Quantitative Autofluorescence Imaging of A375 Human Melanoma Cell Samples: A Pilot Study

Introduction: Skin cancer is one of the most common types of malignancy worldwide. Human skin naturally contains several endogenous fluorophores, as potential sources that can emit inherent fluorescence, called intrinsic autofluorescence (AF). The melanin endogenous fluorophore in the basal cell layer of the epidermis seems to have a strong autofluorescence signal among other ones in the skin. This pilot study aimed to investigate the feasibility of the detection of autofluorescence signals in the A375 human melanoma cell line in the cell culture stage using the FluoVision optical imaging system.Methods: The human skin melanoma cell line (A375) donated as a gift from Switzerland (University Hospital Basel) was cultured. For the imaging of the A375 human melanoma cell sample in this pilot study, the FluoVision optical imaging device (Tajhiz Afarinan Noori Parseh Co) was applied. The proposed clustering image processing code was developed based on the K-mean segmentation method, using MATLAB software (version 16).Results: The quantification of color pixels in the color bar along with the intensity score of the autofluorescence signal ranged between 0 and 70 was written in the image processing code execution and a threshold higher than 40%, proportional to the ratio of autofluorescent cells. The percentage of the signal of A375 autofluorescent melanoma cells in the 3 studied cell samples was calculated as 3.11%±0.6.Conclusion: This imaging method has the advantage of no need for fluorophore labels over the existing fluorescence imaging methods, and it can be regarded as one of the important choices of label-free imaging for this A375 melanoma cell line containing the intrinsic endogenous fluorophore in cell studies. DOI: 10.34172/jlms.2021.0

Experimental Study on Plasmodium berghei, Anopheles Stephensi, and BALB/c Mouse System: Implications for Malaria Transmission Blocking Assays

Background:Plasmodium berghei is a rodent malaria parasiteand has been very valuable means in the progress of our understanding of the essential molecular and cellular biology of the malaria parasites. Availability of hosts such as mice and vectors such as Anopheles stephensi has made this parasite a suitable system to study the parasite-host and vector-parasite relationships. Numerous studies have described life cycle and parameters influencing maintenance of the parasite within the mice or the mosquito. In this paper we revealed more details and have addressed some parameters and points influence maintenance of various life stages of the parasite (merozoites, macrogametocytes, ookinetes, oocysts and sporozoites) in the laboratory model P.berghei–A.stephensi-BALB/c mouse. This study helps understanding the biology of vertebrate-parasite and mosquito-malaria interactions that may aid in the development of a new generation of drug/vaccine and vector-based measures for malaria control

