CDR grafting and site-directed mutagenesis approach for the generation and affinity maturation of Anti-CD20 nanobody.

BACKGROUND: Recently, new and advanced techniques have been adopted to design and produce nanobodies, which are used in diagnostic and immunotherapy treatments. Traditionally, nanobodies are prepared from camelid immune libraries that require animal treatments. However, such approaches require large library sizes and complicated selection procedures. The current study has employed CDR grafting and site-directed mutagenesis techniques to create genetically engineered nanobodies against the tumor marker CD20 (anti-CD20 nanobodies) used in leukemia treatment. METHODS AND RESULTS: In this study, we utilized the swapping method to graft CDRs from the VH Rituximab antibody to VHH CDRs. We aimed to enhance the binding affinity of the nanobodies by substituting the amino acids (Y101R-Y102R-Y107R) in the VHH-CDR3. To assess the binding capacity of the mutated nanobodies, we conducted an ELISA test. Moreover, through flow cytometry analysis, we compared the fluorescence intensity of the grafted CD20 and mutant nanobodies with that of the commercially available human anti-CD20 in Raji cells. The results showed a significant difference in the fluorescence intensity of the grafted nanobodies and mutant nanobodies when compared to the commercially available human anti-CD20. CONCLUSION: The approach we followed in this study makes it possible to create multiple anti-CD20 nanobodies with varying affinities without the need for extensive selection efforts. Additionally, our research has demonstrated that computational tools are highly reliable in designing functional nanobodies.

Novel and deleterious nucleotide variations in the HAND1 gene probably affect miRNA target sites and protein function in pediatric patients with congenital heart disease.

BACKGROUND: Congenital heart disease (CHD) is the most prevalent developmental defect and principal cause of infant mortality and affects cardiac and large blood vessel structures in approximately 1% of live births worldwide. To date, numerous studies have related critical genetic dysfunctions to the pathogenesis of CHDs. However, the genetic basis underlying CHD remains largely unknown. In the present study, we investigated the association of nucleotide variations in coding and noncoding regions of the HAND1 gene with the risk of CHD. The HAND1 gene, encoding a helix-loop-helix transcription factor, is particularly relevant for mechanisms underlying CHD since it plays a significant role in heart development. METHODS AND RESULTS: The genomic DNA of 150 unrelated pediatric patients with CHD was screened by PCR-SSCP and direct sequencing. Four novel and heterozygous missense mutations were identified in the first exon, with three causing amino acid substitutions (p.Val149Met, p.Tyr142His, and p.Leu146Met). In-silico analysis also indicated their deleterious impact on protein structure and function. In addition, we identified five novel nucleotide variants in the 3'UTR region (c.*461, c.*342, c.*529, c.*448, c.*593), potentially altering the target sites of miRNAs. These changes include the loss of certain target sites and the acquisition of new ones. CONCLUSIONS: These findings confirm the phenotypic association between CHDs and HAND1 mutations and can pave the way for developing new preventive and therapeutic strategies.

Uncovering the extent of dementia prevalence in Iran: a comprehensive systematic review and meta-analysis.

BACKGROUND: Dementia is one of the major causes of disability and dependency among older people worldwide. The formation of an aging population in Iran can be associated with societal problems, including age-related disorders such as dementia. This study aimed to estimate the prevalence of dementia& Alzheimer disease in adults aged 60 years or older and it's its geographical distribution in Iran. METHODS: A systematic review and meta-analysis study included articles published in both English and Persian languages and utilized various databases including: Google Scholar, PubMed, Web of Science, Magiran, and thesis database of medicine universities up to December 2022. The pooled prevalence was calculated using random effects models. The prevalence was reported separately for different geographical locations and types of area sampling, and age adjustment was performed for the selected studies. All statistical analyses were conducted using metaprop package in STATA version 17. The I2 statistic was applied to assess heterogeneity. RESULTS: The meta-analysis considered nine relevant studies that were carried out up to 2023 in Iran. The study found that the prevalence of dementia in central and east counties was estimated to be 0.14 (95% CI; 0.04-0.31), while in western counties, the prevalence was estimated to be 0.1 (95%CI; 0.01-0.27). The estimated overall crude prevalence of dementia was estimated at 0.14 (95% CI; 0.03-0.31). Estimated prevalence-based health centers sampling and hospital-based studies were 0.02 (95% CI; 0.02-0.03), 0.05 (95% CI 0.06-0.11), respectively. One study used nursing home sampling as the sampling method, and the estimated prevalence was 0.43 (95%CI 0.38-0.49). CONCLUSION: This is the first systematic review and meta-analysis of the prevalence of dementia's disease up to 2023 in Iran. The estimated overall prevalence of dementia is lower than the reported prevalence in European countries and similar to other Asian countries.

Propolis as an adjunctive therapy for treatment of uncomplicated cystitis in women: A randomized double-blind placebo-controlled trial.

