Prostaglandin-H-synthase (PGHS)-1 and -2 microtiter assays for the testing of herbal drugs and in vitro inhibition of PGHS-isoenzyms by polyunsaturated fatty acids from Platycodi radix

In order to test inhibition of prostaglandin-H-synthase-1 and -2 (PGHS-1 and -2) by plant extracts, we have established two enzyme based in vitro assays with enzyme immunoassay (EIA) evaluation. The assays have been evaluated with known synthetic inhibitors and with plant extracts. In a screening of traditionally used Chinese herbs for anti-inflammatory activity, a series of n-hexane and dichloromethane extracts showed significant inhibitory effect in comparison with the known specific PGHS-2 inhibitors NS-398 (IC(50) = 2.6 microM) and nimesulide (IC(50) = 36 microM). The lipophilic extracts of the Chinese drug Jiengeng, the dried roots of Platycodon grandiflorum (Jacq.) A. DC. (Campanulaceae), showed good inhibitory activity against both PGHS isoenzymes. The directly prepared DCM-extract exhibited better activity against PGHS-2 (IC(50) = 4.0 microg/ml) than against PGHS-1 (IC(50) = 17.6 microg/ml). We identified fatty acids as main active constituents and quantified them. Linoleic acid showed the highest content (ca. 20% of the dried extract) and a high and preferential PGHS-2 inhibitory activity (IC(50) (PGHS-1) = 20 microM; IC(50) (PGHS-2) = 2 microM). The comparison of the concentration of linoleic acid and the inhibitory activity of the direct DCM-extract showed, that linoleic acid is mainly responsible for the in vitro activity of the extract on PGHS-2.     

Assay and biochemical characterization of angiotensin-I-converting enzyme in cerebrospinal fluid

A spectrofluorimetric method was adapted for determination of angiotensin-I-converting enzyme (ACE) in untreated native cerebrospinal fluid (CSF). Benzyloxycarbonyl-phenylalanyl-histidyl-leucine was applied as enzyme substrate. The biochemical behavior of ACE of CSF (CACE) was studied. The pH optimum was found to be 8.0 using borax phosphate buffer. The determination of Km was 10.7 +/- 3.3 (SD) mumol/l. ACE could be blocked by 8-hydroxyquinoline (100%) and phenanthroline (80%), but only slight inhibition was observed by teprotide (43%), EDTA (40%) and captopril (31%). The influence of various drugs on CACE was also tested. The different biochemical behavior of CACE compared to serum ACE suggested that an isoenzyme exists in CSF. CACE has been found to be elevated in neuroimmunological and inflammatory disorders of the nervous system compared to levels in healthy controls.     

Modification of the coronary artery disease risk associated with the presence of traditional risk factors by insertion/deletion polymorphism of the ACE gene

Cigarette smoking, hypercholesterolemia, and obesity influence the renin-angiotensin system (RAS) functions including an increased synthesis of the angiotensin I converting enzyme (ACE). Thus in the present work we explore the interactions of the ACE gene insertion/deletion (I/D) polymorphism and traditional risk factors. The study cohort included 341 subjects composed of 172 patients with angiographically confirmed CAD and 169 blood donors without a history of cardiovascular diseases. The I/D polymorphism was genotyped using polymerase chain reaction (PCR) methodology. To determine the interactions between the ACE genotypes and traditional risk factors the epidemiologic approach was used (4 x 2 tables and the synergy measures). The frequency of the DD genotype was significantly higher in patients than in controls (33.7% versus 21.3%, odds ratio [OR] = 1.88, 95% CI; 1.13-3.15, p = 0.010), but greater differences were found in males (35.7% versus 20.5%, OR = 2.15, 95% CI: 1.14-4.04, p = 0.010). We found a synergy of the DD genotype with smoking (SI = 1.88, SIM = 1.22), total cholesterol > or =5 mmol/l (SI = 2.12, SIM = 1.31) and elevated low density lipoprotein (LDL) cholesterol level (> or =3 mmol/l) (SI = 1.78, SIM = 1.14). The presence of the D allele (DD + ID subjects) also increased the risk of coronary artery disease (CAD) associated with the presence of elevated total cholesterol and LDL cholesterol (SI = 1.69, SIM = 1.18, in both cases), elevated level (> or =1.7 mmol/l) of triacylglycerols (SI = 1.81, SIM = 1.18) and overweight/obesity (SI = 4.25, SIM = 2.36). In each case the estimated CAD risk was greater than that predicted by assuming the additivity of effects (the risk increased from 69% for the D allele - total cholesterol interaction to 325% for the D allele - overweight/obesity). The statistical significance was also confirmed by a multiplicative model of synergy. The DD genotype/D allele of the ACE gene increases the risk of CAD associated with the presence of traditional risk factors.     

