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Introduction: The human Y chromosome contains genes that are essential for spermatogenesis specially those that are located on Yq11.23 in a rejoin called azoospermia factor locus (AZF). Deletions in these genes may result in nonobstructive azoospermia and oligozoospermia. Widely different frequencies of such deletions (0-55%) have been reported from different populations. The aim of this study was to establish the prevalence of microdeletions on the Y chromosome in infertile Iranian males with azoospermia. Materials/Methods: Multiplex polymerase chain reaction (PCR) was applied using several sequence-tagged site (STS) primer sets, in order to determine Y chromosome microdeletions in 75 infertile males. Results: Microdeletions in AZFa, AZFb and AZFc (DAZ gene) regions were detected in 10 of the 75 (13.3%) infertile males with normal karyotypes, which seems to be one of the greatest reported frequencies. Conclusion: Our findings suggest that knowing the genetic cause for male infertility will be informative before performing intracytoplasmic sperm injection or testicular sperm extraction treatment. [ABSTRACT FROM AUTHOR]

Objective: Pharmacopoeias regulate the manufacture of potentised pharmaceutical preparations used in different branches of complementary and integrative medicine. The physicochemical properties and biological activity of these preparations are often investigated in preclinical research, yet no guidelines for experimental research currently exist in this area. The present PrePoP guidelines aim to provide recommendations to promote high-quality, statistically sound, and reproducible preclinical research on potentised preparations., Methods: Input was gathered from researchers nominated by the relevant scientific societies using a simplified Delphi consensus approach covering the most relevant aspects of basic research methodology in the field including appropriate controls, sample preparation and handling, and statistics. After three rounds of feedback, a consensus was finally reached on the most important aspects and considerations for conducting high-quality research on potentised preparations., Results: We present a series of recommendations on a range of topics including experimental controls, system stability, blinding and randomisation, environmental influences, and procedures for the preparation of potentised samples and controls, and we address some specific challenges of this research field., Conclusion: This expert consensus process resulted in a robust set of methodological guidelines for research on potentised preparations and provides a valuable framework that will inform and improve the quality of subsequent research in this emerging field., Please Cite This Article as: Tournier AL, Bonamin LV, Buchheim-Schmidt S, Cartwright S, Dombrowsky C, Doesburg P, Holandino C, Kokornaczyk MO, van de Kraats EB, López-Carvallo JA, Nandy P, Mazón-Suástegui JM, Mirzajani F, Poitevin B, Scherr C, Thieves K, Würtenberger S, Baumgartner S. Scientific guidelines for preclinical research on potentised preparations manufactured according to current pharmacopoeias-the PrePoP guidelines. J Integr Med. 2024; 22(5): 533-544., (Copyright © 2024 Shanghai Yueyang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine. Published by Elsevier B.V. All rights reserved.)

Purpose: This article deals with producing ultra-diluted compounds (UHDs) prepared from Iranian calendula's characteristic and endemic species. It compares their chemical, biological and biochemical characteristics with the commercial sample of calendula species (grown in the Alps). In the following, these UHDs have been used to improve the quality of germination and growth and reduce contamination of rice ( Oryza sativa ) seeds in the laboratory environment., Methods: High-performance thin-layer chromatography (HPTLC) is used to isolate the active compounds. On the separated results, antioxidant and antibacterial were identified directly on the plate (Bio-autographic method). Direct on the plate)DESI mass spectrometry was used to identify the active compounds., Results: The HPTLC reveals that the chromatogram of native C. percica and C. officinalis extract is the most similar to the commercial compounds. The highest antioxidant activity is related to C. officinalis . The best antibacterial activity of the extracts against Staphylococcus aureus and Escherichia coli belongs to C. officinalis and C. tripterocarpa . Rutin, quercitrin, β-campstrole and di- o -caffeoylquinic acid, which are among the flavonoid and terpenoid categories were identified as active compounds. The prepared UHDs from native calendula are biologically more effective than the commercial ones in increasing seed germination efficiency, improving rooting quality and reducing contamination., Conclusion: Using UHDs increases the production of photosynthetic pigments the root length and the number of lateral roots. Also, the amount of protein, gibberellic acid and abscisic acid in seedlings treated using native UHDs of C. officinalis (native or commercial) is higher than the others., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Hassan Rezadoost reports financial support was provided by 10.13039/501100003968Iran National Science Foundation (INSF) [Project No. 99012038]. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

