The global distribution of Pistacia is correlated to its adaptability to environmental conditions... more The global distribution of Pistacia is correlated to its adaptability to environmental conditions and mechanisms that had driven the genus to the current unique narrow latitudinal belt in between 10° North and 45° North. The current geostatisitcal distribution maps of the genus are shown and the derived probability maps over a period between 121 Kyr before present and the year 2100 were calculated. The tolerance of Pistacia trees to harsh climate conditions was related to leaf phenology, evergreeness vs deciduousness, which has led to geographic classification of the genus in two corresponding sections that corroborate recent molecular genetic studies. The deciduous trees are more tolerant to extreme climate conditions (−26°C to 46°C) than the evergreen species (−8°C to 41°C), except Pistacia lentiscus, which occurs at a max. temperature of 45°C. The close spatial distribution of the later species and the deciduous ones may have been conducive in further evolution of the genus. Base...
Acute myeloid leukemia (AML) is an aggressive hematological malignancy characterized by uncontrol... more Acute myeloid leukemia (AML) is an aggressive hematological malignancy characterized by uncontrolled proliferation of immature myeloid progenitors. Here, we report the in vitro antileukemic effects of the sesquiterpene thioalkaloid-enriched fraction of the Nuphar lutea leaf extract (NUP) and a purified thioalkaloid 6,6′-dihydroxythiobinupharidine (DTBN). Treatment with 0.3–10 µg/mL NUP caused a dose- and time-dependent reduction in proliferation and viability of human AML cells (KG-1a, HL60 and U937). This was associated with apoptosis induction manifested by annexin-V/propidium iodide binding as well as cleavage of caspases 8, 9, and 3 as well as poly (ADP-ribose) polymerase. Caspase-dependence of the apoptotic effect was confirmed using the pan-caspase inhibitor Q-VD-OPH. NUP induced significant biphasic changes in the cytosolic levels of reactive oxygen species (ROS) compared to untreated cells—a decrease at early time points (2–4 h) and an increase after a longer incubation (24 ...
Determining the ploidy and geneticdiversity of a germplasm is necessarybefore initiating breeding... more Determining the ploidy and geneticdiversity of a germplasm is necessarybefore initiating breeding or geneticstudies. This study was conducted tocharacterize the ploidy level of 57 naturalpopulations of orchardgrass (Dactylisglomerata L.) collected from the ranges ofThrace region of Turkey and the diversityamong populations based on RAPD (RandomAmplified Polymorphic DNA) markers. Flowcytometry was used to determine nuclear DNAcontent (pg 2C-1 = DNA content of adiploid somatic nucleus) of 6 plants foreach population. Nuclear DNA contents werecorrelated to ploidy level with root tipchromosome counts on selected plants. Onthe basis of this study, mean nuclear DNAcontent of orchardgrass was determined as9.57 ± 0.33 (with 95% confidenceinterval) while all the plants used inchromosome counting were determined to betetraploid, with 2n = 28 mitoticchromosomes, suggesting that diploidorchardgrass plants are likely very rare orabsent in ranges of Thrace region ofTurkey. In the RAPD assay, over 40polymorphic fragments were generated whichallowed some populations to bedistinguished from the rest by uniquemarkers. A cluster analysis was performedusing Nei's (1972) genetic distance indexwith an unweighted pair group method witharitmetic mean (UPGMA). The clusteranalysis indicated that there is a highlevel of gene flow among naturalorchardgrass populations and thereforegenes distributed quite homogeneouslythrough out the region. The results of thisstudy can be useful in the development ofDactylis germ plasm collectionstrategies in Thrace region for breedingpurpose.
Several anti-leishmanial drugs of choice are of plant origin. Many of the available drugs against... more Several anti-leishmanial drugs of choice are of plant origin. Many of the available drugs against the disease are toxic and in certain cases parasite drug resistance is developed. The development of new compounds is urgently required.
