Guideline adherence for empirical treatment of pneumonia and patient outcome. Treating pneumonia in the Netherlands.

INTRODUCTION: According to the Dutch guidelines, severity of community acquired pneumonia (CAP) (mild, moderate-severe, severe) should be based on either PSI, CURB65 or a 'pragmatic' classification. In the last mentioned, the type of ward of admission, as decided by the treating physician, is used as classifier: no hospital admission is mild, admission to a general ward is moderate-severe and admission to an intensive care unit (ICU) is severe CAP. Empiric antibiotic recommendations for each severity class are uniform. We investigated, in 23 hospitals, which of the three classification systems empirical treatment of CAP best adhered to, and whether a too narrow spectrum coverage (according to each of the systems) was associated with a poor patient outcome (in-hospital mortality or need for ICU admission). PATIENTS AND METHODS: Prospective observational study in 23 hospitals. RESULTS: 271 (26%) of 1047 patients with CAP confirmed by X-ray were categorised in the same severity class with all three classification methods. Proportions of patients receiving guideline-adherent antibiotics were 62.9% (95% CI 60.0-65.8%) for the pragmatic, 43.1% (95% CI 40.1-46.1%) for PSI and 30.5% (95% CI 27.8-33.3%) for CURB65 classification. 'Under-treatment' based on the pragmatic classification was associated with a trend towards poor clinical outcome, but no such trend was apparent for the other two scoring systems. CONCLUSIONS: Concordance between three CAP severity classification systems was low, implying large heterogeneity in antibiotic treatment for CAP patients. Empirical treatment appeared most adherent to the pragmatic classification. Non-adherence to treatment recommendations based on the PSI and CURB65 was not associated with a poor clinical outcome.

[Can the thick drop/smear examination for malaria be replaced by a rapid diagnostic test in first intention? The Mayotte experience]. / Peut-on remplacer en première intention le frottis/goutte épaisse par un test de diagnostic rapide pour le diagnostic du paludisme ? L'expérience de Mayotte.

SETTINGS: Malaria is a public health problem in the French island of Mayotte (160,000 inhabitants) in the Indian Ocean. In the late 1990, resistance to chloroquine greatly increased, and so did the number of malaria cases, so that a new health policy had to be adopted. Since 2001, the initial smear/thick drop examination, the results of which took too long to obtain, has systematically been replaced by a rapid diagnosis test (Optimal IT Diamed) in all hospitals and public health centers. METHOD: Epidemiological data of malaria on the island was collected and a prospective study was made from March 2005 to February 2006, on two sites (the emergency department of the main hospital and a rural health centre) on all patients presenting with malaria (104 and 139 cases respectively). RESULTS: The first Optimal IT test diagnosed the condition accurately in 88 and 96% of the cases, respectively. Every time symptoms would persist after negative test results and an Optimal IT test was repeated within three days, the parasitemia level was low (0.08 to 0.66%). Very low parasitemia level was very likely to account for a false negative (test result). CONCLUSIONS: These results concerning malaria (and its epidemiological data) in Mayotte show that the initial use of an Optimal IT test instead of the thin/thick blood smear results in a faster management of patients with malaria, although the Optimal IT test is slightly less sensitive and requires training/practice.

Electrocardiographical case. Young woman with epilepsy.

A 24-year-old Chinese woman was referred for evaluation of palpitations. She had a background history of epilepsy. A 12-lead electrocardiogram (ECG) showed prolonged rate-corrected QT of 600 ms and a T-U complex. The ECG findings with her typical history suggest a diagnosis of congenital long QT syndrome. Diagnosis and treatment options are discussed.

The reelin pathway components disabled-1 and p35 in gangliogliomas--a mutation and expression analysis.

