Search

In this work, a new control scheme for synchronization of AC microgrids with upstream power grid is presented. The effects of V2Gs (vehicle to grid) dynamics on synchronization process is studied. This new control approach is based on the optimal fractional calculus and has been developed for synchronization of the microgrid. The V2Gs effect on the dynamics of the microgrid is analyzed through small signal stability and simulations. This effect is also considered in synchronization process by considering a PHEV-dominated-microgrid. The proposed control scheme is a coordinated control of distributed resources and provides a soft and reliable synchronization for microgrid. In the proposed control scheme, the fractional order proportional-integral-derivative (FOPID) controllers have optimally been tuned and implemented using the genetics algorithm (GA). The simulation results confirm the effectiveness of the proposed control strategy in soft and swift synchronization of the microgrid.

Subinhibitory concentrations (subMIC) of antibiotics, although not able to kill bacteria, can modify their physicochemical characteristics and may interfere with some bacterial functions. This study aimed to investigate the effect of subMIC of imipenem and piperacillin on the transcriptional expression of virulence-related genes toxA and exoS in Pseudomonas auroginosa. Five clinical isolates of P. aeruginosa were screened for the presence of toxA and exoS genes and MICs of imipenem and piperacillin were determined using broth macrodilution. The expression levels of toxA and exoS at subMIC concentrations of antibiotics were measured by real-time PCR. Our results showed that the expression of toxA decreased at all subinhibitory concentrations of imipenem, especially at concentrations 2, 4 and 8 mg/L (p 0.05). Further studies will be required to assess whether subMIC of imipenem can improve the outcomes of severe and serious infections caused by P. aeruginosa. Keywords: Exoenzyme S, exotoxin A, Pseudomonas aeruginosa, Subinhibitory concentrations, Imipenem, Piperacillin

Background& Purpose: The Purpose of this study was to evaluate the effect of specific type of exposure, visiting the stressful event location, on PTSD symptoms are caused by the war. Methods & Material: This is an intervention study and data are measured as before and after. Twenty five people with PTSD symptoms were selected based on inclusion criteria. The intervention was exposure to traumatic event place that caused PTSD, For this purpose each of these people were sent to combat zones. PTSD check list (PCL) before exposure, 2 weeks and 3 months after that, was completed by the participants and then Paired t- test was used to compare of 3 measurements as before, after and follow-up 3 months after exposure and analysed with SPSS /16 software. Results: The effect of this exposure on overall symptoms of PTSD was significant (p0.05). Conclusion: Exposure to the traumatic event place decreased the overall symptoms of PTSD and also Re experience and Hyper arousal symptoms.

This research was carried out for 10 successive weeks (2 weeks for adaptation and 8 weeks for culture) in summer 2008 at Abzisytan Shrimp Hatchery Center in Bushehr province. The effects of dietary nucleotides at 5 levels including 0.0% (control), 0.15%, 0.25%, 0.35% and 0.5% on the growth performance and whole-body composition of Epinephelus coioides was investigated. Juvenile fish specimens with average weight of 10.70±0.29g were fed by experimental diets for 8 weeks. The trial was carried out in 300 liter fiberglass tanks. The highest protein content was observed in 0.15% diet nucleotide and the highest fat in 0.15% and 0.25% diet nucleotide which showed significant difference with th

Background and objectives: Due to the high cancer mortality rates and side effects of different types of cancer treatments, discovering effective treatments without or with fewer side effects is the main purpose of many researchers all around the world. Plants play an important role in the discovery of new drugs. Iran owns rich and varied vegetation but the majority of these plants have not yet undergone chemical, pharmacological and toxicological studies. In the present study, some species from East Azarbaijan province of Iran were evaluated for cytotoxicity effects. Methods: Total methanol extract of 29 plants from 18 families were screened for their cytotoxic activities. The inhibition of cell growth for these extracts was evaluated against MCF-7, A-549, Hep-G2, HT-29 and MDBK cell lines. Their 50% inhibitions of growth (IC50) were determined by MTT assay. Moreover, cytotoxic evaluation of different fractions (ether de petrol, chloroform and methanol) of the most potent species was performed. Results: Total extracts and fractions of Bryonia aspera, Centaurea salicifolia, Cuscuta chinensis, Ecbalium elaterium, Gypsophila ruscifolia, Ononis spinosa exhibited potent cytotoxic activity against one or more of the cell lines. Three of the mentioned total extracts presented cytotoxicity effects exclusively against HT-29 cells. Also three fractions (one ether de petrol and two chloroform fractions) demonstrated selective cytotoxicity effects against MCF-7cells. Conclusion: It was concluded that these 6 potent species were proper candidates for identification and isolation of active ingredients with cytotoxic effects and further studies about these species are recommended.

Background and objectives: Cancer is the third leading cause of mortality and morbidity in Iran and kills out about 30000 people a year. The drugs used to treat cancer have many side effects. So developing novel and effective drugs with lower side effects has a great importance. Located in the north-west of Iran, Ardebil province has a rich plant diversity which the biological effects of most of them are unknown. In this study, the cytotoxic effects of 18 plants collected from Ardebil have been investigated. Methods: The cytotoxic effect of total methanol extract and petroleum ether, chloroform and methanol fractions were investigated by using MTT assay against 4 selected human cancer cell lines MCF7, HepG-2, A-549, HT-29 and also a normal cell line, MDBK. Results: Total methanol extract and chloroform fraction of the aerial parts of Onopordon acanthium showed significant cytotoxic effect against all cell lines with 12.51≤ IC50 ≤ 55.72 and 6.49≤IC50≤31.5, respectively. Conclusion: The plant could be a candidate for further investigations on its cytotoxicity mechanism and also identification of active ingredients.

Abstract Background and aim: With an aging population, considering the factors affecting the quality of life more than ever is necessary. The aim of current research was to investigate the effectiveness of cognitive existential therapy on quality of life of elderly people. Methods: The current research is semi experimental with pre and post test with control group. Statistical population of research consists of all elderly people in Kahrizak nursing homes. In the first phase, the participants were selected through purposive sampling method and after responding to the quality of life questionnaire and obtaining score for enter to research they were divided in two groups of experimental and control (N = 12 per group) using random sampling method. The experimental group participated in 10 sessions of group counseling based on cognitive- existential approach and control group received no intervention. The gathered data were analyzed using covariance analysis. Results: There was no difference between pre-test and control groups, but the mean scores of post-test experimental and control groups were statistically significant. and cognitive group therapy improves quality of life is (p=0.001). Therefore it seems that cognitive-existential group therapy increase quality of life of elderly people. Conclusion: Cognitive Existential Group therapy utilizes concepts such as death, meaning, cognitive distortions and responsibility could increase the level ofquality of life of elderly people. Thus interventions based on this approach could be useful in improving the quality of life.

