Asian Pacific Journal of Tropical Biomedicine
Issue 5,2018 Table of Contents
2018(5):237-244.
DOI: 10.4103/2221-1691.233004
Abstract:
Human respiratory system is harbored by a vast array of transient and normal microbial flora. A number of pathogenic viruses were diagnosed from samples in different occasions from mild to severe infections of respiratory tract. Molecular methods were developed for detection of these viruses during last two decades. Nucleic acid amplification methods were introduced for rapid and accurate diagnosis of pathogenic viruses. Multiplex detection systems were employed to identify a panel of pathogenic viruses, which requires specialized kits and instruments in some cases. This review summarizes different types of molecular approaches which were developed with time and applied for the detection of pathogenic viruses associated with infections of the respiratory system.
Human respiratory system is harbored by a vast array of transient and normal microbial flora. A number of pathogenic viruses were diagnosed from samples in different occasions from mild to severe infections of respiratory tract. Molecular methods were developed for detection of these viruses during last two decades. Nucleic acid amplification methods were introduced for rapid and accurate diagnosis of pathogenic viruses. Multiplex detection systems were employed to identify a panel of pathogenic viruses, which requires specialized kits and instruments in some cases. This review summarizes different types of molecular approaches which were developed with time and applied for the detection of pathogenic viruses associated with infections of the respiratory system.
Bruno Rafael Barboza
,
Bárbara Rafaela da Silva Barros
,
Bárbara de Azevedo Ramos
,
Maiara Celine de Moura
,
Thiago Henrique Napole?o
,
Maria Tereza dos Santos Correia
,
Luana Cassandra Breitenbach Barroso Coelho
,
Iranildo José da Cruz Filho
,
Ana Maria Souto Maior
,
Túlio Diego da Silva
,
Leylianne de Cássia Rodrigues Nerys
,
Edson Renan Barros de Santana
,
Cláudia Sampaio de Andrade Lima
,
Virgínia Maria Barros de Lorena
,
Cristiane Moutinho Lagos de Melo
2018(5):245-253.
DOI: 10.4103/2221-1691.233005
Abstract:
Objective: To evaluate antimicrobial and antioxidant properties of saline extract from Tithonia diversifolia leaves by phytochemical bioprospecting, and investigate its safety against animal cells. Methods: The saline extract was prepared, with NaCl (0.15 M), by constant stirring of the dried and pulverized leaves, followed by volume reduction by lyophilization. The extract was phytochemical characterized using ultra-performance liquid chromatography, and total phenol and flavonoid analysis also was performed. The antioxidant capacity was determined through DPPH radical, the antimicrobial property was evaluated against standard bacteria and fungi, and the viability assays were performed against mice splenocytes. Results: Fifteen compounds were identified belonging to two main classes terpenoids and phenolics. The extract showed 22.185 mg GAE/g of total phenolic compounds and 3.220 mg QE/g of flavonoid. Moreover, extract showed higher antioxidant ability similar to butylated hydroxytoluene a standard molecule [(3.042±0.019) mg AAE/g and (4.12±0.10) mg AAE/g to saline extract and butylated hydroxytoluene, respectively]. The antimicrobial assays demonstrated that the extract had a significant antifungal potential against Candida species and could be used with safety against mice splenocytes, in concentrations lower than 50 μg/mL, promoting higher proliferation in these cells. Conclusions: Saline extract from Tithonia diversifolia leaves presents potential antioxidant, antifungal properties and induces immunostimulation in mice splenocytes.
Objective: To evaluate antimicrobial and antioxidant properties of saline extract from Tithonia diversifolia leaves by phytochemical bioprospecting, and investigate its safety against animal cells. Methods: The saline extract was prepared, with NaCl (0.15 M), by constant stirring of the dried and pulverized leaves, followed by volume reduction by lyophilization. The extract was phytochemical characterized using ultra-performance liquid chromatography, and total phenol and flavonoid analysis also was performed. The antioxidant capacity was determined through DPPH radical, the antimicrobial property was evaluated against standard bacteria and fungi, and the viability assays were performed against mice splenocytes. Results: Fifteen compounds were identified belonging to two main classes terpenoids and phenolics. The extract showed 22.185 mg GAE/g of total phenolic compounds and 3.220 mg QE/g of flavonoid. Moreover, extract showed higher antioxidant ability similar to butylated hydroxytoluene a standard molecule [(3.042±0.019) mg AAE/g and (4.12±0.10) mg AAE/g to saline extract and butylated hydroxytoluene, respectively]. The antimicrobial assays demonstrated that the extract had a significant antifungal potential against Candida species and could be used with safety against mice splenocytes, in concentrations lower than 50 μg/mL, promoting higher proliferation in these cells. Conclusions: Saline extract from Tithonia diversifolia leaves presents potential antioxidant, antifungal properties and induces immunostimulation in mice splenocytes.
