Detection of Adenoviruses from clinical samples in bone marrow transplant patients by nested PCR (polymerase chain reaction)

Adenoviruses are recognized as common human pathogens that are responsible for a wide variety of infectious syndromes. Bone marrow transplant patients are prone to life threatening opportunistic infections like adenoviruses. The nested polymerase chain reaction has provided an alternative, sensitive diagnostic method for detection of Adenoviruses. In this study we developed PCR from hexon genes as rapid diagnostic method of Advs infections on different clinical samples. Adenovirus infections was defined as the presence of DNA in the blood, urine, stool, or respiratory lavage from bone marrow transplant patients. Two sets of primers (Group specific primers and internal primers) were required to optimize the PCR protocol. This highly sensitive method could detect different types of Advs in two separate sets of PCR. Therfore,DNA amplification in BMT patients would be valuable screening way to evaluate bone marrow transplant recipients. Early detection of Advs by PCR assay is important to asymptomatic infections or preventing aggressive antiviral thearapy

The Development a Polymerase Chain Reaction (PCR) to detect C.Pneumoniae and C.Psittaci

     C.pneumoniae and C.psittaci both cause respiratory infections in human. detection  of  these  organisms  in  tissue  culture  is  difficult  and serological  testing  is unreliable. There  are  no  sensitive  and  reliable  tests  for  the  detection  of  these  organisms. The  polymerase  chain reaction  (PCR  )  has  provided  an alternative  diagnostic  method  for the detection of these fastidious  organisms. The aim of this study was to develop a PCR to detect C.pneumoniae and C.psittaci from clinical samples. The PCR was optimized in a series of experiments. To determine  if  the  optimized  PCR  could  be applied  to  clinical samples, mock  positive specimen  were  produced by  adding  chlamydiae  to  throat  swab  from  healthy  adults. The DNA was extracted by phenol/chloroform. When tested by PCR all the throat swabs were negative. However,  when  diluted  and  retested,  many  of the  swabs  were  positive  and 10 IFU  of C.psittaci  and  100  IFU  of C.pneumoniae  could  be   detected.  This  experiment  indicates  that  inhibitor  to  PCR  are  found  in  throat  swab  and  further  work  is  needed   on  specimen  preparation.   The PCR was optimized in a series of experiments.    The  optimal  conditions  were  to  use  a two  segment  PCR  with  68°c  annealing  and  polymerization  temperature, 2.0mM  Mgcl2,  0.2μM  primers  and  40  cycles. The  PCR   was  highly  sensitive  and  could  detect  one  inclusion  forming  unit  with  both  C.pneumoniae   and  C.psittaci  strains.  Two  human  strains  and  one   nonhuman   strain  of  C.pneumoniae   and   two  avian   strains   and  three  mammalian   strains   of C.psittaci   were   used   to  determine   the specificity  of PCR. The PCR detected all these different strains.   C.trachomatis   strains   were not detected.  Various bacterial  strains, fungi  DNA,  and  human  DNA  were  negative  in  PCR  and no  amplification  DNA  was  found   in  negative  controls

T cell expansion from umbilical cord blood without thymic stroma cells after stimulation with SCF, IL-7, AND IL-2

We analyzed in vitro expansion and differentiation of T progenitor cells from umbilical cord blood in the absence of thymic epithelium. The expansion setup is performed in the presence of SCF, IL-7 , and IL -2 with autologous serum .Using CBMCs as initial source , we compared the growth kinetics of several cell populations in either whole CBMC or  CD34+ -enriched-, as well as in CD3CD4CD8-depleted expansion assays by FACS analysis. After 11 days of culture, cell increase values were about 7 fold for CD3+, 6 fold for CD3+CD4+, 7 fold for CD3+CD8+, 4fold for CD3+CD56, 6fold for CD56+, and 0.2 fold for CD34+. We characterized the developmental state of these cell populations by RT –PCR analysis of the lymphoid differentiation markers RAG-1 and pre T-Alpha. In all samples , transcripts of both markers could be detected from day 0 though day 11, however , in case of pre – T-Alpha,  nested PCR  was always required , indicating lower expression . These findings; therefore, demonstrate that T-cell differentiation events (as opposed to mere expansion) do occur in stroma cell free expansion assays

