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Introduction: Endocrine disorders of sex steroid hormones may adversely affect men's sexual function. Aim. To provide expert opinions/recommendations concerning state-of-the-art knowledge for the pathophysiology, diagnosis and treatment of endocrinologic sexual medicine disorders., Methods: An International Consultation in collaboration with the major urology and sexual medicine associations assembled over 200 multidisciplinary experts from 60 countries into 17 committees. Committee members established specific objectives and scopes for various male and female sexual medicine topics. The recommendations concerning state-of-the-art knowledge in the respective sexual medicine topic represent the opinion of experts from five continents developed in a scientific and debate process. Concerning the Endocrine committee, there were eight experts from seven countries., Main Outcome Measure: Expert opinions/recommendations are based on grading of evidence-based medical literature, extensive internal committee discussion over 2 years, public presentation and deliberation., Results: Hypogonadism is a clinical and biochemical syndrome characterized by a deficiency in serum androgen levels which may decrease sexual interest, quality of erections and quality of life. Biochemical investigations include testosterone and either bioavailable or calculated free testosterone; prolactin should be considered when hypogonadism has been documented. If clinically indicated, androgen therapy should maintain testosterone within the physiological range avoiding supraphysiologic values. Digital rectal examination and determination of serum prostate specific antigen values are mandatory prior to therapy and regularly thereafter. Androgen therapy is usually long-term requiring regular follow-up, frequent monitoring of blood levels and beneficial and adverse therapeutic responses., Conclusions: Safe and effective treatments for endocrinologic sexual medicine disorders examined by prospective, placebo-controlled, multi-institutional clinical trials are needed.

Aim: To explore the barriers faced by general practitioners (GPs) in the management of patients with erectile dysfunction (ED)., Methods: This was a qualitative analysis of focus group discussions and in-depth interviews involving 28 Malaysian GPs., Results: GPs' perception of ED being not a serious condition was a major determinant of their prescribing practice. Doctor's age (younger), gender (female), short consultation time and lack of experience were cited as barriers. The GPs' prescribing habits were heavily influenced by the feedback from the first few patients under treatment, the uncertainty of etiology of ED without proper assessment and the profit margin with bulk purchase. Other barriers include Patients' coexisting medical conditions, older age, lower socio-economic status, unrealistic expectations and inappropriate use of the anti-impotence drugs. Cardiovascular side effects and cost were two most important drug barriers., Conclusion: The factors influencing the management of ED among the general practitioners were multiple and complex. An adequate understanding of how these factors (doctors, patients and drugs) interact can assist in the formulation and implementation of strategies that encourage GPs to identify and manage ED patients.

The characteristic clinicoradiological findings of multiple sclerosis and acute disseminated encephalomyelitis (ADEM), demonstrating a recurrent progressive course in the former and monophasicity in the latter associated with multiple discrete white matter lesions with variable enhancement on MRI, are not a diagnostic challenge. On the other hand, the less typical radiological presentation of a solitary tumefactive demyelinating lesion mimics a neoplasm, and often necessitates a biopsy. Nonetheless, histopathological examination is an imperfect gold standard and the recognition of certain imaging features may facilitate the correct diagnosis.

Aim: Apart from conventional epidural analgesia (EA), the combined spinal-epidural (CSE) modality is fast becoming a popular technique for treating labour pain. In this study, we investigated the differences in the patient profile and outcome between CSE and EA for labour pain in KK Women's and Children's Hospital., Methodology: Data pertaining to 1,532 healthy parturients who had received either CSE or EA for labour pain during a six-month period was systematically collected by using a specially designed form. Multiple logistic regression analysis was used to determine the independent predictors of patient satisfaction and the relation of parturient factors on the choice of block. The side effects and the outcome of labour were also compared., Results: CSE accounted for 80% of all neuraxial blocks performed for labour analgesia (vs 20% for EA). Anaesthesiologists were more inclined to using CSE than EA for multiparous parturients (OR 2.03, p<0.01) in a more painful (OR=1.61, p=0.03) and advanced stage of labour (OR=1.12, p=0.03). The need for supplemental analgesics was greater for EA (p<0.01). Patient satisfaction was higher for CSE (OR=1.77, p<0.026). CSE had a higher risk of pruritus (29% vs 14%, p<0.01) but lower risk of post block neural deficits (0% vs 2%, p<0.01) than EA. No difference in the mode of delivery was detected between the two groups., Conclusion: CSE is a safe and good alternative to EA as a technique of neuraxial block for labour analgesia.

Two patients who had acute pancreatitis subsequently developed characteristic appearance on urography of smooth extrinsic narrowing and medial deviation of the right ureter suggestive of retroperitoneal fibrosis (RPF) resulting in ureteric obstruction. Both these patients had clinical, biochemical and sonographic evidence of acute pancreatitis. CT scan of the abdomen performed on the second patient also documented acute pancreatitis. Intravenous urograms were consistent with ureteric obstruction due to retroperitoneal fibrosis. Both cases were treated conservatively. They were well after an average of 20 months. These 2 cases illustrate the uncommon association between pancreatitis and RPF.