This Study used a semi experimental method as the pretest – posttest and to evaluate the effectiveness of cognitive-existential therapy on depression of parent with children having cancer. Methods: For this purpose 30 parents with children having cancer (24 to 54 years old) were divided randomly into experimental and control groups. Both groups were evaluated with Beck depression Inventory–II (1996), before the intervention. The experimental group received group cognitive-existential therapy in 12 sessions, each lasting 90 minutes (2 sessions per week), but no psychological intervention was applied in control group. At the end of intervention, both groups were reassessed. Data using SPSS software for statistical analysis of covariance test were analyzed. Results indicate significant differences in rates of depression between the two groups in favor of experimental group. Conclusion: cognitive-existential therapy can be effective in reducing depression in parent of children with cancer.

Familial adenomatous polyposis (FAP) is a genetic disease causing hundreds of premalignant polyps in affected persons and is an ideal model to study transitions of early precancer states to colorectal cancer (CRC). We performed deep multiomic profiling of 93 samples, including normal mucosa, benign polyps and dysplastic polyps, from six persons with FAP. Transcriptomic, proteomic, metabolomic and lipidomic analyses revealed a dynamic choreography of thousands of molecular and cellular events that occur during precancerous transitions toward cancer formation. These involve processes such as cell proliferation, immune response, metabolic alterations (including amino acids and lipids), hormones and extracellular matrix proteins. Interestingly, activation of the arachidonic acid pathway was found to occur early in hyperplasia; this pathway is targeted by aspirin and other nonsteroidal anti-inflammatory drugs, a preventative treatment under investigation in persons with FAP. Overall, our results reveal key genomic, cellular and molecular events during the earliest steps in CRC formation and potential mechanisms of pharmaceutical prophylaxis., (© 2024. The Author(s).)

Immune therapeutics holds great promise in the treatment of type 1 diabetes (T1D). Nonetheless, their progress is hampered by limited efficacy, equipoise, or issues of safety. To address this, a novel and specific nanodelivery platform for T1D that targets high endothelial venules (HEVs) presented in the pancreatic lymph nodes (PLNs) and pancreas is developed. Data indicate that the pancreata of nonobese diabetic (NOD) mice and patients with T1D are unique in their expression of newly formed HEVs. Anti-CD3 mAb is encapsulated in poly(lactic-co-glycolic acid)-poly(ethylene glycol) nanoparticles (NPs), the surfaces of which are conjugated with MECA79 mAb that recognizes HEVs. Targeted delivery of these NPs improves accumulation of anti-CD3 mAb in both the PLNs and pancreata of NOD mice. Treatment of hyperglycemic NOD mice with MECA79-anti-CD3-NPs results in significant reversal of T1D compared to those that are untreated, treated with empty NPs, or provided free anti-CD3. This effect is associated with a significant reduction of T effector cell populations in the PLNs and a decreased production of pro-inflammatory cytokine in the mice treated with MECA79-anti-CD3-NPs. In summary, HEV-targeted therapeutics may be used as a means by which immune therapeutics can be delivered to PLNs and pancreata to suppress autoimmune diabetes effectively., (© 2023 Wiley-VCH GmbH.)

Background: Multiple sclerosis (MS) is a neurological disease in which the myelin lining the central nervous system is damaged. The complex and unpredictable nature of MS disease makes the diagnosis process more difficult for the patient. The aim of this study was to review the lived experiences of patients with multiple sclerosis when receiving the diagnosis., Methods: We followed the guidelines of the Preferred Reporting Items for Systematic Reviews (PRISMA) statement. A systematic search was performed using four databases including PubMed, Scopus, Web of Science, and psych info on April 2022., Results: We found 537 articles. After Applying relevant exclusion criteria removing duplicate and irrelevant articles, 13 studies were included in our systematic review after the abstract and full-text screening. Our findings collected data from 10 sub-themes in the following 3 themes to capture patients' experiences after receiving the diagnosis. These included: Emotional reactions to receiving the diagnosis; Communication with health professionals and knowledge about MS and Fear of being different., Conclusion: It is important to understand patients' experiences with the disease and identifying problems to help and support families, patients, and health care personnel's. Therefore, it is necessary to design or implement therapeutic interventions for patients at the time of receiving the diagnosis to reduce psychological problems., Competing Interests: Declaration of Competing Interest The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Objective: the aim of this systematic review was to summarize the coping strategies utilized by people with multiple sclerosis (pwMS) and the association of these strategies with mood symptoms, quality of life, fatigue and disability in pwMS., Methods: search in the literature was performed across electronic databases of PubMed, Web of Science, and Scopus. No time constraint was applied for searching across the databases. Manual search was also performed on the list of references of the retrieved papers. Two authors independently evaluated the retrieved citations based on predetermined inclusion criteria in two screening stages. This systematic review followed Joanna Briggs Institute (JBI) guidelines for cross-sectional research. The findings related to the coping patterns against MS as well as the relationship between the patterns and MS symptoms were extracted and synthesized., Findings: A total of 2390 records were identified. After removing duplicates and based on screening, 39 papers met the eligibility criteria for qualitative synthesis, and they were evaluated in terms of quality of study. When comparing genders, women used more religious coping, social support seeking, and positive coping compared to men. Depression and anxiety were associated with maladaptive and emotion-oriented coping strategies. The patients who used positive and problem solving oriented coping strategies had a better score of quality of life. Disability and fatigue were associated with applying avoidance and maladaptive coping strategies., Conclusion: the findings of this review suggest that understanding the relationship between coping strategies and quality of life as well as the symptoms related to the disease is important to understand development of psychological interventions adopted by specialists to support these patients., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest relating to this study., (Copyright © 2022. Published by Elsevier B.V.)