Environmental Science and Pollution Research, 2009
Background, aim, and scope The project was set to construct an extensive wetland in the southernm... more Background, aim, and scope The project was set to construct an extensive wetland in the southernmost region of Israel at Kibbutz Neot Smadar (30°02′45″ N and 35°01′ 19″ E). The results of the first period of monitoring, summary, and perspectives are presented. The constructed wetland (CW) was built and the subsequent monitoring performed in the framework of the Southern Arava Sustainable Waste Management Plan, funded by the EU LIFE Fund. The specific aims were: (1) To end current sewage disposal and pollution of the ground, the aquifer, and the dry river bed (wadi) paths by biologically treating the sewage as part of the creation of a sustainable wetland ecosystem. (2) Serve as an example of CW in the Negev highlands and the Arava Valley climates for neighboring communities and as a test ground for plants and building methods appropriate to hyper arid climate. (3) Serve as an educational resource and tourist attraction for groups to learn about water reuse, recycling, local wildlife and migrating birds, including serving the heart of a planned Ecological-Educational Bird Park. This report is intended to allow others who are planning similar systems in hyper arid climates to learn from our experience. Materials and methods The project is located in an extreme arid desert with less than 40 mm of rain annually and temperature ranges of −5°C to +42°C. The site receives 165-185 m 3 of municipal and agricultural wastes daily, including cowshed and goat wastes and winery outflow.
Aims The ecological, evolutionary and genetic bases of population differentiation in a variable e... more Aims The ecological, evolutionary and genetic bases of population differentiation in a variable environment are often related to the selection pressures that plants experience. We compared differences in several growth-and defence-related traits in two isolated populations of Acacia raddiana trees from sites at either end of an extreme environmental gradient in the Negev desert. † Methods We used random amplified polymorphic DNA (RAPD) to determine the molecular differences between populations. We grew plants under two levels of water, three levels of nutrients and three levels of herbivory to test for phenotypic plasticity and adaptive phenotypic plasticity. † Key Results The RAPD analyses showed that these populations are highly genetically differentiated. Phenotypic plasticity in various morphological traits in A. raddiana was related to patterns of population genetic differentiation between the two study sites. Although we did not test for maternal effects in these long-lived trees, significant genotype × environment (G × E) interactions in some of these traits indicated that such plasticity may be adaptive. † Conclusions The main selection pressure in this desert environment, perhaps unsurprisingly, is water. Increased water availability resulted in greater growth in the southern population, which normally receives far less rain than the northern population. Even under the conditions that we defined as low water and/or nutrients, the performance of the seedlings from the southern population was significantly better, perhaps reflecting selection for these traits. Consistent with previous studies of this genus, there was no evidence of trade-offs between physical and chemical defences and plant growth parameters in this study. Rather, there appeared to be positive correlations between plant size and defence parameters. The great variation in several traits in both populations may result in a diverse potential for responding to selection pressures in different environments.
Background, aim, and scope The project was set to construct an extensive wetland in the southernm... more Background, aim, and scope The project was set to construct an extensive wetland in the southernmost region of Israel at Kibbutz Neot Smadar (30°02′45″ N and 35°01′ 19″ E). The results of the first period of monitoring, summary, and perspectives are presented. The constructed wetland (CW) was built and the subsequent monitoring performed in the framework of the Southern Arava Sustainable Waste Management Plan, funded by the EU LIFE Fund. The specific aims were: (1) To end current sewage disposal and pollution of the ground, the aquifer, and the dry river bed (wadi) paths by biologically treating the sewage as part of the creation of a sustainable wetland ecosystem. (2) Serve as an example of CW in the Negev highlands and the Arava Valley climates for neighboring communities and as a test ground for plants and building methods appropriate to hyper arid climate. (3) Serve as an educational resource and tourist attraction for groups to learn about water reuse, recycling, local wildlife and migrating birds, including serving the heart of a planned Ecological-Educational Bird Park. This report is intended to allow others who are planning similar systems in hyper arid climates to learn from our experience. Materials and methods The project is located in an extreme arid desert with less than 40 mm of rain annually and temperature ranges of −5°C to +42°C. The site receives 165-185 m 3 of municipal and agricultural wastes daily, including cowshed and goat wastes and winery outflow.