Angiotensin-converting enzyme and anorexia nervosa

Angiotensin-converting enzyme (ACE) activity was measured in 10 patients with anorexia nervosa, 6 with hyperthyroid Graves' disease, and 7 with primary hypothyroidism. Patients with anorexia nervosa had a low serum ACE activity (9.8 +/- 2.2 IU/l), as compared to findings in normal subjects (13.4 +/- 3.5 IU/l) (P less than 0.05). Patients with hyperthyroid Graves' disease had high serum ACE activity (23.7 +/- 5.8 IU/l), as compared to levels in normal subjects (P less than 0.01), and patients with primary hypothyroidism tended to have low serum ACE activity (10.1 +/- 1.8 IU/l), compared to the normal subjects (P less than 0.1). Following weight gain (before; 71.3 +/- 10.2% of ideal body weight, after; 88.7 +/- 5.6% of ideal body weight), serum ACE activity in patients with anorexia nervosa reverted to within the normal range (13.8 +/- 3.5 IU/l), and serum T3 concentration was restored to the normal range (before; 0.7 +/- 0.2 ng/ml, after; 1.1 +/- 0.3 ng/ml). In these patients, ACE activity correlated with the per cent of ideal body weight (P less than 0.05). These data suggest that, in underweight subjects with anorexia nervosa, decreased serum ACE activities may relate to emaciation.     

Familial resemblance of plasma angiotensin-converting enzyme level: the Nancy Study.

Plasma angiotensin I-converting enzyme (ACE) activity has been measured in a sample of 87 healthy families participating in a study of cardiovascular risk factors. The mean +/- SD levels of plasma ACE were 34.1 +/- 10.7, 30.7 +/- 10.4 and 43.1 +/- 17.2 units/liter in fathers (n = 87), mothers (n = 87) and offspring (n = 169), respectively. Plasma ACE was uncorrelated with age, height, weight, or blood pressure in the parents, but a negative correlation with age was observed in offspring (r = -.32). The age-adjusted familial correlations of plasma ACE were .038, .166, .323 and .303 for spouses, father-offspring, mother-offspring, and siblings, respectively. The results of the genetic analysis suggest that a major gene may affect the interindividual variability of plasma ACE, with different codominant effects in parents and offspring. According to this model, the major gene effect accounts for 4.8, 4.0, and 10.8 units/liter of the overall mean and for 29%, 29% and 75% of the variance of age-adjusted ACE in fathers, mothers, and offspring, respectively. The estimate of the probability of the less frequent allele is .26, and the major gene effect is approximately twice as great in high homozygotes than in heterozygotes and in offspring than in parents. The results of this study demonstrate the occurrence of a familial resemblance of plasma ACE activity in healthy families and suggest that this observation can be explained by the segregation of a major gene.     