Three essays on family structure and school dropout among adolescents

This dissertation is a collection of three essays on high school dropout by adolescents and the roll of their siblings in this risky behavior. Dropping out of high school can scar the individual for his entire labor supply period with lower earnings, higher unemployment, and in turn incur considerable social and welfare costs to the economy. The data used in this study is from National Longitudinal Survey of Youth 1997, which has information on about 8,900 individuals aged 12-17 in the year 1997 who are followed up every year since. I use the first 10 rounds of this survey. The first essay in this dissertation identifies the influence of older siblings on their younger siblings’ decision to leave school before graduation. It starts by representing the significant adverse effect of birth order on school completion outcome of the teen. The results indicate that teens with older siblings are more at risk of dropping out of school. Having these initial estimates of the siblings’ effect, I look at the behavior of the siblings pairs in the in a subset of 519 pairs from families with two children for which the data is available on both siblings’ outcome and characteristics. I address the question of the inherent endogeneity by using family fixed effects models on these pairs, and also by implementing instrumental variables technique. I exploit the older sibling’s specific characteristics as an instrumental variable for his outcome in the younger sibling's equation. The sibling specific characteristic used as instruments are: the older sibling’s gender, the unemployment rate when the older sibling was 16 years old, the mother’s age at his birth, and the intactness of his family when he was 14 years old. The results of the estimation using this set of instruments show positive and significant effect on early school leaving of the younger sisters. The second essay utilizes a survival model to determine the timing pattern of teenager’s decision to drop out. Preventing and intervening in school dropping out in teens require the knowledge of the timing and pattern of occurrence of this act. I use nonparametric, semiparametric and parametric hazard models to explain the factors affecting the age onset of high school dropping out. Alongside with other socioeconomic factors, this chapter reinforces the roll of siblings on reducing the starting age of this risky behavior in the younger siblings. The results show that a teenager who has an older sibling is more likely to stop schooling at a younger age. In teenage boys who have an older sibling the hazard of dropping out is about 3 times as much depending on the choice of the hazard model for the time to first dropout. The shared frailty among siblings of the same family that contaminates the estimates is measured for the parametric and semiparametric duration models using the expected-maximization algorithm for the clustered data. The impact of shared frailty is ruled out in the case of Cox semiparametric models, but had been evident in some of the parametric models used in this chapter. The third essay develops a binary choice interaction model with finite number of agents to characterize the peer effect of siblings on the strategic choices of the teenager. This dynamic model incorporates the attractiveness of imitating the behavior of the peers inside the family. The model measures the strategic complementarity between the choice of the teenager and the choice of his or her siblings, after controlling for shared family fixed effects and utilizing the lagged dependent variable to reduce the unobserved contextual and correlated effect. This model finds significant social interaction effects between siblings, and specifically siblings that are closer in age to one another. The broad policy implication of this research is to indicate another important channel through which the strategic planning to reduce school dropout rate could be directed. School dropout prevention programs can put more emphasis on the first order or lower order children in multi-kid families to utilize the existing spillover effect on the younger siblings. Also considering this spillover effect, parents’ investment in raising a more scholarly firstborn might help them get an additional indirect return to their investment benefiting the other kids in the family.

Emergence of plasmid-mediated quinolone-resistant determinants in Klebsiella pneumoniae isolates from Tehran and Qazvin provinces, Iran

Background. Plasmid-mediated quinolone resistance is an increasing clinical concern, globally. The major objective of the present study was to identify the qnr-encoding genes among the quinolone non-susceptible K. pneumoniae isolates obtained from two provinces in Iran. Methods. A total of 200 K. pneumoniae isolates were obtained from hospitals of Qazvin and Tehran, Iran. The identification of bacterial isolates was carried out by standard laboratory methods and API 20E strips. Susceptibility to quinolone compounds were examined by standard Kirby-Bauer disk diffusion method according to the CLSI guideline. PCR and sequencing were employed to detect qnrA, qnrB and qnrS-encoding genes. Results. Of 200 K. pneumoniae isolates, 124 (62%) were nonsusceptible to quinolone compounds among those 66 (53.2%) and 58 (46.8%) isolates showed high and low-level quinolone resistance rates, respectively. Out of 124 quinolone non-susceptible isolates, qnr-encoding genes were present in 49 (39.5%) isolates with qnrB1 (30.6%) as the most dominant gene followed by qnrB4 (9.7%), and qnrS1 (1.6%) either alone or in combination. Conclusions. This study, for the first time, revealed the high appearance of qnrB1, qnrS1 and qnrB4 genes among the clinical isolates of K. pneumoniae in Iran. Therefore, the application of proper infection control measures and well-established antibiotic administration guideline should be strictly considered within our medical centers

Diagnosing Malaria Cases Referred to the Malaria Reference Laboratory in Tehran University of Medical Science, Iran