In this study, a proper and reliable fluorometric method is introduced for screening acetylcholinesterase (AChE) and its inhibitors, using carbon quantum dots (CQDs) as the signal reporter. Pure, S-doped, and P-doped CQDs, were synthesized and their recoverable fluorescence quenching properties were observed, when exposed to Hg 2+ , Cu 2+ , and Fe 3+ quenching ions, respectively. The study on the recovery of their emission showed that after the introduction of another guest substance with a stronger affinity to the quenching ions, their fluorescence is restored. The Design Expert software was employed to compare the performance of the three CQDs, as fluorescent probes, based on their quenching efficiency and the percentage of their emission recovery in the presence of AChE and acetylthiocholine (ATCh). Based on the statistical analysis, among the studied CQDs, S-doped CQD was the most suitable candidate for sensor designing. The detection mechanism for the proposed S-doped CQD-based sensor is as follows: The strong binding of Cu 2+ ions to carboxyl groups of S-doped CQD quenches the fluorescence signal. Then, hydrolysis of ATCh into thiocholine (TCh) in the presence of AChE causes fluorescence recovery, due to the stronger affinity of Cu 2+ to the TCh, rather than the CQD. Finally, in the presence of malathion and chlorpyrifos inhibitors, AChE loses its ability to hydrolyze ATCh to TCh, so the fluorescence emission remains quenched. Based on the proposed detection technique, the designed sensor showed detection limits of 1.70 ppb and 1.50 ppb for malathion and chlorpyrifos, respectively., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Objectives: Hemodialysis (HD) patients suffer more sleep problems (poor sleep quality and restless leg syndrome [RLS]). Complementary therapy, especially massage with aromatherapy oil is one of the non-pharmacological treatment options with less adverse effects than routine methods. The purpose of this study was to determine the effects of foot massage with of lavender and orange essential oil on HD patients' sleep quality and RLS., Methods: This is a double blind randomized controlled trial on 105 HD patients was conducted at a large educational hospital in Iran, Hamadan province between January and September 2017. Patients divided into three groups with random allocation (35 participants per groups in lavender, orange, and control group). Foot massage during HD with lavender and orange essential oil was administered to the patients three times a week for three weeks, and every massage lasted half an hour. The control group received routine care. Before the intervention, the end of the first, second, and third weeks Pittsburgh Sleep Quality Index (PSQI) and RLS questionnaire were completed for all three groups. Data were statistically analyzed with Independent Samples t-test, chi-square test and repeated-measures analysis of variance (ANOVA) by SPSS version 16., Results: The mean score of sleep quality and RLS in the intervention groups were significantly different compare with the control group in all three time of data collecting (p<0 001)., Conclusions: Aromatherapy prepared with lavender oil and sweet orange may be recommended to increase sleep quality and RLS level of the HD patients., (© 2021 Walter de Gruyter GmbH, Berlin/Boston.)

Graphene quantum dots (GQDs) are known as emerging sub-10 nm nanoparticles (NPs), which are in fact few-layered pieces of graphene, capable of emitting blue fluorescence, when exposed to 360 nm UV light. Understanding the details of the interaction between GQDs and lipase can serve as a critical step for improving the biological outcome of GQD-derived drug-delivery and diagnosis systems. The interaction occurs in the form of surface adsorption, which can subsequently influence the physicochemical properties of both the NP and the protein. Hence, a systematic approach was taken here to optimize the GQDs' synthesis conditions in order to achieve the highest possible quantum yield (QY). Furthermore, to understands the influence of the interaction of GQDs and lipase, on both the activity of lipase and the emission intensity of GQDs, various incubation conditions were tested to achieve optimized conditions over central composite design algorithm by Design-Expert®, using response surface methodology. The results show that the GQDs fabricated by thermal decomposition of citric acid at 160°C, with a heating duration of 55 min, obtain almost three times higher QY than the highest values reported previously. The best enzymatic activity after the formation of the hard corona, as well as the highest fluorescent emission, were achieved at GQD-to-enzyme ratios within the rage of 23-25%, at temperatures between 41 and 42°C, for 6-8 min. In the aforementioned condition, the enzyme retains 91-95% of its activity and the NP preserves about 80-82% of its fluorescence intensity after incubation., (© 2020 Wiley Periodicals, Inc.)

Background: Studies have revealed a strong association between mutations of CFTR gene and the congenital bilateral absence of the vas deferens (CBAVD), but the role of this gene in other types of male infertility is still unclear. The purpose of this study was to investigate the frequency of the most common mutations of the CFTR gene (DF508, G542X, N1303K, G551D, and W1282X) in a population of infertile men with nonobstructive azoospermia (NOA) and CBAVD in Iran., Methods: Blood samples were obtained from 50 NOA, 50 CBAVD, and 100 normal males (control). Genomic DNA was isolated from whole blood leukocytes, and the presence of common mutations of the CFTR gene was assessed by an amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Restriction fragment length polymorphism (PCR-RFLP) was also used to analyze IVS8-Tn polymorphism., Results: It was found that 16%, 8%, and 8% of patients with CBAVD were heterozygote for DF508, G542X, and N1303K, respectively. The frequency of the 5T allele was 34% and higher than the normal group (p < 0.001). None of the common CFTR gene mutations were detected in NOA patients, and no significant difference was found in the distribution of the 5T allele between the NOA patients and the control group (5 vs. 3 p = 0.721)., Conclusion: Based on the present case-control study, the CFTR gene mutations and IVS8-Tn polymorphisms are correlated with CBAVD; however, extensive investigations are necessary to determine the exact relationship between the gene mutations and other forms of male infertility.