The current research is designed to investigate the effect of propolis supplementation on the clinical manifestations in women suffering from uncomplicated cystitis. In this randomized double-blind, placebo-controlled trial, 120 women with uncomplicated cystitis were selected and randomly assigned into two groups to receive two 500 mg capsules of propolis or placebo daily for 7 days along with ciprofloxacin (250 mg). Clinical symptoms including hematuria, urinary frequency, dysuria, suprapubic pain, and urgency, as well as bacteriuria, were assessed before and after the intervention. After supplementation, participants in the intervention group had significantly fewer days of urinary frequency (p < 0.001), dysuria (p = 0.005), and urgency (p = 0.03). However, there was no significant difference between the two groups regarding hematuria and suprapubic pain (p > 0.05). Furthermore, the severity of bacteriuria decreased significantly in both groups. In conclusion, it seems that propolis supplementation in women with uncomplicated cystitis could improve urinary frequency, dysuria, and urgency. However, further clinical trials should be conducted to fully understand the effects of propolis in women suffering from uncomplicated cystitis.

Effects of a Spiritual Care Program on Body Image and Resilience in Patients with Second-Degree Burns in Iran.

This research was conducted with the aim of investigating the effect of a spiritual care program (SCP), on the body image and resilience of second-degree burn patients in Iran. In this semi-experimental study, 60 patients with second-degree burns were selected in the Tehran Mottahari Burns Hospital, Iran in 2022. These patients were divided through random allocation into two intervention and control groups. The patients of both groups filled out Beck's self-esteem Test (BSCT) and Connor-Davidson Resilience Scale (CD-RISC) before, immediately, and 3 months after the intervention. In the intervention group, SCP was performed for eight sessions over two weeks. Firstly, the 4D Model of Spiritual Health and Well-Being was examined, then two one-hour sessions were held with each patient about each dimension, with a total of four dimensions including eight sessions, in which these dimensions were examined and discussed. There was no difference in the score of body image (P = 0.326) and resilience (P = 0.597) before intervention in the control and intervention groups. However, the mean of body image score in the intervention group immediately (109.56 ± 4.86) and 3 months after (109.16 ± 6.06) the intervention was significant, respectively. Also, the mean of resilience score in the intervention group was significant immediately (85.86 ± 6.78) and 3 months after the intervention (85.73 ± 6.27). It is suggested for healthcare providers to use the SCPs to reduce the recovery time of burned patients and reduce complications and treatment costs.

Autophagy, a critical element in the aging male reproductive disorders and prostate cancer: a therapeutic point of view.

Autophagy is a highly conserved, lysosome-dependent biological mechanism involved in the degradation and recycling of cellular components. There is growing evidence that autophagy is related to male reproductive biology, particularly spermatogenic and endocrinologic processes closely associated with male sexual and reproductive health. In recent decades, problems such as decreasing sperm count, erectile dysfunction, and infertility have worsened. In addition, reproductive health is closely related to overall health and comorbidity in aging men. In this review, we will outline the role of autophagy as a new player in aging male reproductive dysfunction and prostate cancer. We first provide an overview of the mechanisms of autophagy and its role in regulating male reproductive cells. We then focus on the link between autophagy and aging-related diseases. This is followed by a discussion of therapeutic strategies targeting autophagy before we end with limitations of current studies and suggestions for future developments in the field.

Significant hypomethylation of MMP9 gene promoter in patients with systemic lupus erythematosus.

OBJECTIVE: Scholars are exploring novel diagnostic and prognostic biomarkers with higher sensitivity and specificity for systemic lupus erythematosus (SLE). In this regard, DNA methylation alterations have aroused attention. The association between the dysfunction of MMP9 and TNFAIP3 genes and SLE has been previously demonstrated. Therefore, in this study, we investigated the methylation level of MMP9 and TNFAIP3 promoters in peripheral blood mononuclear cells (PBMCs) of SLE patients and healthy controls. METHODS: Eighty Iranian SLE patients and 77 healthy individuals were enrolled. The methylation quantification endonuclease-resistant DNA (MethyQESD) method was used to assess methylation levels of MMP9 and TNFAIP3 in extracted DNA of PBMCs. To quantify the diagnostic utility of the promoter methylation level of these genes, the receiver operating characteristic (ROC) curve was constructed. RESULTS: MMP9 promoter was significantly hypomethylated in SLE patients compared with healthy people (p < 0.001), while there was no significant difference in terms of TNFAIP3 promoter methylation levels (p = 0.167). Also, this differential MMP9 methylation was observed in patients with renal involvement and patients without renal involvement (42.07 ± 25.73 vs 56.74 ± 29.71, p = 0.007). ROC analyses indicated that the diagnostic power of the MMP9 promoter methylation level for SLE was 0.839 [95% CI (0.781-0.911)]. Moreover, MMP9 methylation level was negatively correlated with creatinine and anti-dsDNA concentration and positively correlated with C3 and C4 levels. CONCLUSION: The results of this study highlight the application of MMP9 methylation level in PBMCs of SLE patients as a diagnostic biomarker.

Satralizumab, Novel Interleukine-6 Inhibitor for Preventing Descending Thoracic Aorta Aneurysm Development.