Simple and rapid determination of histamine in food using a new histamine dehydrogenase from Rhizobium sp

A colorimetric enzyme assay for the quantitative analysis of histamine in food has been developed using a new histamine dehydrogenase (HDH) from Rhizobium sp. The HDH specifically catalyzes the oxidation of histamine but not other biogenic amines such as putrescine and cadaverine. The principle of our photometric assay is as follows. The HDH catalyzes the oxidative deamination of histamine in the presence of 1-methoxy PMS (electron carrier), which converts WST-8 (tetrazolium salt) to a formazan. This product is measured in the visible range at 460 nm. The correlation between the histamine level and absorbance was acceptable, ranging from 0 to 96 microM with histamine standard solutions, corresponding to 0 to 30 microM of the reaction solution (r = 1.000, CV = 1.0% or less). Assays of canned tuna (in oil and soup) and raw tuna with 45-675 micromol/kg histamine added showed good recoveries of 96-113, 98-108, and 100-106%. The histamine contents of a commercial canned tuna and fish meal containing histamine at high concentrations were determined using the new method and other reference methods (HPLC method, Association of Official Analytical Chemists official method, and two commercial enzyme immunoassay test kits). This simple and rapid enzymatic method is as reliable as the conventional methods.     

Total phenolics concentration and antioxidant potential of extracts of medicinal plants of Pakistan

Thirty-seven plant organs, traditionally used as drugs, collected in Pakistan, were extracted with 70% acetone and analyzed for their total phenolics concentration and antioxidant potential. Seven extracts showed more than 85% inhibition of lipid peroxidation in vitro as compared with blank. Butylated hydroxytoluene (BHT) (IC50 = 233.6 microg/l +/- 28.3) was the strongest antioxidant in our test system. The IC50 results indicate that the extracts of Nymphaea lotus L. flowers, Acacia nilotica (Linn.) Delile beans, Terminalia belerica Roxb. fruits, and Terminalia chebula Retz. (fruits, brown) were stronger antioxidants than alpha-tocopherol, while Terminalia chebula Retz. (fruit coat), Terminalia chebula Retz. (fruits, black) and Ricinus communis L. leaves were weaker antioxidant extracts than alpha-tocopherol and BHT. Total phenolics concentration, expressed as gallic acid equivalents, showed close correlation with the antioxidant activity. High performance liquid chromatographic analysis with diode array detection at 280 nm, of the seven extracts indicated the presence of hydroxybenzoic acid derivatives, hydroxycinnamic acid derivatives, flavonol aglycones and their glycosides as main phenolics compounds. This information, based on quick screening methods, enables us to proceed towards more detailed chemical and pharmacological understanding of these plant materials.     

A gold nanoparticle-mediated enzyme bioreactor for inhibitor screening by capillary electrophoresis

A facile protocol to prepare highly effective and durable in-line enzyme bioreactors inside capillary electrophoresis (CE) columns was developed. To demonstrate the methodology, l-glutamic dehydrogenase (GLDH) was selected as the model enzyme. GLDH was first immobilized onto 38-nm-diameter gold nanoparticles (GNPs), and the functionalized GNPs were then assembled on the inner wall at the inlet end of the CE capillary treated with polyethyleneimine (PEI), producing an in-line GLDH bioreactor. Compared with a GLDH bioreactor prepared by immobilizing GLDH directly on PEI-treated capillary, the GNP-mediated bioreactor showed a higher enzymatic activity and a much better stability. The in-capillary enzyme bioreactor was proven to be very useful for screening of GLDH inhibitors deploying the GLDH-catalyzed α-ketoglutaric acid reaction. The screening assay was preliminarily validated by using a known GLDH inhibitor, namely perphenazine. A Z′ factor value of 0.95 (n = 10) was obtained, indicating that the screening results were highly reliable. Screening of GLDH inhibitors present in medicinal plant extracts by the proposed method was demonstrated. The inhibition percentages were found to be 53% for Radix scutellariae, 45% for Radix codonopsis, 37% for Radix paeoniae alba, and 0% for the other 22 extracts tested at a concentration of 0.6 mg extract/ml.     