Resistance to antibiotics is an emerging and growing threat. To address this threat, attempts are being made by researchers to identify the Volatile Organic Compounds (VOCs) of bacteria. It is believed that unique combinations could be found among the VOCs produced by each microorganism. The current study aimed to identify and compare the VOCs of antibiotic-resistant and standard strains of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae. A polymer of divinylbenzene /carboxen /polydimethylsiloxane was applied for absorption of volatile compounds in headspace bacterial samples in form of a solid phase micro-extraction fiber holder. Gas chromatography-mass spectrometry technique was used for identification of volatile compounds. The analysis of the VOCs indicated that some VOCs appeared only in standard strains while others were common only among resistant strains. Exclusive VOCs to a specific strain were also detected. This study demonstrated that resistant strains of bacteria produced VOCs that were different from those of the standard strains. In addition, VOCs released by bacteria after passing the logarithmic growth phase showed no significant differences. The identification of VOCs can be a precise way to differentiate bacterial species, also it can be said that the VOCs produced by different pathogenic microorganisms can be the suitable biomarkers for their detection.

Background : Diagnoses  of  respiratory  tract  infections  usually happen  in  the  late  phase  of  the  disease  and  usually  result  in  reduction  of  the  pathogen  load after broad-spectrum  antibiotic  therapy,  but  not  in eradication of the pathogen.  The  development  of a  non-invasive,  fast,  and  accurate  method  to  detect  pathogens  has  always  been  of  interest  to  researchers  and  clinicians  alike.  Previous studies have shown that bacteria produce organic gases.  The  current  study  aimed  to  identify  the  volatile  organic  compounds  (VOCs)  produced  by three  respiratory  tract  pathogens,  including  Staphylococcus  aureus ,  Escherichia  coli  and  Candida  albicans. Methods : The  VOCs  produced  were identified by gas chromatography-mass spectrometry (GC-MS), with  prior  collection  of  microbial  volatile  compounds  using  solid  phase  microextraction  (SPME)  fiber.  The volatile compounds were collected by obtaining bacterial headspace samples. Results : Results  showed  that  these  three  organisms  have  various  VOCs,  which  were  analyzed  under  different  conditions.  By ignoring common VOCs, some species-specific VOCs could be detected.  The most important VOC of E. coli was Indole, also some important VOCs produced by S. aureus   were 2,3-Pentandione,  cis-Dihydro-α-terpinyl  acetate,  1-Decyne,  1,3-Heptadiene-3-yne,  2,5-dimethyl  Pyrazine,  Ethyl  butanoate  and  Cyclohexene,4-ethenyl  furthermore,  most  of  identified  compounds  by  C.  albicans are  alcohols. Conclusions : The  detection  of  VOCs  produced  by  infectious  agents  maybe  the  key  to  make   a  rapid  and  precise  diagnosis  of  infection,  but  more  comprehensive  studies  must  be  conducted  in this  regard., Competing Interests: No competing interests were disclosed.

Background: Diagnoses  of  respiratory  tract  infections  usually happen  in  the  late  phase  of  the  disease  and  usually  result  in  reduction  of  the  pathogen  load after broad-spectrum  antibiotic  therapy,  but  not  in eradication of the pathogen.  The  development  of a  non-invasive,  fast,  and  accurate  method  to  detect  pathogens  has  always  been  of  interest  to  researchers  and  clinicians  alike.  Previous studies have shown that bacteria produce organic gases.  The  current  study  aimed  to  identify  the  volatile  organic  compounds  (VOCs)  produced  by three  respiratory  tract  pathogens,  including  Staphylococcus  aureus ,  Escherichia  coli  and  Candida  albicans. Methods: The  volatile  organic  compounds  (VOCs)  produced  were identified by gas chromatography-mass spectrometry(GC-MS), with  prior  collection  of  microbial  volatile  compounds  using  solid  phase  microextraction  (SPME)  fiber.  The volatile compounds were collected by obtaining bacterial headspace samples., Results: Results  showed  that  these  three  organisms  have  various  VOCs,  which  were  analyzed  under  different  conditions.  By ignoring common VOCs, some species-specific VOCs could be detected.  The most important VOC of E. coli was Indole, also some important VOCs produced by S. aureus   were 2,3-Pentandione,  cis-Dihydro-α-terpinyl  acetate,  1-Decyne,  1,3-Heptadiene-3-yne,  2,5-dimethyl  Pyrazine,  Ethyl  butanoate  and  Cyclohexene,4-ethenyl  furthermore,  most  of  identified  compounds  by  C.  albicans are  alcohols., Conclusions: The  detection  of  VOCs  produced  by  infectious  agents  maybe  the  key  to  make   a  rapid  and  precise  diagnosis  of  infection,  but  more  comprehensive  studies  must  be  conducted  in this  regard., Competing Interests: Competing interests: No competing interests were disclosed.