Background: The number of malaria cases is declining worldwide; however, it remains as a serious health problem. Diagnosing unusual cases is the most im­portant issue to manage the problem. This study designed to describe the number of falciparum and vivax malaria infected patients referred to Malaria Reference Labora­tory in Tehran University of Medical Science from 2000 to 2012. Methods: A retrospective study was conducted based on the collected question­naires from each patient referred to the laboratory. Diagnosing results and demo­graphic information for positive cases were analyzed using SPSS software. Problem­atic cases were evaluated for any difficulties in diagnosis or in clinical signs. Scanning and molecular methods were performed whenever there was an atypical case referred to the laboratory. Some of the samples had various difficulties for diagnosing such as presence of fussed gametocytes and schizonts of Plasmodium falciparum in peripheral blood and CCHF like hemoragic disorders. Results: Plasmodium vivax caused a large proportion of the cases (76.1%) in con­trast with P. falciparum that included smaller proportion (22.8%) and the rest (1.1) belonged to mixed infection. Most of the positive cases (69.6%) were belonged to Afghani people. Men (94.6%) showed more infection than women (5.4%), moreo­ver the most infection (44.5%) was seen at a range of 21-30 yr. Conclusion: In the case of existing atypical issues to diagnose, it is needed to per­form more precise microscopical examination beyond the current standard condi­tions. Sometimes molecular method is required to verify the exact agent of the dis­ease

Monitoring the Response of Plasmodium vivax to Chloroquine and Uncomplicated P. falciparum to Artesunate-fansidar Antimalarials in Southeastern Iran

Background: For many years, malaria was a major life-threatening parasitic infection in Iran. Although malaria elimination program is implementing in the country, still some cases annually are reported from malaria-endemic areas. Methods: This study was conducted in five malaria endemic districts of Sistan and Baluchistan Province, southeastern Iran, neighboring Afghanistan and Pakistan countries. Overall, 170 and 38 vivax malaria and falciparum malaria infected patients were enrolled in the study from 2013-2014. All the cases were selected according to criteria of the WHO guideline for in vivo drug sensitivity tests in malaria parasites. Evaluation of drug sensitivity test was conducted with some modifications. Results: The patients with vivax malaria responded to the regimen of chloroquine in 37.4(±15.9), 40(±13.8) and 42(±17.7) h for Pakistani, Iranian and Afghani nationalities respectively based on MPCT evaluation. The results showed a considerable difference between them and Iranian subjects. MPCT for the patients with falciparum malaria was calculated as 28(±18.05), 26(±12.03) and 36(±16.9) h for Iranian, Pakistani and Afghani nationalities respectively. There was a marginally significant difference between Afghani and other nationalities and between males and females. Conclusion: Treatment of all the patients resulted in ACPR and MPCT of P. vivax showed that the parasite became more sensitive to chloroquine than previous years in studied areas

Determination of Asymptomatic Malaria among Afghani and Pakistani Immigrants and Native Population in South of Kerman Province, Iran

Background: This study was proposed to monitor the situation of asymptomatic malaria among the native population and Afghani and Pakistani immigrants in Kahnooj and Ghale-Ganj districts from Kerman Province, Southeastern Iran. Methods: A number of 180 and 120 individuals from Kahnooj and Ghale-Ganj respectively were registered and considered based on a cross-sectional surveillance method. From 300 registered cases, 200 individuals (66.7%) were selected among Afghani and Pakistani immigrants and the rest (33.3%) were native resident individuals. All samples were processed with employing microscopical examination, Rapid Diagnostic Tests (RDTs) and Semi- nested Multiplex PCR techniques. Results: None of the samples collected from native residents showed any malaria parasite, but among Afghani immigrants, one asymptomatic vivax malaria was detected in a 12 yr old girl with 280 parasites per microliter of blood. Moreover, one symptomatic vivax malaria was detected from a Pakistani immigrant with 47560 parasites per microliter of blood. All results obtained via microscopical method, confirmed by RDTs and PCR techniques. Conclusion: To achieve the malaria elimination program different studies are needed that to be performed. Monitoring the asymptomatic malaria in all over the malaria endemic areas especially among the immigrant individuals is the most crucial necessity.

