This article examines the cytotoxic effects of perezone and its isomer isoperezone on human leukemia cells. The key findings are:
1. Both perezone and isoperezone induce cytotoxicity in a concentration-dependent manner through both caspase-dependent and caspase-independent mechanisms.
2. The compounds cause changes in cell size, granularity, phosphatidylserine translocation, mitochondrial membrane potential disruption, and do not induce apoptosis inducing factor.
3. Perezone showed greater cytotoxic effects than isoperezone, but the mechanisms of cell death induction differed between the two compounds depending on concentration.
1) The study identifies the 11-kDa protein psoriasin as the principal bactericidal factor against E. coli on human skin. Psoriasin is produced by keratinocytes and found on the skin surface.
2) Experiments showed psoriasin inhibits E. coli growth through zinc sequestration. Zinc is required for psoriasin's anti-E. coli activity.
3) Psoriasin expression is focused in areas exposed to bacteria, such as sebaceous glands, and is upregulated in response to E. coli and proinflammatory cytokines.
This document presents a literature review and proposed study on the effectiveness of volatiles produced by jasmonic acid (JA) seed treatment in tomato plants. The review discusses JA structure and biosynthesis, its role in direct and indirect plant defense responses, and interplant signaling via volatile organic compounds. It aims to determine if JA can induce plants to produce volatiles capable of signaling to other plants. The proposed study will examine effects of JA-treated and adjacent untreated tomato plants on the behavior of green peach potato aphids and two-spotted ladybirds using a y-tube olfactory assay.
The two documents discuss recent discoveries about membrane protein dynamics and degradation. The first describes tracking ATP splitting in the transport protein MsbA, revealing that it splits off one phosphate group. The second found that a protease recognizes and degrades aberrant membrane proteins directly. Understanding these processes provides insight into diseases like cystic fibrosis and neurodegeneration.
Innovative research is done mainly by taxpayer funded research – government and universities funded by the NIH usually in universities and government labs and now in smaller biotech companies and then they license those to big drug companies.
Studies of in vitro antioxidant and cytotoxic activities of extracts and isol...Alexander Decker
This study evaluated the in vitro antioxidant and cytotoxic activities of extracts and isolated compounds from Parinari curatellifolia. The ethyl acetate and methanol extracts showed moderate antioxidant activity in the DPPH radical scavenging assay, with IC50 values of 13.47 μg/mL and 5.667 μg/mL, respectively. In cytotoxicity testing against cervical cancer cells (HeLa cell line), the extracts and pure compounds displayed dose-dependent cytotoxic effects. The ethyl acetate extract and compounds C7 and C8 showed the most potent cytotoxic activities, with IC50 values below 100 μg/mL. The results provide support for the traditional use of P. curatellifolia in cancer treatment and indicate that further investigation
This study analyzed gene expression profiles in potato plants subjected to cold, heat, or salt stress using a potato cDNA microarray containing over 11,000 clones. Plants were harvested at 3, 9, and 27 hours after stress initiation and tissue was collected from roots and leaves. A total of 3,314 clones showed significant up- or down-regulation in response to at least one stress condition. These included transcription factors, signaling proteins, transporters, and heat shock proteins. Clones were separated into stress-specific and shared response groups. The study identified gene expression changes indicative of the type of abiotic stress applied.
The Moss- Physcomitrella patens : A Novel Model System for Plant Development...Senthil Natesan
The moss P.patens has been used as a versatile experimental model organism for the past 80 years and it falls in the division Bryophyta. Being relatively simple in morphology and it generates only few tissues that contain limited number of cell fates (Mark leech et al., 1993), it is extensively being undertaken by plant biologists on both basic and applied strategies covering major areas namely developmental biology, evolution, systems biology, biotechnology ,biodiversity etc., As the organism is predominantly haploid in nature, it is evident that it could allow to develop insights on straight forward investigation/understanding of plant systems and hence, it is amenable for genetic and molecular level studies (Cove., 2009). The availability of complete genome sequence information, genetic and physical map ease the utilization of P.patens in all most all the fields of biology (Yasuko Kamisugi., 2008). It is the only land plant with an efficient system of homologous recombination in its nuclear DNA by which specific gene targeting could be achieved and the strategy of loss of function mutants can be generated by RNAi approach. In this context, the functional genomics of P.patens helps in identifying novel genes which could be employed in metabolic engineering and stress tolerance like drought, salt and osmotic stress thus helps in improving the crop plant performance. Moreover, it serves as a valuable platform for the production of recombinant pharmaceuticals (Anna K. Beike., 2010) Thus, the transfer of novel genes from P.patens has a greater biotechnological impact and may help in better public acceptance. Hence, this presentation aims to confine the advantages of this model plant in plant molecular research by discussing its efficiency in reproduction, range of technologies applied, some interesting characteristic features behind this model and chosen case studies will establish the model behaving as a well-versed medium for wide variety of approaches
1) The study identifies the 11-kDa protein psoriasin as the principal bactericidal factor against E. coli on human skin. Psoriasin is produced by keratinocytes and found on the skin surface.
2) Experiments showed psoriasin inhibits E. coli growth through zinc sequestration. Zinc is required for psoriasin's anti-E. coli activity.
3) Psoriasin expression is focused in areas exposed to bacteria, such as sebaceous glands, and is upregulated in response to E. coli and proinflammatory cytokines.
This document presents a literature review and proposed study on the effectiveness of volatiles produced by jasmonic acid (JA) seed treatment in tomato plants. The review discusses JA structure and biosynthesis, its role in direct and indirect plant defense responses, and interplant signaling via volatile organic compounds. It aims to determine if JA can induce plants to produce volatiles capable of signaling to other plants. The proposed study will examine effects of JA-treated and adjacent untreated tomato plants on the behavior of green peach potato aphids and two-spotted ladybirds using a y-tube olfactory assay.
The two documents discuss recent discoveries about membrane protein dynamics and degradation. The first describes tracking ATP splitting in the transport protein MsbA, revealing that it splits off one phosphate group. The second found that a protease recognizes and degrades aberrant membrane proteins directly. Understanding these processes provides insight into diseases like cystic fibrosis and neurodegeneration.
Innovative research is done mainly by taxpayer funded research – government and universities funded by the NIH usually in universities and government labs and now in smaller biotech companies and then they license those to big drug companies.
Studies of in vitro antioxidant and cytotoxic activities of extracts and isol...Alexander Decker
This study evaluated the in vitro antioxidant and cytotoxic activities of extracts and isolated compounds from Parinari curatellifolia. The ethyl acetate and methanol extracts showed moderate antioxidant activity in the DPPH radical scavenging assay, with IC50 values of 13.47 μg/mL and 5.667 μg/mL, respectively. In cytotoxicity testing against cervical cancer cells (HeLa cell line), the extracts and pure compounds displayed dose-dependent cytotoxic effects. The ethyl acetate extract and compounds C7 and C8 showed the most potent cytotoxic activities, with IC50 values below 100 μg/mL. The results provide support for the traditional use of P. curatellifolia in cancer treatment and indicate that further investigation
This study analyzed gene expression profiles in potato plants subjected to cold, heat, or salt stress using a potato cDNA microarray containing over 11,000 clones. Plants were harvested at 3, 9, and 27 hours after stress initiation and tissue was collected from roots and leaves. A total of 3,314 clones showed significant up- or down-regulation in response to at least one stress condition. These included transcription factors, signaling proteins, transporters, and heat shock proteins. Clones were separated into stress-specific and shared response groups. The study identified gene expression changes indicative of the type of abiotic stress applied.
