Differential gene expression analysis reveals the fast-growth mechanisms in Melia dubia at different stand ages.

BACKGROUND: Melia dubia Cav. is a fast-growing multipurpose tree suitable for agroforestry and has been widely cultivated for wood-based industries, particularly pulp and paper production. Despite its high economic value in India, there is a lack of information regarding the molecular mechanism driving its fast-growth. Therefore, this study aimed to elucidate the molecular mechanisms responsible for fast-growth by expression analysis of selective candidate genes. METHODS AND RESULTS: Initially, growth traits were assessed, including tree height and diameter at breast height (DBH), across three different ages (one-year-old, two-year-old, and three-year-old) of M. dubia plantations. Tree volume based on tree height and DBH, was also calculated. The analysis of annual tree height increment revealed that the second-year plantation exhibited the higher increment, followed by first and third years. In contrast, DBH was maximum in third-year plantation, followed by the second and first years. Similarly, annual tree volume increment showed a similar trend with DBH that maximum in the third year, followed by second and first years. Furthermore, a differential gene expression analysis was performed using qRT-PCR on four genes such as Phloem Intercalated with Xylem (PXY), Clavata3/Embryo Surrounding Region-Related 41 (CLE41), 1-aminocyclopropane-1-carboxylic acid synthase (ACS-1) and Hemoglobin1 (Hb1) for downstream analysis. The relative gene expression showed up-regulation of CLE41, ACS-1, and Hb1 genes, while the PXY gene was downregulated across the tree ages. Interestingly, a positive association was observed between tree growth and the expression of the selected candidate genes. CONCLUSION: Our results pave the way for further research on the regulatory mechanisms of genes involved in fast-growth and provide a basis for genetic improvement of Melia dubia.

Whole plant response of Pongamia pinnata to drought stress tolerance revealed by morpho-physiological, biochemical and transcriptome analysis.

BACKGROUND: Pongamia is considered an important biofuel species worldwide. Drought stress in the early growth stages of Pongamia influences negatively on the germination and seedling development. Due to lack of cultivar stability under drought stress conditions, establishment of successful plantation in drought hit areas becomes a major problem. To address this issue, drought stress response of four Pongamia genotypes was studied at morphological, physio-chemical and transcriptome levels. METHODS AND RESULTS: Drought stress was levied by limiting water for 15 days on three months old seedlings of four genotypes. A significant effect of water stress was observed on the traits considered. The genotype NRCP25 exhibited superior morpho-physiological, biochemical drought responses. Also, the genotype had higher root length, photosynthetic pigments, higher antioxidant enzymes and solute accumulation compared to other genotypes. In addition, transcript profiling of selected drought responsive candidate genes such as trehalose phosphate synthase 1 (TPS1), abscisic acid responsive elements-binding protein 2 (ABF2-2), heat shock protein 17 (HSP 17 kDa), tonoplast intrinsic protein 1 (TIP 1-2), zinc finger homeodomain protein 2 (ZFP 2), and xyloglucan endotransglucolase 13 (XET 13) showed only up-regulation in NRCP25. Further, the transcriptome responses are in line with key physio-chemical responses exhibited by NRCP25 for drought tolerance. CONCLUSIONS: As of now, there are no systematic studies on Pongamia drought stress tolerance; therefore this study offers a comprehensive understanding of whole plant drought stress responsiveness of Pongamia. Moreover, the results support important putative trait indices with potential candidate genes for drought tolerance improvement of Pongamia.

Head and neck involvement with histoplasmosis; the great masquerader.

INTRODUCTION: Head and neck involvement with histoplasmosis usually occurs as a part of the disseminated illness. There are no pathognomic features of the upper aerodigestive tract involvement and the lesion may mimic a host of other conditions. The current report presents our experience with head and neck histoplasmosis in a non-endemic tertiary care center. MATERIALS AND METHODS: We present a case of disseminated histoplasmosis with oral symptoms and lesions as the chief complaints. A 10 years' retrospective institutional database search was undertaken to identify the patients with histoplasmosis affecting head and neck region treated at our institution. The demographic and treatment details of the patients were reviewed. RESULTS: In addition to the index patient, four more patients (two with gingivobuccal and one each with nasal and laryngeal histoplasmosis) were found. Out of the five patients, only one patient was found to have underlying immunosuppression. All of the patients were diagnosed with biopsy showing typical appearance of the intracellular organism. All the patients were satisfactorily treated with systemic antifungal treatment. CONCLUSION: Upper aerodigestive tract involvement with histoplasmosis can present as an intriguing clinical puzzle. A high index of suspicion is needed and biopsy is the gold standard for the diagnosis. Intravenous Liposomal Amphotericin B and oral Itraconazole are standard treatment agents of choice and are highly efficacious in achieving cure.