Interaction between Chitosan and Chloroquine against Plas-modium berghei and P. falciparum using In-Vivo and In-Vitro Tests

Background: The use of antimalarial drugs with number of compounds in combination form may potentiate each other's activity. Methods: This study was conducted in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2018. It was based on two methods including in vivo and in vitro tests with aim of considering interaction between chitosan and chloroquine against Plasmodium berghei and P. falciparum parasites using different ratios of the agents with ED50s and IC50s baselines. Results: Administrating 10 and 20 mg/kg (mouse body weight) of chitosan alone to the P. berghei –infected mice up to 4 successive days resulted in 37% and 45% inhibition of P. berghei respectively, while employing the compound with chloroquine in combination form with ratios of 90/10 and 70/30 (chloroquine/chitosan) had a considerable potentiation including 71.58% and 83.85% inhibition effectiveness against P. berghei. Moreover, 20 mg/L (CCM) concentration of chitosan alone could eliminate 69.55% of P. falciparum in culture medium while in combination with chloroquine in ratios of 90/10 (chloroquine/chitosan) had considerable potentiation including 79.14% inhibition effectiveness. Mean survival time of those mice received combination therapy in ratios of 90/10 and 70/30 (chloroquine/chitosan) was longer than those took up mono therapy of either chloroquine or chitosan based on their ED50s doses. Conclusion: Interaction between chloroquine and chitosan showed considerable potentiation in combination form against either P. berghei or P. falciparum using in vivo and in vitro tests respectively. Meanwhile, interaction between the above mentioned agents resulted in a notable survival time for those P. berghei-infected mice treated with the combination

Endoparasites of Wild Rodents in Southeastern Iran

Background: This study was aimed to collect wild rodents for endoparasites determination in some parts of Sistan and Baluchistan Province, southeastern Iran nearby Pakistan and Afghanistan countries. Methods: A total of 100 wild rodents were captured alive with cage traps. Various samples were collected from blood and feces, also impression smear prepared from different organs. The samples were prepared by formalin-ether or stained with Giemsa, after that were examined under microscope. Results: All the caught rodents (47 Tatera indica, 44 Meriones hurriana, 5 Gerbilus nanus and 4 Meriones libycus) were studied for endoparasites emphasizing to their zoonotic aspects. Endoparasites including Spirurida, Hymenolepis diminuta, Hymenolepis nana feraterna, Trichuris trichiura, Skerjabino taenia, Trichostrongylus spp, Entamoeba muris, Chilomastix mesnili and Leishmania spp were parasitologically identified. Conclusion: Among 9 genera or species of the identified parasites at least 5 of them have zoonotic and public health importance

Antimalarial activity of Alcoholic Extract of Curcuma longa and Heracleum persicum on Cultivated Plasmodium falciparum 3D7 strain: Antimalarial Effect of C. longa and H. persicum

Introduction: Plasmodium falciparum causes the most fatal form of malaria in human. At present, the common treatments are not effective enough and the incidence of drug resistance is being increased in malarious areas. Therefore, presenting the novel methods for therapeutic purposes assumes significant importance.  Recent studies indicated that aqueous or alcoholic extracts of Curcuma longa and Heracleum persicum showed a broad spectrum of anti-microorganisms activity. In this in vitro study the effects of C. longa and H. persicum extracts were assessed on P. falciparum since there has been limited clinical research into their effectiveness on Malaria. Materials and Methods:  The alcoholic extracts of H. persicum and C. longa were prepared in 10-1, 10-3 and 10-5 mg/ml dilutions. These solutions were tested on P. falciparum with 10% parasitemia in RPMI 1640 medium with 10% hematocrit. Each of dilutions was examined in triplicate and the inhibitory effect of the solutions on parasites was measured via determining the average parasitemia and their schizont rate. Finally, the results were analyzed using SPSS software. Results: The rate of parasitemia declined in three different dilutions of both H. persicum and C. longa. The mean of antiplasmodial inhibitory activity of herbs was 83.23±2.47% in H.persicum and 99.91±0.0% in C.longa. Moreover, all dilutions of both H.persicum and C.longa showed significant effect on decreasing of schizont percentage in comparison with control group (P-value<0.05). Conclusions: the present study indicated that alcoholic extracts of C. longa and H. persicum possess acceptable antiplasmodial effects and could be developed as valuable alternatives to ineffective antimalarial drugs. These results support the claims of recent studies that C. longa and H. persicum, have considerable antimicrobial activities. Considering notable in vitro antiplasmodial efficacy of C.longa and H.persicum, further studies with in vivo method is recommended