The Moss- Physcomitrella patens : A Novel Model System for Plant Development...Senthil Natesan
The moss P.patens has been used as a versatile experimental model organism for the past 80 years and it falls in the division Bryophyta. Being relatively simple in morphology and it generates only few tissues that contain limited number of cell fates (Mark leech et al., 1993), it is extensively being undertaken by plant biologists on both basic and applied strategies covering major areas namely developmental biology, evolution, systems biology, biotechnology ,biodiversity etc., As the organism is predominantly haploid in nature, it is evident that it could allow to develop insights on straight forward investigation/understanding of plant systems and hence, it is amenable for genetic and molecular level studies (Cove., 2009). The availability of complete genome sequence information, genetic and physical map ease the utilization of P.patens in all most all the fields of biology (Yasuko Kamisugi., 2008). It is the only land plant with an efficient system of homologous recombination in its nuclear DNA by which specific gene targeting could be achieved and the strategy of loss of function mutants can be generated by RNAi approach. In this context, the functional genomics of P.patens helps in identifying novel genes which could be employed in metabolic engineering and stress tolerance like drought, salt and osmotic stress thus helps in improving the crop plant performance. Moreover, it serves as a valuable platform for the production of recombinant pharmaceuticals (Anna K. Beike., 2010) Thus, the transfer of novel genes from P.patens has a greater biotechnological impact and may help in better public acceptance. Hence, this presentation aims to confine the advantages of this model plant in plant molecular research by discussing its efficiency in reproduction, range of technologies applied, some interesting characteristic features behind this model and chosen case studies will establish the model behaving as a well-versed medium for wide variety of approaches
By: Edward M. Barbieri and James Shorter
TDP-43 forms cytoplasmic aggregates in degenerating neurons of frontotemporal dementia (FTD) patients.
Laferrière et al. now establish that TDP-43 assemblies from distinct FTD subtypes have different structures,
neurotoxicities, and seeding activities, which correlate with FTD severity. Thus, distinct pathological TDP-43
assemblies akin to prion strains might underpin distinct FTD subtypes.
Webinar presentation made by Dr. Jochen Walter, July 8, 2014 - http://www.alzforum.org/webinars/mutations-impair-trem2-maturation-processing-and-microglial-phagocytosis
2009,planta, nitric oxide ros chitosan, srivastava et alNupur Srivastava
1) The study found that nitric oxide (NO) production occurs downstream of reactive oxygen species (ROS) in guard cells during chitosan-induced stomatal closure in pea plants.
2) Using fluorescent probes, the study showed that chitosan exposure led to increased ROS levels within 5 minutes, while NO levels rose after 10 minutes. Inhibiting ROS prevented the rise in both ROS and NO, but inhibiting NO only prevented the rise in NO.
3) The results suggest that chitosan signaling leads to ROS production, which then triggers NO production, and both ROS and NO are required for chitosan-induced stomatal closure.
Introduction to and Applications of Unison, an Open Source Database for Targe...Reece Hart
Unison is an open source database that stores extensive precomputed proteomic predictions to enable efficient target discovery. It has been used to identify candidates for tumor necrosis factor ligands, helical cytokines, and death fold proteins. This document provides an overview of Unison and examples of its use in mining for immunoreceptor tyrosine inhibitory motif-containing proteins and tumor necrosis factor ligands. It also describes an analysis of NOD2 polymorphisms using Unison's integrated search methods, structure predictions, and visualization tools.
A rapid and efficient assay for extracting dna from fungiCAS0609
The document describes a new rapid method for extracting DNA from fungi in a batch format. The method involves scraping fungal tissue, subjecting it to seven rounds of freeze/thaw lysis using dry ice and boiling water, followed by boiling for 30 minutes and grinding. The Qiagen DNeasy Plant Tissue Kit is then used to purify the extracted DNA. Testing showed this new method allowed effective DNA extraction from multiple fungal isolates simultaneously in a simple and reliable way. It provides researchers an improved technique for obtaining DNA from fungi for molecular assays compared to more specialized or time-consuming existing methods.
Radiation toxicity, plants toxicity after irradiation.Dmitri Popov
1) Programmed cell death (PCD) occurs in plants under stress and involves vacuolar processing enzymes that cleave proteins leading to cell death. 2) PCD plays a role in plant development and is triggered by stresses like radiation exposure. 3) Studies show radiation can induce toxicity in plants by causing hydrogen peroxide production and PCD, and irradiated plants fed to animals have led to decreased survival rates and tumors, likely due to vitamin deficiencies caused by the irradiation.
This document discusses the role of TREM2, a receptor expressed on myeloid cells and microglia, in neurodegenerative diseases. It notes that loss-of-function mutations in TREM2 are associated with Nasu-Hakola disease and increased risk for late-onset Alzheimer's disease. Heterozygous mutations are also linked to frontotemporal dementia and related syndromes, as well as amyotrophic lateral sclerosis and Parkinson's disease. The document describes how FTD-linked mutations impair TREM2 maturation and shedding, inhibiting its transport to the cell surface. This reduces phagocytic functions of microglia, including uptake of amyloid-beta, bacterial particles, and apopt
The document examines the effects of the TACE inhibitor BMS-561392 on APP processing both in vitro and in vivo. In cell cultures expressing APP, BMS-561392 reduced secretion of sAPPα without increasing Aβ production. Conversely, a BACE inhibitor decreased sAPPβ and Aβ without affecting sAPPα. In vivo infusion of BMS-561392 into mouse brains decreased sAPPα levels but did not significantly change steady-state Aβ levels. The findings suggest that under normal conditions, BACE and TACE do not compete for APP as a substrate, though they share localization in the trans-Golgi network. Inhibition of TACE may therefore reduce TNFα levels in
DOI: 10.21276/ijlssr.2016.2.4.18
ABSTRACT- The Jambulor jambolan has been used for many years in traditional Indian medicine as treatment for
various diseases for example, diabetes mellitus, as well as bactericide. This study aimed to determine the conductivity in
different samples of a freeze-dried aqueous extract of Syzygium jambolanum. The readings of the electrical voltages of the
samples subjected to different treatment conditions were carried out in a pH meter. Depending on the performed statistical
analysis could be consttar a significant difference (p <0.001) among all samples analyzed groups. In view of the results
obtained, it can be suggested que the behavior related to changes in the electrical voltage values of the samples of an
aqueous extract of Syzygium jambolum, is related to the presence of molecules derived from flavonoids with antioxidant
properties, which can be molecules anthocyanin, which may be present in the samples of said plant extract Studied. Key-words- Syzygium jambolanum, Anthocyanin, Flavonoids, Atioxidant, Natural extract
Radiation. Plants Immunity. Toxicity of Plants after irradiation.Dmitri Popov
A high level of Plant Vacuolar Protease, activated after moderate and high doses of radiation, possible playing role as radiation toxin and can induce development of Acute Radiation Syndromes in mammals after ingestion.
The document contains 3 short phrases that seem to be instructions for someone named Mr. Yoso to watch a slide show. It provides very little context or detail to summarize further in 3 sentences or less.
The document discusses the changing expectations of consumers and the challenges faced by marketers in engaging with consumers. It notes that consumers now research products across multiple channels and expect to purchase through various channels as well. The document then advocates for a customer engagement approach using tools like Alterian Alchemy to listen to customers, learn from their interactions, understand them as individuals, and speak to them through relevant messaging across channels. This facilitates moving marketing from mass communication to personalized engagement with customers at each stage of the customer lifecycle.