Mohamed Bouhrim
,
Hayat Ouassou
,
Mohamed Choukri
,
Hassane Mekhfi
,
Abderrahim Ziyyat
,
Abdelkhaleq Legssyer
,
Mohammed Aziz
,
Mohamed Bnouham
2018(5):254-260.
DOI: 10.4103/2221-1691.233006
Abstract:
Objective: To investigate the hepatoprotective effect of Opuntia dillenii seed oil (ODSO) on CCl4 provoked liver injury in rat. Methods: Animals were treated orally with ODSO at a concentration of 2 mL/kg, once daily for one week before the first intraperitoneal injection of CCl4, and thereafter the administration of the oil was continued for 7 days until the introduction of the second injection of CCl4. Fourteen hours after the last dose of CCl4, rats were sacrificed, and the relative liver weight, weight gain, alkaline phosphatase, aspartate amino transferase, alanine aminotransferase, direct bilirubin, total bilirubin, triglycerides, total cholesterol, very low density lipoprotein, low density lipoprotein, high density lipoprotein, plasmatic glucose, urea, creatinine, acid uric and malondialdehyde were determined. Results: The significant increase was found in relative liver weight and plasma levels of alanine aminotransferase, aspartate amino transferase, alkaline phosphatase, total bilirubin, direct bilirubin, triglycerides, very low-density lipoprotein, urea, uric acid and malondialdehyde. Likewise, the significant decrease was indicated in the weight gain and the level of glucose plasmatic, and high-density lipoprotein levels in CCl4 produced liver injury in rats were re-established to normal levels when treated with ODSO. While, no change was observed in the total cholesterol, low-density lipoprotein and creatinine in all animals. Conclusions: We conclude that the ODSO has a protective effect on CCl4-mediated liver injury. Hence, we suggest its inclusion as a preventive control of liver disorders.
Objective: To investigate the hepatoprotective effect of Opuntia dillenii seed oil (ODSO) on CCl4 provoked liver injury in rat. Methods: Animals were treated orally with ODSO at a concentration of 2 mL/kg, once daily for one week before the first intraperitoneal injection of CCl4, and thereafter the administration of the oil was continued for 7 days until the introduction of the second injection of CCl4. Fourteen hours after the last dose of CCl4, rats were sacrificed, and the relative liver weight, weight gain, alkaline phosphatase, aspartate amino transferase, alanine aminotransferase, direct bilirubin, total bilirubin, triglycerides, total cholesterol, very low density lipoprotein, low density lipoprotein, high density lipoprotein, plasmatic glucose, urea, creatinine, acid uric and malondialdehyde were determined. Results: The significant increase was found in relative liver weight and plasma levels of alanine aminotransferase, aspartate amino transferase, alkaline phosphatase, total bilirubin, direct bilirubin, triglycerides, very low-density lipoprotein, urea, uric acid and malondialdehyde. Likewise, the significant decrease was indicated in the weight gain and the level of glucose plasmatic, and high-density lipoprotein levels in CCl4 produced liver injury in rats were re-established to normal levels when treated with ODSO. While, no change was observed in the total cholesterol, low-density lipoprotein and creatinine in all animals. Conclusions: We conclude that the ODSO has a protective effect on CCl4-mediated liver injury. Hence, we suggest its inclusion as a preventive control of liver disorders.
2018(5):261-267.
DOI: 10.4103/2221-1691.233007
Abstract:
Objective: To understand the effects of combination treatment of pamidronate with isolated Quercus infectoria semi-purified fraction (QIsm-F) on human foetal osteoblast cell model (hFOB 1.19 cell line) through assessment of Runt related transcription fraction-2 (Runx2) and Osterix (Osx). Methods: The isolation and purification of QIsm-F were conducted by chromatographic technique. In order to assess relative efficacy of QIsm-F to the osteoblast model, the determination of half maximal effective concentration (EC50) was performed by MTT assay. hFOB 1.19 cells were cultured in DMEM F-12 and supplemented with 10% fetal bovine serum along with 1% penicillin-streptomycin incubated in 5% CO2 at 37°C. Expression of Runx2 and Osx was assessed through western blotting and confirmed with immunofluorescence staining. Results: Results of western blot analysis and immunofluorescence staining demonstrated that compared to hFOB 1.19 cells treated with single individual treatment of QIsm-F and control groups, levels of Runx2 and Osx were elevated with higher fluorescence intensity and more rapid proliferation in hFOB 1.19 cells treated with combined treatment of QIsm-F and pamidronate. Conclusions: The finding demonstrates the synergistic effect between osteoporotic drug pamidronate and established QIsm-F. The combination treatment helps increase the efficiency of pamidronate acting on osteoblast cells by stimulating osteoblast proliferation and differentiation via expression of Runx2 and Osx.
