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Streptopodium caricae sp. nov., with a discussion of powdery mildews on papaya, and emended descriptions of the genus Streptopodium and Oidium caricae.

A new powdery mildew infecting papaya (Carica papaya) in Brazil, Streptopodium caricae sp. nov., is described. The species is compared with other anamorphic Erysiphales known to infect papaya: Oidiopsis sicula, Ovulariopsis papayae, Oidium caricae, O. papayae, O. caricicola, O. indicum, O. caricae-papayae, Podosphaera (syn. Sphaerotheca) spp., and Erysiphe spp. An emended description Streptopodium and a key to the anamorphs of powdery mildews on papaya are also presented. A re-examination of the type material of Phyllactinia caricaefolia showed that conidia in this material are dimorphic, indicating that its anamorph does not belong to Ovulariopsis and that the teleomorph is not conspecific with Phyllactinia guttata. Oidium caricae, the common powdery mildew of papaya, was re-examined, recognized as a member of subgenus Pseudoidium, an emended description was prepared, and a new type was indicated. O. papayae was recognized as a synonym of O. caricae, and many of the records of this fungus are considered to be doubtful or incorrect, either omitting a description of the fungus or including a description or illustration of an euodium conidiophore morphology.

Brazil↗

Cyanogenesis in glucosinolate-producing plants: Carica papaya and Carica quercifolia.

(R)-2-(beta-D-Glucopyranosyloxy)-2-phenylacetonitrile (prunasin) was isolated from Carica papaya L. and C. quercifolia (A. St.-Hil.) Hieron. (syn. C. hastata Brign.). Earlier reported presence of cyclopentanoid cyanohydrin glycosides in C. papaya could not be confirmed, and no cyclopentanoid amino acids could be detected in extracts of C. papaya and C. quercifolia. Conversion of [2,3,4,5,6-3H]phenylalanine into tritiated prunasin was demonstrated in both species. On the other hand, when the plants were administered [2-14C]-2-(2'cyclopentenyl)glycine, extracted, and the extracts hydrolyzed with beta-glucosidase (Helix pomatia), formation of labelled cyanide was not observed. The absence of cyclopentanoids, which are typical for the Passifloraceae, and the inability of Carica species to utilize 2-(2'-cyclopentenyl)glycine as a precursor of cyanogenic glycosides are in agreement with the relative phylogenetic position of the Caricaceae and the Passifloraceae. Carica species are thus rare examples of taxa in which glucosinolates and cyanogenic glycosides co-occur, both types of natural products being derived from the same amino acid, phenylalanine.

Glucosinolates↗

Possible immunomodulatory actions of Carica papaya seed extract.

Carica papaya seed extract is currently being marketed as a nutritional supplement with purported ability "to rejuvenate the body condition and to increase energy". The product claims to improve immunity against common infection and body functioning. The present study was initiated to analyze the chemical constituents of the Carica Seed Extract and determine the potential immunomodulatory properties of the different bioactive fractions. These immunomodulatory activities of crude Carica Seed Extract and its bioactive fractions were examined in vitro using lymphocyte proliferation assays and complement-mediated hemolytic assay. Three major observations were made in this study: (1) the crude Carica Seed Extract and two other bioactive fractions significantly enhanced the phytohemagglutinin responsiveness of lymphocytes; (2) none of the Carica Seed Extract (at the concentrations used in this study) was able to protect the lymphocytes from the toxic effects of chromium; and (3) some of the bioactive fractions of Carica Seed Extract were able to significantly inhibit the classical complement-mediated hemolytic pathway. These findings provide evidence for immunostimulatory and anti-inflammatory actions of Carica Seed Extract. No single compound is likely responsible for these activities. Further purification, isolation and characterization of the active components are needed.

Adjuvants, Immunologic↗

Aerobiological and immunochemical studies on Carica papaya L. pollen: an aeroallergen from India.