Cas IIgly induces apoptosis in glioma C6 cells through both caspase-dependent and caspase-independent pathways. When exposed to Cas IIgly, glioma C6 cells showed inhibited proliferation, increased reactive oxygen species formation, and induced apoptosis in a dose-dependent manner. Both the mitochondria-nuclear translocation of apoptosis induction factor and endonuclease G, as well as the fragmentation of nucleosomal DNA and caspase-3 activation were involved in the apoptotic effects of Cas IIgly. Reactive oxygen species formation induced by Cas IIgly may also contribute to its apoptotic effects. Administration of Cas IIgly to glioma-bearing rats reduced tumor size and increased apoptosis.
2011 repeated restraint stress reduces the ig a producing cells in peyers pat...LUVIA ENID SANCHEZ TORRES
1. The study examined the effects of repeated restraint stress on Peyer's patches (PPs), which are lymphoid tissues in the intestine that play a key role in immune responses.
2. It found that repeated restraint stress did not modify the morphological structure of PPs, but it did reduce the total number of lymphocytes, CD8+ T cells, B cells, and plasma cells in PPs.
3. Specifically, restraint stress reduced the number of IgA-producing plasma cells in the dome region of PPs, where these cells are usually most numerous. Since IgA protects against intestinal infections, repeated stress may increase susceptibility to pathogens.
This article examines the effects of repeated restraint stress on the structure and function of the nasal-associated lymphoid tissue (NALT) in mice. The study found that 4 days of restraint stress decreased the percentage of CD3+ and CD4+ T cells in the NALT as well as IgA+ cells and nasal IgA levels, while 8 days of stress increased the numbers of these cells and IgA levels compared to the control group. Repeated restraint stress was found to selectively affect immune cells in the nasal mucosa and basal IgA production.
Este documento presenta la información de contacto y las líneas de investigación de la Dra. Luvia Enid Sánchez Torres en el Posgrado de Inmunología de la Escuela Nacional de Ciencias Biológicas. Sus líneas de investigación incluyen la respuesta inmunológica a la malaria, la participación de la apoptosis en diferentes patologías y la evaluación de la actividad antineoplásica de fitoquímicos y compuestos sintéticos. También incluye cinco de sus publicaciones más recientes sobre estos temas.
This article examines the effects of repeated restraint stress on the structure and function of the nasal-associated lymphoid tissue (NALT) in mice. The study found that 4 days of restraint stress decreased the percentage of CD3+ and CD4+ T cells in the NALT as well as IgA+ cells and nasal IgA levels, while 8 days of stress increased the numbers of these cells and IgA levels compared to the control group. Repeated restraint stress was found to selectively affect immune cells in the nasal mucosa and basal IgA production.
This document summarizes a study that characterized the ecdysone receptor (EcR) gene in the salmon louse (Lepeophtheirus salmonis), an economically important parasite in salmon farming. The researchers isolated and sequenced cDNA of the predicted L. salmonis EcR gene, which encoded a protein highly similar to other arthropod EcRs. In situ analysis showed the EcR transcript is expressed in ovaries, sub-cuticle, and oocytes of adult female lice. Knockdown of EcR using RNA interference terminated egg production, indicating it plays an important role in reproduction and oocyte maturation. This suggests disrupting EcR signaling may provide a way to control louse reproduction and infestation.
The paper proposes 9 hallmarks of aging based on accumulated cellular damage over time: 1) genomic instability, 2) telomere attrition, 3) epigenetic alterations, 4) loss of proteostasis, 5) deregulated nutrient sensing, 6) mitochondrial dysfunction, 7) cellular senescence, 8) stem cell exhaustion, and 9) altered intercellular communication. Some hallmarks like cellular senescence are beneficial at low levels but harmful in excess. The hallmarks represent primary damage, compensatory responses turned pathological over time, and integration of various damage types systemically.
This document summarizes a review article on the reproductive toxic effects of pyrethroid pesticides in non-target species. The review finds that pyrethroid exposure decreases fertility in both sexes by disrupting endocrine function and steroid hormone levels. It decreases sperm counts and increases fetal mortality in females mated with exposed males. Histopathological examination found abnormalities in testes, epididymis and ovaries after pyrethroid exposure. The review concludes that pyrethroids impair reproduction in animals and vitamin E may help prevent some toxic effects due to its antioxidant properties.
- Epigenetic modifications like DNA methylation and histone modifications play an important role in regulating gene expression and cell differentiation.
- Certain transcription factors are asymmetrically distributed during early cell divisions, leading to new patterns of gene expression over generations in response to cellular signaling.
- While epigenetic changes are not always heritable, some studies have found evidence that epigenetic switches can be transmitted from parents to offspring, though these effects may be reversible. This has implications for the inheritance of acquired traits.
By: Edward M. Barbieri and James Shorter
TDP-43 forms cytoplasmic aggregates in degenerating neurons of frontotemporal dementia (FTD) patients.
Laferrière et al. now establish that TDP-43 assemblies from distinct FTD subtypes have different structures,
neurotoxicities, and seeding activities, which correlate with FTD severity. Thus, distinct pathological TDP-43
assemblies akin to prion strains might underpin distinct FTD subtypes.
Webinar presentation made by Dr. Jochen Walter, July 8, 2014 - http://www.alzforum.org/webinars/mutations-impair-trem2-maturation-processing-and-microglial-phagocytosis
2009,planta, nitric oxide ros chitosan, srivastava et alNupur Srivastava
1) The study found that nitric oxide (NO) production occurs downstream of reactive oxygen species (ROS) in guard cells during chitosan-induced stomatal closure in pea plants.
2) Using fluorescent probes, the study showed that chitosan exposure led to increased ROS levels within 5 minutes, while NO levels rose after 10 minutes. Inhibiting ROS prevented the rise in both ROS and NO, but inhibiting NO only prevented the rise in NO.
3) The results suggest that chitosan signaling leads to ROS production, which then triggers NO production, and both ROS and NO are required for chitosan-induced stomatal closure.
Introduction to and Applications of Unison, an Open Source Database for Targe...Reece Hart
Unison is an open source database that stores extensive precomputed proteomic predictions to enable efficient target discovery. It has been used to identify candidates for tumor necrosis factor ligands, helical cytokines, and death fold proteins. This document provides an overview of Unison and examples of its use in mining for immunoreceptor tyrosine inhibitory motif-containing proteins and tumor necrosis factor ligands. It also describes an analysis of NOD2 polymorphisms using Unison's integrated search methods, structure predictions, and visualization tools.
A rapid and efficient assay for extracting dna from fungiCAS0609
The document describes a new rapid method for extracting DNA from fungi in a batch format. The method involves scraping fungal tissue, subjecting it to seven rounds of freeze/thaw lysis using dry ice and boiling water, followed by boiling for 30 minutes and grinding. The Qiagen DNeasy Plant Tissue Kit is then used to purify the extracted DNA. Testing showed this new method allowed effective DNA extraction from multiple fungal isolates simultaneously in a simple and reliable way. It provides researchers an improved technique for obtaining DNA from fungi for molecular assays compared to more specialized or time-consuming existing methods.
Radiation toxicity, plants toxicity after irradiation.Dmitri Popov
1) Programmed cell death (PCD) occurs in plants under stress and involves vacuolar processing enzymes that cleave proteins leading to cell death. 2) PCD plays a role in plant development and is triggered by stresses like radiation exposure. 3) Studies show radiation can induce toxicity in plants by causing hydrogen peroxide production and PCD, and irradiated plants fed to animals have led to decreased survival rates and tumors, likely due to vitamin deficiencies caused by the irradiation.
