21% to silver and 7.3% of the strains were resistant to chlorhexidine. Mercury resistance was shown to be plasmid-mediated in 17 strains and silver resistance in 10 strains. Chlorhexidine resistance was not shown to be transferable.

A bacteriological study of some frozen and nonfrozen foods.

Over a period of 19 months, a total of 331 food samples were submitted to the Food Section of the Bacteriology Division for bacteriological examination. These included 184 samples of frozen seafoods from exporters and 147 samples of fresh, nonfrozen foods from food caterers. The total bacterial count for frozen seafoods ranged from 1 x 10(2) to 2.98 x 10(6) per gm with a mean of 2.14 x 10(5) per gm. Coliforms, Escherichia coli and Staphylococcus aureus were present in 48.9%, 3.3% and 8.2% of the samples examined respectively. Two of the cooked prawn samples showed the presence of Vibrio parahaemolyticus. For the fresh, nonfrozen foods, the total bacterial count ranged from 1 x 10(2) to 3.87 x 10(6) per gm with a mean of 2.58 x 10(5) per gm. The examination also showed that 74.8% were coliform positive, 14.9% were E. coli positive, and 4.8% were S. aureus positive. V. parahaemolyticus was not isolated in any of the samples tested. Other pathogens, namely, Vibrio cholerae, Salmonella and Shigella were not isolated from any of the foods examined. The bacterial levels in these foods were determined and their sanitary and public health significance is discussed.

Occurrence of coliform bacteria other than Escherichia coli in foods.

An evaluation of the BGLB (44 degrees C) confirmation test for the enumeration of E. coli as an index of sanitary quality of some foods was made. Out of 70 samples tested, 43 (61.4%) samples contained E. coli. A total of 347 BGLB gas positive cultures obtained were subjected to the IMViC procedure. It was found that 42.7% of these BGLB gas positives were E. coli and 57.3% were ten other coliform bacteria giving the following IMViC patterns: 31.4% --++, 6.3% -+-+, 4.3% each of +-++ and -+++, 3.2% ++-+, 2.9% ++++, 2% ---+, 1.4% +++-, 1.2% --+- and 0.3% -++-. Some of the possible reasons for obtaining low recovery of E. coli in foods by the BGLB test have been discussed. The various fields to be investigated for the improvement of the test procedure and thus the elimination of false positives have been suggested.

Seroreactivity to syphilis in Malaysian blood donors and expectant mothers.

Seroreactivity to syphilis is high among Malaysian blood donors and expectant mothers indicating a high degree of treponemal infection. Further epidemiological studies are required to ascertain what proportion of these could be syphilis and what porportion yaws. Blood donors hava a higher reactivity rate than expectant mothers, the reasons probably being soical. The titres obtained in the VDRL test appear to have a relation to FTA-ABS reactivity although this is not to say treponematosis can be excluded on the basis of low titre VDRL results.

Antibiotic resistance and R plasmids in coliforms isolated from some Malaysian cooked foods.

Forty samples of Malaysian cooked foods were examined for the presence of antibiotic-resistant coliforms and R plasmids. Twenty seven (68%) of the foods had antibiotic-resistant coliforms and 5 (13%) had R plasmids. Nineteen samples (48%) had total bacterial counts over 10(6) per gm and in 5 samples, no coliforms were detected. Our findings show that cooked food may be one possible way by which R plasmids are spread. The control of the spread of R plasmids is discussed.

Enterotoxin production by Staphylococcus aureus strains isolated from humans, foods and animals in Malaysia.

Enterotoxin production by strains of Staphylococcus aureus isolated from human, food and animal sources was investigated. Of the 130 isolates studied, 27 (20.8%) were found to be enterotoxigenic. The most common enterotoxin detected from human sources was enterotoxin C whereas enterotoxin B occurred more frequently in staphylococcal strains of food origin. The 2 enterotoxigenic strains, from animals isolated from a dog and a goat, produced enterotoxins A and C, respectively. Enterotoxin E was not detected alone from any of the enterotoxigenic strains studied, but occurred together with other enterotoxins. The need to detect enterotoxin in staphylococcal strains and in suspected foods for the confirmation of staphylococcal food poisoning is discussed.

