Invivo and invitro evaluation of antitumor effects of iron oxide and folate core shell-iron oxide nanoparticles / Avaliação in vivo e in vitro dos efeitos antitumorais de nanopartículas de óxido de ferro e óxido de ferro com núcleo de folato

Abstract Nanoparticles are considered viable options in the treatment of cancer. This study was conducted to investigate the effect of magnetite nanoparticles (MNPs) and magnetite folate core shell (MFCS) on leukemic and hepatocarcinoma cell cultures as well as their effect on the animal model of acute myelocytic leukemia (AML). Through current study nanoparticles were synthesized, characterized by various techniques, and their properties were studied to confirm their nanostructure. Invivo study, nanoparticles were evaluated to inspect their cytotoxic activity against SNU-182 (human hepatocellular carcinoma), K562 (human leukemia), and THLE2 (human normal epithelial liver) cells via MTT test. Apoptotic signaling proteins Bcl-2 and Caspase-3 expression were inspected through RT-PCR method. A cytotoxic effect of MNPs and MFCS was detected in previous cell cultures. Moreover, the apoptosis was identified through significant up-regulation of caspase-3, with Bcl-2 down-regulation. Invitro study, AML was induced in rats by N-methyl-N-nitrosourea followed by oral treatment with MNPS and MFCS. Biochemical indices such as aspartate and alanine amino transferases, and lactate dehydrogenase activities, uric acid, complete blood count, and Beta -2-microglubulin were assessed in serum. Immunophenotyping for CD34 and CD38 detection was performed. Liver, kidney, and bone marrow were microscopically examined. Bcl-2 promoter methylation, and mRNA levels were examined. Although, both MNPs and MFCS depict amelioration in biochemical parameters, MFCS alleviated them toward normal control. Anticancer activity of MNPs and MFCS was approved especially for AML. Whenever, administration of MFCS was more effective than MNPs. The present work is one of few studies used MFCS as anticancer agent.

Resumo Nanopartículas são consideradas opções viáveis ​​no tratamento do câncer. Este estudo foi conduzido para investigar o efeito de nanopartículas de magnetita (MNPs) e núcleo de folato de magnetita (MFCS) em culturas de células leucêmicas e de hepatocarcinoma, bem como seu efeito no modelo animal de leucemia mielocítica aguda (LMA). Através do atual estudo, nanopartículas foram sintetizadas, caracterizadas por várias técnicas, e suas propriedades foram estudadas para confirmar sua nanoestrutura. No estudo in vivo, as nanopartículas foram avaliadas para inspecionar sua atividade citotóxica contra células SNU-182 (carcinoma hepatocelular humano), K562 (leucemia humana) e THLE2 (fígado epitelial humano normal) por meio do teste MTT. A expressão das proteínas sinalizadoras apoptóticas Bcl-2 e Caspase-3 foram inspecionadas através do método RT-PCR. Um efeito citotóxico de MNPs e MFCS foi detectado em culturas de células anteriores. Além disso, a apoptose foi identificada por meio de regulação positiva significativa da Caspase-3, com regulação negativa de Bcl-2. No estudo in vitro, a AML foi induzida em ratos por N-metil-N-nitrosoureia seguida por tratamento oral com MNPS e MFCS. Índices bioquímicos como aspartato e alanina aminotransferases e atividades de lactato desidrogenase, ácido úrico, hemograma completo e Beta-2-microglubulina foram avaliados no soro. A imunofenotipagem para detecção de CD34 e CD38 foi realizada. Fígado, rim e medula óssea foram examinados microscopicamente. A metilação do promotor Bcl-2 e os níveis de mRNA foram examinados. Embora tanto os MNPs quanto os MFCS representem uma melhora nos parâmetros bioquímicos, o MFCS os aliviou em direção ao controle normal. A atividade anticâncer de MNPs e MFCS foi aprovada especialmente para AML. Sempre, a administração de MFCS foi mais eficaz do que MNPs. O presente trabalho é um dos poucos estudos que utilizou o MFCS como agente anticâncer.

Invivo and invitro evaluation of antitumor effects of iron oxide and folate core shell-iron oxide nanoparticles