Focal epilepsies in young patients are frequently associated with differentiated glioneuronal tumours. Dysplastic neurones represent a characteristic neuropathological feature of gangliogliomas, the most common entity encountered in this group. Here, we have analysed two major components of the reelin pathway involved in neuronal migration and cortical development, that is, p35 and disabled-1 (dab1), in gangliogliomas. Genomic structures of human dab1 and p35 were identified 'in silico' using the HTGS databank, NCBI BLAST 2.1. DNA sequence analysis was carried out in gangliogliomas obtained from 29 epilepsy patients vs. peripheral blood DNA from non-affected control individuals (n = 100). Gene expression of dab1 and p35 was determined by real-time RT-PCR (reverse transcriptase polymerase chain reaction) in gangliogliomas (n = 14) vs. non-neoplastic central nervous system tissue (n = 20). The human dab1 gene contains 13 coding exons and is located on chromosome 1p31-32. A single coding exon constitutes the human p35 gene, which is located on chromosome 17q11.2. A novel homologueous genomic region on chromosome 2 has to be taken into account for future studies on p35. One ganglioglioma patient showed a unique polymorphism in the p35 gene. The single base exchange (C to A) at nucleotide 904 of the p35 cDNA (GenBank X80343, start ATG, codon 302) results in a leucine-isoleucine amino acid substitution. No mutations of the dab1 and p35 genes in gangliogliomas were observed. However, significantly lower levels of dab1 and p35 gene transcripts were detected in gangliogliomas compared to controls (dab1 28.24%, t-test P < 0.001; p35 21.28%, t-test P < 0.001, in gangliogliomas vs. controls). Our data suggest that mutational events of dab1 and p35 are not involved in the molecular pathogenesis of gangliogliomas. A potential functional role of these developmentally regulated genes for the formation of epileptogenic glioneuronal lesions remains to be elucidated.

Ethnic variations in female vulnerability after an acute coronary event.

OBJECTIVE: To determine the ethnic variation of short and long term female vulnerability after an acute coronary event in a population of Chinese, Indians, and Malays. DESIGN: Population based registry. PATIENTS: Residents of Singapore between the ages of 20-64 years with coronary events. Case identification and classification procedures were modified from the MONICA (monitoring trends and determinants in cardiovascular disease) project. MAIN OUTCOME MEASURES: Adjusted 28 day case fatality and long term mortality. RESULTS: From 1991 to 1999, there were 16 320 acute coronary events, including 3497 women. Age adjusted 28 day case fatality was greater in women (51.5% v 38.6%, p < 0.001), with a larger sex difference evident among younger Malay patients. This inequality between the sexes was observed in both the pre-hospitalisation and post-admission periods. Among hospitalised patients, women were older, were less likely to have suffered from a previous Q wave or anterior wall myocardial infarction, and had lower peak creatine kinase concentrations. Case fatality was higher among women, with adjusted hazard ratios of 1.64 (95% confidence interval (CI) 1.43 to 1.88) and 1.50 (95% CI 1.37 to 1.64) for 28 day and mean four year follow up periods. There were significant interactions of sex and age with ethnic group (p = 0.017). The adjusted hazards for mortality among Chinese, Indian, and Malay women versus men were 1.30, 1.71, and 1.96, respectively. The excess mortality among women diminished with age. CONCLUSION: In this multiethnic population, both pre-hospitalisation and post-admission case fatality rates were substantially higher among women. The sex discrepancy in long term mortality was greatest among Malays and in the younger age groups.

Transpositional behaviour of an Ac/Ds system for reverse genetics in rice.