Flow Mediated Dilatation and Progression of Abdominal Aortic Aneurysms.

OBJECTIVE/BACKGROUND: Biomarker(s) for prediction of the future progression rate of abdominal aortic aneurysms (AAA) may be useful to stratify the management of individual patients. AAAs are associated with features of systemic inflammation and endothelial dysfunction. Flow mediated dilatation (FMD) of the brachial artery is a recognised non-invasive measurement for endothelial function. We hypothesised that FMD is a potential biomarker of AAA progression and reflects the temporal changes of endothelial function during AAA progression. METHODS: In a prospectively recruited cohort of patients with AAAs (Oxford Abdominal Aortic Aneurysm Study), AAA size was recorded by antero-posterior diameter (APD) (outer to outer) on ultrasound. Annual AAA progression was calculated by (ΔAPD/APD at baseline)/(number of days lapsed/365 days). FMD was assessed at the same time as AAA size measurement. Analyses of data were performed in the overall cohort, and further in subgroups of AAA by size (small: 30-39 mm; moderate: 40-55 mm; large: > 55 mm). RESULTS: FMD is inversely correlated with the diameter of AAAs in all patients (n=162, Spearman's r=-.28, p<.001). FMD is inversely correlated with AAA diameter progression in the future 12 months (Spearman's r=-.35, p=.001), particularly in the moderate size group. Furthermore, FMD deteriorates during the course of AAA surveillance (from a median of 2.0% at baseline to 1.2% at follow-up; p=.004), while surgical repair of AAAs (n=50 [open repair n=22, endovascular repair n=28)] leads to an improvement in FMD (from 1.1% pre-operatively to 3.8% post-operatively; p<.001), irrespective of the type of surgery. CONCLUSION: FMD is inversely correlated with future AAA progression in humans. FMD deteriorates during the natural history of AAA, and is improved by surgery. The utility of FMD as a potential biomarker in the context of AAA warrants further investigation.

Delays in the presentation to stroke services of patients with transient ischaemic attack and minor stroke.

BACKGROUND: The UK National Institute for Health and Care Excellence (NICE) guidelines state that carotid endarterectomy should be scheduled within 2 weeks of symptoms. The recent National Stroke Strategy has reduced the time interval to 48 h. This study aimed to review the possible delays. METHODS: This study analysed patients with confirmed transient ischaemic attack (TIA) or minor stroke, referred to a single tertiary centre clinic and followed up 1 month after the event. A questionnaire was used to collect data on the rapid-access clinic pathway, and details of previous medication and treatment. RESULTS: Some 150 patients presented with a confirmed TIA or minor stroke during a 5-month interval (June to October 2014). Fifty-one (34·0 per cent) had a history of TIA or stroke and 35 (23·3 per cent) had undergone an 'index' event in the 5 days before presentation. Forty-five patients (30·0 per cent) experienced a reduction or loss of vision. Of this group, 32 had a deficit in vision only, none of whom attributed these symptoms to a cerebrovascular event. Overall 92 (61·3 per cent) of the 150 patients had a delay in presentation to medical services. Forty-seven (31·3 per cent) had residual symptoms at the clinic appointment. Eighty-eight patients (58·7 per cent) did not think they were having a stroke and 54 (36·0 per cent) were unaware of the National Stroke Strategy (FAST campaign - Face, Arm, Speech, Time). CONCLUSION: Two-thirds of patients were not aware they were having a stroke, one-third were unaware of the FAST campaign and nearly one-third presented with eye symptoms. Inclusion of eye symptoms and reaffirmation of the need to react might avoid unnecessary delays in the presentation of patients with TIA and minor stroke.

Age-specific incidence, risk factors and outcome of acute abdominal aortic aneurysms in a defined population.