This document discusses the role of TREM2, a receptor expressed on myeloid cells and microglia, in neurodegenerative diseases. It notes that loss-of-function mutations in TREM2 are associated with Nasu-Hakola disease and increased risk for late-onset Alzheimer's disease. Heterozygous mutations are also linked to frontotemporal dementia and related syndromes, as well as amyotrophic lateral sclerosis and Parkinson's disease. The document describes how FTD-linked mutations impair TREM2 maturation and shedding, inhibiting its transport to the cell surface. This reduces phagocytic functions of microglia, including uptake of amyloid-beta, bacterial particles, and apopt
The document examines the effects of the TACE inhibitor BMS-561392 on APP processing both in vitro and in vivo. In cell cultures expressing APP, BMS-561392 reduced secretion of sAPPα without increasing Aβ production. Conversely, a BACE inhibitor decreased sAPPβ and Aβ without affecting sAPPα. In vivo infusion of BMS-561392 into mouse brains decreased sAPPα levels but did not significantly change steady-state Aβ levels. The findings suggest that under normal conditions, BACE and TACE do not compete for APP as a substrate, though they share localization in the trans-Golgi network. Inhibition of TACE may therefore reduce TNFα levels in
DOI: 10.21276/ijlssr.2016.2.4.18
ABSTRACT- The Jambulor jambolan has been used for many years in traditional Indian medicine as treatment for
various diseases for example, diabetes mellitus, as well as bactericide. This study aimed to determine the conductivity in
different samples of a freeze-dried aqueous extract of Syzygium jambolanum. The readings of the electrical voltages of the
samples subjected to different treatment conditions were carried out in a pH meter. Depending on the performed statistical
analysis could be consttar a significant difference (p <0.001) among all samples analyzed groups. In view of the results
obtained, it can be suggested que the behavior related to changes in the electrical voltage values of the samples of an
aqueous extract of Syzygium jambolum, is related to the presence of molecules derived from flavonoids with antioxidant
properties, which can be molecules anthocyanin, which may be present in the samples of said plant extract Studied. Key-words- Syzygium jambolanum, Anthocyanin, Flavonoids, Atioxidant, Natural extract
Radiation. Plants Immunity. Toxicity of Plants after irradiation.Dmitri Popov
A high level of Plant Vacuolar Protease, activated after moderate and high doses of radiation, possible playing role as radiation toxin and can induce development of Acute Radiation Syndromes in mammals after ingestion.
The document contains 3 short phrases that seem to be instructions for someone named Mr. Yoso to watch a slide show. It provides very little context or detail to summarize further in 3 sentences or less.
The document discusses the changing expectations of consumers and the challenges faced by marketers in engaging with consumers. It notes that consumers now research products across multiple channels and expect to purchase through various channels as well. The document then advocates for a customer engagement approach using tools like Alterian Alchemy to listen to customers, learn from their interactions, understand them as individuals, and speak to them through relevant messaging across channels. This facilitates moving marketing from mass communication to personalized engagement with customers at each stage of the customer lifecycle.
Cas IIgly induces apoptosis in glioma C6 cells through both caspase-dependent and caspase-independent pathways. When exposed to Cas IIgly, glioma C6 cells showed inhibited proliferation, increased reactive oxygen species formation, and induced apoptosis in a dose-dependent manner. Both the mitochondria-nuclear translocation of apoptosis induction factor and endonuclease G, as well as the fragmentation of nucleosomal DNA and caspase-3 activation were involved in the apoptotic effects of Cas IIgly. Reactive oxygen species formation induced by Cas IIgly may also contribute to its apoptotic effects. Administration of Cas IIgly to glioma-bearing rats reduced tumor size and increased apoptosis.
2011 repeated restraint stress reduces the ig a producing cells in peyers pat...LUVIA ENID SANCHEZ TORRES
1. The study examined the effects of repeated restraint stress on Peyer's patches (PPs), which are lymphoid tissues in the intestine that play a key role in immune responses.
2. It found that repeated restraint stress did not modify the morphological structure of PPs, but it did reduce the total number of lymphocytes, CD8+ T cells, B cells, and plasma cells in PPs.
3. Specifically, restraint stress reduced the number of IgA-producing plasma cells in the dome region of PPs, where these cells are usually most numerous. Since IgA protects against intestinal infections, repeated stress may increase susceptibility to pathogens.
This article examines the effects of repeated restraint stress on the structure and function of the nasal-associated lymphoid tissue (NALT) in mice. The study found that 4 days of restraint stress decreased the percentage of CD3+ and CD4+ T cells in the NALT as well as IgA+ cells and nasal IgA levels, while 8 days of stress increased the numbers of these cells and IgA levels compared to the control group. Repeated restraint stress was found to selectively affect immune cells in the nasal mucosa and basal IgA production.
Este documento presenta la información de contacto y las líneas de investigación de la Dra. Luvia Enid Sánchez Torres en el Posgrado de Inmunología de la Escuela Nacional de Ciencias Biológicas. Sus líneas de investigación incluyen la respuesta inmunológica a la malaria, la participación de la apoptosis en diferentes patologías y la evaluación de la actividad antineoplásica de fitoquímicos y compuestos sintéticos. También incluye cinco de sus publicaciones más recientes sobre estos temas.
This article examines the effects of repeated restraint stress on the structure and function of the nasal-associated lymphoid tissue (NALT) in mice. The study found that 4 days of restraint stress decreased the percentage of CD3+ and CD4+ T cells in the NALT as well as IgA+ cells and nasal IgA levels, while 8 days of stress increased the numbers of these cells and IgA levels compared to the control group. Repeated restraint stress was found to selectively affect immune cells in the nasal mucosa and basal IgA production.
This document summarizes a study that characterized the ecdysone receptor (EcR) gene in the salmon louse (Lepeophtheirus salmonis), an economically important parasite in salmon farming. The researchers isolated and sequenced cDNA of the predicted L. salmonis EcR gene, which encoded a protein highly similar to other arthropod EcRs. In situ analysis showed the EcR transcript is expressed in ovaries, sub-cuticle, and oocytes of adult female lice. Knockdown of EcR using RNA interference terminated egg production, indicating it plays an important role in reproduction and oocyte maturation. This suggests disrupting EcR signaling may provide a way to control louse reproduction and infestation.
The paper proposes 9 hallmarks of aging based on accumulated cellular damage over time: 1) genomic instability, 2) telomere attrition, 3) epigenetic alterations, 4) loss of proteostasis, 5) deregulated nutrient sensing, 6) mitochondrial dysfunction, 7) cellular senescence, 8) stem cell exhaustion, and 9) altered intercellular communication. Some hallmarks like cellular senescence are beneficial at low levels but harmful in excess. The hallmarks represent primary damage, compensatory responses turned pathological over time, and integration of various damage types systemically.
This document summarizes a review article on the reproductive toxic effects of pyrethroid pesticides in non-target species. The review finds that pyrethroid exposure decreases fertility in both sexes by disrupting endocrine function and steroid hormone levels. It decreases sperm counts and increases fetal mortality in females mated with exposed males. Histopathological examination found abnormalities in testes, epididymis and ovaries after pyrethroid exposure. The review concludes that pyrethroids impair reproduction in animals and vitamin E may help prevent some toxic effects due to its antioxidant properties.
- Epigenetic modifications like DNA methylation and histone modifications play an important role in regulating gene expression and cell differentiation.
- Certain transcription factors are asymmetrically distributed during early cell divisions, leading to new patterns of gene expression over generations in response to cellular signaling.
- While epigenetic changes are not always heritable, some studies have found evidence that epigenetic switches can be transmitted from parents to offspring, though these effects may be reversible. This has implications for the inheritance of acquired traits.