BACKGROUND: Carica papaya L. is a fruit yielding tree, wildly grown or cultivated in the tropics and subtropics. Its pollen grain has been reported to be airborne and cause immunoglobulin E (IgE)-mediated hypersensitivity. OBJECTIVE: To conduct long-term aerobiological study on Carica pollen, along with aeroallergenic particles originating from it and to identify vis-a-vis characterize an important IgE-reactive component present in this pollen. METHODS: The seasonal and diurnal periodicities of airborne C. papaya pollen were recorded in a 5-year survey using a Burkard volumetric sampler. The allergenic potential was studied by skin prick tests, IgE-enzyme-linked immunosorbent assay (ELISA) and also by aeroallergen immunoblotting. The total pollen extract was fractionated by Sephacryl S-200 column, and out of the eluted five fractions, the maximum IgE-reactive fraction (as found in ELISA inhibition) was resolved into five major subfractions in reverse-phase high-performance liquid chromatography (RP-HPLC). The subfraction with optimum IgE reactivity was studied by activity gel, native and nonreducing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The homogeneity of the isolated protein fraction was checked by crossed immunoelectrophoresis with rabbit antisera and IgE reactivity was confirmed by ELISA inhibition and immunoblotting using individual patient sera. RESULTS: The Carica pollen occurred in the air round the year with peaks during January and September-October. Among a patient population of 1000, skin-test results showed 27.8% +1 level and 5.6% +2/+3 level reactions. In aeroallergen immunoblotting of exposed Burkard tape segments, the detected allergen spots showed a significant correlation with airborne pollen count recorded. The pollen extract elicited loss of IgE reactivity when treated with reducing agent-like beta-mercaptoethanol and heat, but showed six IgE-reactive components in nonreducing IgE-immunoblot. The fraction 1 eluted from Sephacryl S-200 column showed highest IgE reactivity and resolved into five major components in RP-HPLC. Out of these, the fraction showing optimum IgE reactivity in IgE-ELISA inhibition and immunoblotting with patient antisera, elicited esterase activity and found to be a homogenous protein of 100 kDa. CONCLUSION: Carica papaya tree contributes significantly to the aeropollen and aeroallergen load of the suburban outskirts of Calcutta metropolis, India. The pollen extract contains an important IgE-reactive protein component of 100 kDa molecular weight with esterase activity.

Adult↗

Genetic diversity of Carica papaya as revealed by AFLP markers.

Genetic relationships among Carica papaya cultivars, breeding lines, unimproved germplasm, and related species were established using amplified fragment length polymorphism (AFLP) markers. Seventy-one papaya accessions and related species were analyzed with nine EcoRI-MseI primer combinations. A total of 186 informative AFLP markers was generated and analyzed. Cluster analysis suggested limited genetic variation in papaya, with an average genetic similarity among 63 papaya accessions of 0.880. Genetic diversity among cultivars derived from the same or similar gene pools was smaller, such as Hawaiian Solo hermaphrodite cultivars and Australian dioecious cultivars with genetic similarity at 0.921 and 0.912, respectively. The results indicated that self-pollinated hermaphrodite cultivars were as variable as open-pollinated dioecious cultivars. Genetic diversity between C. papaya and six other Carica species was also evaluated. Carica papaya shared the least genetic similarity with these species, with an average genetic similarity of 0.432; the average genetic similarity among the six other species was 0.729. The results from AFLP markers provided detailed estimates of the genetic variation within and among papaya cultivars, and supported the notion that C. papaya diverged from the rest of Carica species early in the evolution of this genus.

Carica↗

Antibacterial effects of Carica papaya fruit on common wound organisms.