A collection of transposon Ac/ Ds enhancer trap lines is being developed in rice that will contribute to the development of a rice mutation machine for the functional analysis of rice genes. Molecular analyses revealed high transpositional activity in early generations, with 62% of the T0 primary transformants and more than 90% of their T1 progeny lines showing ongoing active transposition. About 10% of the lines displayed amplification of the Ds copy number. However, inactivation of Ds seemed to occur in about 70% of the T2 families and in the T3 generation. Southern blot analyses revealed a high frequency of germinal insertions inherited in the T1 progeny plants, and transmitted preferentially over the many other somatic inserts to later generations. The sequencing of Ds flanking sites in subsets of T1 plants indicated the independence of insertions in different T1 families originating from the same T0 line. Almost 80% of the insertion sites isolated showing homology to the sequenced genome, resided in genes or within a range at which neighbouring genes could be revealed by enhancer trapping. A strategy involving the propagation of a large number of T0 and T1 independent lines is being pursued to ensure the recovery of a maximum number of independent insertions in later generations. The inactive T2 and T3 lines produced will then provide a collection of stable insertions to be used in reverse genetics experiments. The preferential insertion of Ds in gene-rich regions and the use of lines containing multiple Ds transposons will enable the production of a large population of inserts in a smaller number of plants. Additional features provided by the presence of lox sites for site-specific recombination, or the use of different transposase sources and selectable markers, are discussed.

Treatment of heart failure--role of biventricular pacing for heart failure not responding well to drug therapy.

OBJECTIVE: Patients with heart failure may have conduction abnormalities in up to 30%, further aggravating cardiac output. Drugs worsen these abnormalities and resynchronisation therapy with biventricular pacing improves cardiac function by effecting a more coordinated and efficient ventricular contraction. We report here the technique of biventricular pacing and its results. METHODOLOGY: Patients with NYHA Class III to IV heart failure, widened QRS (> or = 130 ms) complex on the ECG and impaired LVEF < or = 40% were enrolled. RESULTS: Biventricular pacing was performed in 29 patients (26 males, three females) from August 1999 to December 2001. The mean age of the patients was 59.6 +/- 12.8 years and 62% had underlying ischemic heart disease. All were in NYHA class III or more. Twenty-three had LBBB, four RBBB and two had widened paced QRS complex. The QRS duration was 161 +/- 21 ms and LVEF was 22 +/- 8%. All the left ventricular leads were implanted successfully. The procedure time was 167.0 +/- 79.6 mins and the fluoroscopy time was 43.8 +/- 41.4 mins. There were no significant complications. The NYHA class improved from a mean of 3.1 to 2.0 and exercise time from 252 +/- 95 seconds to 392 +/- 152 seconds at six months post implant (p=0.049). On follow-up (one month to 28 months), 25 (86%) patients had improvement in heart failure symptoms and 26 (90%) of the patients remained alive. CONCLUSION: Biventricular pacing can be safely performed and results in improvement in symptoms and exercise tolerance in heart failure patients with ventricular dyssynchrony not responding to drug therapy.

A case series of sotalol-induced torsade de pointes in patients with atrial fibrillation--a tale with a twist.

INTRODUCTION: Sotalol is a potent antiarrhythmic often used in patients with atrial fibrillation. However, it has been associated with a risk of provoking other potentially dangerous arrhythmias, especially if used in high doses and in patients with uncorrected electrolyte imbalance or impaired renal and cardiac function. CLINICAL PICTURE: We present 4 patients with atrial fibrillation treated with sotalol who developed torsade de pointes due to marked prolongation of the QT interval. While 1 patient had renal failure, all had normal left ventricular function. One patient had been treated with sotalol for more than 10 months before developing torsade de pointes precipitated by hypokalaemia, while another had tolerated sotalol for a 3-month period before the drug was discontinued, and only developed torsade de pointes when the drug was restarted 2 years later. Significantly, the doses used in all patients were relatively low, in contrast to most other reported cases where higher doses were used. CONCLUSION: As with all antiarrhythmic therapy, these cases illustrate the need for close follow-up of patients treated with sotalol, even if relatively low doses are used. In addition, patients who had previously tolerated the drug well are still susceptible to its proarrhythmic effects.

Immediate and mid-term safety and efficacy of single lead VDD pacemakers for patients with atrioventricular block and normal sinus node function--a single centre experience.