This qualitative study aimed to examine cultural differences in knowledge, attitudes and practices related to erectile dysfunction (ED) utilizing focus group discussion. Six focus groups consisting of 66 men, 45-70-y-old were conducted-two Malay groups (n=18), two Chinese groups (n=25) and two Indian groups (n=23). Participants were purposely recruited from the general public on a voluntary basis with informed consent. Transcripts were analyzed using qualitative data analysis software ATLASti. The Malay and Chinese traditional remedies for preventing or treating ED are commonly recognized among all races. Many have a negative perception of someone with ED. Malay and Chinese men tended to blame their wife for their problem and thought that the problem might lead to extra-marital affairs, unlike the Indian men who attributed their condition to fate. Malays would prefer traditional medicine for the problem. The Chinese felt they would be more comfortable with a male doctor whilst this is not so with the Malays or Indians. Almost all prefer the doctor to initiate discussion on sexual issues related to their medical condition. There is a need for doctors to consider cultural perspectives in a multicultural society as a lack of understanding of this often contributes to an inadequate consultation.

Riemerella anatipestifer, a gram-negative bacillus, is the causative agent of duck septicemia, a disease which could incur much economic loss in the duck industry. An indirect enzyme-linked immunosorbent assay (ELISA) has been developed to facilitate early detection of R. anatipestifer infection in ducks. The antigen used was a recombinant 41 kDa N-terminal fragment (rP45N') of a newly characterized R. anatipestifer potential surface protein, P45, which was expressed in Escherichia coli as an N-terminal GST fusion protein. The rP45N'-based ELISA successfully detected P45 antibodies in the sera of 20 ducks immunized with bacterin preparations of R. anatipestifer serotypes 1, 10 15, 19 and the ATCC11845 strain. Antibodies to P45 were also detected in the sera of 25% (75/296) of White Pekin ducks which were imported into Singapore from three different farms. Successful discrimination was obtained between sera from infected ducks and that of specific-pathogen free ducks (p<0.01). The rP45N'-GST antigen did not cross-react with antibodies in sera from guinea pigs which were infected with other gram-negative and gram-positive bacterial pathogens, including Aeromonas hydrophila, Citrobacter freundii, E. coli, Klebsiella pneumoniae, Pastuerella multocida, Proteus mirabilis, Salmonella spp., Serratia maccescens, Shigella sonnei and Yersinia enterocolitica. In addition, the DNA sequence encoding P45 was detected in R. anatipestifer serotypes 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 14, 15, 16, 17, 18, 19 and the ATCC11845 strain, suggesting that P45 is probably also universally expressed in these R. anatipestifer serotypes. Thus, the ELISA described is applicable to the detection of R. anatipestifer infection in ducks.

Riemerella anatipestifer is responsible for exudative septicemia in ducks. The genetic determinant of the CAMP cohemolysin, cam, from a strain of R. anatipestifer was cloned and expressed in Escherichia coli. Chromosomal DNA from serotype 19 strain 30/90 was used to construct a gene library in pBluescript II SK(-) vector in E. coli XL-1-Blue strain. The clones containing recombinant plasmids were screened for the CAMP reaction with Staphylococcus aureus. Those that showed cohemolysis were chosen for further analysis by sequencing. One of these clones, JFRA8, was subcloned to identify the smallest possible DNA fragment containing the CAMP cohemolysin determinant, which was located on a 3,566-bp BamHI-BstXI fragment which specified a 1,026-bp open reading frame. Clones containing recombinant plasmids carrying cam obtained by PCR cloning into E. coli M15 strain secreted an active CAMP cohemolysin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analyses confirmed that the recombinant strain expressed a protein with a molecular mass of 37 kDa and that strains from serotypes 1, 2, 3, 5, 6, and 19 expressed the cohemolysin. The deduced amino acid sequence showed high homology to those of O-sialoglycoprotein endopeptidases. Hydrolysis of radioiodinated glycophorin A confirmed that Cam is a sialoglycoprotease.

Parapharyngeal space tumours account for only 0.5 per cent of all head and neck tumours. Due to their inherent location, they present with varied non-specific signs and symptoms, resulting in a delay in diagnosis and unnecessary procedures, such as a 'tonsillectomy' or 'incision and drainage' of a 'quinsy'. Thirty-one patients, operated on over an 18-year period (1981 to 1998), in the Department of Otolaryngology, Singapore General Hospital, are presented. Their ages ranged from 21 to 86 years, with a mean of 52 years, with equal sex distribution. The commonest aetiology was a deep lobe of parotid tumour (44 per cent), followed by neurilemmomas (18 per cent), there was only one paraganglioma. The transcervical and transparotid approaches were the commonest used. The mean surgical time was three hours, mean hospital stay was 5.3 days and post-operative complications were minimal. The average follow-up time was 5.6 years. Although parapharyngeal space tumours are uncommon, recognizing them would enable the correct sequence of investigations, instead of unnecessary procedures resulting in an increased morbidity for the patient.