Abstract Nanoparticles are considered viable options in the treatment of cancer. This study was conducted to investigate the effect of magnetite nanoparticles (MNPs) and magnetite folate core shell (MFCS) on leukemic and hepatocarcinoma cell cultures as well as their effect on the animal model of acute myelocytic leukemia (AML). Through current study nanoparticles were synthesized, characterized by various techniques, and their properties were studied to confirm their nanostructure. Invivo study, nanoparticles were evaluated to inspect their cytotoxic activity against SNU-182 (human hepatocellular carcinoma), K562 (human leukemia), and THLE2 (human normal epithelial liver) cells via MTT test. Apoptotic signaling proteins Bcl-2 and Caspase-3 expression were inspected through RT-PCR method. A cytotoxic effect of MNPs and MFCS was detected in previous cell cultures. Moreover, the apoptosis was identified through significant up-regulation of caspase-3, with Bcl-2 down-regulation. Invitro study, AML was induced in rats by N-methyl-N-nitrosourea followed by oral treatment with MNPS and MFCS. Biochemical indices such as aspartate and alanine amino transferases, and lactate dehydrogenase activities, uric acid, complete blood count, and Beta -2-microglubulin were assessed in serum. Immunophenotyping for CD34 and CD38 detection was performed. Liver, kidney, and bone marrow were microscopically examined. Bcl-2 promoter methylation, and mRNA levels were examined. Although, both MNPs and MFCS depict amelioration in biochemical parameters, MFCS alleviated them toward normal control. Anticancer activity of MNPs and MFCS was approved especially for AML. Whenever, administration of MFCS was more effective than MNPs. The present work is one of few studies used MFCS as anticancer agent.

Resumo Nanopartículas são consideradas opções viáveis no tratamento do câncer. Este estudo foi conduzido para investigar o efeito de nanopartículas de magnetita (MNPs) e núcleo de folato de magnetita (MFCS) em culturas de células leucêmicas e de hepatocarcinoma, bem como seu efeito no modelo animal de leucemia mielocítica aguda (LMA). Através do atual estudo, nanopartículas foram sintetizadas, caracterizadas por várias técnicas, e suas propriedades foram estudadas para confirmar sua nanoestrutura. No estudo in vivo, as nanopartículas foram avaliadas para inspecionar sua atividade citotóxica contra células SNU-182 (carcinoma hepatocelular humano), K562 (leucemia humana) e THLE2 (fígado epitelial humano normal) por meio do teste MTT. A expressão das proteínas sinalizadoras apoptóticas Bcl-2 e Caspase-3 foram inspecionadas através do método RT-PCR. Um efeito citotóxico de MNPs e MFCS foi detectado em culturas de células anteriores. Além disso, a apoptose foi identificada por meio de regulação positiva significativa da Caspase-3, com regulação negativa de Bcl-2. No estudo in vitro, a AML foi induzida em ratos por N-metil-N-nitrosoureia seguida por tratamento oral com MNPS e MFCS. Índices bioquímicos como aspartato e alanina aminotransferases e atividades de lactato desidrogenase, ácido úrico, hemograma completo e Beta-2-microglubulina foram avaliados no soro. A imunofenotipagem para detecção de CD34 e CD38 foi realizada. Fígado, rim e medula óssea foram examinados microscopicamente. A metilação do promotor Bcl-2 e os níveis de mRNA foram examinados. Embora tanto os MNPs quanto os MFCS representem uma melhora nos parâmetros bioquímicos, o MFCS os aliviou em direção ao controle normal. A atividade anticâncer de MNPs e MFCS foi aprovada especialmente para AML. Sempre, a administração de MFCS foi mais eficaz do que MNPs. O presente trabalho é um dos poucos estudos que utilizou o MFCS como agente anticâncer.

Invivo and invitro evaluation of antitumor effects of iron oxide and folate core shell-iron oxide nanoparticles.

Nanoparticles are considered viable options in the treatment of cancer. This study was conducted to investigate the effect of magnetite nanoparticles (MNPs) and magnetite folate core shell (MFCS) on leukemic and hepatocarcinoma cell cultures as well as their effect on the animal model of acute myelocytic leukemia (AML). Through current study nanoparticles were synthesized, characterized by various techniques, and their properties were studied to confirm their nanostructure. Invivo study, nanoparticles were evaluated to inspect their cytotoxic activity against SNU-182 (human hepatocellular carcinoma), K562 (human leukemia), and THLE2 (human normal epithelial liver) cells via MTT test. Apoptotic signaling proteins Bcl-2 and Caspase-3 expression were inspected through RT-PCR method. A cytotoxic effect of MNPs and MFCS was detected in previous cell cultures. Moreover, the apoptosis was identified through significant up-regulation of caspase-3, with Bcl-2 down-regulation. Invitro study, AML was induced in rats by N-methyl-N-nitrosourea followed by oral treatment with MNPS and MFCS. Biochemical indices such as aspartate and alanine amino transferases, and lactate dehydrogenase activities, uric acid, complete blood count, and Beta -2-microglubulin were assessed in serum. Immunophenotyping for CD34 and CD38 detection was performed. Liver, kidney, and bone marrow were microscopically examined. Bcl-2 promoter methylation, and mRNA levels were examined. Although, both MNPs and MFCS depict amelioration in biochemical parameters, MFCS alleviated them toward normal control. Anticancer activity of MNPs and MFCS was approved especially for AML. Whenever, administration of MFCS was more effective than MNPs. The present work is one of few studies used MFCS as anticancer agent.