BACKGROUND: Contemporary population-based data on age-specific incidence and outcome from acute abdominal aortic aneurysm (AAA) events are needed to understand the impact of risk factor modification and demographic change, and to inform AAA screening policy. METHODS: In a prospective population-based study (Oxfordshire, UK, 2002-2014), event rates, incidence, early case fatality and long-term outcome from all acute AAA events were determined, both overall and in relation to the four main risk factors: smoking, hypertension, male sex and age. RESULTS: Over the 12-year interval, 103 incident acute AAA events occurred in the study population of 92,728 (men 72·8 per cent; 59·2 per cent 30-day case fatality rate). The incidence per 100,000 population per year was 55 in men aged 65-74 years, but increased to 112 at age 75-84 years and to 298 at age 85 years or above. Some 66·0 per cent of all events occurred in those aged 75 years or more. The incidence at 65-74 years was highest in male smokers (274 per 100,000 population per year); 27 (96 per cent) of 28 events in men aged less than 75 years occurred in ever-smokers. Mean(s.d.) age at event was lowest in current smokers (72·2(7·2) years), compared with that in ex-smokers (81·2(7·0) years) and never-smokers (83·3(7·9) years) (P < 0·001). Hypertension was the predominant risk factor in women (diagnosed in 93 per cent), with 20 (71 per cent) of all 28 events in women occurring in those aged 75 years or above with hypertension. The 30-day case fatality rate increased from 40 per cent at age below 75 years to 69 per cent at age 75 years or more (P = 0·008). CONCLUSION: Two-thirds of acute AAA events occurred at age 75 years or above, and more than 25 per cent of events were in women. Taken with the strong associations with smoking and hypertension, these findings could have implications for AAA screening.

First Report of Apple Top Working Disease Caused by Viruses (Apple stem grooving virus, Apple chlorotic leaf spot virus, and Apple stem pitting virus) in Apple in India.

Top working of apple (Malus domestica Borkh.) trees of old, unproductive, and less preferred cultivars with the newly introduced spur type commercial cultivars has become a common practice with many growers in the northwestern Himalayan region of India. Typical viral symptoms of curling, puckering, and necrosis on leaves were observed with an incidence of 80% on Red Chief, Super Chief, Scarlet Spur, Schillet Spur, Washington Red Delicious, and many other newly introduced cultivars during surveys conducted in May and June 2009. Leaf samples from top worked trees were tested for the presence of Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), and Apple stem pitting virus (ASPV) by employing biological detection (herbaceous and woody indicators), double antibody sandwich (DAS)-ELISA), and reverse transcriptase (RT)-PCR based detection. Mechanical transmission to herbaceous indicators produced chlorotic lesions on Chenopodium quinoa and C. amaranticolor, whereas marginal necrosis was induced on Phaseolous vulgaris within 9 to 21 days after sap inoculations. All three viruses, i.e., ASGV, ASPV, and ACLSV, were detected from these herbaceous indicators in DAS-ELISA (BIOREBA AG, Switzerland). Furthermore, symptoms similar to those observed in orchards were produced when the test budwood was inoculated onto the woody indicator (M. pumila 'Spy 227') plant by double grafting, grafting cum budding, and double budding methods within time periods ranging from 4 months in double grafting, 5 months in double budding, to 1 year 4 months in the grafting cum budding method. The presence of all three viruses was confirmed by DAS-ELISA again in Spy 227 woody indicator. PCR detection was carried out by using the coat protein gene specific primers (ASGV5641 [forward], ASGV6396 [reverse]; ACLSV6784 [forward], ACLSV7365 [reverse] [2]; ASP-C [sense], ASP-A [anti-sense] [1]) of all the viruses detected through ELISA. The amplified products were cloned, sequenced, and deposited in NCBI (GenBank Accessions KC110892 for ASGV, KC154859 for ASPV, and KC154862 for ACLSV). BLASTn analysis showed the ASGV isolate had 97 to 98% sequence identity with Indian (FM204881) and Brazilian (AF438409) ASGV isolates. The ASPV and ACLSV isolates had 98% and 99% sequence identity with Chinese (JF895517) and Japanese (AB326230) isolates, respectively. To the best of our knowledge, this is the first report of apple top working disease associated with ASGV, ASPV, and ACLSV infection in commercial cultivars of apple from India and seems to be a serious threat for growing virus-free healthy stocks in orchards. Top working disease in apple associated with ASGV, ASPV, and ACLSV viruses has been reported from Japan (3,4). References: (1) J. K. Kundu et al. Plant Prot. Sci. 39:88, 2003. (2) O. Nickel et al. Fitopatol. Brasil. 26:655, 2001. (3) H. Yanase. Bull. Fruit Tree Res. Stn., Japan Ser. C 1:47, 1974. (4) H. Yanase et al. Acta Hortic. 44:221, 1975.