بررسی تاثیر همزمان ملاتونین، ال-کارنتین و آتورواستاتین بر میزان بروز آپوپتوز در اووسیت‌های کشت داده شده موش سوری در محیط آزمایشگاه (In vitro)

چكيده: مقدمه: ناباروري يك بيماري شايع است كه حدود 10 الی 15 درصد زوج ها را تحت تاثير قرار مي‌دهد (آمار سال 2020). اين درحالي است كه تقريبا 50 درصد از زوج هايي كه IVF را شروع مي‌كنند، بدون فرزند باقي مي‌مانند. عوامل مختلفي در پيش‌بيني حاملگي توسط IVF تاثير دارد كه يكي از آن‌ها كيفيت تخمك است. ملاتونين، ال – كارنتين و آتورواستاتین، به عنوان عوامل مهاركننده راديكال آزاد، آنتي اكسيدان و ضد آپوپتوز به بهبودي رشد تخمك كمك مي‌كنند. لذا در مطالعه‌ي حاضر اثرات احتمالي تركيب اين سه عامل آنتي‌اكسيدان بر آپوپتوز تخمك موش مورد بررسي قرار گرفت. روش: 40 راس موش سوري ماده نژاد NMRI به صورت تصادفي به پنج گروه تقسيم شدند. در هر گروه تحريك تخمك گذاري با 5/7 واحد PMSG انجام شد، و پس از 24 ساعت موش‌ها به روش كشش گردني كشته شده و تخمك‌ها از تخمدان‌هاي موش خارج شدند و با تركيب ملاتونين و ال – كارنتين و آتورواستاتین درمان شدند. در نهایت آپوپتوز تخمك‌ها با استفاده از ميكروسكوپ معکوس و آزمون TUNEL ارزيابي شدند و داده‌ها با استفاده از نرم افزار آماري SPSS و آزمون آناليز ANOVA و TUKEYدر سطح معني‌داري P<0.05 آناليز شدند. يافته ها: نتايج نشان داد تعداد تخمك‌هاي بالغ شده در تمام گروه‌هاي مورد مطالعه نسبت به گروه كنترل افزايش داشته و بيشترين افزايش بلوغ در گروه ترکیبی (ملاتونين + ال – كارنتين + آتورواستاتین) و 83 درصد بود. همچنين نتايج TUNEL نشان داد كه كمترين ميزان آپوپتوز نیز در گروه ترکیبی (ملاتونين+ ال – كارنتين + آتورواستاتین) و10 درصد بود. نتيجه گيری: نتایج نشان داد از نظر آماری تفاوت معناداری از نظر بلوغ و آپوپتوز در گروه‌های درمان شده با آنتی‌اکسیدان وجود دارد. اطلاعات به دست آمده استفاده از آنتی اکسیدان‌ها، ترجیحا به صورت ترکیبی (ملاتونين+ ال – كارنتين+ آتورواستاتین) را توصیه می‌کند. واژگان كليدي: بلوغ آزمايشگاهي (IVM)، ملاتونين، ال – كارنتين، آتورواستاتین، تخم

Improving milk donation behavior through an educational intervention based on the theory of planned behavior: study protocol of a cluster randomized controlled trial

Abstract Background Breastfeeding mothers’ decisions to donate their milk is influenced by their knowledge, attitudes, and subjective norms such as their family’s opinions on milk donation. In Iran, women have favorable knowledge and positive attitudes toward breastfeeding, but they lack sufficient information and education about milk banks. With respect to current childbearing policies, an increase in the number of infants who will be admitted to neonatal intensive care units is expected. Accordingly, improving milk donation behavior, which requires mothers’ intention and ability to donate breast milk, is important for infants’ survival and well-being. Therefore, this study aims to evaluate whether an educational program based on the theory of planned behavior affects breastfeeding mothers’ decisions to donate their breast milk. Methods This cluster randomized controlled trial will be conducted in health centers affiliated to Tehran University of Medical Sciences and will enroll 66 breastfeeding mothers (intervention, n = 33; control, n = 33). After collecting baseline data, the intervention group will begin receiving a 60-minute weekly educational program based on Theory of Planned Behavior components for four weeks. The first follow-up assessment will be conducted immediately after the intervention, and the final one will be conducted 12 weeks later. The primary outcomes are the number of mothers who have donated their breast milk and changes in breastfeeding mothers’ knowledge, attitude, subjective norms, and perceived behavioral control about donor human milk and intention to donate milk. Discussion This trial will evaluate if a well-designed educational program can improve breastfeeding mothers’ knowledge and promote their behavioral factors regarding milk donation and lead to a significant increase in the number of potential milk donors. Trial registration irct.ir (IRCT20230124057203N1) registered February 14, 2023