Although the use of silver nanoparticles (AgNPs) has substantial benefits, their entrance into the environment, food chain, and human body and their toxicity have come under serious scrutiny. Multiple noncovalent attractive forces between AgNPs and bio-macromolecules are responsible for immediate corona formation upon exposure to biological tissue. Here, the influence of AgNPs with neuro-enzyme Acetylcholinesterase (AChE) was investigated. AgNPs to enzyme ratio had an effect on the enzyme and features of the treated samples. It was also observed that time increments had a positive effect on the size of AgNPs and caused an increase in their initial size. In other words, smaller AgNPs resulted in size increments after interaction with enzymes, while the larger ones showed size decrements. The nano-crystalline AgNPs were identified in x-ray powder diffraction analyses before and after treatment with AChE. The (220) crystalline plane is related to the internal crystallinity of cubic Ag. The results show that the interaction between AChE and AgNPs could lead not only to a decrease in AChE activity, but also to a reduction in the crystallinity and stability of AgNPs. The circular dichroism demonstrates that the secondary structure of AChE also declined after 30 min of incubation with AgNPs at 37 °C., (Copyright © 2017. Published by Elsevier Inc.)

Male factor infertility is involved in almost half of all infertile couples. Lack of the ejaculated sperm owing to testicular malfunction has been reported in 6-10% of infertile men, a condition named nonobstructive azoospermia (NOA). In this study, we investigated untargeted metabolomic profiling of the seminal plasma in NOA men using gas chromatography-mass spectrometry and advance chemometrics. In this regard, the seminal plasma fluids of 11 NOA men with TESE-negative, nine NOA men with TESE-positive and 10 fertile healthy men (as a control group) were collected. Quadratic discriminate analysis (QDA) technique was implemented on total ion chromatograms (TICs) for identification of discriminatory retention times. We developed multivariate classification models using the QDA technique. Our results revealed that the developed QDA models could predict the classes of samples using their TIC data. The receiver operating characteristic curves for these models were >0.88. After recognition of discriminatory retention time's asymmetric penalized least square, evolving factor analysis, correlation optimized warping and alternating least squares strategies were applied for preprocessing and deconvolution of the overlapped chromatographic peaks. We could identify 36 discriminatory metabolites. These metabolites may be considered discriminatory biomarkers for different groups in NOA., (Copyright © 2017 John Wiley & Sons, Ltd.)

Recent investigations have demonstrated that normobaric hyperoxia induces neuroprotection against ischemic injury. The aim of study was to determine the survey of HO (hyperoxia) preconditioning on brain lipidome.The animals were assigned into three groups, the first experimental group was exposed to 95% inspired HO for 4 h /day for six consecutive days. The second experimental group considered as the control group and was exposed to 21% oxygen as room air (RA) in the same chamber. The third group acted as sham, which was under the stress of surgery condition without ischemia. The first two groups were divided into 2 subgroups, intact (without any surgery) and middle cerebral artery occlusion- operated (MCAO). Twenty-four hours after exposure to hyperoxia, MCAO subgroups were subjected to 60 min of right middle cerebral artery occlussion. After 24 h reperfusion, infarct volume (IV) and neurological deficit score (NDS) were assessed in MCAO subgroup. Brain lipidomics were measured in the intact subgroup. Preconditioning with HO significantly reduced NDS and IV and elevated the level of phosphatidylethanolamine (PE), sphingomyelin (SM), cholesterol ester (CE), cholesterol (Chol), phosphatidylcholine (PC), triglyceride (TG) and cerebroside (CB) in the brain as compared with the control (sham and RA). HO preconditioning, significantly decreased the brain ceramide (Cer) and lyso- phosphatidylcholine (Lyso-PC or LPC) levels. Preconditioning with HO decreases brain ischemia injury via changes in brain lipidomics and significantly decreases the brain ceramides (CER).Although more studies are required to explain the mechanisms of time course of neuroprotection, HO preconditioning partly decreases brain ischemia injury via changes in brain lipidome.

Background: Plants are considered as promising sources of new antibacterial agents as well as bioassay guided fractionation., Objectives: In the present work, the antibacterial properties, especially against methicillin-resistant Staphylococcus aureus (MRSA), of Bromus inermis inflorescence was studied, using the bioassay guided fractionation as well as the bio-autographic method., Materials and Methods: The plant organic extract was prepared via maceration in methanol, followed by the fractionation using n-hexane. The extracts were subjected for minimum inhibitory concentrations (MICs) against some human pathogenic bacteria via standard broth micro-dilution assay. Thereafter, a bio-autographical method was applied using the high performance thin layer chromatography (HPTLC) coupled with agar overlay assays for the primary characterization and identification of bioactive substance (s)., Results: Through the bioassay guided fractionation method, the greatest antibacterial activities were related to the n-hexane extract. It was also revealed that the effective anti-MRSA agent of the assessed plant was a relatively polar substance with an MIC value of about 8 μg/mL against the tested MRSA strain (in comparison with the MIC value of 32 μg/mL for chloramphenicol)., Conclusions: As a result of the full range UV-Vis scanning of the responsible band in the HPTLC experiments (200-700 nm), the flavonoid was the most imaginable natural compound.