Increased use of laparoscopy in acute right iliac fossa pain - is it good for patients?

AIM: The use of laparoscopy, with or without appendicectomy, is becoming more common in the management of acute right iliac fossa (RIF) pain, but little is known of the 'unintended' consequences of this change. This study aimed to evaluate the impact of increased use of laparoscopy on the number and type of patients treated surgically and on the rate of negative appendicectomy. METHOD: A prospective audit was carried out of admissions to a teaching hospital over two, 3-month periods during 2007 and 2008. The management, investigations and outcome of patients presenting with RIF pain were studied. RESULTS: Admissions were stable over the two time-periods. There was a significant increase in the number of laparoscopic operations performed, from 22.5% (14/62) in 2007 to 85.7% (72/84) in 2008 (P < 0.0001), and the percentage of patients undergoing surgery rose from 55.4% (n = 62) in 2007 to 71.2% (n = 84) in 2008 (P < 0.01). In 2008, female patients were more likely to have surgery, an increase from 37.1% to 66.2% (P < 0.001), and were more likely to have a laparoscopic procedure, an increase from 50% to 98% (P < 0.0001). The rate of histologically confirmed appendicitis did not increase significantly (50/122 vs 57/118; P = 0.25), but the number of patients with a normal appendix either left in situ because it was macroscopically normal or found to be histologically normal following excision, increased significantly, from 9.01% in 2007 to 21.2% in 2008 (P < 0.01). The diagnostic value of pelvic ultrasound decreased from 75.6% of examinations in 2007 to 54.5% in 2008 (P = 0.039). CONCLUSION: An increase in laparoscopic procedures has resulted in more operations in women, an associated higher negative appendicectomy rate and decreased usefulness of pelvic ultrasound. Increased use of laparoscopy needs to be balanced against the diagnostic benefits of 'negative' laparoscopy.

The role of cell salvage autotransfusion in abdominal aortic aneurysm surgery.

OBJECTIVE: Abdominal aortic aneurysm (AAA) repairs, both elective and rupture, are associated with significant blood loss often requiring transfusion. Cell-salvage autotransfusion has been developed to reduce the need for allogeneic blood. We review the literature to delineate the role of cell salvage in reducing allogeneic blood use in open AAA repairs. METHODS: A systematic search of the English-language literature was performed using the PubMed, Embase and Cochrane databases up to August 2010. RESULTS: Twenty-three studies were identified. Whilst some data are conflicting, cell salvage appears to reduce overall use and exposure to allogeneic blood, and reduces length of intensive care unit and hospital stay after elective AAA repairs. There may be additional benefit by combining cell salvage with other blood-conservation techniques. Use of cell salvage in ruptured AAA repairs consistently reduced blood-product requirements. CONCLUSIONS: Cell salvage appears to reduce blood-product use in both elective and rupture AAA repairs. Owing to the heterogeneity in methodology of published data, further study may be required before cell salvage becomes standard practice in open AAA repairs.

Prevalence and Epidemiological Pattern of Abdominal Aortic Aneurysms in Africa: A Systematic Review.