This paper highlights women's perceptions of sildenafil citrate (Viagra, Pfizer). It is based on a qualitative study on perceptions of erectile dysfunction in the Malaysian multicultural society. Six focus groups were conducted, consisting of 69 women, aged between 40 and 70 years, recruited from the general public and who had given informed consent. The findings revealed that the women were aware of erectile dysfunction and Viagra. Due to their concern about the negative aspects of Viagra, the Chinese and Malay traditional methods of treatment were commonly mentioned. The women from three ethnic groups viewed the possibility of their husband starting to take Viagra with lots of suspicion, mistrust and fear. They would prefer their husband discussing with them the issue of resorting to taking Viagra. The Chinese and Indian women perceived that if a man takes Viagra, it will boost his ego and he will feel more manly. Indian women felt that a man taking Viagra is proof of his love for his wife. The Malay women felt that a man would be ashamed and have a low self-esteem if he were to resort to taking Viagra. Although Viagra is meant for the male, understanding of women's perception of it is beneficial for a couple's sexual relationship.

The generation of protective immunity against Riemerella anatipestifer infection in ducks were investigated by immunizations with recombinant glutathione sulfatransferase (GST) fusion's proteins of OmpA, a 42kDa major outer membrane protein, and P45N', a 41kDa N-terminal fragment of a newly identified 45kDa potential surface protein from R. anatipestifer. The DNA encoding OmpA and P45N' were isolated from R. anatipestifer serotype 15 (field strain 110/89) and serotype 19 (reference strain 30/90), respectively. Immunoblotting and ELISA results showed that the purified recombinant proteins induced the production of antibodies in immunized ducks. However, neither was protective against subsequent challenge with the virulent serotype 15 strain, 34/90. All the five ducks immunized with formalinized R. anatipestifer strain 34/90 survived the challenge with the homologous strain whereas six out of seven ducks in the non-immunized control group died within a week following the challenge.

Purpose: a) To evaluate the effect of adding 1.25 mg of bupivacaine to intrathecal fentanyl on the duration of analgesia in an Asian population and b) to examine if the baricity of the local anesthetic at this dose has any bearing on the duration and quality of block., Methods: Forty-eight parturients in early labour received combined spinal epidural (CSE) analgesia to evaluate a) the effect of adding 1.25 mg of bupivacaine to intrathecal (IT) fentanyl 25 microg on the duration of analgesia and b) the effect of baricity of intrathecal local anesthetic on the duration and quality of the block. Patients were randomly allocated to receive: IT fentanyl 25 microg plus normal saline (Group f, n=16), IT fentanyl 25 microg plus plain bupivacaine 1.25 mg (Group f+pb, n=16) and IT fentanyl 25 microg plus heavy bupivacaine 1.25 mg (Group f+hb, n=16). The two components of the IT injectate (total of 2.25 mL) were given sequentially., Results: Group f+hb had the lowest sensory dermatomal block (T7 vs T4 (Group f), T5 (Group f+pb), P <0.01). There were no differences in the duration of analgesia and incidence of side effects among the groups., Conclusion: We found no advantage of adding 1.25 mg bupivacaine to fentanyl 25 microg. At this dose, the baricity of bupivacaine has no effect on the duration of analgesia.

Purpose: The study was aimed primarily at comparing the duration of analgesia produced by intrathecal fentanyl 25 microg with sufentanil 5 microg when added to bupivacaine 1.25 mg as the initial component of the combined spinal epidural (CSE) technique in early labour., Methods: Forty healthy parturients were randomly assigned into two groups to receive either intrathecal sufentanil 5 microg plus bupivacaine 1.25 mg (Group S) or intrathecal fentanyl 25 microg plus bupivacaine 1.25 mg (Group F). Apart from the duration of analgesia, pain scores and side effects were also evaluated., Results: There was no significant difference in the duration of analgesia (mean 109 +/- SD 49 min in Group F vs 118 +/- 54 min in Group S, P=0.9). Group F had a more rapid onset of analgesia (P <0.05) and a higher cephalad block (median T4 vs T7, P <0.05) in the first 30 min after the block. No difference in the side effects was detected., Conclusion: Fentanyl 25 microg is a good alternative to sufentanil 5 microg when added to bupivacaine 1.25 mg for early labour analgesia.

Introduction: Between 0.1% and 0.9% of women develop pregnancy complications which require admission to an intensive therapy unit. The aim of this study was to review all obstetric admissions to the intensive therapy unit at the KK Women's and Children's Hospital from 1998 to 1999 with respect to indications for admission, interventions employed and clinical outcome., Method: The medical records of all obstetric patients admitted to the intensive therapy unit during the 2-year period were analysed retrospectively. Subjects were included if they were admitted during pregnancy up to 42 days postpartum., Results: There were 31,725 deliveries in our hospital during the study period of which there were 239 admissions to the intensive therapy unit. Of these, 42% were Malays, 41% Chinese, 12% Indians and 5% other races. 65% stayed 1 day, 24% 2 days, 7% 3 days and 4% more than 3 days. The patients' ages ranged from 18 to 44 years. The indications for admission were hypertension (50%), haemorrhage (24%), respiratory insufficiency (10%), neurological problems (11%) and sepsis (3%). Intervention-wise, 43% of patients required vasoactive infusions, 35% had arterial line placement, 22% central venous pressure monitoring, 21% ventilatory support and 2% pulmonary artery catheter placement. The maternal mortality and stillbirth rates were 1.3% and 3.7% of intensive therapy unit admissions, respectively., Conclusion: The admission rate to the intensive therapy unit in our institution was 0.73% of all deliveries during the 2-year study period. Hypertensive disease and haemorrhage were the predominant admitting diagnoses.