Lymph nodes (LNs) are the critical sites of immunity, and the stromal cells of LNs are crucial to their function. Our understanding of the stromal compartment of the LN has deepened recently with the characterization of nontraditional stromal cells. CD41 (integrin αIIb) is known to be expressed by platelets and hematolymphoid cells. We identified two distinct populations of CD41 + Lyve1 + and CD41 + Lyve1 - cells in the LNs. CD41 + Lyve1 - cells appear in the LN mostly at the later stages of the lives of mice. We identified CD41 + cells in human LNs as well. We demonstrated that murine CD41 + cells express mesodermal markers, such as Sca-1, CD105 and CD29, but lack platelet markers. We did not observe the presence of platelets around the HEVs or within proximity to fibroblastic reticular cells of the LN. Examination of thoracic duct lymph fluid showed the presence of CD41 + Lyve1 - cells, suggesting that these cells recirculate throughout the body. FTY720 reduced their trafficking to lymph fluid, suggesting that their egress is controlled by the S1P1 pathway. CD41 + Lyve1 - cells of the LNs were sensitive to radiation, suggestive of their replicative nature. Single cell RNA sequencing data showed that the CD41 + cell population in naïve mouse LNs expressed largely stromal cell markers. Further studies are required to examine more deeply the role of CD41 + cells in the function of LNs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Dai, Uehara, Li, LaBarre, Banouni, Ichimura, Lee-Sundlov, Kasinath, Sullivan, Ni, Barone, Giannini, Bahmani, Sage, Patsopoulos, Tsokos, Bromberg, Hoffmeister, Jiang and Abdi.)

For material modeling and discovery, synthetic microstructures play a critical role as digital twins. They provide stochastic samples upon which direct numerical simulations can be conducted to populate material databases. A large ensemble of simulation data on synthetic microstructures may provide supplemental data to inform and refine macroscopic material models, which might not be feasible from physical experiments alone. However, synthesizing realistic microstructures with realistic microstructural attributes is highly challenging. Thus, it is often oversimplified via rough approximations that may yield an inaccurate representation of the physical world. Here, we propose a novel deep learning method that can synthesize realistic three-dimensional microstructures with controlled structural properties using the combination of generative adversarial networks (GAN) and actor-critic (AC) reinforcement learning. The GAN-AC combination enables the generation of microstructures that not only resemble the appearances of real specimens but also yield user-defined physical quantities of interest (QoI). Our validation experiments confirm that the properties of synthetic microstructures generated by the GAN-AC framework are within a 5% error margin with respect to the target values. The scientific contribution of this paper resides in the novel design of the GAN-AC microstructure generator and the mathematical and algorithmic foundations therein. The proposed method will have a broad and substantive impact on the materials community by providing lenses for analyzing structure-property-performance linkages and for implementing the notion of 'materials-by-design'., (© 2022. The Author(s).)

Background and Aim: Aging is a stage during which stressful events occur frequently. The method of coping with this stress can play an important role in an elderly's mental health. The present study aimed at investigating the role of coping strategies in stressful life events associated with depression., Method: The present study was a correlational one conducted by structural equation analysis. As many as 841 elderly people were selected from the general population by adopting a cluster sampling method. Beck's Depression Inventory as well as strategies of coping with stress and stressful events were applied., Results: The model test, analyzed by AMOS by using path analysis, indicated that age is positively associated with stressful life events and depression. The "stressful life events" variable is directly and indirectly associated with depression. The emotion-focused coping strategies were positively associated with depression, and problem-focused coping strategies were negatively associated with depression., Conclusion: As an individual grows older and experiences stressful life events, his/her depression increases. Elderly people with problem-focused coping strategies are likely to experience less depression., Competing Interests: There are no conflicts of interest., (Copyright: © 2021 Journal of Family Medicine and Primary Care.)

Intratumoral immunotherapy is an emerging modality for the treatment of solid tumors. Toll-like receptor (TLR) agonists have shown promise for eliciting immune responses, but systemic administration often results in the development of adverse side effects. Herein, we investigate whether localized delivery of the TLR agonist, resiquimod (R848), via platelet membrane-coated nanoparticles (PNP-R848) elicits antitumor responses. The membrane coating provides a means of enhancing interactions with the tumor microenvironment, thereby maximizing the activity of R848. Intratumoral administration of PNP-R848 strongly enhances local immune activation and leads to complete tumor regression in a colorectal tumor model, while providing protection against repeated tumor re-challenges. Moreover, treatment of an aggressive breast cancer model with intratumoral PNP-R848 delays tumor growth and inhibits lung metastasis. Our findings highlight the promise of locally delivering immunostimulatory payloads using biomimetic nanocarriers, which possess advantages such as enhanced biocompatibility and natural targeting affinities.

Cervical cancer is the second most common leading cause of women's death due to cancer worldwide, about 528,000 patients' cases and 266,000 deaths per year, related to human papillomavirus (HPV). Peptide-based vaccines being safe, stable, and easy to produce have demonstrated great potential to develop therapeutic HPV vaccine. In this study, the major histocompatibility complex (MHC) class I, class II T cell epitopes of HPV16-E7 were predicted. Therefore, we designed a plan to find the most effective peptides to prompt appropriate immune responses. For this purpose, retrieving protein sequences, conserved region identification, phylogenic tree construction, T cell epitope prediction, epitope-predicted population coverage calculation, and molecular docking were performed consecutively and most effective immune response prompting peptides were selected. Based on different tools index, six CD8+ T cells and six CD4+ epitopes were chosen. This combination of 12 epitopes created a putative global vaccine with a 95.06% population coverage. These identified peptides can be employed further for peptide analysis and can be used as a peptide or poly-epitope candidates for therapeutic vaccine studies to treat HPV-associated cancers., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© Springer Nature B.V. 2020.)

Specific delivery platforms for drugs to the kidney and diagnostic agents to renal cell carcinoma (RCC) constitute urgent but unfulfilled clinical needs. To address these challenges, we engineered nanocarriers that interact selectively for the first time with proximal tubule epithelial cells (PTECs) in the kidney and with RCC through the interplay between lambda light chains (LCs) attached to PEGylated polylactic-co-glycolic acid (PLGA) nanoparticles and the membrane protein megalin. Systemic administration of these light chain-conjugated nanoparticles (LC-NPs) to mice resulted in their specific retention by megalin-expressing PTECs for seven days. Repetitive dosing of LC-NPs demonstrated no renal toxicity. LC-NPs also localized selectively to megalin-expressing RCC tumors in mice. Moreover, we confirmed that both the primary tumor and lymph node metastases of human RCC express megalin, reinforcing the potential of LC-NPs for clinical use. Thus, LC-NPs can contribute potentially to improving the management of both non-oncologic and oncologic renal disorders., Competing Interests: Conflict of Interest All authors declare no conflict of interests.