Morphomolecular and pathological study of Scaphanocephalus sp. in new host Siganus argenteus in the Arabian Gulf.

In this study, Scaphanocephalus parasite (Platyhelminthes: Heterophyidae) metacercariae were found in Siganus argenteus (forktail rabbitfish or streamlined spinefoot) in the Arabian Gulf of Jubail province, Saudi Arabia. The findings may constitute new host and locality records for this parasite. Based on the number of black spots containing parasite cysts per fish, our study indicated that Siganus argenteus had high infection intensities of encysted metacercariae belonging to the genus Scaphanocephalus. Of the 3500 S. argenteus specimens examined, 800 (22.9%) showed multiple black cysts over the entire body surface, including the membranous parts of fins, while none were seen on the internal organs. The prevalence of infection was highest in summer (June-August) (8.8%). The excysted metacercariae differed morphologically from previously identified Scaphanocephalus spp. Molecular analysis of rDNA showed 100% identity with an unnamed Scaphanocephalus sp. reported in Caribbean fish. Therefore, our findings may indicate a new species of this previously rarely recorded fish parasite. The histopathological examination revealed that the encysted parasites were restricted to the dermal layer of the skin and surrounded by melanophores and a fibrous connective tissue capsule, with focal myositis and Zenker's necrosis in the underlying muscle tissue. The characteristically 'winged' parasite was clearly observed within the cysts. The high prevalence of Scaphanocephalus infection in siganid fish we detected requires further epidemiological study.

Analysis of reproductive biology and spawning season of the pink ear emperor Lethrinus lentjan, from marine ecosystem

A total of 593 samples of Lethrinus lentjan (Lacepede, 1802) were collected from the Red Sea, Jeddah, Saudi Arabia, to study their productive biology and spawning season of the local population. Sampling was carried out on a monthly basis for a period of one year. The monthly sex ratios indicated that females were dominant throughout the study period, with an overall male:female sex ratio of 1:7.98, although males were larger than females. The highest monthly performance maturation index (PMI), as well as the male and female gonadosomatic index (GSI) and ovarian maturation rate (OMR) were observed in February and March. Histological examination of the gonads confirmed the process of sexual transformation in this fish species, wherein individuals mature first as female, and then change sex to male (protogynous hermaphroditism). Histological sections also showed that the sexual maturation of males of L. lenjtan comprised three main stages, while the sexual development of females could be classified into four main stages. Extended spawning in the form of batches released during different months throughout the year were recorded for this fish species, with the main spawning season in February and March, and an additional, shorter spawning season in September.(AU)

Analysis of reproductive biology and spawning season of the pink ear emperor Lethrinus lentjan, from marine ecosystem

A total of 593 samples of Lethrinus lentjan (Lacepede, 1802) were collected from the Red Sea, Jeddah, Saudi Arabia, to study their productive biology and spawning season of the local population. Sampling was carried out on a monthly basis for a period of one year. The monthly sex ratios indicated that females were dominant throughout the study period, with an overall male:female sex ratio of 1:7.98, although males were larger than females. The highest monthly performance maturation index (PMI), as well as the male and female gonadosomatic index (GSI) and ovarian maturation rate (OMR) were observed in February and March. Histological examination of the gonads confirmed the process of sexual transformation in this fish species, wherein individuals mature first as female, and then change sex to male (protogynous hermaphroditism). Histological sections also showed that the sexual maturation of males of L. lenjtan comprised three main stages, while the sexual development of females could be classified into four main stages. Extended spawning in the form of batches released during different months throughout the year were recorded for this fish species, with the main spawning season in February and March, and an additional, shorter spawning season in September.

Oxidative Stress of in-Ovo Ochratoxin A Administered during Chick Embryonic Development

The present work was carried out to study the effect of in-ovo injection of ochratoxin A (OTA) as an oxidative stress and its consequences on hepatic and kidney functions, thyroid activity, and histological examination of brain and liver in chicken embryos and subsequently in the hatching chicks. On the 10th day of incubation, one hundred and sixty-two fertile eggs were randomly divided into two equal treatments. Control treatment, (injected by 50 µl sodium carbonate) and OTA treatment (injected by 12.5 ng OTA dissolved in 50 µl sodium carbonate). OTA treatement group significantly reduced glutathione (GSH) and significantly increased thiobarbituric acid reactive substances production (TBARS) in embryonic and hatched chicks regarding livers, spleen, bursa of Fabricius, heart, and brain as an indicator of oxidative stress. OTA injection increased TBARS and decreased GSH levels in both allantoic and amniotic fluids. On the 14th and 16th days of incubation and at the hatch, a significant lower concentration in cholesterol and higher concentrations of alanine amino transferase, aspartate amino transferase, alkaline phosphatase, gamma glutamyl transferase, acid phosphatase enzymes activities and triglycerides in the hepatic tissues of the OTA group were observed. Histological examination of OTA group of brain and liver tissues showed some degenerative changes through the experimental period. In conclusion, in-ovo OTA treated had teratogenic and embryotoxic effects.(AU)