The purpose of the study was to investigate antibacterial activity of ripe and unripe Carica papaya on selected micro-organisms. Cultures of micro-organisms were routinely maintained in nutrient agar slants at 4 degrees C. Extracts of immature, mature and ripe Carica papaya fruit were obtained by separately grinding factions of the epicarp, endocarp and seeds and filtering them through gauze. Sensitivity tests were conducted by adding 0.06 ml of extract to agar wells (6 mm diameter) prepared from 20 ml agar seeded with 10(6) cells/ml suspension of one of the eight organisms per plate. The inoculated plates were allowed to equilibrate at 4 degrees C for 1 hour, incubated at 37 degrees C for 24 hours, and zones of inhibition measured in millimetres. Anti-bacterial activity was expressed in terms of the radius of zone of inhibition. Seed extracts from the fruit showed inhibition in the following order: B cereus > E coli > S faecalis > S aureus > P vulgaris > S flexneri. No significant difference was found in bacterial sensitivity between immature, mature and ripe fruits. No inhibition zone was produced by epicarp and endocarp extracts. Carica papaya seeds contain anti-bacterial activity that inhibits growth of gram-positive and gram-negative organisms. Observed activity was independent of stage of fruit maturity. Carica papaya has antibacterial effects that could be useful in treating chronic skin ulcers to promote healing.

Anti-Bacterial Agents↗

Hypotriglyceridaemic activity of Ficus carica leaves in experimental hypertriglyceridaemic rats.

A model of hypertriglyceridaemia in rats is described, which was used to investigate the hypolipidaemic effect of an intraperitoneal (i./p.) administration of a Ficus carica leaf decoction. Hypertriglyceridaemia was induced in rats following the protocol: a fasting period of 22 h, 2 h of oral (p.o.) administration of 20% emulsion of longchain triglycerides (LCT emulsion), both repeated once. The plasma triglyceride and total cholesterol levels obtained 2 h after the protocol were 5.7 +/- 2.5 mmol/L (p < 0.0001 vs basal levels) and 1.7 +/- 0.3 mmol/L, respectively, n = 10. The new model was used to test the hypotriglyceridaemic effect of a single dose of Ficus carica (fig tree) leaf decoction administered i./p. (50 g dry wt/kg body wt). After the i.p. injection of serum saline (control group, n = 10) or Ficus carica extract (group A, n = 10), plasma triglyceride levels in the control group and group A were 5.9 +/- 2.9 mmol/L and 5.5 +/- 2.9 mmol/L just after the LCT emulsion protocol; 4.7 +/- 2.7 mmol/L and 0.9 +/- 0.4 mmol/L, p < 0.005, 60 min after the LCT protocol; and 3.6 +/- 2.9 mmol/L and 1.0 +/- 0.4 mmol/L, p < 0.05, 90 min after the LCT protocol. The plasma total cholesterol levels, which were not modified in our experimental model, showed no significant differences in relation to baseline levels in the presence or absence of Ficus carica treatment either. The clearly positive results suggest the presence in the fig leaf decoction of a compound or compounds that influence lipid catabolism.

Animals↗

The micromorphology and protein characterization of rubber particles in Ficus carica, Ficus benghalensis and Hevea brasiliensis.

Rubber biosynthesis takes place on the surface of rubber particles. These particles are surrounded by a monolayer membrane in which the rubber transferase is anchored. In order to gain better insight into whether rubber particles from different plant species share common structural characteristics, the micromorphology of rubber particles from Ficus carica, Ficus benghalensis, and Hevea brasiliensis was examined by electron microscopy. Rubber particles of all three species were spherical in shape, and the size of rubber particles of H. brasiliensis was much smaller than those of F. carica and F. benghalensis. In addition, investigations were undertaken to compare the cross-reactivity of the antibody raised against either the H. brasiliensis small rubber particle protein (SRPP) which is suggested to be involved in rubber biosynthesis, or the cis-prenyltransferase (CPT) which has an activity similar to rubber transferase. Both western analysis and TEM-immunogold labelling studies showed that rubber particles of F. carica and F. benghalensis do not contain the SRPP. None of the rubber particles in F. carica, F. benghalensis and H. brasiliensis contained the CPT, suggesting that the CPT itself could not catalyse the formation of high molecular weight rubber. These results indicate that rubber particles in the three different plant species investigated share some degree of similarity in architecture, and that the SRPP and CPT themselves are not the core proteins necessary for rubber biosynthesis.

Blotting, Western↗

Isolation and preliminary characterization of the cysteine-proteinases from the latex of Carica candamarcensis Hook.