Cross-sectional study of ocular optical components interactions in emmetropes

Purpose of the paper was to evaluate ocular optical components (OOC) interactions in a large number of emmetropes. A cross-sectional study of 1,000 emmetropes, aged from 18-40 years, has been conducted. Complete ophthalmological examination, corneal radius (CR) measurement, keratometry and echobiometry of both eyes were performed. The highest correlation of OOC was that of axial length (Ax) with vitreal body (CV) on both eyes (r = 0.79 for the right eye (RE); r = 0.81 for the left eye (LE)). The axial length had a positive correlation with the anterior chamber depth (ACD) on both eyes as well, but the coefficient was very low (r = 0.29 for the RE; r = 0.32 for the LE). The only negative correlation Ax had on both eyes was with the lens (L) (r = -0.17 for the RE; r = -0.19 for the LE). Keratometry of the horizontal (K1) and vertical meridian (K2) showed a negative correlation with CV and Ax on both eyes (for K1 r = -0.64 for CV r = -0.54 for Ax; for K2 r = -0.67 for CV r = -0.68 for Ax). CR had a positive correlation with Ax (r = 0.74) and CV (r = 0.79). It showed a negative correlation with L (r = -0.58). CV had a high, positive correlation with Ax (r = 0.72 for the RE; r = 0.75 for the LE). The correlation with K1 and K2 was negative. Our study showed that the axial length, keratometry, corneal radius, lens thickness and vitreal body were the most important OOC that correlated with each other following a pattern in our group of emmetropes. They interacted in such a way that in the subjects with axial length above the average value, the vitreal body was longer but the lens was thinner and the cornea was of less power. This could explain at least one of the mechanisms of emmetropization.     

Angiotensin-converting enzyme ID polymorphism and fitness phenotype in the HERITAGE Family Study.

It has been suggested that genetic variation in the angiotensin-converting enzyme (ACE) gene is associated with physical performance. We studied the association between the ACE insertion (I)/deletion (D) polymorphism and several fitness phenotypes measured before and after 20 wk of a standardized endurance training program in sedentary Caucasian (n = 476) and black (n = 248) subjects. Phenotypes measured were oxygen uptake (VO(2)), work rate, heart rate, minute ventilation, tidal volume, and blood lactate levels during maximal and submaximal [50 W and at 60 and 80% of maximal VO(2) (VO(2 max))] exercise and stroke volume and cardiac output during submaximal exercise (50 W and at 60% VO(2 max)). The ACE ID polymorphism was typed with the three-primer PCR method. Out of 216 association tests performed on 54 phenotypes in 4 groups of participants, only 11 showed significant (P values from 0.042 to 0. 0001) associations with the ACE ID polymorphism. In contrast to previous claims, in Caucasian offspring, the DD homozygotes showed a 14-38% greater increase with training in VO(2 max), VO(2) at 80% of VO(2 max), and all work rate phenotypes and a 36% greater decrease in heart rate at 50 W than did the II homozygotes. No associations were evident in Caucasian parents or black parents or offspring. Thus these data do not support the hypothesis that the ACE ID polymorphism plays a major role in cardiorespiratory endurance.     

The determination of potassium concentration in vitreous humor by low pressure ion chromatography and its application in the estimation of postmortem interval

An analytical method was developed for the determination of potassium in vitreous humor by low pressure ion chromatography (LPIC). Experimental conditions for LPIC analysis were optimized. High sensitivity and selectivity were obtained using this method. The LOD and LOQ were 1 and 2 mmol l(-1), respectively. The linearity was demonstrated from 2 to 20 mmol l(-1). The intra- and inter-day precision (CV) based on three concentrations was less than 5.0%. It was a simple and fast method to measure potassium and was suitable for evaluating the postmortem interval (PMI) in relatively well-preserved bodies. Sixty-two samples from medical-legal autopsies with known PMI were analyzed. A linear correlation equation for potassium concentration in the vitreous humor and PMI was established: [K(+)]=0.1702PMI+5.5678, r=0.8692.     