Background: Vitamin D inhibits cell proliferation via the vitamin D receptor (VDR), which may affect breast cancer risk. This study aimed to investigate the association of ApaI and TaqI polymorphisms of the VDR gene with breast cancer risk which followed by stratified analysis., Materials and Methods: A case-control study was conducted on 150 breast cancer patients and 150 healthy controls. VDR ApaI and TaqI genotyping were performed by PCR-RFLP. Some demographic and pathologic features of patients were extracted from their archived files and then were analyzed by genotypes distributions., Results: For ApaI polymorphism, our data showed a significant difference between the patient and healthy groups for mutant allele carriers compared with those with AA genotype. Besides, statistical analysis showed that there was a significant association between the C allele and the increased risk of breast cancer. For TaqI polymorphism, statistical analysis revealed that there was a significant association between CC genotype and increased risk of breast cancer. Also, there was a significant association between the C allele and the increased risk of breast cancer. In a preliminary study, stratified analysis based on the size of tumor and lymph node metastasis revealed no significant association between two ApaI and TaqI variations and these parameters., Conclusions: Based on our results, the VDR ApaI and TaqI variations could be considered as genetic risk factors for breast cancer. However, further studies with a larger sample size are required to obtain more accurate outcomes, especially in stratified analysis.

Objective: Despite the fact that a practical definition of addiction recovery is necessary to conduct an appropriate intervention and research, this concept is still vague and there is no consensus over its meaning and how to measure it. Thus, this study aimed to define and clarify this concept based on the available literature. Method : The theoretical part of Schwartz_Barcott and Kim's Hybrid Model of concept analysis was used to analyze the concept of "Addiction Recovery." To find the relevant literature, an electronic search on valid databases was conducted using keywords related to the concept of addiction recovery. Medlib, IranMedex, Magiran, SID, Irandoc, Google Scholar, PubMed, Web of Science, Medline, Scopus, Pro Quest, CINAHL, Science Direct, Ovid, and Wiley databases were searched up to December 2018 without a time limitation using the following keywords: "Substance use disorders", "Drug use", "Recovery", "Opioids", "Addiction treatment", "Dependency", "Rehabilitation", Remission", "Concept analysis", "Restore", "Definition", "Meaning", and "Conceptualization". The Conventional content analysis was used on selected research articles. Results: From a total of 9520 articles, 39 were reviewed and analyzed. Five attributes were selected, including the process of change, being holistic, being client-centric, learning healthy coping, and being multistage. Antecedents are organized into 2 interacting categories: personal and social resources. Personal resources refer to the person, his/her addiction, and the treatment characteristics, while social resources refer to the family, the community, and the context resources. Addiction recovery leads to sustained abstinence, improved physical and psychological health, improved quality of life and satisfaction, meaningful living, and citizenship. Conclusion: These findings may form a basis for the theories, scales, and criteria for the assessment of addiction recovery and will be useful in clinical practices and research. Also, these findings could help health care professionals to understand the concept of addiction recovery, which is important in improving the recovering person in all aspects of rehabilitation. We will report the implementation and analytical phase of this research project, namely, "the addiction recovery concept analysis" in Iran., (Copyright © Psychiatry & Psychology Research Center, Tehran University of Medical Sciences.)

Background: Meaning in life is one of the psychological domains that is most severely affected in patients with life-threatening illnesses. The importance of meaning-making mandates the development of reliable tools to assess this construct. Steger's Meaning in Life Questionnaire (MLQ) is one of the most valid and reliable instruments that determines the search for and presence of meaning in life. The present study was conducted to provide psychometric data on the MLQ in a sample of patients with life-threatening illnesses., Methods: The MLQ was completed by 301 patients (aged 20-80 years) diagnosed with life-threatening illnesses (cancer and multiple sclerosis) and referred to hospitals. Confirmatory factor analysis and Pearson's correlation test were used to determine the construct validity of the questionnaire., Results: The confirmatory factor analysis supported the original two-factor model of the MLQ, comprised of the presence of meaning (five items) and search for meaning (five items). The responses to the MLQ did not differ by sociodemographic factors. Most importantly, contrary to previous findings, the correlation between the two subscales, i.e., search for meaning and presence of meaning, was significant and positive., Conclusion: The results showed that the MLQ is a valid and reliable measure for assessing meaning in life that can be applied in research on meaning in life among other patient populations., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2020 Maede Naghiyaee et al.)

Background: Paraoxonase 1 (PON1), a multifactorial antioxidant enzyme, has a defensive role against oxidative stress, which is believed to contribute to cancer development. This study aimed to investigate the association of PON1-L55M functional polymorphism with breast cancer risk., Material and Methods: In the experimental study, blood samples were collected from 150 healthy women controls and 150 breast cancer subjects. The L55M genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism., Results: Our analysis showed that the genotypes distribution is in Hardy-Weinberg equilibrium for both case and control groups. Our data revealed that there are significant associations between PON1-L55M polymorphism and breast cancer risk in homozygote (OR= 2.13, 95%CI= 1.14-4.00, p= 0.018), dominant (OR= 1.72, 95%CI= 1.07-2.76, p= 0.024), and allelic (OR= 1.55, 95%CI= 1.12-2.15, p= 0.008) models., Conclusions: Our results suggest that the PON1-L55M genetic variation could be a genetic risk factor for breast cancer risk and it could be considered as a molecular biomarker for screening of susceptible women., .