Systemic tetrahymenosis caused by the protozoan parasite Tetrahymena spp. is a serious problem in... more Systemic tetrahymenosis caused by the protozoan parasite Tetrahymena spp. is a serious problem in guppy (Poecilia reticulata) farms worldwide. There is no therapeutic solution for the systemic form of this disease. Guppies severely infected with Tetrahymena spp. were imported by a commercial ornamental fish farm and brought to our laboratory. Tetrahymena sp. (Tet-NI) was isolated and in vitro cultured. Isolates maintained in culture for different time periods (as reflected by different numbers of passages in culture) were analyzed-Tet-NI 1, 4, 5 and 6, with Tet-NI 1 being cultured for the longest period (about 15 months, 54 passages) and Tet-NI 6 for the shortest (2.5 months, 10 passages). Controlled internal infection was successfully achieved by IP injection with most isolates, except for Tet-NI 1 which produced no infection. The isolate Tet-NI 6 induced the highest infection rates in internal organs (80% vs. 50% and 64% for Tet-NI 4 and 5, respectively) and mortality rates (67% vs. 20% and 27% for Tet-NI 4 and 5, respectively, and 6.7% for Tet-NI 1). The correlation between pathogenicity and Tetrahymena enzymatic activity was studied. Electrophoretic analyses revealed at least two bands of gelanolytic activity in Tet-NI 4 and 5, three bands in Tet-NI 6, and no activity in Tet-NI 1. Total inhibition of gelanolytic activity was observed after pretreatment of Tet-NI 6 with E-64, a highly selective cysteine protease inhibitor. Using hemoglobin as a substrate, Tet-NI 6 had two bands of proteolytic activity and no bands were observed in Tet-NI 1. A correlation was observed between pathogenicity and acid phosphatase activities (analyzed by commercial fluorescence kit) for Tet-NI 1 and Tet-NI 6.
The jojoba, Simmondsia chinensis, is a characteristic desert plant native to the Sonoran desert. ... more The jojoba, Simmondsia chinensis, is a characteristic desert plant native to the Sonoran desert. The jojoba meal after oil extraction is rich in protein. The major jojoba proteins were albumins (79%) and globulins (21%), which have similar amino acid compositions and also showed a labile thrombininhibitory activity. SDS-PAGE showed two major proteins at 50 kDa and 25 kDa both in the albumins and in the globulins. The 25 kDa protein has trypsin-and chymotrypsin-inhibitory activities. In vitro digestibility of the globulins and albumins resembled that of casein and soybean protein concentrates and was increased after heat treatment. The increased digestibility achieved by boiling may be attributed to inactivation of the protease inhibitors and denaturation of proteins.
In this study, twenty-eight South Indian medicinal plants were screened for their anti-fungal act... more In this study, twenty-eight South Indian medicinal plants were screened for their anti-fungal activity against six species of fungi (Trichophyton mentagrophytes, T. rubrum, T. soudanense, Candida albicans, Torulopsis glabrata, and C. krusei). Three plant species extracts, Celastrus paniculatus, Eriodendron anfractuosum and Ficus glomerata showed inhibitory activity. An aqueous extract of galls of Terminalia chebula showed inhibitory effects on three dermatophytes (Trichophyton spp.) and three yeasts (Candida spp.). Seeds extract of T. chebula inhibited only the growth of T. glabrata. An aqueous extract of T. chebula showed inhibitory effects higher than those measured in ethanol extracts. It is therefore suggested that tannins are plausible candidates for the anti-dermatophytic effects of T. chebula. Chebulinic acid, a known tannin of T. chebula was tested and found not inhibitory, thus a search for the active compound is needed.
Phytochemicals exert antiviral activity and may play a potential therapeutic role in hepatitis C ... more Phytochemicals exert antiviral activity and may play a potential therapeutic role in hepatitis C virus (HCV) infection. In this work, we aimed to isolate NS3 inhibitors from traditional Indian medicinal plants that were found, in our earlier study, to inhibit HCV NS3 protease activity and to evaluate their potential to inhibit HCV replication. A potent inhibitory effect of NS3 catalytic activity was obtained with Embelia ribes plant extracts. Quercetin, a ubiquitous plant flavonoid, was identified as the active substance in the fractioned extract. It was found to inhibit NS3 activity in a specific dose-dependent manner in an in vitro catalysis assay. Quercetin inhibited HCV RNA replication as analysed in the subgenomic HCV RNA replicon system. It also inhibited HCV infectious virus production in the HCV infectious cell culture system (HCVcc), as analysed by the focus-forming unit reduction assay and HCV RNA real-time PCR. The inhibitory effect of quercetin was also obtained when using a model system in which NS3 engineered substrates were introduced in NS3-expressing cells, providing evidence that inhibition in vivo could be directed to the NS3 and do not involve other HCV proteins. Our work demonstrates that quercetin has a direct inhibitory effect on the HCV NS3 protease. These results point to the potential of quercetin as a natural nontoxic anti-HCV agent reducing viral production by inhibiting both NS3 and heat shock proteins essential for HCV replication.