The increasing use of silver nanoparticles, (AgNPs), will inevitably result in their release into the environment and thereby cause the exposure to plants. It was claimed that using AgNPs is a safe and efficient method to preserve and treat agents of disease in agriculture. This study tries to understand the protein populations and sub-populations and follow up environmental AgNPs stresses. To accomplish these, the action of homemade spherical AgNPs colloidal suspension against Oryza sativa L. was investigated by a proteomic approach (2-DE and NanoLC/FT-ICR MS identification). Twenty-eight responsive (decrement/increment in abundance) proteins were identified. Proteomic results revealed that an exposure of O. sativa L., root with different concentrations of AgNPs resulted in an accumulation of protein precursors, indicative of the dissipation of a proton motive force. The identified proteins are involved in oxidative stress tolerance, Ca(2+) regulation and signaling, transcription and protein degradation, cell wall and DNA/RNA/protein direct damage, cell division and apoptosis. The expression pattern of these proteins and their possible involvement in the nontoxicity mechanisms were discussed., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Emerging technologies in functional genomics and proteomics provide a way of achieving high-throughput analyses, understanding effects on protein populations and sub-populations and follow up environmental stresses. To accomplish these, the action of homemade spherical Silver nanoparticles colloidal suspension (AgNPs) against Bacillus thuringiensis (isolate from Oryza sativa L. rhizosphere) was investigated by a proteomic approach (2-DE and NanoLC/FT-ICR MS identification). Thirty four responsive (up/down regulated) proteins were identified. Proteomic results revealed that an exposure of B. thuringiensis cells with different concentrations of AgNPs resulted in an accumulation of envelope protein precursors, indicative of the dissipation of a proton motive force. Identified proteins are involved in oxidative stress tolerance, metal detoxification, transcription and elongation processes, protein degradation, cytoskeleton remodeling and cell division. The expression pattern of these proteins and their possible involvement in the nontoxicity mechanisms were discussed., (© 2013 Elsevier Inc. All rights reserved.)

The endoplasmic reticulum (ER)-resident protein kinase PERK is a major component of the unfolded protein response (UPR), which promotes the adaptation of cells to various forms of stress. PERK phosphorylates the α subunit of the translation initiation factor eIF2 at serine 51, a modification that plays a key role in the regulation of mRNA translation in stressed cells. Several studies have demonstrated that the PERK-eIF2α phosphorylation pathway maintains insulin biosynthesis and glucose homeostasis, facilitates tumor formation and decreases the efficacy of tumor treatment with chemotherapeutic drugs. Recently, a selective catalytic PERK inhibitor termed GSK2656157 has been developed with anti-tumor properties in mice. Herein, we provide evidence that inhibition of PERK activity by GSK2656157 does not always correlate with inhibition of eIF2α phosphorylation. Also, GSK2656157 does not always mimic the biological effects of the genetic inactivation of PERK. Furthermore, cells treated with GSK2656157 increase eIF2α phosphorylation as a means to compensate for the loss of PERK. Using human tumor cells impaired in eIF2α phosphorylation, we demonstrate that GSK2656157 induces ER stress-mediated death suggesting that the drug acts independent of the inhibition of eIF2α phosphorylation. We conclude that GSK2656157 might be a useful compound to dissect pathways that compensate for the loss of PERK and/or identify PERK pathways that are independent of eIF2α phosphorylation.