Inflammation occurring within the transplanted organ from the time of harvest is an important stimulus of early alloimmune reactivity and promotes chronic allograft rejection. Chronic immune-mediated injury remains the primary obstacle to the long-term success of organ transplantation. However, organ transplantation represents a rare clinical setting in which the organ is accessible ex vivo , providing an opportunity to use nanotechnology to deliver therapeutics directly to the graft. This approach facilitates the directed delivery of immunosuppressive agents (ISA) to target local pathogenic immune responses prior to the transplantation. Here, we have developed a system of direct delivery and sustained release of mycophenolate mofetil (MMF) to treat the donor organ prior to transplantation. Perfusion of a donor mouse heart with MMF-loaded PEG-PLGA nanoparticles (MMF-NPs) prior to transplantation abrogated cardiac transplant vasculopathy by suppressing intragraft pro-inflammatory cytokines and chemokines. Our findings demonstrate that ex vivo delivery of an ISA to donor organs using a nanocarrier can serve as a clinically feasible approach to reduce transplant immunity.

Background: Nanomedicine offers an excellent opportunity to tackle treatment-refractory malignancies by enhancing the delivery of therapeutics to the tumor site. High endothelial venules (HEVs) are found primarily in lymph nodes or formed de novo in peripheral tissues during inflammatory responses. They express peripheral node addressin (PNAd), which is recognized by the monoclonal antibody MECA79., Methods: Here, we demonstrated that HEVs form de novo in human pancreatic ductal adenocarcinoma (PDAC). We engineered MECA79 coated nanoparticles (MECA79-NPs) that recognize these ectopic HEVs in PDAC., Findings: The trafficking of MECA79-NPs following intravenous delivery to human PDAC implanted in a humanized mouse model was more robust than non-conjugated NPs. Treatment with MECA79-Taxol-NPs augmented the delivery of Paclitaxel (Taxol) to the tumor site and significantly reduced the tumor size. This effect was associated with a higher apoptosis rate of PDAC cells and reduced vascularization within the tumor., Interpretation: Targeting the HEVs of PDAC using MECA79-NPs could lay the ground for the localized delivery of a wide variety of drugs including chemotherapeutic agents. FUND: National Institutes of Health (NIH) grants: T32-EB016652 (B·B.), NIH Cancer Core Grant CA034196 (L.D.S.), National Institute of Allergy and Infectious Diseases grants R01-AI126596 and R01-HL141815 (R.A.)., (Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.)

The targeted delivery of therapeutic drugs to lymph nodes (LNs) provides an unprecedented opportunity to improve the outcomes of transplantation and immune-mediated diseases. The high endothelial venule is a specialized segment of LN vasculature that uniquely expresses peripheral node addressin (PNAd) molecules. PNAd is recognized by MECA79 mAb. We previously generated a MECA79 mAb-coated microparticle (MP) that carries tacrolimus. Although this MP trafficked to LNs, it demonstrated limited therapeutic efficacy in our transplant model. Here, we have synthesized a nanoparticle (NP) as a carrier of anti-CD3, and optimized the conjugation strategy to coat the NP surface with MECA79 mAb (MECA79-anti-CD3-NP) to enhance LN accumulation. As compared with nonconjugated NPs, a significantly higher quantity of MECA79-NPs accumulated in the draining lymph node (DLN). Many MECA79-NPs underwent internalization by T cells and dendritic cells within the LNs. Short-term treatment of murine cardiac allograft recipients with MECA79-anti-CD3-NP resulted in significantly prolonged allograft survival in comparison with the control groups. Prolonged graft survival following treatment with MECA79-anti-CD3-NP was characterized by a significant increase in intragraft and DLN Treg populations. Treg depletion abrogated the prolongation of heart allograft survival. We believe this targeted approach of drug delivery could redefine the methods of administering immune therapeutics in transplantation.

Recent studies in cancer research have focused intensely on the antineoplastic effects of immune checkpoint inhibitors. While the development of these inhibitors has progressed successfully, strategies to further improve their efficacy and reduce their toxicity are still needed. We hypothesized that the delivery of anti-PD-1 antibody encapsulated in PLGA nanoparticles (anti-PD-1 NPs) to the spleen would improve the antitumor effect of this agent. Unexpectedly, we found that mice treated with a high dose of anti-PD-1 NPs exhibited significantly higher mortality compared with those treated with free anti-PD-1 antibody, due to the overactivation of T cells. Administration of anti-PD-1 NPs to splenectomized LT-α-/- mice, which lack both lymph nodes and spleen, resulted in a complete reversal of this increased mortality and revealed the importance of secondary lymphoid tissues in mediating anti-PD-1-associated toxicity. Attenuation of the anti-PD-1 NPs dosage prevented toxicity and significantly improved its antitumor effect in the B16-F10 murine melanoma model. Furthermore, we found that anti-PD-1 NPs undergo internalization by DCs in the spleen, leading to their maturation and the subsequent activation of T cells. Our findings provide important clues that can lead to the development of strategies to enhance the efficacy of immune checkpoint inhibitors.

The contribution of the kidney-draining lymph node (KLN) to the pathogenesis of ischemia-reperfusion injury (IRI) of the kidney and its subsequent recovery has not been explored in depth. In addition, the mechanism by which repetitive IRI contributes to renal fibrosis remains poorly understood. Herein, we have found that IRI of the kidney is associated with expansion of high endothelial venules (HEVs) and activation of fibroblastic reticular cells (FRCs) in the KLN, as demonstrated by significant expansion in the extracellular matrix. The lymphotoxin α signaling pathway mediates activation of FRCs, and chronic treatment with lymphotoxin β receptor-immunoglobulin fusion protein (LTβr-Ig) resulted in marked alteration of the KLN as well as augmentation of renal fibrosis. Depletion of FRCs reduced T cell activation in the KLN and ameliorated renal injury in acute IRI. Repetitive renal IRI was associated with senescence of FRCs, fibrosis of the KLN, and renal scarring, which were ameliorated by FRC administration. Therefore, our study emphasizes the critical role of FRCs in both the initiation and repair phases of injury following IRI of the kidney.

Purpose of Review: Organ transplantation is a life-saving procedure and the only option for patients with end-organ failure. Immune therapeutics have been key to the success of organ transplantation. However, immune therapeutics are still unable to eliminate graft rejection and their toxicity has been implicated in poorer long-term transplant outcomes. Targeted nanodelivery has the potential to enhance not only the therapeutic index but also the bioavailability of the immune therapeutics. One of the key sites of immune therapeutics delivery is lymph node where the priming of immune cells occur. The focus of this review is on nanomedicine research to develop the targeted delivery of immune therapeutics to lymph nodes for controlling immune activation., Recent Findings: As nanomedicine creates its niche in clinical care, it provides novel immunotherapy platforms for transplant recipients. Draining lymph nodes are the primary loci of immune activation and represent a formidable site for delivery of wide variety of immune therapeutics. There have been relentless efforts to improve the properties of nanomedicines, to have in-depth knowledge of antigen and drug loading, and, finally, to explore various routes of passive and active targeted delivery to lymph nodes., Summary: The application of nanotechnology principles in the delivery of immune therapeutics to the lymph node has created enormous excitement as a paradigm shifting approach that enables targeted delivery of a gamut of molecules to achieve a desired immune response. Therefore, innovative strategies that improve their efficacy while reducing their toxicity are among the highest unmet needs in transplantation.

