Comparative proteomic profiling of Leishmania tropica: investigation of a case infected with simultaneous cutaneous and viscerotropic leishmaniasis by 2-dimentional electrophoresis and mass spectrometry

Background: Viscerotropic leishmaniasis caused by Leishmania tropica poses a significant prob­lem in the diagnosis and treatment management. Since differential gene expression is more im­portant in outcome of the infection, we employed proteomic approach to identify potential pro­teins involved in visceralization of L. tropica. Methods: The proteomes profiling of L. tropica isolated from cutaneous and visceral tissues of one host were compared by 2-DE/MS proteomics study. Moreover, the transcript level of some identified proteins was confirmed using real-time RT-PCR. Results: Of the 700 protein spots that were detected reproducibly on each gel, 135 were found to be differentially expressed (P≤ 0.05). Most of responsive proteins in visceral isolate changed in less abundant compared to cutaneous isolate. Among differentially expressed proteins, 56 proteins were confidently identified and classified according to the biological process. The larg­est groups consist of proteins involved in carbohydrate metabolism and protein synthesis. Most of the identified proteins, which implicated in energy metabolism, cell signaling and virulence were down-regulated, whereas some proteins that have a role in protein folding, antioxidant defense and proteolysis were up-regulated in visceral form. Moreover, the transcript level of some identified proteins such as co-chaperon was confirmed using real-time RT-PCR. Conclusion: L. tropica probably uses different mechanisms for survival and multiplication in viscera to establish viscerotropic leishmaniasis. The current study provides some clues into the mechanisms underlying the dissemination of L. tropica

Comparative proteomic profiling of Leishmania tropica: investigation of a case infected with simultaneous cutaneous and viscerotropic leishmaniasis by 2-dimentional electrophoresis and mass spectrometry

Background: Viscerotropic leishmaniasis caused by Leishmania tropica poses a significant prob­lem in the diagnosis and treatment management. Since differential gene expression is more im­portant in outcome of the infection, we employed proteomic approach to identify potential pro­teins involved in visceralization of L. tropica. Methods: The proteomes profiling of L. tropica isolated from cutaneous and visceral tissues of one host were compared by 2-DE/MS proteomics study. Moreover, the transcript level of some identified proteins was confirmed using real-time RT-PCR. Results: Of the 700 protein spots that were detected reproducibly on each gel, 135 were found to be differentially expressed (P≤ 0.05). Most of responsive proteins in visceral isolate changed in less abundant compared to cutaneous isolate. Among differentially expressed proteins, 56 proteins were confidently identified and classified according to the biological process. The larg­est groups consist of proteins involved in carbohydrate metabolism and protein synthesis. Most of the identified proteins, which implicated in energy metabolism, cell signaling and virulence were down-regulated, whereas some proteins that have a role in protein folding, antioxidant defense and proteolysis were up-regulated in visceral form. Moreover, the transcript level of some identified proteins such as co-chaperon was confirmed using real-time RT-PCR. Conclusion: L. tropica probably uses different mechanisms for survival and multiplication in viscera to establish viscerotropic leishmaniasis. The current study provides some clues into the mechanisms underlying the dissemination of L. tropica

Global burden of 288 causes of death and life expectancy decomposition in 204 countries and territories and 811 subnational locations, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