Silver nanoparticles (AgNPs) are widely used as antibacterial and antifungal agents in agriculture. Nevertheless, these nanoparticles with newborn properties pose a potential risk to the environment, Due to contact with crops and bacteria that are beneficial to the soil. This study is based on the examination of the phytotoxic effects of AgNPs on Oryza sativa L. and some of its rhizosphere bacteria, by physiological and biochemical assays. In order to study the complex interaction of the AgNPs life expectancy that are mixed with culture medium, the incubation time for the fresh mixture, 7, 14 and 21 days old of AgNPs, on the seedlings growth was investigated. Results indicated that plant's response to the treatment with AgNPs affected on the cell wall, and that with an increase in its concentration (up to 60 μg/mL). The obtained results of transmission electron microscopy (TEM) exhibited that those particles not only penetrated the cell wall, but they could also damage the cell morphology and its structural features. AgNPs treatment up to 30 μg/mL accelerated root growth and at 60 μg/mL was able to restrict a root's ability to grow. The 30 μg/mL treatment had significant effect on root branching and dry weight. In contrast, shoot growth was more susceptible to the effects of AgNPs treatment. The root content for total soluble carbohydrates and starch demonstrated that despite stable starch content, total soluble carbohydrates showed the tendency to significantly decline in response to AgNPs. However, induction of root branching and photosynthetic pigments can attributed to AgNPs stress based on evidence from the production of the reactive oxygen species (ROS) and local root tissue death. Nine isolates of the genus Bacillus selected and identified according to morphological and chemotaxonomic methods. The AgNPs treatment revolutionized the populations of bacteria as Bacillus thuringiensis SBURR1 was totally eliminated, and Bacillus amyloliquefaciens SBURR5 became the most populated one. Images from an electron microscope and the leakage of reducing sugars and protein through the bacterial membrane, similarly confirmed the "pit" formation mechanism of the AgNPs. Moreover the hypothesis from the growth curve study demonstrated that AgNPs may damage bacterium cell wall and transform them to protoplasts., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: Recent studies suggest that olive extracts suppress inflammation and reduce stress oxidative injury. We have attempted to determine the effect of dietary olive leaf extract (OLE) on brain lipidomics in rat stroke model., Methods: Five groups, each consisting of 12 male Wistar rats, were studied. First and second groups (control, and sham) received distilled water, while three treatment groups received oral olive leaf extract (OLE) for 30 days (50, 75 and 100 mg/kg/day, respectively). Two hours after the last dose, each main group was subdivided to Middle cerebral artery occlusion-operated and intact subgroup for assessment of neuropathology (neurologic deficit scores and infarct volume), brain lipid analysis and brain glutathione levels, respectively., Results: OLE increased the brain cholesterol ester, cholesterol, cerebroside and phosphatidylcholine levels in doses of 50, 75 and 100 mg/kg/day. OLE increased the brain triglyceride levels in doses of 75 and 100 mg/kg/day and OLE reduced the brain ceramide levels in doses of 50, 75 and 100 mg/kg/day in comparison to the control group (p<0.05)., Conclusion: Although further studies are needed, it seems that the mechanism of OLE-induced ischemic tolerance in rats is partly associated with changes in brain lipids level., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

In shoot cultures of Thymus daenensis, hyperhydricity syndrome promoted by benzyladenine (BA) is characterised by the development of chlorophyll-deficient shoots with a high water content and reduced growth that is less differentiated. By removing the BA from the culture medium, the hyperhydricity was reversed, and the reversion toward a normal growth in vitro was more efficient in shoots treated with 5 μM of salicylic acid (SA), showing a significant increase in chlorophyll b after 4 weeks of culture. In the present study, the effect of salicylic acid on the reversion of shoot hyperhydricity was investigated at the level of the free, soluble and insoluble conjugated polyamine content. In T. daenensis micropropagated shoots, the level of polyamines was high, with a predominance of putrescine. BA, which triggered hyperhydricity, caused a reduction of the polyamine (PA) content by one-half due to a decrease in the putrescine content and insoluble conjugated PAs that were not detected in the hyperhydric shoots. In the reverted shoots, changes of the free polyamines, spermidine and, more notably, spermine, were shown. The spermine content doubled after 4 weeks of culture, and its amount was the same as that found in normal shoots, suggesting that free spermine could be particularly involved in the reversion of hyperhydricity. In the SA-reverted tissues, the PA pattern was marked with a transient increase of free putrescine, spermidine and spermine and an enhancement of soluble conjugated spermine. This transitory SA-dependent amplification of PAs was concomitant with a remarkable transient increase of H(2)O(2), suggesting that SA may be implicated in PA signalling pathways for tissue differentiation during the reversion of hyperhydricity in T. daenensis., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)

Aerial parts essential oil of Nepeta betonicifolia and N. saccharata were obtained by hydrodistillation and analysed by GC-FID and GC-MS. Thirty-three and eighteen components represented 97.9% and 98.2% of the total oils identified, respectively. Main compounds of the oil of N. betonicifolia were 4aα,7β,7aα-nepetalactone (42.0%), germacrene D (6.0%), triplal (5.2%), 1-nor-bourbonanone (4.0%) and 1,8-cineole (3.2%). The principal constituents of the essential oil of N. saccharata were found to be 4aβ,7α,7aβ-nepetalactone (66.9%), germacrene D (12.9%), sabinene (6.5%) and trans-caryophyllene (3.3%). The radical scavenging capacity (RSC) of methanol extracts and chloroform, butanol and water subfractions of aerial parts of N. betonicifolia and N. saccharata were evaluated by using DPPH, FRAP and ABTS assays. TPC of each extract was measured using Folin-Ciocalteau. The antioxidant activity of the butanolic subfractions of both plants was higher than other extracts examined.