We investigated the molecular mechanism of apoptosis induced by novel jatropha-6(17),11E-diene class derivatives, compounds A, B, and C that were extracted from Euphorbia osyridea Boiss, in the ovarian cancer cell lines Caov-4 and OVCAR-3. The OVCAR-3 and Caov-4 cell lines were treated with different concentrations of these compounds. Cytotoxicity was evaluated using MTT, clonogenic survival assay, and flow cytometry assays. The production of reactive oxygen species (ROS), mitochondrial membrane potential (Δ Ψm ), and the activity of caspase 3 and 9 were evaluated. Compounds A, B, and C reduced cell viability in a dose-dependent manner (P < 0.05). The IC 50 values were calculated as 46.27 ± 3.86, and 38.81 ± 3.30 μmol/L for compound A, 36.48 ± 3.18 and 42.59 ± 4.50 μmol/L for compound B, and 85.86 ± 6.75 and 75.65 ± 2.56 μmol/L for compound C against the Caov-4 and OVCAR-3 cell lines, respectively. Apoptosis evaluation showed that jatrophane derivatives increase both early and late apoptosis (P < 0.01). These compounds also increased ROS generation, Δ Ψm , and the activity of caspase 3 and 9 in the treated cells. These results showed that compounds A and B have significant inhibitory effects on OVCAR-3 and Caov-4 proliferation and induction of apoptosis.

Ischemia-reperfusion injury (IRI) evokes intragraft inflammatory responses, which markedly augment alloimmune responses against the graft. Understanding the mechanisms underlying these responses is fundamental to develop therapeutic regimens to prevent/ameliorate organ IRI. Here, we demonstrate that IRI results in a marked increase in mitochondrial damage and autophagy in dendritic cells (DCs). While autophagy is a survival mechanism for ischemic DCs, it also augments their production of interleukin (IL)-6. Allograft-derived dendritic cells (ADDCs) lacking autophagy-related gene 5 (Atg5) showed higher death rates posttransplantation. Transplanted ischemic hearts from CD11cCre/Atg5 conditional knockout mice showed marked reduction in intragraft expression of IL-6 compared with controls. To antagonize the effect of IL-6 locally in the heart, we synthesized novel anti-IL-6 nanoparticles with capacity for controlled release of anti-IL-6 over time. Compared with systemic delivery of anti-IL-6, localized delivery of anti-IL-6 significantly reduced chronic rejection with a markedly lower amount administered. Despite improved allograft histology, there were no changes to splenic T cell populations, illustrating the importance of local IL-6 in driving chronic rejection after IRI. These data carry potential clinical significance by identifying an innovative, targeted strategy to manipulate organs before transplantation to diminish inflammation, leading to improved long-term outcomes., (© 2017 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Context: Chronic myelogenous leukemia (CML) is a chronic malignancy of myeloid linage associated with a significant increase in granulocytes in bone marrow and peripheral blood. CML diagnosis is based on detection of Philadelphia chromosome and "Abelson murine leukemia viral oncogene homolog" (ABL)-"breakpoint cluster region protein" fusions (ABL-BCR fusions)., Aims: In this study, patients with CML morphology were studied according to ABL-BCR fusions and the relationship between the fusions and peripheral blood cell changes was examined., Materials and Methods: All patients suspected to chronic myeloproliferative disorders in Lorestan Province visiting subspecialist hematology clinics who were confirmed by oncologist were studied over a period of 5 years. After completing basic data questionnaire, blood samples were obtained with informed consent from the patients. Blood cell count and morphology were investigated and RNA was extracted from blood samples. cDNA was synthesized from RNA and ABL-BCR fusions including b3a2 and b2a2 (protein 210 kd or p210), e1a2 (protein 190 kdor p190), and e19a2 (protein 230 kdor p230) were studied by multiplex reverse transcription polymerase chain reaction method. Coexistence of e1a2 and b2a2 (p210/p190) fusions was also studied. The prevalence of mutations and their correlation with the blood parameters were statistically analyzed., Results: Of 58 patients positive for ABL-BCR fusion, 18 (30.5%) had b2a2 fusion, 37 (62.71%) had b3a2 fusion and three (3.08%) had e1a2 fusion. Coexistence of e1a2 and b2a2 (p210/p190) was not observed. There was no significant correlation between ABL-BCR fusions and white blood cell count, platelet count, and hemoglobin concentration., Conclusions: The ABL-BCR fusions in Lorestan Province were similar to other studies in Iran, and b3a2 fusion had the highest prevalence in the studied patients studied.

Aim: To investigate if there is an association between M235T polymorphism of angiotensinogen gene and myocardial infarction (MI) risk and perform a meta-analysis and an in silico approach., Methods: This case-control study included 340 participants (155 MI patients and 185 controls) examined at Kashan University of Medical Sciences (Kashan, Iran) between 2013 and 2015. Meta-analysis included 25 studies with 6334 MI patients and 6711 controls. Bioinformatics tools were applied to evaluate the impact of M235T polymorphism on angiotensinogen function and structure., Results: Genetic association study revealed a significant association between TT genotype (odds ratio [OR] 2.08, 95% confidence interval [CI] 1.08-4.00, P=0.029) and T allele (OR 1.45, 95% CI 1.06-1.99, P=0.021) and MI risk. Meta-analysis also revealed a significant association between M235T polymorphism and MI risk in allelic (OR 1.55, 95% CI 1.10-2.18, P=0.012) and recessive (OR 1.69, 95% CI 1.13-2.53, P=0.010) models within Asian population. In silico-analysis revealed that M235T fundamentally changed the function of angiotensinogen (score 32; expected accuracy 66%)., Conclusions: Our study suggests that M235T polymorphism might be a helpful biomarker for screening of susceptible individuals for MI in Asian population.