Ninety-one patients were randomly allocated to one of two groups. Group A was induced with a single vital capacity breath of 6% (end-tidal) sevoflurane in nitrous oxide-oxygen (2 : 1 l.min-1), whereas group B was induced with intravenous fentanyl 1 microg.kg-1 + propofol 2 mg.kg-1 followed by nitrous oxide-oxygen (2 : 1 l.min-1) and sevoflurane. Induction was considered to have been achieved when the bispectral index value decreased to below 70. Mean induction time in group A (95.2 s, 95% CI 88.5-101.9 s) was longer than group B (70.3 s, 95% CI 66.3-74.3 s; p < 0.0001). Mild coughing was more common in group A, but relative hypotension was more common in group B. There was no difference in the emergence times. Thirty minutes after emergence, there was no difference in the incidence of adverse effects, with the exception of essentially mild abdominal pain which was more frequent in group A.

Reconstruction surgery for a female to male transsexual usually involves mastectomy, hysterectomy and creating an aesthetically appealing neophallus. We have successfully inserted an inflatable prosthesis using the AMS CX prosthesis in a 45 year old transsexual, who had a large bulky neophallus constructed from the anterior abdominal subcutaneous fat, about 9 years ago. The single cylinder CX prosthesis was well anchored to the symphysis pubis using a dacron windsock tubing, the activation pump was placed in the dependent pouch of the right labium and the reservior in the usual perivesical space. The patient subsequently had debulking procedure using liposuction to create a more aesthetic and functional phallus. To date, the inflatable neophallus prosthesis is functioning well.

Objectives: To evaluate the efficacy, safety, and tolerability of oral sildenafil in Asian men with erectile dysfunction of various causes (organic, psychogenic, or mixed) and of more than 6 months' duration., Methods: In this double-blind, parallel-group trial conducted at eight centers in Malaysia, the Philippines, and Singapore, 254 men, 26 to 78 years old, were randomized to 12 weeks of sildenafil or placebo taken as needed 1 hour before anticipated sexual activity. Initially, the sildenafil (n = 127) or matching placebo (n = 127) dose was 50 mg but could be increased to 100 mg or decreased to 25 mg because of a lack of efficacy or intolerance, respectively. Efficacy was assessed by the 15-question International Index of Erectile Function, patients' event logs of sexual activity, and a global efficacy question about erections., Results: The two primary efficacy variables relating to achievement and maintenance of an erection sufficient for sexual intercourse, as assessed by the mean scores for International Index of Erectile Function question 3 (4.22 versus 2.59) and question 4 (4.15 versus 2.41), were both significantly higher with sildenafil than with placebo (P <0.0001). In addition, the five separate International Index of Erectile Function domains of sexual function, the percentage of successful intercourse attempts, and the global efficacy assessment of erections revealed significantly greater treatment effects in favor of sildenafil (P <0.0001 versus placebo for all variables). Treatment-related adverse events occurred in 22.8% of patients who received sildenafil and in 10.2% of those who received placebo., Conclusions: Sildenafil is an effective and well-tolerated treatment for Asian men with erectile dysfunction of broad-spectrum etiology.

Endomyces fibuliger is a yeast used in the production of Chinese rice wine. It secretes enzymes such as glucoamylase, alpha-amylase and acid protease. Very little is known of the genetics of E. fibuliger. In order to develop a transformation system for this yeast, orotidine-5'-phosphate decarboxylase mutant strains were obtained and characterized. Transformation of the E. fibuliger ura3 mutant F1 with an integrative plasmid that carried the wild-type URA3 gene of E. fibuliger gave complementation of this mutation. The E. fibuliger gene encodes the orotidine-5'-phosphate decarboxylase enzyme consisting of 266 amino acid residues with a 69.4% sequence identity with orotidine-5'-phosphate decarboxylase of Saccharomyces cerevisiae. Our finding that E. fibuliger URA3 complements the ura3 mutation in S. cerevisiae confirms that the URA3 gene of E. fibuliger encodes a protein that exerts a similar function.

The ompA gene, encoding the 42-kDa major antigenic outer membrane protein OmpA of Riemerella anatipestifer, the etiololgical agent of septicemia anserum exsudativa, was cloned and expressed in Escherichia coli. Recombinant OmpA displayed a molecular mass similar to that predicted from the nucleotide sequence of the ompA gene but lower than that observed in total cell lysates of R. anatipestifer. The ompA gene showed a conserved C-terminal region comprising the OmpA-like domain and a variable N-terminal region. This structure is similar to those of the analogous outer membrane proteins of several gram-negative bacteria. However, OmpA of R. anatipestifer contains six EF-hand calcium-binding domains and two PEST regions, which distinguish it from other outer membrane proteins. The occurrence of these motifs in OmpA suggests a possible role in virulence for this protein. The ompA gene is present in the R. anatipestifer type strain and in all serotype reference strains. However, it exhibits some minor genetic heterogeneity among different serotypes, which seems not to affect the strong antigenic characteristics of the protein. OmpA is a conserved and strong antigenic determinant of R. anatipestifer and hence is suggested to be a valuable protein for the serodetection of R. anatipestifer infections, independent of their serotype.