Objective: To understand the effects of combination treatment of pamidronate with isolated Quercus infectoria semi-purified fraction (QIsm-F) on human foetal osteoblast cell model (hFOB 1.19 cell line) through assessment of Runt related transcription fraction-2 (Runx2) and Osterix (Osx). Methods: The isolation and purification of QIsm-F were conducted by chromatographic technique. In order to assess relative efficacy of QIsm-F to the osteoblast model, the determination of half maximal effective concentration (EC50) was performed by MTT assay. hFOB 1.19 cells were cultured in DMEM F-12 and supplemented with 10% fetal bovine serum along with 1% penicillin-streptomycin incubated in 5% CO2 at 37°C. Expression of Runx2 and Osx was assessed through western blotting and confirmed with immunofluorescence staining. Results: Results of western blot analysis and immunofluorescence staining demonstrated that compared to hFOB 1.19 cells treated with single individual treatment of QIsm-F and control groups, levels of Runx2 and Osx were elevated with higher fluorescence intensity and more rapid proliferation in hFOB 1.19 cells treated with combined treatment of QIsm-F and pamidronate. Conclusions: The finding demonstrates the synergistic effect between osteoporotic drug pamidronate and established QIsm-F. The combination treatment helps increase the efficiency of pamidronate acting on osteoblast cells by stimulating osteoblast proliferation and differentiation via expression of Runx2 and Osx.
2018(5):268-272.
DOI: 10.4103/2221-1691.233008
Abstract:
Objective: To synthesize silver nanoparticles using silver nitrate by a green technique which involves different compositions of aqueous leaf extracts of Azadirachta indica (neem) and Ocimum sanctum (tulsi). Methods: Their shape and size were determined using transmission electron microscopy and UV-visible spectroscopy. Their antiplasmodial activity was studied using the malarial parasite strain (Plasmodium falciparum, 3D7). The parasite strain (3D7) was collected and revived in vitro using Trager and Jensen method in RPMI 1640 medium for 7-8 cycles. Half maximal effective concentration values were calculated by nonlinear regression analysis. Results: Transmission electron microscopy results confirmed the formation of silver nanoparticles with size ranging from 4.74-39.32 nm and their size differs by varying the concentrations from 20% to 100% of neem extract in neem and tulsi extracts. It was observed that samples B and C showed half maximum effective concentration of about 0.3 M. Conclusions: It can be easily established that the aqueous leaf extracts of neem and tulsi in combination can be a good source for synthesis of silver nanoparticles with small size possessing appreciable antiplasmodial activity.
Objective: To synthesize silver nanoparticles using silver nitrate by a green technique which involves different compositions of aqueous leaf extracts of Azadirachta indica (neem) and Ocimum sanctum (tulsi). Methods: Their shape and size were determined using transmission electron microscopy and UV-visible spectroscopy. Their antiplasmodial activity was studied using the malarial parasite strain (Plasmodium falciparum, 3D7). The parasite strain (3D7) was collected and revived in vitro using Trager and Jensen method in RPMI 1640 medium for 7-8 cycles. Half maximal effective concentration values were calculated by nonlinear regression analysis. Results: Transmission electron microscopy results confirmed the formation of silver nanoparticles with size ranging from 4.74-39.32 nm and their size differs by varying the concentrations from 20% to 100% of neem extract in neem and tulsi extracts. It was observed that samples B and C showed half maximum effective concentration of about 0.3 M. Conclusions: It can be easily established that the aqueous leaf extracts of neem and tulsi in combination can be a good source for synthesis of silver nanoparticles with small size possessing appreciable antiplasmodial activity.
2018(5):273-278.
DOI: 10.4103/2221-1691.233009
Abstract:
Objective: To evaluate the larvicidal efficacy of crude and fractionated extracts of Dracaena loureiri endocarp against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus mosquitos. Methods: Larvicidal activity was tested according to World Health Organization standard protocol. The third-stage larvae of each mosquito species were exposed to various concentrations of Dracaena loureiri crude extract and six groups of Dracaena loureiri fractionated extracts (RC-DT 009–014). Larval mortality rates were observed after 24 h and 48 h of exposure. Then, a computerized probit analysis of the mortality data was performed to determine lethal concentration 50 (LC50) and lethal concentration 90 values. Results: Anopheles minimus larvae (24-h LC50 77.88 mg/L) had the highest susceptibility to crude extract, whereas others (Aedes aegypti, 24-h LC50 224.73 mg/L; Aedes albopictus, 24-h LC50 261.75 mg/L; and Culex quinquefasciatus, 24-h LC50 282.86 mg/L) were significantly less susceptible. The most effective groups of fractionated extracts were RC-DT 012 and RC-DT 013. The mosquito species most susceptible to fractionated extracts was Culex quinquefasciatus, with 24-h LC50 values of 0.66 and 0.94 mg/L for RC-DT 012 and RC-DT 013, respectively. Conclusions: The larvicidal activity of fractionated extracts is more effective than that of crude extract against all tested mosquito species. For the most effective alternative larvicide, purification and a phytochemical constituent analysis must be performed.