Introduction: Hopefulness is one of the most significant predictors of adaptation in hemodialysis patients, and plays a vital role in the recovery process. In contrast to hopefulness, depression is a frequent psychological reaction of the hemodialysis treatment with many negative consequences. The current research was designed to examine the effect of cognitive-existential treatment on the level of hopefulness and depression in hemodialysis patients., Materials & Methods: This quasi-experimental research included 22 female patients suffering from chronic kidney failure disease undergoing hemodialysis treatment for at least 3 months. The patients were randomly assigned into two groups of experimental and control conditions. The experimental group received a combination of treatment including some elements of "existentialism" philosophy and a "cognitive" approach designed for the Iranian population. The treatment protocol lasted for 12 sessions of 90 minutes twice per week prior to the entry of the patient to the dialysis session.  Miller's hope scale and BDI-II-21 were employed to collect the data. Statistical analysis was performed on the data using analysis of covariance by SPSS: 16 software., Results: The result of the analysis indicated that there was a significant improvement in hopefulness level and decrease in depression of the patients in the experiment condition (P<0.01)., Conclusion: The result of analysis showed that cognitive-existential treatment resulted in the increase of hopefulness and decrease level of depression in the hemodialysis patients suffering from chronic kidney failure.

Ovarian cancer remains the dominant cause of death due to malignancies of the female reproductive system. The capability to identify and remove all tumors during intraoperative procedures may ultimately reduce cancer recurrence, and lead to increased patient survival. The objective of this study is to investigate the effectiveness of an optical nano-structured system for targeted near infrared (NIR) imaging of ovarian cancer cells that over-express the human epidermal growth factor receptor 2 (HER2), an important biomarker associated with ovarian cancer. The nano-structured system is comprised of genome-depleted plant-infecting brome mosaic virus doped with NIR chromophore, indocyanine green, and functionalized at the surface by covalent attachment of monoclonal antibodies against the HER2 receptor. We use absorption and fluorescence spectroscopy, and dynamic light scattering to characterize the physical properties of the constructs. Using fluorescence imaging and flow cytometry, we demonstrate the effectiveness of these nano-structures for targeted NIR imaging of HER2 receptors in vitro. These functionalized nano-materials may provide a platform for NIR imaging of ovarian cancer.

Objectives: Lipocalin2 (Lcn2) gene is highly expressed in response to various types of cellular stresses. The precise role of Lcn2 has not been fully understood yet. However, it plays a key role in controlling vital cellular processes such as proliferation, apoptosis and metabolism. Recently it was shown that Lcn2 decreases senescence and increases proliferation of mesenchymal stem cells (MSC) with finite life span under either normal or oxidative stress conditions. However, Lcn2 effects on immortal cell line with infinite proliferation are not defined completely. Materials and., Materials and Methods: HEK-293 cells were transfected with recombinant pcDNA3.1 containing Lcn2 fragment (pcDNA3.1-Lcn2). Expression of lipocalin2 in transfected cells was evaluated by RT-PCR, real time RT-PCR, and ELISA. Different cell groups were treated with H2O2 and WST-1 assay was performed to determine their proliferation rate. Senescence was studied by β-galactosidase and gimsa staining methods as well as evaluation of the expression of senescence-related genes by real time RT-PCR., Results: Lcn2 increased cell proliferation under normal culture condition, while the proliferation slightly decreased under oxidative stress. This decrease was further found to be attributed to senescence., Conclusion: Our findings indicated that under harmful conditions, Lcn2 gene is responsible for the regulation of cell survival through senescence.

The regenerative potential of mesenchymal stem cells (MSCs) is impaired by cellular senescence, a multi factorial process that has various functions. However, pathways and molecules involved in senescence have not been fully identified. Lipocalin 2 (Lcn2) has been the subject of intensive research, due to its contribution to many physiological and pathophysiological conditions. The implication of Lcn2 has been reported in many conditions where senescence also occurs. In the present study, we evaluated the role of Lcn2 in the occurrence of senescence in human bone marrow-derived mesenchymal stem cells (hB-MSCs) under oxidative conditions. When hB-MSCs were genetically engineered to over-express Lcn2 (MSC-Lcn2) and exposed to H2O2, the proliferation rate of the cells increased. However, the number of colonies and the number of cells that made up each colony in both MSC-V and MSC-Lcn2 cells decreased compared to those cultivated under normal conditions. Our results revealed that over-expression of recombinant Lcn2 in hB-MSCs decreases senescence induced by H2O2 treatment. Senescent cells were observed in aged hB-MSCs; however, no alteration in the expression level of Lcn2 was detected compared to earlier passages. Finally, a higher amount of Lcn2 protein was detected in the plasma of the elderly than in young people. Our findings suggest that Lcn2 might restore the health and regeneration potential of MSCs by decreasing senescence.

Background and Objectives: Ovarian cancer remains the deadliest malignancy of the female reproductive system. The ability to identify and destroy all ovarian tumor nodules may have a termendous impact on preventing tumor recurrence, and patient survival. The objective of this study is to investigate the effectiveness of a nano-structured system for combined near infrared (NIR) fluorescence imaging of human epidermal growth factor receptor-2 (HER2) over-expression, as a biomarker of ovarian cancer cells, and photothermal destruction of these cells in vitro., Materials and Methods: The nano-structured system consists of the near infrared dye, indocyanine green (ICG), encapsulated within poly(allylamine) hydrochloride chains cross-linked ionically with sodium phosphate. The surface of the construct is functionalized by covalently attached polyethylene glycol, and monoclonal antibodies against HER2 using reducitve amination methods. We use dynamic light scattering, and absorption and fluorescence spectroscopy for phyiscal characterization of the constructs. Flow cytometry and fluorescence microscopy are used to investigate molecular targeting and imaging capabilities of the constructs against SKOV3 and OVCAR3 ovarian cancer cell lines, which have relatively high and low expression levels of the HER2 receptor, respectively. Continuous NIR laser irradiation at 808 nm is used to investigating the utility of the constructs in mediating photothermal destruction of SKOV3 cells., Results: Flow cytometry results indicate that the functionalized nano-constructs are more effective in targeting the HER2 receptor than non-encapsulated ICG and non-functionlaized constructs (P < 0.005). Fluorescence microscopic images show the capaiblity of the functionalized constructs in NIR imaging of HER2 overexpression. The functionalized nano-constructs are also capable of inducing a significantly greater increase in photothermal destruction of SKOV3 cells than free ICG and non-functionalized constructs (P < 0.005)., Conclusion: We have demonstrated the efficacy of polymeric nano-structured constructs loaded with ICG, and functionalized with the monoclonal antibodies, as thernaostic materials for targted molecular NIR imaging of the HER2 receptor overexpression on ovarian cancer cells, and photothermal destruction of these cells. These nanoparticles may prove useful towards intraoperative detection, imaging, and phototherapy of small ovarian cancer nodules., (© 2014 Wiley Periodicals, Inc.)