INTRODUCTION: Single lead atrial synchronous ventricular pacing (VDD) is increasingly being used in place of conventional dual chamber pacing (DDD) for patients with atrioventricular block and preserved sinus node function. Compared to the latter, VDD pacemakers provide similar haemodynamic benefits derived from atrial synchronous pacing, with the added benefit of an easier implant procedure. OBJECTIVE: To review the use, safety and efficacy of VDD pacing in a single tertiary referral centre. MATERIALS AND METHODS: A review of all patients with atrioventricular block and normal sinus node function implanted with a VDD pacemaker over a 2-year period at a local tertiary cardiac referral centre. Data on complications, atrial sensing performance and maintenance of atrioventricular synchrony during implant and at subsequent follow-up visits were obtained from a prospectively maintained registry and analysed. RESULTS: Forty-one patients (17 males, 24 females) with a mean age of 72 +/- 9 years received VDD pacing for various forms of high-grade atrioventricular block. The average implantation time was 46.8 +/- 17.1 minutes, and a pneumothorax in 1 patient was the only complication. Electrical measurements at implantation and subsequent follow-up visits revealed an initial rapid decrease in atrial signal amplitude (mean atrial P wave at implant 3.1 +/- 1.1 mV, predischarge 1.9 +/- 1.3 mV) which began to stabilise after 3 months, reaching a mean atrial P wave value of 1.3 +/- 0.3 mV at 24 months. The atrial sensing performance (percentage of atrial synchronous ventricular complexes) was 97% over a mean follow-up period of 9.9 months. Four patients (10%) developed paroxysmal atrial tachyarrhythmias. Sinus node dysfunction was not observed in any of our patients during the follow-up period. CONCLUSION: In patients with atrioventricular block and preserved sinus node function, single lead VDD pacing is safe and effective in maintaining a physiological atrial synchronous pacing mode.

Constrictive pericarditis--a rare but important cause of recurrent cardiac failure: a case report.

Constrictive pericarditis (CP) is an uncommon cardiac disease which is often difficult to diagnose because of its vague and myriad clinical presentations. We report a case of a middle-aged lady who had non-specific symptoms and signs for six years before she was eventually diagnosed to have idiopathic constrictive pericarditis. An awareness and understanding of this condition is important, as it is a progressive condition and the likelihood of cure depends very much on its early identification and treatment.

Gender differences in outcome after an acute myocardial infarction in Singapore.

OBJECTIVES: To characterise gender and age-related differences in presentation and outcome after an acute myocardial infarction (AMI). DESIGN: Data were derived retrospectively from the Singapore Myocardial Infarction Registry from 1988 through 1997. This database comprised all AMI cases for ages between 20 and 64 years (group A). For approximately three months a year, data were also collected for all AMI cases above the age of 64 years (group B). There were 13,048 and 4,425 cases in groups A and B respectively. RESULTS: In age - standardised AMI rates, males outnumbered females by a factor of 4.0 and 1.7 for groups A and B respectively.The median age of presentation was higher in females for both age groups being 58 years versus 54 years for group A and 75 years versus 72 years for group B. Younger females had worse survival at 28 days and were more likely to have prior ischaemic heart disease and require resuscitation. They were also more likely to have atypical symptoms. Previous myocardial infarction was not different between the sexes in both groups. Among the older age group, there was no gender difference in prior ischaemic heart disease, 28-day survival and requirement for resuscitation. CONCLUSION: Women who have AMI tend to be older than men. Gender differences are age-specific. Women who are 64 years and below have more atypical symptoms, prior ischaemic heart disease and worse prognosis than men after AMI. These differences are not seen in those over the age of 64.

Clinical outcomes of patients with diabetes mellitus and acute myocardial infarction treated with primary angioplasty or fibrinolysis.