A full-length dehydrin-like cDNA clone was obtained from a Pistacia vera L. cDNA library. The clo... more A full-length dehydrin-like cDNA clone was obtained from a Pistacia vera L. cDNA library. The clone was sequenced and it was found that the deduced protein consists of 230 amino acids with a molecular weight of 25.87 kDa. The glycine rich protein is highly hydrophilic, contains 47.4% charged amino acids and belongs to the K n -type of dehydrins. It comprises five repetitive units, each containing the K-segment diagnostic of dehydrins, but no serine tract (S-segment) and no Y-segment. Temporally, accumulation of the dehydrin transcript coincides with accumulation of the protein, which remains high in the bud scales of the inflorescence throughout winter. The dehydrin protein accumulates in the outer leaves (scales) of the inflorescence bud and in the bark of 1-year-old stems. In the buds it is mostly concentrated in the palisade-like cells underneath the epidermis, in the bud and in the inner bark parenchyma. Immunogold labeling revealed that it is a cytoplasmic protein with no specific organellar localization. It is suggested that the dehydrin of pistachio may have a dual function, a role in drought and cold tolerances, as well as serving as a storage protein.
Seedlings of Pistacia vera L. developed from seeds of two separate populations in Turkmenistan, K... more Seedlings of Pistacia vera L. developed from seeds of two separate populations in Turkmenistan, Kepele and Agachli, were evaluated for their growth potential and genetic polymorphism. Plant growth rate as well as random amplified polymorphic DNA (RAPD) analysis showed distinct differences between the two populations. In plant height growth rate, 17 Agachli accessions were 1.3 times higher on average than that of 10 accessions of Kepele (significant at p 5 0.046) and 1.2 times higher for trunk diameter growth rate (p 5 0.062). Cluster analysis divided most accessions into two main genetic groups according to their geographic origin. The Agachli group was further divided into two subgroups. One Kepele accession (K4), was genetically different from the rest and clustered on a separate outgroup. Two Agachli accessions (A12 and A17) were outside the two main populations clusters. Accessions K9 and K10 from Kepele were exceptional and were clustered in each of the two Agachli's subgroups, indicating a close genetic relationship between the two populations. In addition, high similarity values (0.58-1.00) and small genetic distances reflect plausible gene flow between Kepele and Agachli, which are 100 Km apart. Mantel test revealed significant relationship between the RAPD and the morphological traits matrices, pointing to the genetic basis for the measured differences in the growth rate. Growing the accessions on the same plot, under similar conditions enabled the evaluation of genotypic differences. The combination of morphological traits and molecular markers will further assist in preservation of genetic variability and cultivation of useful genotypes of P. vera L.
We screened thirty-four methanolic plant extracts for inhibition of the constitutive nuclear fact... more We screened thirty-four methanolic plant extracts for inhibition of the constitutive nuclear factor kappaB (NFkappaB) activity by a NFkappaB-luciferase reporter gene assay. Strong inhibition of NFkappaB activity was found in extracts of leaf and rhizome from Nuphar lutea L. SM. (Nuphar). The inhibitory action was narrowed down to a mixture of thionupharidines and/or thionuphlutidines that were identified in chromatography fractions by one- and two-dimensional NMR analysis. Dimeric sesquiterpene thioalkaloids were identified as the major components of the mixture. The Nuphar alkaloids mixture (NUP) showed a dose dependent inhibition of NFkappaB activity in a luciferase reporter gene assay as well as reduction of nuclear NFkappaB subunits expression as tested by western blots and immunohistochemistry. Decreased DNA binding was demonstrated in electro mobility shift assays. NUP inhibited both inducible and constitutive NFkappaB activation and affected the canonical and alternative pathways. Suppression of NFkappaB was not cell type specific. Induction of apoptosis by the alkaloid mixture was demonstrated by time-dependent and dose-dependent cleavage of procaspase-9 and PARP. Synergistic cytotoxicity of the active mixture with cisplatin and etoposide was demonstrated. Overall, our results show that NUP inhibits the NFkappaB pathway and acts as a sensitizer to conventional chemotherapy, enabling the search for its specific target and application against cancer and inflammation.