Pulsed-field gel electrophoresis as an epidemiologic tool in the investigation of laboratory acquired Salmonella typhi infection.

Strains of Salmonella typhi implicated in two separate cases of laboratory acquired infection from patients and the medical laboratory technologists who processed the patients' samples were analysed by pulsed-field gel electrophoresis. Although all four isolates were of bacteriophage type E1, PFGE was able to demonstrate that the strains responsible for the two laboratory acquired cases were not genetically related. The PFGE patterns of the isolates from the MLTs were found to be identical to those of the corresponding patients after digestion with restriction enzyme AvrII. This provided genetic as well as epidemiological evidence for the source of the laboratory acquired infections.

An evaluation of knowledge and awareness of disinfection and sterilization among health care workers.

Awareness of the disinfection and sterilization policy among hospital staff and their knowledge in basic principles and methods of disinfection and sterilization were studied before and after intervention using a self-administered questionnaire. Survey results showed that awareness (56.2%) before intervention was unsatisfactory. The nurses were more aware of the policy than other groups of medical personnel. Those unaware of the policy perform duties from memory or verbal instructions. A significant increase in awareness to 73.3% was observed after intervention (p < 0.05). Knowledge on methods of decontamination, disinfection and sterilization of equipment varies widely from 28.8% to 90.1%. 23.1% were unaware of the temperature used for sterilization while 72.4% did not know how containers of disinfectant should be refill. Only 14.7% knew the recommended method for washing containers. With education improvement was observed. The average knowledge improved from 44.4% to 57.3%. Our results indicated that continuous in-service education is needed to improve, supplement and update knowledge in this field after basic training. In addition orientation programs for new staff should also be aimed at creating awareness and providing information on guidelines and policies related to their duties.

Disinfection: gaps between recommended and actual practice.

Basic practices on disinfection was surveyed in 6 hospitals using an observation and interview checklist. Two surveys were done, one pre-(first survey) and one post-intervention (second survey). The disinfection and sterilization policy of the Ministry of Health was not available in 66 (70.2%) and 12 (13%) of the units in the first and second survey respectively. In the second survey, staff in all the units washed disinfectant containers before refilling compared with 41.5% of the units in the first survey. Dilution of disinfectants not recommended was found to be used in the first survey. Storing cleaned and sterile items in disinfectants, using disinfectant as a substitute for sterilization of autoclavable items and not decontaminating spillages were some of the wrong practices observed. Considerable improvements were made in the second survey. Improper usage of disinfectants was also indicated by failure of the in-use test. Rate of failure of disinfectants in-use decreased from 11.6% in the first survey to 5.0% in the second survey. To ensure proper disinfection practices, a comprehensive training program on disinfection is required for nurses and attendants.

Epidemiology and control of malaria in Malaysia.

In spite of more than 30 years of control activities, malaria continues to be the most important parasitic infection in Malaysia, accounting for 39,189 confirmed cases in 1991, giving an annual parasite incidence rate of 2.2 per 1,000 population. Some factors contributing to the continued transmission of malaria are the development of drug resistant Plasmodium falciparum, changes in vector behavior, and ecological changes due to socio-economic reasons. Malaria parasite rates are higher among the Aborigines, land scheme settlers and those in intimate contact with the jungle, like loggers. There has been no substantial change in the proportion of the three common malaria species responsible for infections, P. falciparum, P. vivax, P. malariae and mixed infections accounting for about 70%, 28%, 1% and 1%, respectively of all infections. Drug resistant P. falciparum is unevenly distributed in Malaysia, but based on clinical experience and in vitro drug sensitivity studies, chloroquine resistance is frequently encountered. There has been clinical and laboratory evidence of resistance to sulfadoxine/pyrimethamine combination as well as quinine, but all these have so far been successfully treated with a combination of quinine and tetracycline. The eradication of the disease is impossible in the near future but there is confidence that with better surveillance techniques and the use of alternative control measures like permethrin impregnated bed-nets to complement existing ones, the target of bringing down the annual parasite incidence to 2 per 1,000 population during the Sixth Malaysian Plan period (1991-1995) can be achieved.