OBJECTIVE: To compare the early and late outcomes of primary percutaneous transluminal coronary angioplasty (PTCA) with fibrinolytic treatment among diabetic patients with acute myocardial infarction (AMI). DESIGN: Retrospective observational study with data obtained from prospective registries. SETTING: Tertiary cardiovascular institution with 24 hour acute interventional facilities. PATIENTS: 202 consecutive diabetic patients with AMI receiving reperfusion treatment within six hours of symptom onset. INTERVENTIONS: Fibrinolytic treatment was administered to 99 patients, and 103 patients underwent primary PTCA. Most patients undergoing PTCA received adjunctive stenting (94.2%) and glycoprotein IIb/IIIa inhibition (63.1%). MAIN OUTCOME MEASURES: Death, non-fatal reinfarction, and target vessel revascularisation at 30 days and one year were assessed. RESULTS: Baseline characteristics were similar in these two treatment groups except that the proportion of patients with Killip class III or IV was considerably higher in those treated with PTCA (15.5% v 6.1%, p = 0.03) and time to treatment was significantly longer (103.7 v 68.0 minutes, p < 0.001). Among those treated with PTCA, the rates for in-hospital recurrent ischaemia (5.8% v 17.2%, p = 0.011) and target vessel revascularisation at one year (19.4% v 36.4%, p = 0.007) were lower. Death or reinfarction at one year was also reduced among those treated with PTCA (17.5% v 31.3%, p = 0.02), with an adjusted relative risk of 0.29 (95% confidence interval 0.15 to 0.57) compared with fibrinolysis. CONCLUSION: Among diabetic patients with AMI, primary PTCA was associated with reduced early and late adverse events compared with fibrinolytic treatment.

Two case reports on incessant left ventricular tachycardia: curative therapy with radiofrequency ablation.

INTRODUCTION: Incessant ventricular tachycardia is a rare arrhythmia which can be life threatening. Treatment with anti-arrhythmic agents may occasionally fail. CLINICAL PICTURE: We report 2 cases of incessant ventricular tachycardia. The first case was a young man with idiopathic left ventricular tachycardia who was in incessant ventricular tachycardia despite treatment with multiple anti-arrhythmic drugs and developed dilated cardiomyopathy. The second case was an asymptomatic girl with the incidental finding of an incessant ventricular tachycardia which originated from the left ventricular outflow tract. TREATMENT AND OUTCOME: Both patients underwent electrophysiologic study and radiofrequency ablation with complete termination of the tachycardia. CONCLUSION: Radiofrequency catheter ablation in experienced centres should be the first-line therapy for incessant ventricular tachycardia.

Dual-site atrial pacing for atrial fibrillation in patients without bradycardia.

Atrial pacing has been shown to delay the onset of atrial fibrillation (AF) when compared with ventricular pacing in patients with sick sinus syndrome. The role for pacing in the control of AF in patients without bradycardia is uncertain. We performed a randomized, crossover, single-blinded study in 22 patients (14 women, aged 63 +/- 10 years) with paroxysmal AF refractory to treatment with oral sotalol (202 +/- 68 mg/day) and no bradycardic indication for pacing. All patients received a dual-chamber pacemaker with 2 atrial pacing leads positioned at the high right atrium and coronary sinus ostium, respectively. Patients were randomized in a crossover fashion to be paced for 12 weeks, either with high right atrial (RA) pacing at 30 beats/min ("Off") or dual-site RA pacing with an overdrive algorithm that maintained atrial pacing at a rate slightly above the sinus rate ("On"). Treatment on resulted in a significantly higher percentage of atrial pacing and a reduction in atrial ectopic frequency than the treatment off period. The time to the first clinical AF recurrence was prolonged (15 +/- 17 to 50 +/- 35 days, p = 0.006), and total AF burden was reduced (45 +/- 34% vs 22 +/- 29%, p = 0.04) in the on-treatment phase. However, there was no difference in AF checklist symptom scores or overall quality-of-life measures. Dual-site RA pacing with continued sinus overdrive prolonged the time to AF recurrence and decreased AF burden in patients with paroxysmal AF. The absence of a major impact on symptom control suggests that pacing should be used as an adjunctive therapy with other treatment modalities for AF.