Introduction: The incidence of abdominal aortic aneurysms (AAAs) in high-income countries has been declining in the last three decades. However, in most low-income and middle-income countries especially in Africa, little is known about its burden. The absence of screening services for AAA in African countries makes it difficult to detect and promptly manage AAA before rupture, which has significant implications for mortality. This study sought to systematically assess the prevalence of AAA amongst patients visiting hospitals in Africa and evaluate its epidemiological pattern. Materials and Methods: A systematic review was performed on the EMBASE, GLOBAL HEALTH, MEDLINE, and PUBMED databases. The review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses statement standards and protocol registered with PROSPERO (CRD42020162214). A data extraction tool was used to get relevant information from these studies. Quality assessment and risk of bias were performed using the Newcastle Ottawa Scale for cross-sectional studies. Results were summarised in tables, figures, and a forest plot. A narrative synthesis approach of the articles was taken. Results: Two hundred and sixty-one studies were identified and after the exclusion of 246, a final 15 were deemed suitable for analysis. A total of 4012 participants were screened for AAA and of these, 129 cases were identified. The prevalence of AAA in these studies ranged from 0.7 to 6.4%. Male participants accounted for 115 (89.1%) of the cases. There was a wide age range (31-72 years) reflective of both its possible infective and degenerative aetiology. AAA was reported to be associated with hypertension, smoking, advanced age, coronary artery disease, and HIV infection. There was no association between AAA and diabetes. Over 50% of cases were identified incidentally. About one-third (23-54%) of the participants presented aortic rupture with a mortality rate ranging between 65 and 72%. Conclusions: AAA prevalence in Africa is probably higher than the current thinking as there is no baseline data to compare with. Aetiologically, AAA was shown to be associated with hypertension, smoking, coronary artery disease, and possibly infectious pathologies like HIV. Large epidemiological studies would help better characterise AAA in this setting. Lastly, efforts targeting the reduction of the risk factors for AAA would go a long way in reducing the burden of AAA.

Introduction: L'incidence des Anévrismes Aortiques Abdominaux (AAA) dans les pays à hauts revenus est en déclin depuis les trois dernières décennies. En revanche, dans la plupart des pays à faibles et moyens revenus, particulièrement en Afrique, le fardeau représenté est peu connu. L'absence de service de dépistage des AAA dans les pays africains en rend la détection difficile ainsi que la gestion immédiate avant rupture, ce qui a des répercussions importantes sur la mortalité. Cette étude cherche à évaluer systématiquement la prévalence des AAA parmi les patients qui visitent les hôpitaux en Afrique et à évaluer son profil épidémiologique. Procédés: Une revue systématique a été réalisée sur les bases de données EMBASE, GLOBAL HEALTH, MEDLINE et PUBMED. La revue a été menée conformément aux normes et au protocole des Éléments de Rapport Préférés pour les Examens Systématiques et les Méta-Analyses (Preferred Reporting Items for Systematic Reviews and Meta-analyses) enregistrés auprès de PROSPERO (CRD42020162214). Un outil d'extraction de données a été utilisé afin d'obtenir des informations pertinentes de ces études. L'évaluation de la qualité et le risque de partialité a été effectuée au moyen de l'Échelle de Newcastle Ottawa pour les études transversales. Les résultats ont été récapitulés dans des tableaux, des graphiques et un "graphique en forêt" (forest plot). Une approche de synthèse narrative des articles a été adoptée. Résultats: Deux cent soixante et une (261) études ont été identifiées et après exclusion de 246, les 15 dernières ont été jugées appropriées pour l'analyse. Un total de 4012 participants ont été dépistés pour des AAA et, parmi ceux-ci, 129 cas ont été identifiés. La prévalence des AAA de ces études s'étendait de 0.7% à 6.4%. Les hommes représentent 115 (89.1%) des cas. Il a été noté une grande amplitude d'âges (31­72 ans) représentative de sa possible étiologie infectieuse comme dégénérative. Les AAA ont été rapportés en association avec l'hypertension, le tabagisme, l'âge avancé, la maladie coronarienne et la séropositivité. Il n'y a pas eu d'association entre les AAA et le diabète. Plus de 50% des cas ont été incidemment identifiés. Environ un tiers (23­54%) a été présenté en tant que rupture aortique avec un taux de mortalité compris entre 65% et 72%. Conclusions: La prévalence des AAA en Afrique est probablement supérieure à l'état actuel de la réflexion étant donné qu'il n'y a pas de données de référence auxquelles la comparer. Étiologiquement, les AAA ont démontré être associés à l'hypertension, au tabagisme, à la maladie coronarienne et éventuellement aux pathologies infectieuses comme le VIH. Des études épidémiologiques de grande envergure permettraient de mieux déterminer les AAA dans un tel cadre. Enfin, les efforts visant à réduire les facteurs de risque pour les AAA contribueraient grandement à réduire le fardeau des AAA.

Profile of Culture Positive Enteric Fever from Bangalore.