The global distribution of Pistacia is correlated to its adaptability to environmental conditions... more The global distribution of Pistacia is correlated to its adaptability to environmental conditions and mechanisms that had driven the genus to the current unique narrow latitudinal belt in between 10° North and 45° North. The current geostatisitcal distribution maps of the genus are shown and the derived probability maps over a period between 121 Kyr before present and the year 2100 were calculated. The tolerance of Pistacia trees to harsh climate conditions was related to leaf phenology, evergreeness vs deciduousness, which has led to geographic classification of the genus in two corresponding sections that corroborate recent molecular genetic studies. The deciduous trees are more tolerant to extreme climate conditions (−26°C to 46°C) than the evergreen species (−8°C to 41°C), except Pistacia lentiscus, which occurs at a max. temperature of 45°C. The close spatial distribution of the later species and the deciduous ones may have been conducive in further evolution of the genus. Base...
Acute myeloid leukemia (AML) is an aggressive hematological malignancy characterized by uncontrol... more Acute myeloid leukemia (AML) is an aggressive hematological malignancy characterized by uncontrolled proliferation of immature myeloid progenitors. Here, we report the in vitro antileukemic effects of the sesquiterpene thioalkaloid-enriched fraction of the Nuphar lutea leaf extract (NUP) and a purified thioalkaloid 6,6′-dihydroxythiobinupharidine (DTBN). Treatment with 0.3–10 µg/mL NUP caused a dose- and time-dependent reduction in proliferation and viability of human AML cells (KG-1a, HL60 and U937). This was associated with apoptosis induction manifested by annexin-V/propidium iodide binding as well as cleavage of caspases 8, 9, and 3 as well as poly (ADP-ribose) polymerase. Caspase-dependence of the apoptotic effect was confirmed using the pan-caspase inhibitor Q-VD-OPH. NUP induced significant biphasic changes in the cytosolic levels of reactive oxygen species (ROS) compared to untreated cells—a decrease at early time points (2–4 h) and an increase after a longer incubation (24 ...
Determining the ploidy and geneticdiversity of a germplasm is necessarybefore initiating breeding... more Determining the ploidy and geneticdiversity of a germplasm is necessarybefore initiating breeding or geneticstudies. This study was conducted tocharacterize the ploidy level of 57 naturalpopulations of orchardgrass (Dactylisglomerata L.) collected from the ranges ofThrace region of Turkey and the diversityamong populations based on RAPD (RandomAmplified Polymorphic DNA) markers. Flowcytometry was used to determine nuclear DNAcontent (pg 2C-1 = DNA content of adiploid somatic nucleus) of 6 plants foreach population. Nuclear DNA contents werecorrelated to ploidy level with root tipchromosome counts on selected plants. Onthe basis of this study, mean nuclear DNAcontent of orchardgrass was determined as9.57 ± 0.33 (with 95% confidenceinterval) while all the plants used inchromosome counting were determined to betetraploid, with 2n = 28 mitoticchromosomes, suggesting that diploidorchardgrass plants are likely very rare orabsent in ranges of Thrace region ofTurkey. In the RAPD assay, over 40polymorphic fragments were generated whichallowed some populations to bedistinguished from the rest by uniquemarkers. A cluster analysis was performedusing Nei's (1972) genetic distance indexwith an unweighted pair group method witharitmetic mean (UPGMA). The clusteranalysis indicated that there is a highlevel of gene flow among naturalorchardgrass populations and thereforegenes distributed quite homogeneouslythrough out the region. The results of thisstudy can be useful in the development ofDactylis germ plasm collectionstrategies in Thrace region for breedingpurpose.
Several anti-leishmanial drugs of choice are of plant origin. Many of the available drugs against... more Several anti-leishmanial drugs of choice are of plant origin. Many of the available drugs against the disease are toxic and in certain cases parasite drug resistance is developed. The development of new compounds is urgently required.