Two directly repeated sequences of the IS elements IS1489v1 and IS1489v2 flank the benzene dioxygenase (bedC1C2BA) and the cis-benzene dihydrodiol dehydrogenase (bedD) genes on the catabolic plasmid pHMT112 in Pseudomonas putida ML2, forming a Class-I-type composite transposon, Tn5542. Both IS1489v1 and IS1489v2 contain an identical 1371-bp open reading frame, tnpA, that is preceded by a possible ribosome binding site. The tnpA gene of IS1489v1 is bound by a pair of 40-bp imperfect inverted repeats while that of IS1489v2 is flanked only by the left inverted repeat. The tnpA gene codes for a putative 53-kDa polypeptide of 456 amino acids bearing similarity to transposases encoded on IS elements of P. alcaligenes, P. aeruginosa, P. stutzeri, and Serratia marcescens. The basic nature of the putative TnpA protein with a deduced pI of 8.93 is typical of IS-encoded transposases. Similar to other IS elements, an outward facing promoter was detected at the right end of IS1489v1. Experiments involving the suicide vector, pKNG101, failed to show transposition of Tn5542., (Copyright 2000 Academic Press.)

The quest for improving and maintaining sexual function has been going on since time immemorial. The advent of an effective oral drug, sildenafil, has brought about unprecedented open discussion on male erectile dysfunction, and gas accelerated the pace of development of new therapies for erectile dysfunction. New knowledge in the physiology of sexual function has enabled researchers to target drug treatment at the whole network of the central nervous system and the numerous cascadic enzymatic reactions leading to relaxation of the corporal smooth muscle. One of the brightest potential applications of future molecular technology in the study of erectile dysfuction is in the utilization of gene therapy.

Riemerella anatipestifer is a gram-negative rod-shaped bacterium associated with epizootic infections in poultry. A total of 35 R. anatipestifer isolates including the type strain ATCC11845T, reference and field strains for 18 different serotypes were characterized by repetitive sequence based-PCR (rep-PCR) with outwardly-directed primers based on the repetitive extragenic palindromic (REP) consensus sequence. This technique was applied by using either extracted genomic DNA or preparation of whole bacterial cells harvested directly from plate cultures. Rep-PCR discriminated the R. anatipestifer isolates into 19 electrophoretic types. DNA fingerprints obtained from rep-PCR of extracted genomic DNA or from preparations of whole cells yielded comparable patterns. Substantial variation was seen among the rep-PCR fingerprints of different serotypes. Moreover, different polymorphisms of the rep-PCR fingerprints were evident among epidemiologically unrelated isolates of the same serotype. These results suggest the presence of repetitive extragenic palindromic-like elements within the genome of R. anatipestifer that can be used in some isolates to discriminate between different strains belonging to the same serotype. Rep-PCR may serve as a useful molecular tool for subtyping R. anatipestifer isolates for epidemiologic investigations. The whole cell procedure offers the advantage of ease of performance requiring only small quantities of cells.

A second and novel cis-benzene dihydrodiol dehydrogenase which is able to dehydrogenate a range of cis-dihydrodiols and other vicinal alcohols has been purified from Pseudomonas putida ML2. The enzyme is a tetramer of a polypeptide of 39 kDa in molecular mass and has a pH optimum of 9.0. Despite having a primary structure that has significant similarity to glycerol dehydrogenases, the kcat/Km value of the enzyme for cis-benzene dihydrodiol is 4300-fold higher compared to glycerol. The apparent Km values of the enzyme for cis-benzene dihydrodiol and glycerol are 0.01 mM and 46 mM, respectively, and 0.22 mM for NAD+.

The recent findings of various mobile catabolic genes have provided some insight into the evolution of microbial degradation systems for xenobiotic compounds. The catabolic genes undergo marked genetic rearrangements due to their presence on transposons or association with mobile genetic elements. Bacterial catabolic transposons fall into three defined structural classes. Class I elements include catabolic genes flanked by two copies of insertion sequences. Class II elements carry short terminal inverted repeats and transpose by the replicative mode in which transposase and resolvase are involved. Conjugative catabolic transposons represent the third class of mobile genetic elements. They carry all the genes required for excision, conjugal transfer to a new host, and integration. This review focuses on the structures, functions and roles of the recently characterized catabolic transposons in bacteria. Also described are the mobile catabolic elements that share structural similarity with the pathogenicity and symbiosis islands.