BACKGROUND: Enteric fever is endemic in India. The aim of this study was to analyse the clinical, laboratory, antibiotic sensitivity profile and response to antibiotics of culture positive enteric fever patients from Bangalore. METHODS: In this retrospective study only culture positive enteric fever patients were taken and their clinical, laboratory, antibiotic sensitivity profile and the clinical response to antibiotics studied. RESULT: Eighty one culture positive enteric fever patients were taken into the study. Presenting symptoms included fever, pain abdomen (18.5%), loose stools (25%), vomiting (33%) and headache (30%). Absolute bradycardia at admission was not found in any of our patients. Normal or low total leucocyte count was seen in 97.5%. Typhoid hepatitis was seen in 8.5%. Salmonella enterica subspecies enterica serovar typhi (S typhi) were isolated in 80% of cases; 83% of all cases showed nalidixic acid resistance. All isolates were sensitive to chloramphenicol and third generation cephalosporins. Ciprofloxacin resistance was found in 19% cases. The time to defervescence in patients treated with ceftriaxone was 4.3 days. There was no statistical difference in time to defervescence in nalidixic acid resistant and sensitive strains. Complications included gastro intestinal bleed and encephalopathy. CONCLUSION: Prevalence of nalidixic acid resistance is high, while clinical resistance to quinolones may be higher than that found in the laboratory which requires detailed study. Chloramphenicol sensitivity has returned and nalidixic acid resistant and sensitive isolates are uniformly sensitive to third generation cephalosporins with no difference in time to defervescence.

Applied Machine Learning for the Prediction of Growth of Abdominal Aortic Aneurysm in Humans.

OBJECTIVE: Accurate prediction of abdominal aortic aneurysm (AAA) growth in an individual can allow personalised stratification of surveillance intervals and better inform the timing for surgery. The authors recently described the novel significant association between flow mediated dilatation (FMD) and future AAA growth. The feasibility of predicting future AAA growth was explored in individual patients using a set of benchmark machine learning techniques. METHODS: The Oxford Abdominal Aortic Aneurysm Study (OxAAA) prospectively recruited AAA patients undergoing the routine NHS management pathway. In addition to the AAA diameter, FMD was systemically measured in these patients. A benchmark machine learning technique (non-linear Kernel support vector regression) was applied to predict future AAA growth in individual patients, using their baseline FMD and AAA diameter as input variables. RESULTS: Prospective growth data were recorded at 12 months (360 ± 49 days) in 94 patients. Of these, growth data were further recorded at 24 months (718 ± 81 days) in 79 patients. The average growth in AAA diameter was 3.4% at 12 months, and 2.8% per year at 24 months. The algorithm predicted the individual's AAA diameter to within 2 mm error in 85% and 71% of patients at 12 and 24 months. CONCLUSIONS: The data highlight the utility of FMD as a biomarker for AAA and the value of machine learning techniques for AAA research in the new era of precision medicine.

An Antisense Pectin Methylesterase Gene Alters Pectin Chemistry and Soluble Solids in Tomato Fruit.

Pectin methylesterase (PME, EC 3.1.11) demethoxylates pectins and is believed to be involved in degradation of pectic cell wall components by polygalacturonase in ripening tomato fruit. We have introduced antisense and sense chimeric PME genes into tomato to elucidate the role of PME in fruit development and ripening. Fruits from transgenic plants expressing high levels of antisense PME RNA showed <10% of wild-type PME enzyme activity and undetectable levels of PME protein and mRNA. Lower PME enzyme activity in fruits from transgenic plants was associated with an increased molecular weight and methylesterification of pectins and decreased levels of total and chelator soluble polyuronides in cell walls. The fruits of transgenic plants also contained higher levels of soluble solids than wild-type fruits. This trait was maintained in subsequent generations and segregated in normal Mendelian fashion with the antisense PME gene. These results indicate that reduction in PME enzyme activity in ripening tomato fruits had a marked influence on fruit pectin metabolism and increased the soluble solids content of fruits, but did not interfere with the ripening process.

Effect of nitrogen starvation on the level of adenosine 3',5'-monophosphate in Anabaena variabilis.