Environmental Science and Pollution Research, 2009
Background, aim, and scope The project was set to construct an extensive wetland in the southernm... more Background, aim, and scope The project was set to construct an extensive wetland in the southernmost region of Israel at Kibbutz Neot Smadar (30°02′45″ N and 35°01′ 19″ E). The results of the first period of monitoring, summary, and perspectives are presented. The constructed wetland (CW) was built and the subsequent monitoring performed in the framework of the Southern Arava Sustainable Waste Management Plan, funded by the EU LIFE Fund. The specific aims were: (1) To end current sewage disposal and pollution of the ground, the aquifer, and the dry river bed (wadi) paths by biologically treating the sewage as part of the creation of a sustainable wetland ecosystem. (2) Serve as an example of CW in the Negev highlands and the Arava Valley climates for neighboring communities and as a test ground for plants and building methods appropriate to hyper arid climate. (3) Serve as an educational resource and tourist attraction for groups to learn about water reuse, recycling, local wildlife and migrating birds, including serving the heart of a planned Ecological-Educational Bird Park. This report is intended to allow others who are planning similar systems in hyper arid climates to learn from our experience. Materials and methods The project is located in an extreme arid desert with less than 40 mm of rain annually and temperature ranges of −5°C to +42°C. The site receives 165-185 m 3 of municipal and agricultural wastes daily, including cowshed and goat wastes and winery outflow.
Aims The ecological, evolutionary and genetic bases of population differentiation in a variable e... more Aims The ecological, evolutionary and genetic bases of population differentiation in a variable environment are often related to the selection pressures that plants experience. We compared differences in several growth-and defence-related traits in two isolated populations of Acacia raddiana trees from sites at either end of an extreme environmental gradient in the Negev desert. † Methods We used random amplified polymorphic DNA (RAPD) to determine the molecular differences between populations. We grew plants under two levels of water, three levels of nutrients and three levels of herbivory to test for phenotypic plasticity and adaptive phenotypic plasticity. † Key Results The RAPD analyses showed that these populations are highly genetically differentiated. Phenotypic plasticity in various morphological traits in A. raddiana was related to patterns of population genetic differentiation between the two study sites. Although we did not test for maternal effects in these long-lived trees, significant genotype × environment (G × E) interactions in some of these traits indicated that such plasticity may be adaptive. † Conclusions The main selection pressure in this desert environment, perhaps unsurprisingly, is water. Increased water availability resulted in greater growth in the southern population, which normally receives far less rain than the northern population. Even under the conditions that we defined as low water and/or nutrients, the performance of the seedlings from the southern population was significantly better, perhaps reflecting selection for these traits. Consistent with previous studies of this genus, there was no evidence of trade-offs between physical and chemical defences and plant growth parameters in this study. Rather, there appeared to be positive correlations between plant size and defence parameters. The great variation in several traits in both populations may result in a diverse potential for responding to selection pressures in different environments.
Background, aim, and scope The project was set to construct an extensive wetland in the southernm... more Background, aim, and scope The project was set to construct an extensive wetland in the southernmost region of Israel at Kibbutz Neot Smadar (30°02′45″ N and 35°01′ 19″ E). The results of the first period of monitoring, summary, and perspectives are presented. The constructed wetland (CW) was built and the subsequent monitoring performed in the framework of the Southern Arava Sustainable Waste Management Plan, funded by the EU LIFE Fund. The specific aims were: (1) To end current sewage disposal and pollution of the ground, the aquifer, and the dry river bed (wadi) paths by biologically treating the sewage as part of the creation of a sustainable wetland ecosystem. (2) Serve as an example of CW in the Negev highlands and the Arava Valley climates for neighboring communities and as a test ground for plants and building methods appropriate to hyper arid climate. (3) Serve as an educational resource and tourist attraction for groups to learn about water reuse, recycling, local wildlife and migrating birds, including serving the heart of a planned Ecological-Educational Bird Park. This report is intended to allow others who are planning similar systems in hyper arid climates to learn from our experience. Materials and methods The project is located in an extreme arid desert with less than 40 mm of rain annually and temperature ranges of −5°C to +42°C. The site receives 165-185 m 3 of municipal and agricultural wastes daily, including cowshed and goat wastes and winery outflow.