Development of theranostic nano-constructs may enable diagnosis and treatment of diseases at high spatial resolution. Some key requirements for clinical translation of such constructs are that they must be non-toxic, non-immunogenic, biodegradable, with extended circulating lifetime. Cell-based structures, particularly those derived from erythrocytes, are promising candidate carrier systems to satisfy these requirements. One particular type of theranostic materials utilize light-sensitive agents that once photo-activated can provide diagnostic imaging capability, and elicit therapeutic effects. Here we demonstrate the first successful engineering of hybrid nano-scale constructs derived from membranes of hemoglobin-depleted erythrocytes that encapsulate the near infrared chromophore, indocyanine green. We show the utility of the constructs as photo-theranostic agents in fluorescence imaging and photothermal destruction of human cells. These erythrocyte-mimicking nano-structures can be derived autologously, and may have broad applications in personal nanomedicine ranging from imaging and photo-destruction of cancerous tissues to vascular abnormalities, and longitudinal evaluations of therapeutic interventions.

Near-infrared nanoconstructs present a potentially effective platform for site-specific and deep tissue optical imaging and phototherapy. We have engineered a polymeric nanocapsule composed of polyallylamine hydrochloride (PAH) chains cross-linked with sodium phosphate and doped with indocyanine green (ICG) toward such endeavors. The ICG-doped nanocapsules were coated covalently with polyethylene glycol (5000 daltons) through reductive amination. We administrated the constructs by tail vein injection to healthy mice. To characterize the biodistribution of the constructs, we performed in vivo quantitative fluorescence imaging and subsequently analyzed the various extracted organs. Our results suggest that encapsulation of ICG in these PEGylated constructs is an effective approach to prolong the circulation time of ICG and delay its hepatic accumulation. Increased bioavailability of ICG, due to encapsulation, offers the potential of extending the clinical applications of ICG, which are currently limited due to rapid elimination of ICG from the vasculature. Our results also indicate that PAH and ICG-doped nanocapsules (ICG-NCs) are not cytotoxic at the levels used in this study.

This article describes the development and the examination of surface coatings that suppress the adhesion between glass surfaces and polymer microspheres. Superparamagnetic doping allowed for exerting magnetic forces on the microbeads. The carboxyl functionalization of the polymer provided the means for coating the beads with polyethylene glycol (PEG) with different molecular weight. Under gravitational force, the microbeads settled on glass surfaces with similar polymer coatings. We examined the efficacy of removing the beads from the glass surfaces by applying a pulling force of ~1.2 pN. The percent beads remaining on the surface after applying the pulling force for approximately 5 s served as an indication of the adhesion propensity. Coating of PEG with molecular weight ranging between 3 and 10 kDa was essential for suppressing the adhesion. For the particular substrates, surface chemistry and aqueous media we used, coatings of 5 kDa manifested optimal suppression of adhesion: that is, only 3% of the microbeads remained on the surface after applying the pulling magnetic force. When either the glass or the beads were not PEGylated, the adhesion between them was substantial. Addition of a noncharged surfactant, TWEEN, above its critical micelle concentrations (CMCs) suppressed the adhesion between noncoated substrates. The extent of this surfactant-induced improvement of the adhesion suppression, however, did not exceed the quality of preventing the adhesion that we attained by PEGylating both substrates. In addition, the use of surfactants did not significantly improve the suppression of bead-surface adhesion when both substrates were PEGylated. These findings suggest that such surfactant additives tend to be redundant and that covalently grafted coatings of PEGs with selected chain lengths provide sufficient suppression of nonspecific interfacial interactions.

Near-infrared (NIR) optically active nanoparticles are promising exogenous chromophores for applications in medical imaging and phototherapy. Since nanoparticles can be rapidly eliminated from the body by cells of the reticuloendothelial system, a thriving strategy to increase their blood circulation time is through surface modification with polyethylene glycol (PEG). We constructed polymeric nanocapsules loaded with indocyanine green (ICG), an FDA-approved NIR dye, and coated with aldehyde-terminated PEG. Using optical absorbance spectroscopy and flow cytometry, we investigated the effect of PEG coating and molecular weight (MW) of PEG [5000 and 30,000 Daltons (Da)] on the phagocytic content of human spleen macrophages incubated with ICG-containing nanocapsules (ICG-NCs) between 15 to 360 min. Our results indicate that surface coating with PEG is an effective method to reduce the phagocytic content of ICG-NCs within macrophages for at least up to 360 min of incubation time. Coating the surface of ICG-NCs with the low MW PEG results in lower phagocytic content of ICG-NCs within macrophages for at least up to 60 min of incubation time as compared to ICG-NCs coated with the high MW PEG. Surface coating of ICG-NCs with PEG is a promising approach to prolong vasculature circulation time of ICG for NIR imaging and phototherapeutic applications.

For more than a century, colorimetric and fluorescence staining have been the foundation of a broad range of key bioanalytical techniques. The dynamics of such staining processes, however, still remains largely unexplored. We investigated the kinetics of fluorescence staining of two gram-negative and two gram-positive species with 3,3'-diethylthiacyanine (THIA) iodide. An increase in the THIA fluorescence quantum yield, induced by the bacterial dye uptake, was the principal reason for the observed emission enhancement. The fluorescence quantum yield of THIA depended on the media viscosity and not on the media polarity, which suggested that the microenvironment of the dye molecules taken up by the cells was restrictive. The kinetics of fluorescence staining did not manifest a statistically significant dependence neither on the dye concentration, nor on the cell count. In the presence of surfactant additives, however, the fluorescence-enhancement kinetic patterns manifested species specificity with statistically significant discernibility.