This document summarizes the causes and consequences of oxidative stress in spermatozoa. It discusses how spermatozoa are vulnerable to oxidative attack due to their lack of antioxidant protection and high levels of polyunsaturated fatty acids. Oxidative stress can damage sperm membranes through lipid peroxidation, suppressing motility. It also causes DNA damage that is associated with miscarriage and developmental abnormalities. While reactive oxygen species are important for sperm capacitation, excessive levels can overwhelm antioxidant defenses and induce apoptosis. The main source of oxidative stress in spermatozoa is mitochondrial reactive oxygen species generation.
Exosomes - Diagnostics and TherapeuticsSumedhaBobade
This document discusses exosomes, which are extracellular vesicles that originate inside cells and are released outside. It provides background on the discovery of exosomes and their structure. Exosomes are 40-150nm in size and have a phospholipid bilayer. They are secreted by various immune cells, epithelial cells, and are present in many body fluids. The document outlines their biogenesis pathway and composition. It discusses the many potential roles and applications of exosomes in diagnostics and therapeutics for diseases like cancer, neurological disorders, and infectious diseases. Exosomes show promise as novel biomarkers for diagnosis and as vehicles for drug delivery.
Nrf2: A Guardian of Healthspan and Gatekeeper of Species LongevityLifeVantage
This document summarizes a symposium discussing the role of the transcription factor Nrf2 in aging and longevity. Nrf2 regulates over 200 genes involved in cytoprotection, metabolism of toxins, oxidative stress response, and protein stability/degradation. Constitutively high expression and activation of Nrf2 and its downstream targets are observed in long-lived species and models of extended lifespan, suggesting Nrf2 plays a critical role in determining species longevity and regulating the aging process by protecting against age-related diseases and stressors. The document hypothesizes that Nrf2 is a "master regulator" of aging.
The document discusses erythropoietic protoporphyria (EPP), a rare blood disorder that can cause light sensitivity, anemia, and fatigue. Researchers from Boston Children's Hospital studied EPP and its symptoms. EPP is caused by a genetic mutation that leads to an excess of the chemical protoporphyrin in the blood and skin. While there is no cure, treatment aims to manage symptoms and improve quality of life. The disorder was proposed as a possible explanation for myths about vampires being sensitive to sunlight.
This document summarizes key findings in the new field of epigenetics. It discusses how environmental factors like diet and stress can induce epigenetic changes that alter gene expression without changing DNA. Specifically, it describes an experiment where changing mother mice's diet resulted in offspring with different physical traits and disease risks. The document also discusses how epigenetic changes can be passed down generations, how the human epigenome is as important as DNA, links between epigenetics and diseases like cancer, and how nurturing behaviors in rats can induce epigenetic changes in offspring's brains that affect stress responses.
1) The study examined potential transgenerational effects of developmental exposure to endocrine-disrupting chemicals (EDCs) like bisphenol A, phthalates, and methoxychlor on female rat reproduction.
2) EDC exposure of pregnant F0 rats and their F1 offspring resulted in accelerated puberty in F1 rats for methoxychlor and DES, and delayed puberty for phthalates. DES also reduced F1 litter size.
3) Further, methoxychlor exposure led to irregular estrous cycles in F1 rats. However, these effects were absent in subsequent unexposed generations F2 and F3, indicating a lack of transgenerational effects.
This study investigates autophagy in neurons and its relationship to Alzheimer's disease pathology. The study finds that:
1) In healthy neurons, autophagosomes are rapidly cleared through fusion with lysosomes, keeping autophagic vacuole levels low.
2) Impeding late stage autophagosome clearance, such as by inhibiting lysosomal proteolysis, causes autophagic vacuoles to accumulate in neurons resembling pathology in Alzheimer's disease.
3) The autophagic pathology observed in Alzheimer's disease likely arises from impaired autophagosome clearance rather than strong induction of autophagy alone.
cell lineage , cell fate - diverse class of cell fate, cell fate in plant meristem, mammalian development cell fate, nutritional effects on epigenetics, epigenetics of plants,
control of cell fate.
The document discusses the importance of DNA replication for life and medicine. DNA replication is essential for cell division and is targeted by chemotherapy drugs to stop the growth of cancer cells. Understanding DNA replication better through research could help develop preventative treatments for diseases and improve medical knowledge.
A Critical Role of Erythropoietin Receptor in Neurogenesis and Post-Stroke Re...johnohab
Erythropoietin (EPO) is the principal growth factor regulating the production of red blood cells. Recent studies demonstrated that
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leads to a specific deficit in post-stroke neurogenesis through impaired migration of neuroblasts to the peri-infarct cortex. Our results
suggest that both EPO and EPOR are essential for early embryonic neural development and that the classical EPOR is important for adult
neurogenesis and for migration of regenerating neurons during post-injury recovery.
This study examined the expression of endothelial nitric oxide synthase (eNOS) in testicular tissue from men with normal spermatogenesis (control group) and men with non-obstructive azoospermia (NOA group). Immunohistochemistry revealed that eNOS was present in Leydig cells, Sertoli cells, immature spermatids and abnormal germ cells in both groups. However, eNOS expression was significantly higher in Leydig cells of the control group and higher in Sertoli cells and abnormal germ cells of the NOA group. The higher eNOS expression in the NOA group suggests it may play a role in the apoptosis of abnormal germ cells and the disruption of sper
Radiation. Plant Toxicity. Inhibitors of radiation toxicity.Dmitri Popov
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This document is a thesis by Jonas Danielson from Lund University in 2010 on the topic of plant Major Intrinsic Proteins (MIPs), also known as aquaporins. It provides an introduction to MIPs and their structure and function. It then focuses on plant MIPs, discussing the large family of MIP isoforms in plants, their evolution and diversity across plant species. It aims to expand knowledge of plant MIP diversity and the roles of different subfamilies and isoforms using both traditional molecular biology approaches and comparative genetics methods.
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Andreas Schleicher presents PISA 2022 Volume III - Creative Thinking - 18 Jun...EduSkills OECD
Andreas Schleicher, Director of Education and Skills at the OECD presents at the launch of PISA 2022 Volume III - Creative Minds, Creative Schools on 18 June 2024.
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These slides walk through the story of 1 Samuel. Samuel is the last judge of Israel. The people reject God and want a king. Saul is anointed as the first king, but he is not a good king. David, the shepherd boy is anointed and Saul is envious of him. David shows honor while Saul continues to self destruct.
Temple of Asclepius in Thrace. Excavation resultsKrassimira Luka
The temple and the sanctuary around were dedicated to Asklepios Zmidrenus. This name has been known since 1875 when an inscription dedicated to him was discovered in Rome. The inscription is dated in 227 AD and was left by soldiers originating from the city of Philippopolis (modern Plovdiv).
Chapter wise All Notes of First year Basic Civil Engineering.pptxDenish Jangid
Chapter wise All Notes of First year Basic Civil Engineering
Syllabus
Chapter-1
Introduction to objective, scope and outcome the subject
Chapter 2
Introduction: Scope and Specialization of Civil Engineering, Role of civil Engineer in Society, Impact of infrastructural development on economy of country.
Chapter 3
Surveying: Object Principles & Types of Surveying; Site Plans, Plans & Maps; Scales & Unit of different Measurements.
Linear Measurements: Instruments used. Linear Measurement by Tape, Ranging out Survey Lines and overcoming Obstructions; Measurements on sloping ground; Tape corrections, conventional symbols. Angular Measurements: Instruments used; Introduction to Compass Surveying, Bearings and Longitude & Latitude of a Line, Introduction to total station.
Levelling: Instrument used Object of levelling, Methods of levelling in brief, and Contour maps.