Genetic factors including Y chromosome microdeletions and androgen receptor (AR) gene mutations are responsible for male infertility. In the present study, genetic analysis was performed in an infertile Iranian male with azoospermia. Multiplex polymerase chain reaction with 6 sequence-tagged site markers on the Yq11 chromosome revealed no microdeletions in the Y chromosome. Single-strand conformational polymorphism and sequencing analyses detected a 1510C→A transversion in exon 1 of the AR gene, which resulted in a p.Pro504Thr substitution in the transactivation domain of the protein. The present study suggested that mutations in the AR gene might be responsible for some cases of idiopathic infertility, and therefore, molecular analyses may be useful for genetic counseling of candidates with regard to the use of assisted reproductive techniques.

Antibacterial activity of silver nanoparticles (AgNPs) was investigated using Staphylococcus aureus PTCC1431 as a model of Gram-positive bacteria. The mechanism of antibacterial activity of AgNPs was then studied by analyzing the growth, morphology, and molecular variations in the cell wall. Experimental data showed that AgNPs at a concentration of 4 μg/ml completely inhibited bacterial growth. Transmission electron microscopy results confirmed cell wall damage produced by AgNPs as well as accumulation of AgNPs in the bacterial membrane. Meanwhile, the AgNP-treated bacteria were monitored by circular dichroism to reveal peptidoglycan variations. Some degree of variation in the α-helix position of the peptide chain was observed. Moreover, increasing the AgNP concentration to 8 μg/ml resulted in release of muramic acid (MA) into the medium, which could be attributed to cell wall distraction. A gas chromatography-tandem mass spectrometry analysis and release of MA, as a bacterial indicator, showed that glycan strands may also be decomposed as a result of AgNP treatment., (Copyright © 2011 Institut Pasteur. Published by Elsevier SAS. All rights reserved.)

The therapeutic effects of anticancer drugs would highly improve if problems with low water solubility and toxic adverse reactions could be solved. In this work, a full factorial experimental design was used to develop a polymeric nanoparticulate delivery system as an alternative technique for anticancer drug delivery. Nanoparticles containing tamoxifen citrate were prepared and characterized using an O/W emulsification-solvent evaporation technique and different analytical methods. Scanning Electron Microscopy (SEM), particle size analysis and High Pressure Liquid Chromatography (HPLC) were used for characterization of nanoparticles. Nanoparticles' characteristics including size, size distribution, drug loading and the efficiency of encapsulation were optimized by means of a full factorial experimental design over the influence of four different independent variables and desirability function using Design-Expert software. The resulting tamoxifen loaded nanoparticles showed the best response with particle sizes less than 200 nm, improved encapsulation efficiency of more than 80% and the optimum loading of above 30%. The overall results demonstrate the implication of desirability functionin experimental design as a beneficial approach in nanoparticle drug delivery design.

In this study, a simple, rapid, and highly efficient liquid-phase microextraction method based on solidification of floating organic droplet was coupled with high performance liquid chromatography-photo diode array detection (HPLC-PDA) for determination of ketoconazole, clotrimazole, and miconazole as antifungal drugs. Central composite design (CCD) was used for optimization of several factors affecting the extraction efficiency. The optimized conditions were established to be 550 rpm for stirring rate, 35 min for extraction time, 57 °C for extraction temperature, 8.5 for solution pH, 10 μl for organic solvent volume, and 7% (w/v) of NaCl for ionic strength. Limit of detections (LODs) of the extraction method ranged from 0.01 to 0.1 μg L(-1) and the linear dynamic ranges (LDRs) ranged from 0.1 to 300 μg L(-1) for the three antifungal drugs. Relative standard deviations (RSDs) of the proposed method were 5-11%. Preconcentration factors in the range of 306-1350 were obtained at extraction time of 35 min. Finally, performance of the proposed method was evaluated for the extraction and determination of the drugs' levels in microgram per liter in samples and satisfactory results were obtained., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Introduction: Recently, there have been growing attention on the modification and optimisation of new extraction and quantification methods, caused by the lack of environmentally friendly methodologies for the extraction of phytochemicals from complex matrices. In the case of pharmaceutical compounds, not only the extraction procedure but also the analysis method should be efficient, precise, fast and easy., Objectives: The essential pharmaceutical characteristics and trace concentration of withanolides led us to modify and optimise the previously reported extraction and quantification procedure for withaferin A (WA) as a candidate for withanolides., Material and Methods: The WA from the air-dried aerial part of Withania somnifera Dunal. was extracted using a microwave-assisted extraction (MAE) technique. Four variables affecting the extraction procedure were optimised using the central composite design approach. The method of high-performance thin-layer chromatography assay was validated and applied for the quantification of each experiment., Results: The optimum values of factors were: extraction time (150 s), extraction temperature (68°C) and 17 mL of methanol : water in the ratio 25 : 75 as extracting solvent. The solvent system consisted of ethyl acetate : toluene : formic acid : 2-propanol (7.0 : 2.0 : 0.5 : 0.5, v/v/v/v), and densitometric scanning at 220 nm was applied for the analysis. The dynamic linear range, LOD, LOQ and recovery with the inter-day, and intra-day RSDs of the developed method indicated the validity of the method., Conclusion: A pressurised MAE method for extracting WA from the plant's aerial part was optimised using factorial-based design. The net effect of time, temperature, solvent volume and its ratio suggests that the yield of WA increases until each factor reaches its optimum value, and decreases with further increase in temperature or solvent ratio., (Copyright © 2010 John Wiley & Sons, Ltd.)