Systemic tetrahymenosis caused by the protozoan parasite Tetrahymena spp. is a serious problem in... more Systemic tetrahymenosis caused by the protozoan parasite Tetrahymena spp. is a serious problem in guppy (Poecilia reticulata) farms worldwide. There is no therapeutic solution for the systemic form of this disease. Guppies severely infected with Tetrahymena spp. were imported by a commercial ornamental fish farm and brought to our laboratory. Tetrahymena sp. (Tet-NI) was isolated and in vitro cultured. Isolates maintained in culture for different time periods (as reflected by different numbers of passages in culture) were analyzed-Tet-NI 1, 4, 5 and 6, with Tet-NI 1 being cultured for the longest period (about 15 months, 54 passages) and Tet-NI 6 for the shortest (2.5 months, 10 passages). Controlled internal infection was successfully achieved by IP injection with most isolates, except for Tet-NI 1 which produced no infection. The isolate Tet-NI 6 induced the highest infection rates in internal organs (80% vs. 50% and 64% for Tet-NI 4 and 5, respectively) and mortality rates (67% vs. 20% and 27% for Tet-NI 4 and 5, respectively, and 6.7% for Tet-NI 1). The correlation between pathogenicity and Tetrahymena enzymatic activity was studied. Electrophoretic analyses revealed at least two bands of gelanolytic activity in Tet-NI 4 and 5, three bands in Tet-NI 6, and no activity in Tet-NI 1. Total inhibition of gelanolytic activity was observed after pretreatment of Tet-NI 6 with E-64, a highly selective cysteine protease inhibitor. Using hemoglobin as a substrate, Tet-NI 6 had two bands of proteolytic activity and no bands were observed in Tet-NI 1. A correlation was observed between pathogenicity and acid phosphatase activities (analyzed by commercial fluorescence kit) for Tet-NI 1 and Tet-NI 6.
The jojoba, Simmondsia chinensis, is a characteristic desert plant native to the Sonoran desert. ... more The jojoba, Simmondsia chinensis, is a characteristic desert plant native to the Sonoran desert. The jojoba meal after oil extraction is rich in protein. The major jojoba proteins were albumins (79%) and globulins (21%), which have similar amino acid compositions and also showed a labile thrombininhibitory activity. SDS-PAGE showed two major proteins at 50 kDa and 25 kDa both in the albumins and in the globulins. The 25 kDa protein has trypsin-and chymotrypsin-inhibitory activities. In vitro digestibility of the globulins and albumins resembled that of casein and soybean protein concentrates and was increased after heat treatment. The increased digestibility achieved by boiling may be attributed to inactivation of the protease inhibitors and denaturation of proteins.
In this study, twenty-eight South Indian medicinal plants were screened for their anti-fungal act... more In this study, twenty-eight South Indian medicinal plants were screened for their anti-fungal activity against six species of fungi (Trichophyton mentagrophytes, T. rubrum, T. soudanense, Candida albicans, Torulopsis glabrata, and C. krusei). Three plant species extracts, Celastrus paniculatus, Eriodendron anfractuosum and Ficus glomerata showed inhibitory activity. An aqueous extract of galls of Terminalia chebula showed inhibitory effects on three dermatophytes (Trichophyton spp.) and three yeasts (Candida spp.). Seeds extract of T. chebula inhibited only the growth of T. glabrata. An aqueous extract of T. chebula showed inhibitory effects higher than those measured in ethanol extracts. It is therefore suggested that tannins are plausible candidates for the anti-dermatophytic effects of T. chebula. Chebulinic acid, a known tannin of T. chebula was tested and found not inhibitory, thus a search for the active compound is needed.
Phytochemicals exert antiviral activity and may play a potential therapeutic role in hepatitis C ... more Phytochemicals exert antiviral activity and may play a potential therapeutic role in hepatitis C virus (HCV) infection. In this work, we aimed to isolate NS3 inhibitors from traditional Indian medicinal plants that were found, in our earlier study, to inhibit HCV NS3 protease activity and to evaluate their potential to inhibit HCV replication. A potent inhibitory effect of NS3 catalytic activity was obtained with Embelia ribes plant extracts. Quercetin, a ubiquitous plant flavonoid, was identified as the active substance in the fractioned extract. It was found to inhibit NS3 activity in a specific dose-dependent manner in an in vitro catalysis assay. Quercetin inhibited HCV RNA replication as analysed in the subgenomic HCV RNA replicon system. It also inhibited HCV infectious virus production in the HCV infectious cell culture system (HCVcc), as analysed by the focus-forming unit reduction assay and HCV RNA real-time PCR. The inhibitory effect of quercetin was also obtained when using a model system in which NS3 engineered substrates were introduced in NS3-expressing cells, providing evidence that inhibition in vivo could be directed to the NS3 and do not involve other HCV proteins. Our work demonstrates that quercetin has a direct inhibitory effect on the HCV NS3 protease. These results point to the potential of quercetin as a natural nontoxic anti-HCV agent reducing viral production by inhibiting both NS3 and heat shock proteins essential for HCV replication.