Low levels of adenosine 3',5'-monophosphate (cyclic AMP) were detected in the cyanobacterium Anabaena variabilis using a protein binding assay and two radioisotopic labelling methods. The basal concentration of intracellular cyclic AMP ranged from 0.27 pmol/mg protein in A. variabilis Kutz grown under heterotrophic conditions to 1.0--2.7 pmol/mg protein in A. variabilis strain 377 grown autotrophically. Extracellular cyclic AMP was found to comprise as much as 90% of the total cyclic AMP in rapidly growing cultures. When A. variabilis strain 377 was starved of nitrogen, a 3--4-fold increase in intracellular cyclic AMP was observed during the 24 h period coincident with early heterocyst development.

Reduction in Pectin Methylesterase Activity Modifies Tissue Integrity and Cation Levels in Ripening Tomato (Lycopersicon esculentum Mill.) Fruits.

Pectin methylesterase (PME, EC 3.1.1.11) is an ubiquitous enzyme in the plant kingdom; however, its role in plant growth and development is not yet understood. Using transgenic tomato (Lycopersicon esculentum Mill.) fruits that show more than 10-fold reduction in PME activity because of expression of an antisense PME gene, we have investigated the role of PME in tomato fruit ripening. Our results show that reduced PME activity causes an almost complete loss of tissue integrity during fruit senescence but shows little effect on fruit firmness during ripening. Low PME activity in the transgenic fruit pericarp modified both accumulation and partitioning of cations between soluble and bound forms and selectively impaired accumulation of Mg2+ over other major cations. Decreased PME activity was associated with a 30 to 70% decrease in bound Ca2+ and Mg2+ in transgenic pericarp. Levels of soluble Ca2+ increase 10 to 60%, whereas levels of soluble Mg2+ and Na+ are reduced by 20 to 60% in transgenic pericarp. Changes in cation levels associated with lowered PME activity do not affect the rate of respiration or membrane integrity of fruit during ripening. Overall, these results suggest that PME plays a role in determining tissue integrity during fruit senescence, perhaps by regulating cation binding to the cell wall.

Pectin Methylesterase Isoforms in Tomato (Lycopersicon esculentum) Tissues (Effects of Expression of a Pectin Methylesterase Antisense Gene).

We have identified two major groups of pectin methylesterase (PME, EC 3.1.1.11) isoforms in various tissues of tomatoes (Lycopersicon esculentum). These two groups exhibited differential immuno-cross-reactivity with polyclonal antibodies raised against tomato fruit PME or flax callus PME and differences in their accumulation patterns in tissues of wild-type and transgenic tomato plants expressing a PME antisense gene. The group I isoforms with isoelectric points (pls) of 8.2, 8.4, and 8.5 are specific to fruit tissue, where they are the major forms of PME activity. The group II PME isoforms, with pl values of 9 and above, are observed in both vegetative and fruit tissues. The group I isoforms cross-react with polyclonal antibodies raised to a PME isoform purified from fruit, whereas the group II isoforms cross-react with antibodies to a PME purified from flax callus. Expression of a fruit-specific PME anti-sense gene impairs accumulation of the group I PME isoforms, with no apparent effect on the accumulation of the group II PME isoforms. The absence of any noticeable effects on growth and development of transgenic plants suggests that the group I PME isoforms are not involved in plant growth and development and may play a role under special circumstances such as cell separation during fruit ripening.

Impaired Wound Induction of 3-Deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) Synthase and Altered Stem Development in Transgenic Potato Plants Expressing a DAHP Synthase Antisense Construct.

Potato (Solanum tuberosum L.) cells were transformed with an antisense DNA construct encoding part of 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase (EC 4.1.2.15), the first enzyme of the shikimate pathway, to examine the role(s) of this protein in plant growth and development. Chimeric DNA constructs contained the transcript start site, the first exon, and part of the first intron of the shkA gene in antisense or sense orientations under the control of the cauliflower mosaic virus 35S promoter. Some, but not all, of the transgenic plants expressing antisense DAHP synthase RNA showed reduced levels of wound-induced DAHP synthase enzyme activity, polypeptide, and mRNA 12 and 24 h after wounding. No alteration in the wound induction of DAHP synthase gene expression was observed in transgenic potato tubers containing the chimeric sense construct. Reduced steady-state levels of DAHP synthase mRNA were observed in stem and shoot tip tissue. Some plants with the chimeric antisense construct had reduced stem length, stem diameter, and reduced stem lignification.