Glucocorticoid-inducible gene expression in rice.

We have studied the use of a glucocorticoid receptor-based inducible gene expression system in the monocotyledonous model plant rice (Oryza sativa L.). This system, originally developed by T. Aoyama and N.-H. Chua [(1997) Plant J 11: 605-612], is based on the chimaeric transcriptional activator GVG, consisting of the yeast Gal4 DNA-binding domain, the VP16 activation domain and the glucocorticoid receptor domain. For application in rice, we designed an optimized binary vector series (pINDEX) and tested this with the beta-glucuronidase (gusA) reporter gene. GUS expression was tightly controlled and relatively low concentrations (1-10 microM) of the glucocorticoid hormone dexamethasone (DEX) were able to induce GUS activities to levels comparable to those conferred by the strong cauliflower mosaic virus (CaMV) 35S promoter. DEX was taken up efficiently by the roots of tissue-cultured plantlets or mature plants in hydroponic culture, and induced GUS activity throughout the whole plant. DEX-induced GUS expression patterns were consistent in all lines and their T1 progeny. The phenotype of tissue-cultured rice plantlets was not affected when inductions with 10-100 microM DEX were limited to 1-4 days or when 2-week inductions were performed with 1 microM DEX, which was already sufficient to reach near-maximal GUS activity. However, 2-week inductions with 10 microM DEX caused growth retardation and developmental defects. As the severity of these effects varied between different lines, we could select lines with a mild phenotype for future use as activator lines in crosses with 'target' plants.

The differential diagnosis of supraventricular tachycardia using clinical and electrocardiographic features.

INTRODUCTION: Noninvasive determination of the mechanism of supraventricular tachycardia (SVT) is useful in order to decide on the appropriate mode of therapy. The aim of this study was to evaluate the usefulness of the clinical and electrocardiographic features in diagnosing the type of SVT. METHODS: Design--A retrospective review of case records and electrocardiograms (ECG) of patients with definitive diagnosis of the mechanism of supraventricular tachycardia (SVT) made during electrophysiological study (EPS) and catheter ablation. Setting--A tertiary referral centre for electrophysiological studies and radiofrequency catheter ablation of arrhythmias. Sample--One hundred consecutive patients with SVT who had EPS and catheter ablation at our institution. Chief Outcome--Comparison of clinical and ECG pacemeters among 3 different types of SVT, namely atrioventricular nodal reentrant tachycardia (AVNRT), atrioventricular reentrant tachycardia (AVRT) and atrial tachycardia (AT). RESULTS: There were 68 atrioventricular nodal reentrant tachycardia (AVNRT), 26 atrioventricular reentrant tachycardia (AVRT) and 6 atrial tachycardia (AT). AVRT had the earliest mean age of presentation at 26.8 +/- 11.9 years. Sex and age of onset of symptoms alone were, however, not valuable in diagnosing the type of SVT. P waves were more discernible in AVRT and AT than in AVNRT (69%, 67% and 44% respectively, P = 0.071). AVNRT had the shortest mean RP' interval (86.3 +/- 47.6 msec), while AT had the longest (187 +/- 80.6 msec, P < 0.0001). Conversely for mean P'R interval, AT had the shortest (125 +/- 30 msec), AVNRT had the longest (262.7 +/- 73.7 msec, P = 0.001). AVNRT had the smallest mean RP':P'R ratio (0.6 +/- 0.9), while AT had the largest (2.2 +/- 0.6, P = 0.001). The presence of pseudo r' in V1 and pseudo s' in II/III/aVF was diagnostic of AVNRT with a specificity of 90% and 100%, respectively, and positive predictive value of 97% and 100%, respectively. CONCLUSION: Pseudo s' in II/III/aVF is highly predictive of AVNRT. Measurement of RP' and PR' interval and ratio are also useful in determining the SVT mechanism.

Electrocardiographic case: diagnosis of acute myocardial infarction in the presence of left bundle branch block.