Chapter 4
Buildings: Selection of site for Buildings, Layout of Building Plan, Types of buildings, Plinth area, carpet area, floor space index, Introduction to building byelaws, concept of sun light & ventilation. Components of Buildings & their functions, Basic concept of R.C.C., Introduction to types of foundation
Chapter 5
Transportation: Introduction to Transportation Engineering; Traffic and Road Safety: Types and Characteristics of Various Modes of Transportation; Various Road Traffic Signs, Causes of Accidents and Road Safety Measures.
Chapter 6
Environmental Engineering: Environmental Pollution, Environmental Acts and Regulations, Functional Concepts of Ecology, Basics of Species, Biodiversity, Ecosystem, Hydrological Cycle; Chemical Cycles: Carbon, Nitrogen & Phosphorus; Energy Flow in Ecosystems.
Water Pollution: Water Quality standards, Introduction to Treatment & Disposal of Waste Water. Reuse and Saving of Water, Rain Water Harvesting. Solid Waste Management: Classification of Solid Waste, Collection, Transportation and Disposal of Solid. Recycling of Solid Waste: Energy Recovery, Sanitary Landfill, On-Site Sanitation. Air & Noise Pollution: Primary and Secondary air pollutants, Harmful effects of Air Pollution, Control of Air Pollution. . Noise Pollution Harmful Effects of noise pollution, control of noise pollution, Global warming & Climate Change, Ozone depletion, Greenhouse effect
Text Books:
1. Palancharmy, Basic Civil Engineering, McGraw Hill publishers.
2. Satheesh Gopi, Basic Civil Engineering, Pearson Publishers.
3. Ketki Rangwala Dalal, Essentials of Civil Engineering, Charotar Publishing House.
4. BCP, Surveying volume 1
Gender and Mental Health - Counselling and Family Therapy Applications and In...PsychoTech Services
A proprietary approach developed by bringing together the best of learning theories from Psychology, design principles from the world of visualization, and pedagogical methods from over a decade of training experience, that enables you to: Learn better, faster!
Gender and Mental Health - Counselling and Family Therapy Applications and In...
Phymed50801[1]
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Phytomedicine 17 (2010) 614–620
Contents lists available at ScienceDirect
Phytomedicine
journal homepage: www.elsevier.de/phymed
Perezone and its isomer isoperezone induce caspase-dependent and
caspase-independent cell death
L.E. Sanchez-Torres a,n, J.A. Torres-Martınez a, M. Godınez-Victoria a,b, J.-M. Omar c, B. Velasco-Bejarano c
´ ´ ´
a
´ ´ ´ ´ ´
Departamento de Inmunologıa, Escuela Nacional de Ciencias Biologicas-IPN, Prol. Carpio y Plan de Ayala s/n, Colonia Santo Tomas, Mexico, D.F., C.P. 11340, Mexico
b
´ ´ ´ ´az ´ ´
Departamento de Bioquımica y Seccion de Estudios de Posgrado e Investigacion, Escuela Superior de Medicina-IPN, Salvador Dı Miron y Plan de San Luis, Colonia Santo Tomas,
´ ´
Mexico, D.F., C.P. 11340, Mexico
c
´ ´ ´ ´ ´ ´ ´
Departamento de Ciencias Quımicias, Seccion Quımica Organica Facultad de Estudios Superiores Cuautitlan, Universidad Nacional Autonoma de Mexico, Av. 1 de mayo S/N,
´ ´ ´
Cuautitlan Izcalli, Estado de Mexico, C.P. 54740, Mexico
a r t i c l e in f o a b s t r a c t
In this study, we investigated the cytotoxic effect of perezone, a constituent isolated from the roots of
Keywords: Perezia spp. and of its synthetic isomer isoperezone on the K562 human leukemia cell line. Perezone
Perezone showed greater cytotoxic effect than isoperezone but both compounds were found to induce
Isoperezone cytotoxicity trough a caspase-dependent and a caspase-independent mechanisms; important changes
Cytotoxic in their light scattering properties, phosphatidylserine translocation and mitochondrial membrane
Sesquiterpenes potential disruption were detected by cytometry. The mechanism of death induction of each compound
Cell death showed interesting concentration-dependent differences. Neither compound induced the apoptosis
inducing factor.
& 2009 Elsevier GmbH. All rights reserved.
Introduction with more accurate descriptions of the biochemical and cellular
parameters that are actually measured. This is because, there are
The therapeutic goal of cancer treatment has been to trigger events that are not exclusive to one type of cell death (Kroemer
tumor selective cell death. Until now, agents inducing apoptosis et al., 2009). In the present report, the recommendations of the
remain the main chemotherapeutic drugs in medical oncology NCCD were taken into account.
(Lefranc et al., 2007; Ghobrial et al., 2005; Howley and Fearnhead, There is growing interest in the ability of phytochemicals to
2008). Apoptosis plays an essential role in regulating develop- prevent and/or cure chronic diseases such as cancer (Howells
ment, growth, immune response and eliminating redundant or et al., 2007). The widespread occurrence and function of quinones
abnormal cells in organisms (Reviewed in Lockshin and Zakeri, in diverse biological systems with particular reference to their
2007 and Lamkanfi et al., 2007). The induction and execution of role in oxygen activation have been used to know the mechanisms
apoptosis require the cooperation of a series of molecules and significance of oxidation–reduction reactions and the energy
including signal molecules, receptors, enzyme and gene regulat- conserving systems in living cells (Nohl et al., 1986). Perezone
ing proteins (Howells et al., 2007; Fan et al., 2005). In addition to (2-(1,5-dimethyl-4-hexenyl)-3-hydroxymethyl-p-benzoquinone),
apoptosis, other types of death have been described. Cell death a sesquiterpene quinone, is recognized as the first secondary
has been classified according to morphological appearance, metabolite isolated in crystalline form in the New World, by Rıo ´
enzymological criteria, functional aspects or immunological ´
de la Loza in 1852 from roots of plants of the genus Perezia (Rıo de
characteristics. Due to the complexity of the subject, the la Loza, 1852). Several pharmacological effects of perezone have
Nomenclature Committee on Cell Death (NCCD) has proposed ´
been reported (Enrıquez et al., 1980; Alarcon-Aguilar et al., 1997;
unified criteria for the definition of cell death and recommends to ˜ ˜
De La Pena et al., 2001; Burgueno-Tapia et al., 2008). Interesting,
avoid expressions like ‘‘percentage apoptosis’’ and to replace them perezone promotes the release of intramitochondrial Ca2 + , thus,
maintaining ATP production during experimental reperfusion
(Tellez et al., 1999), and induce Ca2 + efflux by inducing collapse
´
Abbreviations: AIF, apoptosis inducing factor; DMSO, dimethylsulfoxide; FBS, of the membrane potential and oxidation of pyridine nucleotides,
fetal bovine serum; FITC, fluorescein isothiocyanate; FSC, forward scatter; SSC, producing a cytotoxic effect through an impairment in Ca2 +
side scatter; GSH, glutathione; PE, phycoerythrin; PI, propidium iodide; Rho 123,
rhodamine 123; TP, transition pore
homeostasis (Cuellar et al., 1987). It is clear that perezone acts as
n
Corresponding author. Tel./fax: +52 55 57296300x62489. an electron sink which results in inhibition of electron transport
´
E-mail address: luviasanchez@hotmail.com (L.E. Sanchez-Torres). (Carabez and Sandoval, 1988), but its cytotoxic activity has not
0944-7113/$ - see front matter & 2009 Elsevier GmbH. All rights reserved.