The introduction of foreign organic hydrocarbons into the environment in recent years, as in the widespread use of antibiotics, has resulted in the evolution of novel adaptive mechanisms by bacteria for the biodegradation of the organic pollutants. Plasmids have been implicated in the catabolism of many of these complex xenobiotics. The catabolic genes are prone to undergo genetic rearrangement and this is due to their presence on transposons or their association with transposable elements. Most of the catabolic transposons have structural features of the class I (composite) elements. These include transposons for chlorobenzoate (Tn5271), chlorobenzene (Tn5280), the newly discovered benzene catabolic transposon (Tn5542), and transposons encoding halogenated alkanoates and nylon-oligomer-degradative genes. Transposons for the catabolism of toluene (Tn4651, Tn4653, Tn4656) and naphthalene (Tn4655) belong to class II (Tn3 family) elements. Many catabolic genes have been associated with insertion sequences, which suggests that these gene clusters could be rapidly disseminated among the bacterial populations. This greatly expands the substrate range of the microorganisms in the environment and aids the evolution of new and novel degradative pathways. This enhanced metabolic versatility can be exploited for and is believed to play a major part in the bioremediation of polluted environments.

We have studied the antiemetic efficacy of droperidol alone, and in combination with metoclopramide in first trimester termination of pregnancy in day surgery. The aim was to determine whether the addition of metoclopramide could further reduce the incidence of postoperative nausea and vomiting (PONV) but avoid excessive sedation. Group I (control, n = 40) received i.v. droperidol 0.625 mg at induction. Group II (study, n = 40) received i.v. droperidol 0.625 mg and i.v. metoclopramide 10 mg at induction. The incidence of nausea at 1 and 2 hours postoperatively was 23% and 10% in group I, and 5% and nil in group II respectively. The difference in the incidence of nausea was significant at p < 0.05 at one hour but not at two hours postoperatively. No patients vomited. There was no difference in the sedation and pain score between them. We did not observe any significant side effects attributable to either drug. All patients were discharged home within 3 hours. We conclude that in the prevention of PONV, the combination of metoclopramide and droperidol is superior to the use of droperidol alone at one hour but not at two hours postoperatively.

Riemerella anatipestifer, the causative agent of septicemia anserum exsudativa (also called new duckling disease), belongs to the family Flavobacteriaceae of gram-negative bacteria. We determined the DNA sequences of the rrs genes encoding the 16S rRNAs of four R. anatipestifer strains by directly sequencing PCR-amplified rrs genes. A sequence similarity analysis confirmed the phylogenetic position of R. anatipestifer in the family Flavobacteriaceae in rRNA superfamily V and allowed fine mapping of R. anatipestifer on a separate rRNA branch comprising the most closely related species, Bergeyella zoohelcum, as well as Chryseobacterium balustinum, Chryseobacterium indologenes, and Chryseobacterium gleum. The sequences of the rrs genes of the four R. anatipestifer strains varied between 0.5 and 1.0%, but all of the strains occupied the same position on the phylogenetic tree. In general, differences in rrs genes were observed among R. anatipestifer strains, even within a given serotype, as shown by restriction fragment length polymorphism of PCR-amplified rrs genes.

We gave sedation for the conduct of lumbar epidural analgesia using intravenous ketamine 0.3 mg.kg-1 with intravenous midazolam 2 mg. Forty adult Chinese females undergoing major gynaecological laparotomies had epidural catheter inserted before general anaesthesia, 20 of whom were given ketamine and midazolam (study group) and the other 20 acted as control. During the conduct of the epidural, the pain and anxiety scores in the study group were significantly less than the control group (P < 0.05). Patients were significantly more sedated in the study group (P < 0.05). All the patients in the study group were satisfied and would consent to future epidural versus 75% in the control group (significant at P < 0.05). Ninety per cent of patients in the study group had amnesia but none in the control group. Pain experienced during the epidural was the reason for refusal of future epidural. We did not observe any emergence phenomenon or cardiovascular stimulation. There was a statistically significant decrease in the pulse oximetry oxygen saturation (SpO2) in the study group but none required oxygen supplementation. We concluded that pain caused by the conduct of epidural did decrease the patient's acceptance rate to future epidural, and the combined use of intravenous ketamine 0.3 mg.kg-1 and midazolam 2 mg provided adequate sedation, analgesia, anxiolysis and amnesia to significantly increase the acceptance rate without any significant side effects.

The catabolic plasmid pHMT112 in Pseudomonas putida ML2 contains the bed gene cluster encoding benzene dioxygenase (bedC1C2BA) and a NAD+-dependent dehydrogenase (bedD) required to convert benzene into catechol. Analysis of the nucleotide sequence upstream of the benzene dioxygenase gene cluster (bedC1C2BA) revealed a 1,098-bp open reading frame (bedD) flanked by two 42-bp direct repeats, each containing a 14-bp sequence identical to the inverted repeat of IS26. In vitro translation analysis showed bedD to code for a polypeptide of ca. 39 kDa. Both the nucleotide and the deduced amino acid sequences show significant identity to sequences of glycerol dehydrogenases from Escherichia coli, Citrobacter freundii, and Bacillus stearothermophilus. A bedD mutant of P. putida ML2 in which the gene was disrupted by a kanamycin resistance cassette was unable to utilize benzene for growth. The bedD gene product was found to complement the todD mutation in P. putida 39/D, the latter defective in the analogous cis-toluene dihydrodiol dehydrogenase. The dehydrogenase encoded by bedD) was overexpressed in Escherichia coli and purified. It was found to utilize NAD+ as an electron acceptor and exhibited higher substrate specificity for cis-benzene dihydrodiol and 1,2-propanediol compared with glycerol. Such a medium-chain dehydrogenase is the first to be reported for a Pseudomonas species, and its association with an aromatic ring-hydroxylating dioxygenase is unique among bacterial species capable of metabolizing aromatic hydrocarbons.