BACKGROUND Regular, detailed reporting on population health by underlying cause of death is fundamental for public health decision making. Cause-specific estimates of mortality and the subsequent effects on life expectancy worldwide are valuable metrics to gauge progress in reducing mortality rates. These estimates are particularly important following large-scale mortality spikes, such as the COVID-19 pandemic. When systematically analysed, mortality rates and life expectancy allow comparisons of the consequences of causes of death globally and over time, providing a nuanced understanding of the effect of these causes on global populations. METHODS The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 cause-of-death analysis estimated mortality and years of life lost (YLLs) from 288 causes of death by age-sex-location-year in 204 countries and territories and 811 subnational locations for each year from 1990 until 2021. The analysis used 56 604 data sources, including data from vital registration and verbal autopsy as well as surveys, censuses, surveillance systems, and cancer registries, among others. As with previous GBD rounds, cause-specific death rates for most causes were estimated using the Cause of Death Ensemble model-a modelling tool developed for GBD to assess the out-of-sample predictive validity of different statistical models and covariate permutations and combine those results to produce cause-specific mortality estimates-with alternative strategies adapted to model causes with insufficient data, substantial changes in reporting over the study period, or unusual epidemiology. YLLs were computed as the product of the number of deaths for each cause-age-sex-location-year and the standard life expectancy at each age. As part of the modelling process, uncertainty intervals (UIs) were generated using the 2·5th and 97·5th percentiles from a 1000-draw distribution for each metric. We decomposed life expectancy by cause of death, location, and year to show cause-specific effects on life expectancy from 1990 to 2021. We also used the coefficient of variation and the fraction of population affected by 90% of deaths to highlight concentrations of mortality. Findings are reported in counts and age-standardised rates. Methodological improvements for cause-of-death estimates in GBD 2021 include the expansion of under-5-years age group to include four new age groups, enhanced methods to account for stochastic variation of sparse data, and the inclusion of COVID-19 and other pandemic-related mortality-which includes excess mortality associated with the pandemic, excluding COVID-19, lower respiratory infections, measles, malaria, and pertussis. For this analysis, 199 new country-years of vital registration cause-of-death data, 5 country-years of surveillance data, 21 country-years of verbal autopsy data, and 94 country-years of other data types were added to those used in previous GBD rounds. FINDINGS The leading causes of age-standardised deaths globally were the same in 2019 as they were in 1990; in descending order, these were, ischaemic heart disease, stroke, chronic obstructive pulmonary disease, and lower respiratory infections. In 2021, however, COVID-19 replaced stroke as the second-leading age-standardised cause of death, with 94·0 deaths (95% UI 89·2-100·0) per 100 000 population. The COVID-19 pandemic shifted the rankings of the leading five causes, lowering stroke to the third-leading and chronic obstructive pulmonary disease to the fourth-leading position. In 2021, the highest age-standardised death rates from COVID-19 occurred in sub-Saharan Africa (271·0 deaths [250·1-290·7] per 100 000 population) and Latin America and the Caribbean (195·4 deaths [182·1-211·4] per 100 000 population). The lowest age-standardised death rates from COVID-19 were in the high-income super-region (48·1 deaths [47·4-48·8] per 100 000 population) and southeast Asia, east Asia, and Oceania (23·2 deaths [16·3-37·2] per 100 000 population). Globally, life expectancy steadily improved between 1990 and 2019 for 18 of the 22 investigated causes. Decomposition of global and regional life expectancy showed the positive effect that reductions in deaths from enteric infections, lower respiratory infections, stroke, and neonatal deaths, among others have contributed to improved survival over the study period. However, a net reduction of 1·6 years occurred in global life expectancy between 2019 and 2021, primarily due to increased death rates from COVID-19 and other pandemic-related mortality. Life expectancy was highly variable between super-regions over the study period, with southeast Asia, east Asia, and Oceania gaining 8·3 years (6·7-9·9) overall, while having the smallest reduction in life expectancy due to COVID-19 (0·4 years). The largest reduction in life expectancy due to COVID-19 occurred in Latin America and the Caribbean (3·6 years). Additionally, 53 of the 288 causes of death were highly concentrated in locations with less than 50% of the global population as of 2021, and these causes of death became progressively more concentrated since 1990, when only 44 causes showed this pattern. The concentration phenomenon is discussed heuristically with respect to enteric and lower respiratory infections, malaria, HIV/AIDS, neonatal disorders, tuberculosis, and measles. INTERPRETATION Long-standing gains in life expectancy and reductions in many of the leading causes of death have been disrupted by the COVID-19 pandemic, the adverse effects of which were spread unevenly among populations. Despite the pandemic, there has been continued progress in combatting several notable causes of death, leading to improved global life expectancy over the study period. Each of the seven GBD super-regions showed an overall improvement from 1990 and 2021, obscuring the negative effect in the years of the pandemic. Additionally, our findings regarding regional variation in causes of death driving increases in life expectancy hold clear policy utility. Analyses of shifting mortality trends reveal that several causes, once widespread globally, are now increasingly concentrated geographically. These changes in mortality concentration, alongside further investigation of changing risks, interventions, and relevant policy, present an important opportunity to deepen our understanding of mortality-reduction strategies. Examining patterns in mortality concentration might reveal areas where successful public health interventions have been implemented. Translating these successes to locations where certain causes of death remain entrenched can inform policies that work to improve life expectancy for people everywhere. FUNDING Bill & Melinda Gates Foundation