doi:10.1016/j.phymed.2009.12.011
3. Author's personal copy
ARTICLE IN PRESS
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L.E. Sanchez-Torres et al. / Phytomedicine 17 (2010) 614–620 615
been evaluated yet in the view of its possible use as anti-neoplasic ´
perezone according to protocol previously described (Martınez
agent. In order to clarify the mechanism of the sesquiterpenes et al., 2008). The complete 1H and 13C NMR data assignment was
perezone- and isoperezone-induced cell death, modification in made by comparison with previous data reported (Burgueno- ˜
cell size and granularity, translocation of phosphatidylserine, ´
Tapia and Joseph-Nathan, 1997; Martınez et al., 2008).
activation of caspase-3, mitochondrial potential and the presence
of the inducing apoptosis factor (AIF) in the cytoplasm were
evaluated. Cultures
Perezone and isoperezone were dissolved in a minimum
Materials and methods concentration of dimethylsulfoxide (DMSO, Sigma) and added to
the culture medium at final concentrations of 6.25, 12.5, 25, 50
Cell culture and 100 mM for 24 h. The final concentration of DMSO in the
culture medium was 0.1%. As a positive control of apoptosis K562
K562 cells were cultured in RPMI 1640 medium with 10% heat- were culture with cisplatin (CISP, 30 mg/ml). Treated and
inactivated fetal bovine serum (FBS), 100 units/ml penicillin, untreated (controls) cells were grown in 24-well plates with
100 mg/ml streptomycin and 2 mM L-glutamine. Cells were 5 Â 105 cells/well under the same conditions.
routinely grown at 37 1C in a humidified atmosphere of 5% CO2.
Perezone and isoperezone Viability assay
Perezone (Fig. 1a) was isolated from the dry roots of Perezia In order to investigate if the tested compounds were cytotoxic,
´
spp. as described previously (Enrıquez et al., 1980). The non- cells treated for 24 h were evaluated in their viability by Trypan
natural derivative isoperezone (Fig. 1b) was obtained from blue exclusion analysis.
Fig. 1. Chemical structure of perezone (a) and isoperezone (b).
120
Perezone
100 Isoperezone
*
*
80
*
Viability (%)
60 *
40
*
20
*
*
0
RPMI DMSO CISP 12,5 25 50 100
Controls Concentration (μM)
Fig. 2. Perezone and isoperezone induce cell death. Cytotoxicity induced by different concentrations of perezone and isoperezone was evaluated in K562 cells treated for
24 h by the Trypan blue exclusion assay. As a positive control K562 were culture with cisplatin (CISP, 30 mg/ml). Data represent the mean 7 SD of three different
experiments.
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Phosphatidylserine translocation V-fluorescein isothiocyanate (FITC) and propidium iodide (PI)
according to the manufacturer’s instructions (Caltag Laboratories).
To investigate whether the compound-induced decrease in cell Samples were acquired on a FACScalibur flow cytometer (Becton
viability is accompanied with phosphatidylserine translocation, ´,
Dickinson, San Jose California, USA) and analyzed using Cellquest
cells treated as described above were dual stained with Annexin software (Becton Dickinson). A total of 10,000 cells were recorded.
RPMI DMSO
Controls
CISP
30 μg/ml
Perezone Isoperezone
6.25 μM
12.5 μM
25 μM
50 μM
100 μM
Fig. 3. Perezone and isoperezone induce changes in FSC and SSC parameters. Cells untreated (RPMI and DMSO) and treated for 24 h with cisplatin (CISP, 30 mg/ml) or with
perezone or isoperezone at different concentrations (6.25, 12.5, 25, 50 and 100 mM) were analyzed in their FSC and SSC parameters by flow cytometry. Representative dot
plots are shown of at least three performed (arrow, lower FSC parameter).
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Active caspase-3 washed with PBS and incubated with 20 mg/mL Rhodamine
123 (Rho123, Sigma) at room temperature for 20 min.
To evaluate if the cells death induced was caspase-dependent, After washing cells twice, Rho123-related fluorescence was
cells treated as described previously and untreated cells were recorded and analyzed using a FACSCalibur cytometer (Becton
stained with a FITC-conjugated anti-active caspase-3 antibody Dickinson). Viable cells retain the Rho123 and apoptotic cells
according to the manufacturers specifications (Pharmingen). The release the dye and are detected as cells with reduced fluores-
fluorescent signal intensity was recorded and analyzed in a cence. Results are shown as percentage of cells without Rho123
FACSCalibur flow cytometer (Becton Dickinson). For each sample retention.
10,000 events were collected.
AIF determination
Mitochondrial membrane potential
In order to evaluate if the AIF factor was involved in the cell
To evaluate if the induced cell death on K562 cells altered the death induced by perezone or isoperezone, a PE-labeled anti-AIF
mitochondrial membrane potential, cells were treated with antibody was used. Cells were processed according the manu-
perezone or isoperezone for 24 h. After culture, K562 cells were facturer’s specifications (Pharmingen). Cells were acquired and
120
Perezone
100 * Isoperezone
*
Annexin V positive cells (%)
* *
*
80 *
60
*
*
40
*
20
0
RPMI DMSO CISP 6,25 12,5 25 50 100
Controls Concentration (μM)
Fig. 4. Perezone and isoperezone induce phosphatidylserine translocation. Cells untreated and treated with cisplatin (CISP, 30 mg/ml) or with perezone or isoperezone at
different concentrations for 24 h were stained with Annexin V and PI and analyzed by flow cytometry. Data represent the mean 7 SD of three different experiments
(nPr 0.05).
Perezone
120
Isoperezone
*
100 * *
Cells with low ΔΨm (%)
*
80
*
60
40
*
20 * *
0
RPMI DMSO CISP 6,25 12,5 25 50 100
Controls Concentration (μM)
Fig. 5. Perezone and isoperezone modify the mitochondrial membrane potential. Cells untreated and treated with cisplatin (CISP, 30 mg/ml) or with perezone or
isoperezone at different concentrations for 24 h were stained with Rho 123 and analyzed by flow cytometry. Data represent the mean 7SD of three different experiments,
(nPr 0.05).
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analyzed in a FACSCalibur flow cytometer (Becton Dickinson). Results and discussion
Results are shown as the percentage of cell with cytoplasmic AIF.
Cytotoxic activity
The results of cell viability after 24 h incubation of the K562
Reproducibility of the results and statistical analysis cell line with the compounds tested at different concentrations
are displayed in Fig. 2. It can be seen that both compounds were
All values were presented as the mean7SD of at least three cytotoxic when added at concentrations of 25, 50 and 100 mM.
independent assays. Statistical analyses were performed by using Perezone had a higher cytotoxic activity than its isomer as it can
SPSS 11.0 software. A P-value equal or less than 0.05 was see clearly at 50 mM. As showed, the vehicle (DMSO) does not
considered statistically significant. affect cell viability at the concentration used.
Fig. 6. Perezone and isoperezone induce cell death through mechanisms dependent and independent of caspases. Cells untreated and treated for 24 h with cisplatin (CISP,
30 mg/ml) or with perezone or isoperezone at different concentrations (6.25, 12.5, 25, 50 and 100 mM) were stained with anti-active caspase-3-FITC antibody and analyzed
by flow cytometry. The data represent the mean 7SD of three different experiments, (nP r0.05), (a). Representative histograms are shown (b).