A respiratory defective pet mutant (W303 delta FUM1) of Saccharomyces cerevisiae deficient in fumarase was transformed with the plasmid construct pG5/ST7. This plasmid contains the entire FUM1 gene with only a single base pair change (ATG-->ATC) confined to the putative second inframe translation initiation codon. While transformation of the fumarase deficient mutant with pG5/ST7 resulted in an elevation of fumarase activity in the mitochondria of the transformed strain, fumarase activity in the cytosol remained negligible his result indicated that the cytosolic fumarase isoenzyme is synthesized exclusively from the second translation initiation codon of FUM1. Results of RACE-PCR of the 5' ends of FUM1 transcripts confirmed that the FUM1 gene synthesizes two distinct species of transcripts. These data provide strong evidence for the synthesis and targeting of two fumarase isomers.

One thousand one hundred and sixty three patients (male-852, female-311) with ureteric calculi requiring intervention were treated between April 1988 to July 1992. Four hundred and eleven cases were treated by ESWL Monotherapy, 414 by stone manipulation plus ESWL, 301 by retrograde ureteroscopic lithotripsy, 36 by percutaneous antegrade ureteroscopic lithotripsy and 1 case by open ureterolithotomy. There were 25 failures of the initial procedures. Only three cases that failed primary procedures required open surgery. Other complications include minor ureteric mucosal perforation (3%), infection (3%), transient moderate to gross haematuria (20%), loin ache (26.4%), irritative urination (34.4%) and low grade fever (30.1%). Current modalities used in the treatment of ureteric calculi produce good results and there is generally no primary role for any open surgery.

The nucleotide sequence of YKE2, a yeast nuclear gene, has been determined. The deduced YKE2 protein has 114 amino acids (12 kDa), shows significant homology with the murine KE2 and contains a putative Leu-zipper motif characteristic of a group of DNA-binding proteins [Landschulz et al., Science 240 (1988) 1759-1764]. Strains in which YKE2 has been disrupted show normal cell growth in glucose and galactose media over the temperature range of 16 to 40 degrees C. Disrupted strains also display normal mating and sporulation abilities. Northern analysis revealed that the transcription of YKE2 is unresponsive to catabolite repression by glucose.

The nucleotide sequence of RPS26, the gene encoding a homologue of ribosomal protein small subunit S26 in Saccharomyces cerevisiae, was determined. The deduced amino-acid sequence showed significant identity with its counterparts from Neurospora crassa, human, rat and Arabidopsis thaliana. Disruption of RPS26 resulted in the formation of micro-colonies, suggesting that it is important for the normal cell growth of S. cerevisiae.

We have constructed a hybrid multicomponent dioxygenase gene cluster in which the bphA1 gene, coding for ISP alpha subunit of biphenyl dioxygenase from Pseudomonas pseudoalcaligenes KF707, has been replaced by the bedC1 gene encoding the corresponding subunit of benzene dioxygenase from P. putida ML2. Escherichia coli cells containing the chimeric dioxygenase acquired the novel capability of producing indigo from indole. Furthermore, when compared to biphenyl dioxygenase, the hybrid dioxygenase enzyme was half as active towards benzene but exhibited only 4% activity when biphenyl was the substrate. The results implicate the ISP alpha subunit to be involved in substrate specificity and activity of the dioxygenase enzyme.

The benzene dioxygenase from Pseudomonas putida ML2 is a multicomponent complex comprising a flavoprotein reductase, a ferredoxin, and a terminal iron-sulfur protein (ISP). The catalytic activity of the isolated complex shows a nonlinear relationship with protein concentration in cell extracts, with the limiting factor for activity in vitro being ferredoxin(BED). The relative levels of the three components were analyzed by using 125I-labelled antibodies, and the functional molar ratio of ISP(BED), ferredoxin(BED), and reductase(BED) was shown to be 1:0.9:0.8, respectively. The concentration of ferredoxin(BED) was confirmed by quantitative electron paramagnetic resonance spectroscopy of the 2Fe-2S centers in ferredoxin(BED) and ISP(BED) of whole cells. These results demonstrate that the ferredoxin(BED) component is a limiting factor in dioxygenase activity in vitro. To determine if it is a limiting factor in vivo, a plasmid (pJRM606) overproducing ferredoxin(BED) was introduced into P. putida ML2. The benzene dioxygenase activity of this strain, measured in cell extracts, was fivefold greater than in the wild type, and the activity was linear with protein concentration in cell extracts above 2 mg/ml. Western blotting (immunoblotting) and electron paramagnetic resonance spectroscopic analysis confirmed an elevated level of ferredoxin(BED) protein and active redox centers in the recombinant strain. However, in these cells, the increased level of ferredoxin(BED) had no effect on the overall rate of benzene oxidation by whole cells. Thus, we conclude that ferredoxin(BED) is not limiting at the high intracellular concentration (0.48 mM) found in cells.