耐低磷水稻基因型筛选指标的研究

采用溶液培养试验,并结合大田试验,研究和探讨了耐低磷水稻基因型的筛选指标.结果表明,溶液培养试验中,在正常供磷和低磷胁迫条件下,在所有调查性状中水稻单株干重都具有较大的基因型间变异(CV分别为21.73%和19.54%).在所有调查性状的相对值中,相对单株干重(低磷胁迫/正常供磷)也具有较大的基因型间变异(CV为19.60%);相关分析表明,相对单株干重与相对根干重、相对株高、相对单株吸磷量、相对地上部磷积累、相对磷利用效率和相对植株磷浓度均呈极显著正相关(P＜0.01).因此,水稻相对单株干重可以作为苗期筛选水稻耐低磷基因型的一个筛选指标.溶液培养试验中水稻的相对单株干重和大田试验中水稻的相对稻谷产量(不施磷/施磷)没有显著相关性,因此溶液培养试验的相对单株干重不能作为评价大田试验中水稻耐低磷能力的指标.低磷营养液培养的水稻体内磷利用效率与缺磷土壤生长的水稻体内磷利用效率呈极显著正相关.因此,直接以低磷营养液培养水稻苗期体内磷利用效率作为筛选指标,然后进行大田试验验证,是一条筛选水稻磷高效利用基因型的有效途径.     

Primary role of angiotensin-converting enzyme-2 in cardiac production of angiotensin-(1-7) in transgenic Ren-2 hypertensive rats

Angiotensin-converting enzyme-2 (ACE2) converts angiotensin II (ANG II) to angiotensin-(1-7) [ANG-(1-7)], and this enzyme may serve as a key regulatory juncture in various tissues. Although the heart expresses ACE2, the extent that the enzyme participates in the cardiac processing of ANG II and ANG-(1-7) is equivocal. Therefore, we utilized the Langendorff preparation to characterize the ACE2 pathway in isolated hearts from male normotensive Sprague-Dawley [Tg((-))] and hypertensive [mRen2]27 [Tg((+))] rats. During a 60-min recirculation period with 10 nM ANG II, the presence of ANG-(1-7) was assessed in the cardiac effluent. ANG-(1-7) generation from ANG II was similar in both the normal and hypertensive hearts [Tg((-)): 510 +/- 55 pM, n=20 vs. Tg((+)): 497 +/- 63 pM, n=14] with peak levels occurring at 30 min after administration of the peptide. ACE2 inhibition (MLN-4760, 1 microM) significantly reduced ANG-(1-7) production by 83% (57 +/- 19 pM, P<0.01, n=7) in the Tg((+)) rats, whereas the inhibitor had no significant effect in the Tg((-)) rats (285 +/- 53 pM, P>0.05, n=10). ACE2 activity was found in the effluent of perfused Tg((-)) and Tg((+)) hearts, and it was highly associated with ACE2 protein expression (r=0.78). This study is the first demonstration for a direct role of ACE2 in the metabolism of cardiac ANG II in the hypertrophic heart of hypertensive rats. We conclude that predominant expression of cardiac ACE2 activity in the Tg((+)) may be a compensatory response to the extensive cardiac remodeling in this strain.     

Angiotensin-converting enzyme activity in ovine bronchial vasculature.