The measurement of polychlorinated biphenyl level in breast milk after delivery, and its association with breast milk fat and mother\'s BMI

Background: Human breast milk contains high levels of fat and toxins. These substances which exist in our country based on many reports. This study aimed to evaluate the concentration of polychlorinated biphenyls (PCBs) in the mother&rsquo;s milk during the first postnatal week. Methods: This study was accomplished on 50 mothers who gave birth to a healthy infant at Vali-e-Asr University Hospital, Tehran University of Medical Sciences, Tehran, Iran, from 2014 through 2015. Within the first postnatal week a 20 cc sample of mother&rsquo;s breast milk was obtained and was sent for laboratory analysis to measure PCB levels via GC-Mass method. Except of PCBs, triglycerides (TG), high-density cholesterol in breast milk was also measured. Also the relation between fat and polychlorinated bi phenyl was assessed. The association between PCBs and body mass index of mothers was investigated. Results: Mean age of mothers was 29.41&plusmn;6.14 year. PCB180 was significantly correlated with TG values, while this correlation was not observed for other types of PCBs. There was no correlation between cholesterol and fat high-density with PCBs. Other PCB isomers 138, 153, 180 and 101, had not correlation with triglyceride, cholesterol and high density fat. There was a significant correlation between increasing weight and the concentration of PCB101, and PCB180 concentrations. It was also observed between increasing weight and the concentration of PCB101 (P=0.033, r=0.293) and concentrations of PCB180 (P=0.034, r=0.428), there was a significant correlation. In our study PCB101, (r=0.278, P=0.048) and PCb28 (r=0.328, P=0.021) were significantly correlated with body mass index (BMI). Conclusion: The present study showed a higher than normal concentration of polychlorinated biphenyls in breast milk. And therefore these compounds will be transferred to the baby. Due to the damaging effects of these toxins, it is recommended additional studies on larger groups and other types of samples (mother blood, cord blood etc.)

association breast milk fat and BMI with levels of pollutantsPCB (polyclorinatedbiphenil)in milk of nursing mothers

Background: Human breast milk contains high levels of fat, toxins, these substances which exist in our country based on many reports.this study aimed to evaluate the concentration of PCBs in the mother&rsquo;s milk during the first postnatal week. Methods: this study was conducted on 50 mothers who gave birth to a healthy infant at Vali-Asr teaching hospital, Tehran University of Medical Sciences, Tehran, Iran, from 2014 through 2015. Within the first postnatal week a 20 cc sample of mother&rsquo;s breast milk was obtained and was sent for laboratory analysis to measure PCB levels via GCMassmethod.Using the CHi-Square test and Man whitney relationship between the mother's milk fat and PBCs was evaluated. Results:Mean age of mothers were 29.41 &plusmn; 6.14 year.PCB180 was significantly correlated with TG values, while this correlation was not observed for other types of PCBs. There was a significant correlation between increasing weight and the concentration of PCB101, and PCB180 concentrations. It was also observed between increasing weight and the concentration of PCB101 (p = 0.033, r = 0.293) and concentrations of PCB180 (p = 0.034, r = 0.428), there was a significant correlation. In our studyPCB101(p=0.048 ,r=0.278) and PCB28(p=0.021, r=0.328) were significantly correlated with BMI.&nbsp; Conclusions:The present study showed a higher-than-normal concentration of PCBs in breast milk. And therefore these compounds will be transferred to the baby. Due to the damaging effects of these toxins, it is recommended measures be taken to solve this problem