The cysteine-proteinase chymopapain from Carica papaya L. is used for chemonucleolysis of damaged human intervertebral spinal discs. The purification of this enzyme is difficult. To overcome these problems, we were looking for a substitute among the cysteine-proteinases of Carica candamarcensis Hook. The latex from unripe fruits was collected in an aqueous solution of methylethanethiolsulfonate to prevent proteolytic activities. The soluble fraction of the lypophilized product provided four enzymatically active peaks (CC-I-CC-IV) during chromatography on CM-Sephadex C-50 in sodium acetate buffer, pH5.0. They could be further purified by rechromatography under similar conditions. The isolated enzymes have been characterized by PAGE, analysis of the Fourier transform infrared spectra, preliminary studies of their specificities as well as a comparison of the N-terminal amino-acid sequences up to position 43. CC-III proved to be glycosylated. CC-I and CC-III from Carica candamarcensis Hook are suggested to correspond to papain and chymopapain from Carica papaya L., respectively.

Amidohydrolases↗

[Studies on anti-HSV effect of Ficus carica leaves].

OBJECTIVE: To study the anti-HSV effect of the extract from the leaves of Ficos carica. METHODS: The effective ingredient was extracted from the leaves of Ficus carica, and the anti-virus effect was observed on Hep-2, BHK21 and PRK cells. RESULTS: The water extract from the leaves of Ficus carica possessed distinct anti-HSV-1 effect. The MTC was 0.5 mg/ml, TDO was 15 mg/ml, and TI was 30.0. It possessed low toxicity and directly killing-virus effect on HSV-1. CONCLUSIONS: The leaves of Ficus carica possess anti-HSV-1 effect, and their application on the area of medicine, food and drugs has expansive foreground.

Animals↗

Chloroform extract of Carica papaya seeds induces long-term reversible azoospermia in langur monkey.

AIM: To evaluate the antifertility activity of the chloroform extract of Carica papaya seeds by oral administration in langur monkey, Presbytis entellus entellus. METHODS: The chloroform extract of Carica papaya seeds, 50 mg/kg/day, was administered orally for 360 days to adult male langur monkeys. The sperm characteristics by light and electron microscopy, the sperm functional tests, the semen biochemistry, the serum testosterone level, the Leydig cell function, and the histology and ultrastructure of testis were determined to evaluate the antifertility activity and the blood biochemistry and hematology, to evaluate the toxicology. RESULTS: The extract gradually decreased the sperm concentration since days 30-60 of treatment with a total inhibition of sperm motility, a decrease in sperm viability and increase in sperm abnormality. Azoospermia was observed after day 90 of treatment and continued during the whole treatment period. Treatment withdrawal resulted in a gradual recovery in these parameters and 150 days later they reverted to nearly the pretreatment values. Morphological observation of the ejaculated sperm by light and scanning electron microscopy showed deleterious changes, particularly on the mid-piece. Sperm functional tests, viz., sperm mitochondrial activity index, acrosome intactness test and hypo-osmotic swelling test scored in the infertile range during treatment and returned to the fertile values 150 days after drug withdrawal. Histology of the testis revealed shrunken tubules, germ cell atrophy and normal Leydig cells. Ultrastructure of the testis showed vacuolization in the cytoplasm of Sertoli cells and germ cells. Loss of cytoplasmic organelles were evident in the spermatocytes and spermatids. Round spermatids showed loss of Golgi bodies, peripheral mitochondria and vacuolated cytoplasm, indicating maturational arrest. Leydig cell functional test indicated a mild inhibition of steroidogenic function. Haematology and serum biochemistry study disclosed no significant toxicological effect and the serum testosterone level was not affected. CONCLUSION: Carica papaya seed extract may selectively act on the developing germ cells, possibly mediated via Sertoli cells, leading to azoospermia.

Animals↗

Abortifacient properties of aqueous extract of Carica papaya (Linn) seeds on female Sprague-Dawley rats.