Angiotensin-converting enzyme (ACE) plays a major role in the metabolism of bradykinin, angiotensin, and neuropeptides, which are all implicated in inflammatory airway diseases. The activity of ACE, which is localized on the luminal surface of endothelial cells (EC), has been well documented in pulmonary EC; however, few data exist regarding the relative activity of ACE in the airway vasculature. Therefore, we measured ACE activity in cultured EC from the sheep bronchial artery and bronchial mucosa (microvascular) and compared it with pulmonary artery EC. The baseline level of total ACE activity (cellular plus secreted) was significantly greater in bronchial microvascular EC (1.24 +/- 0.24 mU/106 cells) compared with bronchial artery EC (0.59 +/- 0.15 mU/106 cells; P < 0.05) and comparable to pulmonary artery EC (1.12 +/- 0.14 mU/106 cells; P > 0.05). Measured ACE activity secreted into culture medium for each cell type was 64-74% of total activity and did not differ among the three EC types (P = 0.17). Hydrocortisone (10 microg/ml; 48-72 h) treatment resulted in a significant increase in ACE activity in bronchial EC. Likewise, TNF-alpha (0.1 ng/ml) treatment markedly increased ACE activity in all cell lysates (P < 0.05). We confirmed the importance of ACE activity in vivo since, at the highest dose of bradykinin studied (10-8 M), bronchial artery pressure at constant flow showed a greater decrease after captopril treatment (36% before vs. 60% after; P = 0.05). These results demonstrate high ACE expression of the bronchial microvasculature and suggest an important regulatory role for ACE in the metabolism of kinin peptides known to contribute to airway pathology.     

Changes in plasma angiotensin-converting enzyme activity and noradrenaline responses to long-term nitric oxide inhibition vary depending on their basal values in chickens

In this study, we investigated the effects of N(omega)-nitro-L-arginine (L-NNA) on arterial blood pressure (BP), plasma noradrenaline (NA) and adrenaline (A) levels and angiotensin-converting enzyme (ACE) activity. L-NNA was applied with tap water (1 mg/ml) from the 3rd to the 8th week of age (group L-NNA1). In Experiment 1, long-term L-NNA application increased BP compared to the control group (group C1) (L-NNA1 = 131.4 +/- 6.3, n = 6; C1 = 82.7 +/- 4.7 mm Hg, n = 7) but decreased plasma noradrenaline and adrenaline levels and ACE activity (NA levels: C1 = 15.5 +/- 0.8, n = 7; L-NNA1 = 8.6 +/- 0.5 ng/ml, n = 7; A levels: C1 = 15.5 +/- 0.8, n = 7; L-NNA1 = 6.0 +/- 0.5 ng/ml, n = 7; ACE activities: C1 = 87.3 +/- 3.1, n = 6; L-NNA1 = 46.2 +/- 1.9 U/l, n = 5). On the other hand, in Experiment 2 (carried out under the same conditions and in age-matched chickens), blood pressure, plasma noradrenaline levels and ACE activity were found to differ in the control group (C2) (BP = 141.4 +/- 15.5 mm Hg, n = 7; NA = 1.1 +/- 0.4 ng/ml, n = 7; ACE = 57.2 +/- 5.3 U/l, n = 7) as compared to C1, while plasma adrenaline levels were similar. In this series, long-term L-NNA application (group L-NNA2) did not change the BP, but surprisingly increased noradrenaline and ACE values (values of L-NNA2: BP = 165.7 +/- 15.6 mm Hg, n = 7; NA = 9.3 +/- 1.3 ng/ml, n = 8; ACE = 149.4 +/- 16 U/l, n = 8) while decreasing plasma adrenaline levels. L-arginine addition to L-NNA treatment completely reversed plasma noradrenaline and ACE activity values. These results indicate the modulatory activity of an L-arginine-NO pathway on adrenaline release as well as on the renin-angiotensin system in chickens.     

Modification of the furanacryloyl-L-phenylalanylglycylglycine assay for determination of angiotensin-I-converting enzyme inhibitory activity

Angiotensin-I-converting enzyme (ACE, EC 3.4.15.1) plays a central role in the regulation of blood pressure in man. The objective of this study was to evaluate and modify the furanacryloyl-L-phenylalanylglycylglycine (FAPGG) assay method for quantification of ACE activity. The fixed time conditions developed for assay of ACE activity were as follows: 0.8 mM FAPGG, 175 + or - 10 units l(-1) ACE, incubation at 37 degrees C for 30 min and enzyme inactivation with 100 mM ethylenediaminetetra-acetic acid (EDTA). Hydrolysis of FAPGG to FAP and GG was quantified by measuring the decrease in absorbance at 340 nm. It was shown that increasing the level ACE activity in the assay from 155 to 221 + or - 15 units l(-1) resulted in a corresponding increase in the apparent IC(50) value for Captopril from 9.10 to 39.40 nM. Similar trends in the apparent IC50 values for a whey protein hydrolysate were obtained. The results demonstrate the requirement for carefully controlling ACE activity levels in the assay in order to obtained comparable and reproducible values for the inhibitory potency of ACE inhibitors.     