Objective: To evaluate the larvicidal efficacy of crude and fractionated extracts of Dracaena loureiri endocarp against Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus mosquitos. Methods: Larvicidal activity was tested according to World Health Organization standard protocol. The third-stage larvae of each mosquito species were exposed to various concentrations of Dracaena loureiri crude extract and six groups of Dracaena loureiri fractionated extracts (RC-DT 009–014). Larval mortality rates were observed after 24 h and 48 h of exposure. Then, a computerized probit analysis of the mortality data was performed to determine lethal concentration 50 (LC50) and lethal concentration 90 values. Results: Anopheles minimus larvae (24-h LC50 77.88 mg/L) had the highest susceptibility to crude extract, whereas others (Aedes aegypti, 24-h LC50 224.73 mg/L; Aedes albopictus, 24-h LC50 261.75 mg/L; and Culex quinquefasciatus, 24-h LC50 282.86 mg/L) were significantly less susceptible. The most effective groups of fractionated extracts were RC-DT 012 and RC-DT 013. The mosquito species most susceptible to fractionated extracts was Culex quinquefasciatus, with 24-h LC50 values of 0.66 and 0.94 mg/L for RC-DT 012 and RC-DT 013, respectively. Conclusions: The larvicidal activity of fractionated extracts is more effective than that of crude extract against all tested mosquito species. For the most effective alternative larvicide, purification and a phytochemical constituent analysis must be performed.
Mohsen Kalantari
,
Mohammad Hossein Motazedian
,
Qasem Asgari
,
Iraj Mohammadpour
,
Aboozar Soltani
,
Kourosh Azizi
2018(5):279-284.
DOI: 10.4103/2221-1691.233010
Abstract:
Objective: To explore the co-detection of natural infection of Trypanosomatidae parasites such as Leishmania and Crithidia in reservoir hosts of leishmaniasis. Methods: Rodent populations were monitored in two endemic foci of cutaneous leishmaniasis of Fars province, southern Iran from March to October 2016. Rodents were trapped alive in several parts of Shiraz and Kharameh cities. Afterwards, their organs were prepared for detection of Leishmania and Crithidia species by molecular, microscopic, and culture methods. Results: Totally, 115 rodents of five species; Tatera indica (T. indica) (85), Rattus rattus (12), Meriones libycus (9), Mus musculus (7), and Rattus norvegicus (2), were trapped alive and their tissue samples were examined using microscopic, cultivation, and molecular assays. Overall, 59 (51.3%) rodents were positive for Leishmania or Crithidia parasites. The highest rate (61.2%; 52/85) of Leishmania infection was related to the T. indica population. The cultivation, and molecular observations showed that two (2.4%; 2/85) of T. indica (foot-pad, and spleen samples) were positive to Crithidia. Conclusions: This is the first report of Crithidia infection in T. indica in Iran. Consequently, more epidemiological and ecological studies are needed to understand the role of Crithidia and Leishmania in T. indica.
Objective: To explore the co-detection of natural infection of Trypanosomatidae parasites such as Leishmania and Crithidia in reservoir hosts of leishmaniasis. Methods: Rodent populations were monitored in two endemic foci of cutaneous leishmaniasis of Fars province, southern Iran from March to October 2016. Rodents were trapped alive in several parts of Shiraz and Kharameh cities. Afterwards, their organs were prepared for detection of Leishmania and Crithidia species by molecular, microscopic, and culture methods. Results: Totally, 115 rodents of five species; Tatera indica (T. indica) (85), Rattus rattus (12), Meriones libycus (9), Mus musculus (7), and Rattus norvegicus (2), were trapped alive and their tissue samples were examined using microscopic, cultivation, and molecular assays. Overall, 59 (51.3%) rodents were positive for Leishmania or Crithidia parasites. The highest rate (61.2%; 52/85) of Leishmania infection was related to the T. indica population. The cultivation, and molecular observations showed that two (2.4%; 2/85) of T. indica (foot-pad, and spleen samples) were positive to Crithidia. Conclusions: This is the first report of Crithidia infection in T. indica in Iran. Consequently, more epidemiological and ecological studies are needed to understand the role of Crithidia and Leishmania in T. indica.
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