Oxidative Stress of in-Ovo Ochratoxin A Administered during Chick Embryonic Development

The present work was carried out to study the effect of in-ovo injection of ochratoxin A (OTA) as an oxidative stress and its consequences on hepatic and kidney functions, thyroid activity, and histological examination of brain and liver in chicken embryos and subsequently in the hatching chicks. On the 10th day of incubation, one hundred and sixty-two fertile eggs were randomly divided into two equal treatments. Control treatment, (injected by 50 µl sodium carbonate) and OTA treatment (injected by 12.5 ng OTA dissolved in 50 µl sodium carbonate). OTA treatement group significantly reduced glutathione (GSH) and significantly increased thiobarbituric acid reactive substances production (TBARS) in embryonic and hatched chicks regarding livers, spleen, bursa of Fabricius, heart, and brain as an indicator of oxidative stress. OTA injection increased TBARS and decreased GSH levels in both allantoic and amniotic fluids. On the 14th and 16th days of incubation and at the hatch, a significant lower concentration in cholesterol and higher concentrations of alanine amino transferase, aspartate amino transferase, alkaline phosphatase, gamma glutamyl transferase, acid phosphatase enzymes activities and triglycerides in the hepatic tissues of the OTA group were observed. Histological examination of OTA group of brain and liver tissues showed some degenerative changes through the experimental period. In conclusion, in-ovo OTA treated had teratogenic and embryotoxic effects.

Toxic Effects of Ochratoxin A on Calcium Metabolism during Chick Embryo Developmentand in Hatched Chicks

ABSTRACT The present study was carried out to study the effect of in-ovo ochratoxin A (OTA) injection in Highline layer eggs on calcium metabolism and blood biochemical parameters of embryos and after the hatch. At day 10 of embryonic development, one hundred and sixty-two fertile eggs were individually weighed and divided into two equal treatments. The first treatment (control) consisted of the individual injection of fertile eggs with 50 µL sodium carbonate. In the second treatment (OTA), fertile eggs were individually injected with 12.5 ng OTA dissolved in 50 µL sodium carbonate. On days 12, 14, and 16 of incubation and at the hatch, serum calcium and inorganic phosphorus concentrations were lower (p 0.05), while sodium, alkaline phosphatase and triiodothyronine concentrations were higher (p 0.05) in the OTA-injected eggs compared with the controls. Serum potassium concentration was not affected (p 0.05) by OTA treatment. Lower calcium and phosphorus levels were determined (p 0.05) in the allantoic fluid of OTA-injected eggs compared with the controls. On days 12, 14, and 16 of incubation and at the hatch, lower whole body and yolk calcium and phosphorus, but not sodium levels, were measured (p 0.05) in the OTA treatment compared with the controls. In conclusion, the injection of eggs with OTA reduced blood calcium and phosphorus levels, which were associated with reduced whole body and yolk content from these electrolytes. Therefore, ochratoxin A had a negative effect on calcium metabolism.(AU)

Toxic Effects of Ochratoxin A on Calcium Metabolism during Chick Embryo Developmentand in Hatched Chicks

ABSTRACT The present study was carried out to study the effect of in-ovo ochratoxin A (OTA) injection in Highline layer eggs on calcium metabolism and blood biochemical parameters of embryos and after the hatch. At day 10 of embryonic development, one hundred and sixty-two fertile eggs were individually weighed and divided into two equal treatments. The first treatment (control) consisted of the individual injection of fertile eggs with 50 µL sodium carbonate. In the second treatment (OTA), fertile eggs were individually injected with 12.5 ng OTA dissolved in 50 µL sodium carbonate. On days 12, 14, and 16 of incubation and at the hatch, serum calcium and inorganic phosphorus concentrations were lower (p 0.05), while sodium, alkaline phosphatase and triiodothyronine concentrations were higher (p 0.05) in the OTA-injected eggs compared with the controls. Serum potassium concentration was not affected (p 0.05) by OTA treatment. Lower calcium and phosphorus levels were determined (p 0.05) in the allantoic fluid of OTA-injected eggs compared with the controls. On days 12, 14, and 16 of incubation and at the hatch, lower whole body and yolk calcium and phosphorus, but not sodium levels, were measured (p 0.05) in the OTA treatment compared with the controls. In conclusion, the injection of eggs with OTA reduced blood calcium and phosphorus levels, which were associated with reduced whole body and yolk content from these electrolytes. Therefore, ochratoxin A had a negative effect on calcium metabolism.