A full-length dehydrin-like cDNA clone was obtained from a Pistacia vera L. cDNA library. The clo... more A full-length dehydrin-like cDNA clone was obtained from a Pistacia vera L. cDNA library. The clone was sequenced and it was found that the deduced protein consists of 230 amino acids with a molecular weight of 25.87 kDa. The glycine rich protein is highly hydrophilic, contains 47.4% charged amino acids and belongs to the K n -type of dehydrins. It comprises five repetitive units, each containing the K-segment diagnostic of dehydrins, but no serine tract (S-segment) and no Y-segment. Temporally, accumulation of the dehydrin transcript coincides with accumulation of the protein, which remains high in the bud scales of the inflorescence throughout winter. The dehydrin protein accumulates in the outer leaves (scales) of the inflorescence bud and in the bark of 1-year-old stems. In the buds it is mostly concentrated in the palisade-like cells underneath the epidermis, in the bud and in the inner bark parenchyma. Immunogold labeling revealed that it is a cytoplasmic protein with no specific organellar localization. It is suggested that the dehydrin of pistachio may have a dual function, a role in drought and cold tolerances, as well as serving as a storage protein.
Seedlings of Pistacia vera L. developed from seeds of two separate populations in Turkmenistan, K... more Seedlings of Pistacia vera L. developed from seeds of two separate populations in Turkmenistan, Kepele and Agachli, were evaluated for their growth potential and genetic polymorphism. Plant growth rate as well as random amplified polymorphic DNA (RAPD) analysis showed distinct differences between the two populations. In plant height growth rate, 17 Agachli accessions were 1.3 times higher on average than that of 10 accessions of Kepele (significant at p 5 0.046) and 1.2 times higher for trunk diameter growth rate (p 5 0.062). Cluster analysis divided most accessions into two main genetic groups according to their geographic origin. The Agachli group was further divided into two subgroups. One Kepele accession (K4), was genetically different from the rest and clustered on a separate outgroup. Two Agachli accessions (A12 and A17) were outside the two main populations clusters. Accessions K9 and K10 from Kepele were exceptional and were clustered in each of the two Agachli's subgroups, indicating a close genetic relationship between the two populations. In addition, high similarity values (0.58-1.00) and small genetic distances reflect plausible gene flow between Kepele and Agachli, which are 100 Km apart. Mantel test revealed significant relationship between the RAPD and the morphological traits matrices, pointing to the genetic basis for the measured differences in the growth rate. Growing the accessions on the same plot, under similar conditions enabled the evaluation of genotypic differences. The combination of morphological traits and molecular markers will further assist in preservation of genetic variability and cultivation of useful genotypes of P. vera L.
We screened thirty-four methanolic plant extracts for inhibition of the constitutive nuclear fact... more We screened thirty-four methanolic plant extracts for inhibition of the constitutive nuclear factor kappaB (NFkappaB) activity by a NFkappaB-luciferase reporter gene assay. Strong inhibition of NFkappaB activity was found in extracts of leaf and rhizome from Nuphar lutea L. SM. (Nuphar). The inhibitory action was narrowed down to a mixture of thionupharidines and/or thionuphlutidines that were identified in chromatography fractions by one- and two-dimensional NMR analysis. Dimeric sesquiterpene thioalkaloids were identified as the major components of the mixture. The Nuphar alkaloids mixture (NUP) showed a dose dependent inhibition of NFkappaB activity in a luciferase reporter gene assay as well as reduction of nuclear NFkappaB subunits expression as tested by western blots and immunohistochemistry. Decreased DNA binding was demonstrated in electro mobility shift assays. NUP inhibited both inducible and constitutive NFkappaB activation and affected the canonical and alternative pathways. Suppression of NFkappaB was not cell type specific. Induction of apoptosis by the alkaloid mixture was demonstrated by time-dependent and dose-dependent cleavage of procaspase-9 and PARP. Synergistic cytotoxicity of the active mixture with cisplatin and etoposide was demonstrated. Overall, our results show that NUP inhibits the NFkappaB pathway and acts as a sensitizer to conventional chemotherapy, enabling the search for its specific target and application against cancer and inflammation.
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Papers by Avi Golan