Background & Objectives: Genetic factors contribute about 10 per cent of male infertility. Among these, genes in azoospermia factor (AZF) region including AZFa, AZFb, AZFc and AZFd on the long arm of Y chromosome are considered most important for spermatogenesis. Deletions in these regions are thought to be involved in some cases of male infertility associated with azoospermia or oligozoospermia. We studied the incidence of AZF deletions among Iranian infertile men with idiopathic non-obstructive azoospermia., Methods: A total of 100 Iranian azoospermic infertile men were selected for the molecular study of Y chromosome microdeletions. The presence of 13 sequence tagged site (STS) markers from AZF region was investigated using multiplex polymerase chain reaction (M-PCR). One hundred fertile men were also studied as control group., Results: Twelve (12%) patients showed Y chromosome microdeletions and among these, deletion in AZFb region was the most frequent (66.67%) followed by AZFc (41.67%), AZFd (33.33%) and AZFa (8.33%), respectively., Interpretation & Conclusions: Because of relatively high incidence of Y chromosome microdeletions among Iranian azoospermic patients, molecular screening may be advised to infertile men before using assisted reproductive treatments.

For successful conservation and breeding of a medicinal species, it is important to evaluate its genetic diversity as well as its content of phytochemical compounds. The aim of the present study was to investigate the genetic variation of Iranian natural populations of W. somnifera and W. coagulans, using the RAPD (random amplified polymorphic DNA) markers, and their withaferin A content. Using 16 RAPD primers, a total of 282 RAPD bands were achieved. The highest and lowest percentages of polymorphism were observed with primers OPAD-15 (100.0%) and OPC-06 (75.0%), respectively. Cluster analysis of the genotypes was performed based on data from polymorphic RAPD bands, using Dice's similarity coefficient and the UPGMA clustering method. Variations in the RAPD results were found to reflect geographical distribution and genetic factors of the plant populations. The HPTLC analysis of the studied samples revealed the presence of withaferin A in W. coagulans and W. somnifera. Moreover, the concentration of withaferin A had a range from 2.2 to 32.5 microg/g DW and was higher in the aerial part than in the root in all used samples. The results of the present study show that there is a high level of variation in the Iranian natural population of Withania, which is significant for conservation and breeding programs to improve production of withaferin A.

1. Alzheimer's disease (AD) is the most common form of dementia in the elderly in which interplay between genes and the environment is supposed to be involved. Mitochondrial DNA (mtDNA) has the only noncoding regions at the displacement loop (D-loop) region that contains two hypervariable segments (HVS-I and HVS-II) with high polymorphism. mtDNA has already been fully sequenced and many subsequent publications have shown polymorphic sites, haplogroups, and haplotypes. Haplogroups could have important implications to understand the association between mutability of the mitochondrial genome and the disease.2. To assess the relationship between mtDNA haplogroup and AD, we sequenced the mtDNA HVS-I in 30 AD patients and 100 control subjects. We could find that haplogroups H and U are significantly more abundant in AD patients (P = 0.016 for haplogroup H and P = 0.0003 for haplogroup U), Thus, these two haplogroups might act synergistically to increase the penetrance of AD disease.

Objectives: Classical galactosemia (McKusick 230400) is an autosomal recessive disorder caused by mutations in the galactose-1-phosphate uridyl transferase (GALT;EC 2.7.7.10) gene., Design and Methods: In the present study, we report molecular analysis of 14 unrelated Iranian galactosemia children with reduced or without GALT activity using PCR-RFLP and SSCP-Sequencing methods., Results: Q188R mutation was the most observed mutation with the allelic frequency of 57.1%. The allelic frequencies for S135L, Y209S, A320T, and K285N were found to be 7.1%, 7.1%, 7.1%, and 3.57% respectively., Conclusions: Our results show that galactosemia is a heterogeneous disorder at the molecular level among the Iranian population.

Thirty-seven unrelated Iranian CF families were screened for the presence of seven common mutations (DeltaF508, G542X, W1282X, G551D, N1303K, 1717-1G-->A, and 621-1G-->T) using ARMS PCR and exons 4 and 7 of the CFTR gene by SSCP method. This study resulted in the identification of 26.8 per cent of all CF alleles: DeltaF508 (16.2 per cent), W1282X (4 per cent), G542X (2.7 per cent), R117H (1.3 per cent), R347H (1.3 per cent), and A120T (1.3 per cent) mutations were detected. To the best of our knowledge, it is the first report of an Asian subject carrying the A120T mutation. Our findings suggest heterogeneity in the Iranian population, stressing the need to draw attention to sequence analysis in order to find population-specific mutations.