Experiments were conducted to investigate the abortifacient potential of aqueous extract of Carica papaya (Linn) seeds in female Sprague-Dawley rats. Oral doses of 100 and 800 mg/kg body weight were administered once a day on days 1-10 post-coitum. No significant differences in total body weight were found in foetuses exposed to these regimes. However, in the group treated with 100 mg/kg body weight, there was a significant increase (p < 0.05) in the implantation sites and foetal weight was significantly decreased (p < 0.05) compared to the controls. No dead or malformed foetuses were found. However, in the group treated with 800 mg/kg body weight, there was obvious vaginal bleeding but no treatment related increase in implantation sites compared with control. There was however, complete resorption of about 30% of the foetuses. The surviving foetuses were stunted when compared with the control but were without any external malformations. The results of the present investigations lead to the clear conclusion that low dose aqueous crude extract of Carica papaya (Linn) seeds does not adversely affect prenatal development. The altered toxicological profile indicates that the abortifacient property is a high dose side effect. The results indicate that Carica papaya toxicity can adversely affect the foetus.

Abortifacient Agents↗

The effect of crude extract of the bark of Carica papaya on the seminiferous tubules of male Sprague-Dawley rats.

The contraceptive potential of an aqueous crude extract of the bark of Carica papaya administered orally is reported at the dose regime of 5 and 10 ml/animal/day for 4 weeks. Although the body weight or the weights of reproductive organs, kidneys, adrenals remained unaltered during the course of treatment, there was significant alteration in the histology of the testis and semen analysis when compared to the intact control group. The seminiferous tubules of rats treated with low doses of Carica papaya for 4 weeks showed no significant histological changes compared with the control. At the high dose concentration of the extract there was disorganisation in some of the seminiferous tubules with arrest of spermatogenesis beyond the level of spermatocytes. There was also widening of the lumen of the tubules. Seminal analysis showed some of the motile spermatozoa with abnormal feature in all the experimental groups. There was a dose-dependent suppression of sperm progressive forward motility which coincided with a decrease in sperm count, viability and a dose-dependent increase in percentage abnormal spermatozoa during the 14-28 days experimental period. Taken together, the present experiment showed that complete loss of fertility is attributed to decline in sperm motility and alteration in their morphology and suggest that the aqueous extracts of Carica papaya bark is safe and could serve as an effective male contraceptive in animals.

Administration, Oral↗

Experimental diabetes treated with ficus carica extract: effect on oxidative stress parameters.

Parameters related to oxidative stress were studied in rats divided into 4 groups: streptozotocin-induced diabetic rats (n=10), diabetic rats who received a single dose of a basic fraction of Ficus carica extract (n=14), diabetic rats who received a single dose of a chloroform fraction of the extract (n=10), and normal rats (n=10). Compared to normal animals, the diabetic animals presented significantly higher values for erythrocyte catalase normalized to haemoglobin levels (1.5+/-0.15 vs. 0.96+/-0.18 microg/mg) and for plasma vitamin E (73.4+/-43.9 vs. 12.0+/-1.6 mg/l), monounsaturated fatty acids (0.219+/-0.118 vs. 0.067+/-0.014 mg/ml), polyunsaturated fatty acids (PUFA, 0.567+/-0.293 vs. 0.175+/-0.040 mg/ml), saturated fatty acids (0.779+/-0.262 vs. 0.401+/-0.055 mg/ml), and linoleic acid (0.202+/-0.086 vs. 0.106 +/-0.014 mg/ml). Both Ficus carica fractions tended to normalize the values of the diabetic animals' fatty acids and plasma vitamin E values. On studying the ratios of vitamins E and A to PUFA (129.4+/-77.5 diabetic and 68.8+/-9.1 microg/mg normal; 37.5+/-20.8 vs. 108.0+/-43.6 microg/mg) and to C18:2 (259.9+/-65.8 vs. 161.0+/-21.3 microg/mg; 68.3+/-37.9 vs. 252.7+/-102.1 microg/mg), we found statistically significant differences as a function of diabetes, with the vitamin E/C18:2 ratio being normalized by the administration of the chloroform fraction (to 152.1+/-80.3 microg/mg) and the vitamin A/C18:2 ratio being raised relative to the untreated diabetic rats by the administration of the basic fraction (91.9+/-14.5 microg/mg). Our work confirms that antioxidant status is affected in the diabetes syndrome, and that Ficus carica extracts tend to normalize it.