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

اثرات صدای بنفش بر روی میزان بیان ژن SLC26A4در بافت حلزون گوش رت

Background and Aims: As one of the most important physical detrimental factors, purple noise can be considered similar to sounds that a wide range of people are exposed to in their workplaces. Also, cochlear tissue samples are the best tissue samples to evaluate gene expression and pathologic studies following noise exposure. Therefore, the aim of this study was to investigate the effects of purple noise on the expression of SLC26A4 gene in cochlear tissue. Methods & material: Totally, 10 male Wistar rats were used in this experimental study. Both N1 and N2 exposure groups were exposed to purple noise with sound pressure level of 115-120 dB and frequency range of 4-20 kHz. Histological tests were performed for pathological studies. Finally, the relative expression of SLC26A4 gene was determined by qRT-PCR technique. All experiments were conducted according to ethical standards of working with laboratory animals. Results: The results showed that the expression of SLC26A4 gene decreased significantly compared to the normal level in both N1 and N2 groups. The results of cochlear tissue pathology showed that the group exposed to purple noise for 6 days had more mechanical damage 7 days after cessation of exposure. Conclusion: Significant decreased expression in SLC26A4 gene and permanent damage to the Reissner and Basal lamina membranes in the cochlear tissue and the auditory nerve ganglion leads to strengthening the incidence of advancing sensorineural hearing impairment and increases the likelihood of metastasis in cochlear tissue.زمینه و اهداف: صدای بنفش را می­توان مشابه اصواتی در نظر گرفت که طیف وسیعی از افراد به عنوان یکی از مهم­ترین عوامل فیزیکی زیان­ آور، در محیط­های کاری خود در تماس با آن هستند. از طرفی، نمونه­ی بافت حلزون گوش بهترین نمونه بافتی جهت بررسی میزان بیان ژن­ و مطالعات آسیب شناسی، پس از تماس با اصوات محسوب می­شوند لذا هدف از این پژوهش، بررسی اثرات صدای بنفش بر روی میزان بیان ژن  SLC26A4در بافت حلزون گوش بود.   مواد و روش­ها: در مجموع، 10 سر رت نر با نژاد ویستار در این مطالعه تجربی استفاده گردید. دو گروه مواجههN1  و N2 در تماس با صدای بنفش (kHz20-4، dB120-115=SPL) قرار گرفتند. آزمایشات بافت شناسی جهت مطالعات آسیب شناسی بافت صورت گرفت و در پایان، میزان بیان نسبی ژن SLC26A4 بوسیله ی qRT-PCR تعیین گردید. کلیه آزمایشات این مطالعه، طبق موازین اخلاقی کار با حیوانات آزمایشگاهی انجام گرفت.   یافته ها: نتایج نشان داد که در هر دو گروه مواجهه N1 و N2 کاهش معنادار میزان بیان ژن SLC26A4 نسبت به سطح نرمال اتفاق افتاده است. نتایج مطالعه­ی آسیب­ شناسی بافت حلزون گوش نشان داد که نمونه ی گروهی که 6 روز با صدای بنفش مواجهه داشت، پس از 7روز از قطع مواجهه صدمات مکانیکی بیشتری دیده بود.   نتیجه­گیری: این سطح از کاهش بیان در ژن SLC26A4 و آسیب دائمی غشای رایسنر و بازال لامینا در بافت حلزون گوش و گانگلیون عصب شنوایی، منجر به تقویت بروز نقص شنوایی حسی-عصبی پیش رونده و افزایش احتمال متاستاز در بافت حلزون گوش می­گردد

Treatment Failure in Cutaneous Leishmaniasis Patients Referred to the School of Public Health, Tehran University of Medical Sciences During 2008–2017

Background: Cutaneous leishmaniasis (CL) is a vector borne disease predominantly found in tropical and subtropical countries, including Iran. For more than 6 decades, pentavalent antimonials have been used successfully worldwide for the treatment of leishmaniasis, but over the past few years, clinical resistance to these medications has increased. In this study, we evaluated CL patients who did not show any desirable responses to the anti-leishmanial treatment within a 10-year period (2008 to 2017). Methods: All patients from different parts of Iran suspected of having cutaneous leishmaniasis, who were referred to the laboratory of leishmaniosis in Tehran University of Medical Sciences from 2008–2017 were parasitological exam­ined. Results: During this period, a total of 1480 suspected CL patients were referred to the laboratory of leishmaniosis. Samples from 655 patients (70.8%) suspected of having CL were positive microscopically. The failure rate in patients treated with anti-leishmaniasis medications for a minimum of three complete treatment periods was 1.83% (12 cases). There was no association between the number and size of skin lesions and patient characteristics. Also, the route of drug administration had no significant effect on the number and size of lesions. Conclusion: In the present study, treatment failure was found in some confirmed CL patients treated with meglu­mine antimoniate. Over the past few years, it seems that had been increased in resistance to these medications. So, a review of the correct implementation of the treatment protocol and/or a combination therapy may be helpful in prevent­ing an increase in the rate of treatment failure

Comparison of p27 Gene Expression of Promastigote and Amastigote Forms of Leishmania major (MRHO/IR/75/ER) by Real-time RT-PCR

Background: Cutaneous leishmaniasis (CL) is one of the world health problems. Leishmania major is the etiological agent of zoonotic cutaneous leishmaniasis (ZCL). Promastigote and amastigote are two morphological forms of Leishmania parasites that express different proteins and p27 is an important gene encoding cytochrome c oxidase (COX) component. P27 gene expresses a 27 kDa protein that essential in ATP synthesis. This study aimed to compare p27 gene expression in promastigote and amastigote forms in Iranian strain of L. major (MRHO/IR/75/ER). Methods: This study was conducted in 2015. Clinical isolates of CL patients from north, center, west and south parts of Iran were collected and identified by PCR-RFLP. After RNA extraction of promastigotes and amastigotes and cDNA synthesis, the expression level of p27 gene was compared by real-time RT-PCR. Results: By comparison of expression level between amastigote and promastigote forms of Iranian strain of L. major, up-regulation of p27 gene (2.73 fold) was observed in amastigotes. Moreover, there was no significant difference in p27 gene expression between L. major isolates. Conclusion: p27 gene and protein can be considered as a target in recombinant vaccine production and treatment process