Benzene dioxygenase, catalyzing the oxidation of benzene to cis-1,2-dihydroxy-cyclohexa-3,5-diene, comprises four polypeptides that are encoded by plasmid pHMT112 of Pseudomonas putida ML2. In this study, the nucleotide (nt) sequences of four genes encoding this enzyme (bedC1C2BA) were determined, and the amino acid (aa) sequences were deduced. The sequence showed significant homology with the chromosomally encoded benzene dioxygenase and toluene dioxygenase genes (73-77% for nt and 83-99% for aa), but not the plasmid-encoded naphthalene dioxygenase genes (20-26% for nt and 32-36% for aa). A conserved motif (Cys-Xaa-His-15-to-17 aa-Cys-Xaa2-His, where Xaa is any aa), proposed to bind the Rieske-type [2Fe-2S] cluster, was identified in the deduced aa sequence of the iron-sulfur proteins. Three regions were also identified in the flavoprotein which are likely to be involved in FAD and NAD+ binding. The gene order of bedC1C2BA is consistent with most ring-hydroxylating dioxygenases isolated from Pseudomonas. However, the G+C content of 47% is in contrast to the high G+C content of the Pseudomonas chromosome (63%) and other Pseudomonas plasmids (57%), and with its unique codon usage preference this suggests that bedC1C2BA originated from a host derived from a different genus.

Pseudomonas putida ML2 contains a large catabolic plasmid, pHMT112, carrying genes that encode the dioxygenase and dehydrogenase involved in the catabolism of benzene via the ortho or beta-ketoadipate pathway. pHMT112 was derived from a larger and less stable plasmid in P. putida ML2 following growth on succinate as carbon and energy source but was, however, stably maintained in P. putida even in the absence of selection for growth on benzene. Cleavage sites for the restriction endonucleases DraI, XbaI, and BamHI were mapped on the plasmid. A region of the plasmid, downstream of the benzene dioxygenase genes (bedC1C2BA), was found to encode the cis-benzene dihydrodiol dehydrogenase gene (bedD). Recombinant Escherichia coli containing bedC1C2BAD genes was found to express benzene dioxygenase and dehydrogenase activity, indicated by the production of catechol when incubated in the presence of benzene. Hybridization using benzene dioxygenase genes as probes was used to construct a restriction map of the 35.5-kb XhoI-DraI fragment on which the bed operon was located.

Hybridization using heterologous dioxygenase gene probes indicated the presence of the genes encoding the enzyme benzene dioxygenase on a 112 kb plasmid from Pseudomonas putida ML2. They were identified as benzene dioxygenase genes (bed ABC1C2) by cloning in Escherichia coli and analysis of expression by Western blotting using antibodies raised to the four polypeptides of purified benzene dioxygenase.

Three hundred and ninety five cases in 358 consecutive patients (male-232, female-126) with renal and ureteric stones were treated with extracorporeal shockwave lithotripsy (ESWL) from March to November 1988. They either had ESWL alone, or in combination with stone manipulation or debulking percutaneous nephrolithotripsy (PCNL). Seventy five percent of the stones were found in the pelvicalyceal system and 25% in the ureter. Seventy-six percent of the stones were less than 25mm size. Two hundred and ninety (79%) cases were followed up to three months. Two hundred and forty nine (85.9%) cases were stone free and 36 (12.4%) had residual sand less than 3mm size. Five (1.7%) cases failed to fragment with ESWL monotherapy and were salvaged by either percutaneous or ureteroscopic intervention. None of the cases required any open surgery intervention.

Sixty eight consecutive cases of percutaneous renal surgery, percutaneous nephrolithotripsy (PCNL), were performed on 64 patients (male-41, female-23) at the Subang Jaya Medical Centre from April 1988 to July 1989. All the cases were done as a one stage procedure. Fifty eight stones were large renal or staghorn and ten were ureteric. Thirty cases (41%) were stone free after PCNL alone. Thirty eight cases had residual fragments needing extracorporeal shockwave lithotripsy (ESWL). Mean operating time was 109.6 +/- 36.0 minutes. Mean hospital stay was 4.5 +/- 1.8 days. At three months follow-up, 86% of the cases were stone free. The remaining had residual sand (less than 3mm). Minor complications occurred in six patients. None required major surgical intervention post PCNL.

Two photosynthetic algal cultures, one Chlorella vulgaris, and the other a Chlorogonium sp., were cultured under light limitations in chemostats. The effects of growth temperature on their energy yield and maintenance energy requirement were studied. It was observed that a lowering in temperature resulted in a lower maximum growth yield from the light energy, Y(G). This was attributed to two reasons. First, at low temperatures there was a change in the algal cell composition with more energy being expended to synthesize a higher biomass protein content. Secondly, at low temperatures, a cyanide-resistant respiratory pathway became operative which led to a decrease in the number of ATP being generated. The maintenance energy coefficient was a function of temperature increasing with decreasing temperature. This might reflect energy wastage by the cell at low temperatures. The maximum specific growth rate dropped with decreasing temperature, and can be described by an Arrhenius type rate-temperature model up to the optimal temperature for growth; i.e., activation energy remained constant.