Animals↗

Activation and inactivation of human factor X by proteases derived from Ficus carica.

We investigated the effect of proteases derived from Ficus carica (common fig) on human blood coagulation. The milky sap (latex) of several Ficus (F.) species contain ficin, which is a mixture of proteases. Ficin derived from Ficus carica shortened the activated partial thromboplastin time and the prothrombin time of normal plasmas and plasmas deficient in coagulation factors, except plasma deficient in factor X (FX) and generated activated FX (FXa) in defibrinated plasma. Chromatographic separation of ficin from Ficus carica yielded six proteolytic fractions with a different specificity towards FX. We isolated two factor X activators with molecular masses of 23.2 and 23.5 kDa, and studied their action on purified human FX. Factor X was converted to activated FXbeta by consecutive proteolytic cleavage in the heavy chain between Leu178 and Asp179, Arg187 and Gly188, and Arg194and Ile195 (FX numbering system) with concomitant release of a carboxy-terminal peptide. The cleavage pattern of FXa degradation products in the light chain was influenced by Ca2+ and Mn2+. These data suggest the haemostatic potency of Ficus proteases is based on activation of human coagulation factor X.

Amino Acid Sequence↗

Primary structure of CC-III, the glycosylated cysteine proteinase from the latex of Carica candamarcensis Hook.

The amino acid sequence of the cysteine proteinase CC-III from the latex of the subtropical species Carica candamarcensis Hook has been determined with the exception of seven residues (pos. 180-186). It was deduced from the sequence analysis of the whole chain and peptides obtained by tryptic, chymotryptic, peptic and thermolysinolytic hydrolysis. CC-III consists of 214 amino acid residues. Out of a total of eight cysteine residues, six are located at positions involved in the formation of the three disulfide bridges stabilizing the structure of papain related enzymes. CC-III from Carica candamarcensis is a glycoprotein with the carbohydrate moiety bound to asparagine at position 44. Out of 210 residues compared with the sequences of the four cysteine proteinases of Carica papaya L., CC-III shares 125 identical ones (59.5%) with papain, 142 (67.6%) with papaya proteinase IV, 146 (69.5%) with papaya proteinase III and 156 (74.3%) with chymopapain. All amino acid residues constituting the active site and subsite S2 in chymopapain are conserved in CC-III with the exception of the substitution Leu157--> Val in the latter. This fact as well as the highest degree of identity between CC-III and chymopapain point to a similar specificity of both enzymes and thus CC-III might be a suitable substitute for chymopapain as a chemonucleolytic agent.

Amino Acid Sequence↗

Human sperm immobilization effect of Carica papaya seed extracts: an in vitro study.

AIM: To examine if the seed extracts of Carica papaya, which showed antispermatogenic/sperm immobilization properties in animal models, could cause human sperm immobilization in vitro. METHODS: Chloroform extract, benzene chromatographic fraction of the chloroform extract, its methanol and ethyl acetate sub-fractions and the isolated compounds from the sub-fractions i.e., ECP 1 & 2 and MCP 1 &2, of the seeds of Carica papaya were used at concentrations of 0.1%, 0.5%, 1% and 2%. Sperm motility was assessed immediately after addition of extracts and every 5 minutes thereafter for 30 minutes. RESULTS: There were dose-dependent spermicidal effects showing an instant fall in the sperm motility to less than 20% at 2% concentration. Isolated compounds ECP 1 & 2 were more effective inducing a motility of less than 10%. Many of the spermatozoa became vibratory on the spot. Total inhibition of motility was observed within 20-25 min at all concentrations of all products. Scanning and transmission electron microscopy revealed deleterious changes in the plasma membrane of the head and mid-piece of spermatozoa. Sperm viability test and the number of abnormal spermatozoa after completion of incubation suggested that the spermatozoa were infertile. The effects were spermicidal but not spermiostatic as revealed by the sperm revival test. CONCLUSION: The results reveal spermicidal activity in vitro of the seed extracts of Carica papaya.

Fruit↗