胰蛋白酶水解β-乳球蛋白获得ACE抑制肽的条件优化

采用三因素二次通用旋转设计和体外检测法,对胰蛋白酶水解β-乳球蛋白获得ACE抑制肽的条件进行优化。结果表明,底物浓度(X1)、温度(X2)、酶与底物的质量比(X3)对ACE抑制率的影响回归方程为:Y=50.62-2.33X1-1.97X2+5.81 X3-3.36X2X3-6.56X22-1.96X32,胰蛋白酶水解β-乳球蛋白获得ACE抑制肽的最优水解条件为:底物质量浓度为60 g/L,水解温度30℃,酶与底物的质量比为5.5%,水解时间6 h,水解产物对ACE抑制活性最大抑制率为53.86%。     

Phenotypic variation of agronomic traits among coyote gourd accessions and their progeny

As a prelude to domestication efforts, variation of agronomic traits was determined among accessions of the polytypic, xerophytic cucurbit, coyote gourd [Cucurbita digitata subsp. digitata (DIG), palmata (PAL), cylindrata (CYL), and cordata (COR)] and among and within their progeny. Oil content in 60 accession seed lots (x = 27.8%, CV 21.4%) was more variable than protein content (x = 33.1%, CV = 13.5%). Punicic acid (c,t,c-9,ll,13-octadecatrienoic acid) levels in seed oils were also variable (x = 12.0%, CV = 20.6%) among accession seed lots. Substantial differences among and within subspecies were noted in 40 progeny lines for fruit/plant (x = 55.2, CV = 47.5%), seed wt/plant (x = 0.89 kg, CV = 51.1%), seed wt/fruit (x = 17.4 g, CV = 39.8%), seed no./fruit (x = 356, CV= 30.8%), 100-seed wt (x = 4.8 g, CV = 21.6%) and fruit diameter (x = 77 mm, CV 4.8%). Correlations among parameters suggested selection for fruit/plant to be the most effective primary strategy for seed yield improvement, but among high fruit-yielders, selection for seed wt/fruit was also important. The two subspecies PAL and CYL exhibited superior seed wts/plant. CYL types matured high fruit-loads, but their fruits were smaller and contained a smaller number of lighter seed than PAL or PAL hybrids. In contrast, PAL and PAL hybrids displayed lower fruit-yields/ plant but their fruits were larger and contained higher seed wts/fruit than their CYL counterparts. Root wt/plant was also variable (x = 1.55 kg, CV = 63.2%). Roots of DIG were larger and less branched than those of other subspecies. Proximate and liquid Chromatographic analyses of selected accession seed lots and controlled crosses failed to reveal advantages for the inclusion of specific types in a breeding program for the development of high oil or high/low punicic acid lines.     

An enhanced chemiluminescence enzyme immunoassay for serum progesterone

A competitive enhanced luminescent enzyme immunoassay for serum progesterone is described, which is based on a 11 alpha-hydroxyprogesterone 11-hemisuccinyl-horseradish peroxidase conjugate and a black polystyrene microtitre plate sensitised with anti-progesterone IgG. Bound label was determined using a mixture of 4-iodophenol, luminol and peroxide, and the light emitted from the wells of the plate quantitated using a luminescent plate reader. The assay was sensitive (detection limit 0.5 pg), precise (CV 2.7 - 9.0% in the concentration range 4.3-67.7 nM) and showed good correlation (r = 0.99) with a conventional radioimmunoassay.