7. Author's personal copy
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L.E. Sanchez-Torres et al. / Phytomedicine 17 (2010) 614–620 619
Modifications in forward and side scatter parameters mitochondrial electron transport inducing the intrinsic pathway
of apoptosis
Cell death induces changes in size and granularity in the cell
which can be detected by analyzing variations in their light scattering
properties by flow cytometry. When K562 cells treated with both
Active caspase-3
compounds were analyzed, important changes in Forward Scatter
(FSC) and Side Scatter (SSC) parameters were detected. Fig. 3 clearly Mitochondrial damage may result in the release of apopto-
shows the modification induced by both perezone and isoperezone in genic molecules to cytosol and further activation of caspases. In
their light-scattering properties. In the particular case of the cells order to evaluate if the cell death process induced by perezone
treated with perezone it can be observed that at 6.25 and 12.5 mM or isoperezone involved a caspase-dependent pathway, we
appeared a new population with lower size (arrow, lower FSC determined by cytometry the percentage of cell with cytoplasmic
parameter) whereas K562 cells showed an increased in granularity active caspase-3, since this enzyme is a key executioner of
(higher SSC parameter) mainly at the highest concentrations (50 and apoptosis and it is activated both in the intrinsic and in the
100 mM). At 25 mM both changes can be detected indicating that, at extrinsic pathway of apoptosis. Fig. 6 shows that the presence of
this concentration, perezone induces two different events. The same the active caspase-3 did not show dose-dependent changes. The
changes were observed in cells treated with isoperezone but the presence of the active enzyme was evident in the lowest
decreased in FSC parameter was also observed at 50 mM. At this concentrations of both compounds but at higher concentrations,
concentration, the increased in granularity was also evident, but the the percentage of positive cells decreased. The results showed
majority of the cells showed this effect when 100 mM was employed. that at lower doses both compounds induced cell death through a
It is important to note that when isoperezone was added at 25 mM, caspase-dependent mechanism. In the particular case of
almost all the cells show a reduced size. The aforementioned changes isoperezone, the caspase-3 participation to induce cell death
are consistent with those mentioned for the apoptotic (lower FSC was evident even at 50 mM. At higher doses, the cell death
with or without changes in SSC parameter) and necrotic (higher FSC induced by both compounds was mediated through a caspase-
and SSC parameters) cell death. As expected, and in correlation with independent pathway mainly in perezone-treated cells. It is
the results obtained with the Trypan blue exclusion assay (Fig. 2), a evident that the position of the hydroxyl substituent in the
proportion of the cisplatin-treated cells showed a clear reduction in isoperezone molecule diminishes the cytotoxic activity of the
the FSC parameter. perezone but favors the caspase-dependent apoptotic cell
death.
Phosphatidylserine translocation
To investigate whether the compound-induced decrease in cell Apoptosis inducing factor (AIF)
viability was accompanied with phosphatidylserine exposure, we
measured Annexin V-FITC/PI positive cells after incubation with Due to at higher concentrations of both compounds, the active
or without perezone or isoperezone at varying concentrations form of the caspase-3 was not detected, we determined the possible
(6.25, 12.5, 25, 50 and 100 mM). Similar to cisplatin treated-K562 participation of AIF in the cell death induced by both compounds at
cells, both compounds had the capacity to induce the shift of higher concentrations. AIF is one of the apoptogenic molecules that
phosphatidylserine from the inner to the outer layer of the can be release from the mitochondria and it is translocated into the
cytoplasmic membrane. The translocation was more profound nucleus during the induction of apoptosis. Within the nucleus, in
with increasing concentrations. As seen in cell viability, perezone cooperation with Endo G, it induces apoptosis without the
showed the more potent activity. Fig. 4 shows that the participation of caspases necessarily. As shown in Fig. 7 the cell
significative increase began at 12.5 mM for both compounds. For death induced by both compounds is not related to the release of AIF
perezone-treated cells at 12.5 mM it was possible to detect at any of the concentration tested. These results suggest that the
Annexin V+ /PI- referred as ‘‘early apoptosis’’, but at higher perezone- or isoperezone-induced cell death through a caspase-
concentrations only Annexin V+ /PI+ (late apoptosis or independent pathway was related to necrosis.
secondary necrosis) was present (data not shown). These
observations showed that the phosphatidylserine translocation
is involved in the cell death induced. This event is related to a
‘‘silent’’ clearance of dying cells in vivo (Ghobrial et al., 2005;
50
Lockshin and Zakeri, 2007).
Perezone
Isoperezone
AIF positive cells (%)
Mitochondrial membrane potential 30
Mitochondria play a critical role in cell death caused by drugs
such as chemotherapeutics. Because mitochondria manifest signs
of outer and/or inner membrane permeabilization when exposed 10
to a variety of cytotoxic drugs as cisplatin, the drug used as a
positive control, we determined the mitochondrial membrane
potential in K562 cells using the fluorescent dye Rho123 and RPMI DMSO CISP 6,25 12,5 25 50 100
-10
analyzed it by flow cytometry. Fig. 5 shows that perezone and Controls Concentration (μM)
isoperezone produced a reduction in the mitochondria membrane
potential in a dose-dependent manner that correlate with the Fig. 7. Cell death induced by perezone and isoperezone is an AIF-independent
event. Cells untreated and treated for 24 h with cisplatin (CISP, 30 mg/ml) or with
cytotoxic activity. Once again perezone had the highest activity in perezone or isoperezone at different concentrations were stained with anti-AIF-PE
comparison with its isomer as it can be seen clearly at 25 mM. Our antibody. Cells were analyzed by flow cytometry. The data represent the
results suggest that both sesquiterpene quinones modulate the mean 7SD of three different experiments (nP r0.05).
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Conclusion ´
Enrıquez, R., Ortega, J., Lozoya, X., 1980. Active components in Perezia roots. J.
Ethnopharmacol. 2, 389–393.
Fan, T.J., Han, L.H., Cong, R.S., Liang, J., 2005. Caspase family proteases and
Perezone showed greater cytotoxic effect than isoperezone but apoptosis. Acta Biochim. Biophys. Sin. (Shanghai) 37, 719–727.
both compounds induce the cell death trough caspase-dependent Ghobrial, I.M., Witzig, T.E., Adjei, A.A., 2005. Targeting apoptosis pathways in
and caspase-independent mechanisms. cancer therapy. CA Cancer J. Clin. 55, 178–194.
Howells, L.M., Moiseeva, E.P., Neal, C.P., Foreman, B.E., Andreadi, C.K., Sun, Y.Y.,
Hudson, E.A., Manson, M.M., 2007. Predicting the physiological relevance of
in vitro cancer preventive activities of phytochemicals. Acta Pharmacol. Sin.
Acknowledgements 28, 1274–1304.
Howley, B., Fearnhead, H.O., 2008. Caspases as therapeutic targets. J. Cell Mol. Med.
´
This research was financially supported by SIP-IPN. Godınez- 12, 1502–1516.
Kroemer, G., Galluzzi, L., Vandenabeele, P., Abrams, J., Alnemri, E.S., Baehrecke,
´
Victoria thanks CONACyT for the scholarship. Luvia Enid Sanchez-
E.H., Blagosklonny, M.V., El-Deiry, W.S., Golstein, P., Green, D.R., Hengartner,
´
Torres, is a fellow of COFAA-IPN and EDI-IPN. Benjamın Velasco ˜
M., Knight, R.A., Kumar, S., Lipton, S.A., Malorni, W., Nunez, G., Peter, M.E.,
Bejarano is a fellow of CONACYT-SNI-08/89717. We thank Alan Tschopp, J., Yuan, J., Piacentini, M., Zhivotovsky, B., Melino, G., 2009.
Larsen for reviewing the use of English in this manuscript. Classification of cell death: recommendations of the Nomenclature Committee
on Cell Death 2009. Cell Death Differ. 16, 3–11.
Lamkanfi, M., Festjens, N., Declercq, W., Vanden Berghe, T., Vandenabeele, P., 2007.
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