The electrocardiographic features associated with acute myocardial infarction (AMI) are often easily recognised. However, interpretation is made more difficult in the presence of confounding patterns such as a left bundle branch block (LBBB). This may result in missed cases which may otherwise have benefited from acute revascularisation therapy. Though not straightforward, the diagnosis of AMI in the presence of LBBB can be made with a reasonable amount of accuracy. We report a case of acute myocardial infarction with LBBB that was appropriately diagnosed and underwent acute revascularisation by angioplasty. A detailed knowledge of the typical electrocardiographic features associated with LBBB, especially the ST segment morphologies, is very important. This will greatly aid recognition of an evolving AMI and help us decide on the most appropriate therapy.

Overexpression of Arabidopsis thaliana SKP1 homologues in yeast inactivates the Mig1 repressor by destabilising the F-box protein Grr1.

The timed destruction of cell cycle regulatory proteins is of key importance in controlling cell cycle progression in eukaryotes. Recently, Skp1 from yeast (Saccharomyces cerevisiae) was shown to play an important role in the ubiquitin-mediated proteolysis of these proteins via the Skp1-Cdc53-F-box (SCF) pathway. Here we describe the fortuitous cloning of cDNAs for two Skp1 homologues from the plant Arabidopsis thaliana on account of their ability to activate reporter gene expression in yeast directed by the cyt-1 element from the promoter of the Agrobacterium tumefaciens T-cyt gene, which is essential for expression of the gene in plants. This element is strikingly similar in sequence to the binding site for the yeast Migl protein, a transcriptional repressor of genes involved in the utilisation of carbohydrates other than glucose. We report that Mig1 protein binds to the cyt-1 element with similar specificity as a previously described plant nuclear protein factor, and that the cyt-1 element is a target for an unknown yeast transcriptional activator when Mig1 itself is inactivated. Interestingly, our data further indicate that A. thaliana Skp1 inactivates Mig1 by destabilising the yeast F-box protein Grr1, which is required for cyclin degradation and is thus involved in control of the cell cycle, and for glucose-regulated gene repression. Our results suggest that the plant counterpart of yeast Skp1 is probably also instrumental in ubiquitin-mediated proteolysis of specific proteins via an SCF-like pathway.

HD-Zip proteins of families I and II from rice: interactions and functional properties.

Proteins of the closely related homeodomain-leucine zipper (HD-Zip) families I and II in plants are putative transcription factors that interact with similar pseudopalindromic DNA recognition sites. We have previously described the Oshox1 gene from rice, which encodes an HD-Zip II protein. To identify further rice HD-Zip proteins, one-hybrid screens were performed in yeast strains containing a HIS3 reporter gene with upstream HD-Zip recognition sites. This resulted in the isolation of six new cDNAs encoding HD-Zip proteins belonging to family I (Oshox4, -5, -6) or family II (Oshox2, -3, -7). In transient assays, using rice suspension-cultured cells transformed by particle bombardment, we showed previously that Oshox1 can transcriptionally repress the activity of reporter gene constructs with upstream HD-Zip binding sites. Here, we confirm the repression properties of Oshox1 by showing that the repression function can be conferred on a heterologous DNA-binding domain. This portable functional domain (residues 1-155) is located proximal to the HD-Zip domain. In yeast, the same region of the Oshox1 protein was found to confer transcriptional activation instead of repression, pointing to the possibility that cell type-specific factors may determine the functional properties of the Oshox1 protein in rice. Like Oshox1, another HD-Zip family II protein (Oshox3) was also found to function as a transcriptional repressor in rice cells. In contrast, two HD-Zip I family proteins (Oshox4 and -5) appeared to act as activators in both rice and yeast cells. Results of two-hybrid assays and electrophoretic mobility shift assays strongly suggest that all HD-Zip proteins of families I and II can form homodimers and also heterodimers with all HD-Zip proteins of the same family. Heterodimerization across the HD-Zip families I and II apparently does not to occur.