BACKGROUND: Descending thoracic aorta aneurysm (dTAA) has increasing incidence and, if left untreated, could lead to death. There is not any study of satralizumab treatment for preventing dTAA formation and progression. MATERIALS AND METHODS: Forty male 10-week-old Rattus norvegicus were enrolled in the experiment. They were divided into four equal groups: dTAA treated with saline (dTAA-P) and dTAA treated with satralizumab (dTAA-S). One of the control groups was treated with saline (C-P), and the other was treated with satralizumab (C-S). Satralizumab and saline were used once every 2 weeks, subcutaneously 120 mg for 4 weeks. dTA diameter was measured at days 0, 3, 7, 14, 21, and 28. RESULTS: IL-6 level was measured on the 7th day that showed significantly increased IL-6 serum level in dTAA-P rats compared to C-P. Maximal dTA diameter (%MAD) was obtained at day 14, which was scientifically matched to the aorta aneurysm definition (>50% increase in diameter). From the seventh day, a significant difference in %MAD was observed between dTAA-P and dTAA-S groups. However, the %MAD of these two groups was significantly higher than control groups till the end of the 28th day. CONCLUSION: Using an IL-6 inhibitor agent to prevent dTAA formation and progression showed promising results. It suggests that using the IL-6 inhibitors in susceptible persons can be considered a lifesaving therapeutic approach.

Oleanolic acid increases the anticancer potency of doxorubicin in pancreatic cancer cells.

Combination therapy is a novel cancer therapy approach that combines two or more chemotherapy drugs. This treatment modality enhances the efficacy of chemotherapy by targeting key pathways in an additive or synergistic manner. Therefore, we investigated the efficacy of combination therapy by widely used chemotherapy drug doxorubicin (DOX) and oleanolic acid (OA) to induction of apoptosis for pancreatic cancer (PC) therapy. The effects of DOX, OA, and their combination (DOX-OA) were investigated on proliferation and viability of PC cell line (PANC-1) by MTT assay. Moreover, migration and invasion of the cancer cells were evaluated by trans-well migration assay and wound healing assay. Flow cytometry and DAPI (4',6-diamidino-2-phenylindole) staining were employed to investigate apoptosis quantification and qualification of the treated cancer cells. Finally, mRNA expression of apoptosis-related genes was assessed by quantitative real-time polymerase chain reaction. Our results demonstrated that the proliferation and metastasis potential of PC cells significantly decreased after treatment by DOX, OA, and DOX-OA. Moreover, we observed an increase in apoptosis percentage in the treated cancer cells. The apoptosis-related gene expression was modified to increase the apoptosis rate in all of the treatment groups. However, the anticancer potency of DOX-OA combination was significantly more than that of DOX and OA treatments alone. Our study suggested that DOX-OA combination exerts more profound anticancer effects against PC cell lines than DOX or OA monotherapy. This approach may increase the efficiency of chemotherapy and reduce unintended side effects by lowering the prescribed dose of DOX.

Type 2 diabetes mellitus and In-hospital Major Adverse Cardiac and Cerebrovascular Events (MACCEs) and postoperative complications among patients undergoing on-pump isolated coronary artery bypass surgery in Northeastern Iran.

BACKGROUND: Diabetes Mellitus (DM) is a rapidly growing disorder worldwide, especially in the Middle East. A higher incidence of coronary artery diseases requiring coronary artery bypass graft (CABG) surgery has been reported in patients with diabetes. We assessed the association between type 2 diabetes mellitus (T2DM) and in-hospital major adverse cardiac and cerebrovascular events (MACCEs) and postoperative complications among patients who underwent on-pump isolated CABG. METHODS: In this retrospective cohort study, we used the data registered for CABG patients from two heart centers in the Golestan province (North of Iran) between 2007 and 2016. The study population included 1956 patients divided into two groups: 1062 non-diabetic patients and 894 patients with diabetes (fasting plasma glucose ≥126 mg/dl or using antidiabetic medications). The study outcome was in-hospital MACCEs, a composite outcome of myocardial infarction (MI), stroke and cardiovascular death, and postoperative complications, including postoperative arrhythmia, acute atrial fibrillation (AF), major bleeding (defined as reoperation due to bleeding), and acute kidney injury (AKI). RESULTS: During the 10-year study period, 1956 adult patients with a mean (SD) age of 59.0 (9.60) years were included. After adjustment for age, gender, ethnicity, obesity, opium consumption, and smoking, diabetes was a predictor of postoperative arrhythmia (AOR 1.30, 95% CI 1.08-1.57; P = 0.006). While it was not a predictor of in-hospital MACCEs (AOR 1.35, 95% CI 0.86, 2.11; P = 0.188), AF (AOR 0.85, 95% CI 0.60-1.19; P = 0.340), major bleeding (AOR 0.80, 95% CI 0.50, 1.30; P = 0.636) or AKI (AOR 1.29, 95% CI 0.42, 3.96; P 0.656) after CABG surgery. CONCLUSION: Findings indicated that diabetes increased the risk of postoperative arrhythmia by 30%. However, we found similar in-hospital MACCEs, acute AF, major bleeding, and AKI following CABG surgery in both diabetic and non-diabetic patients.

Grafted Sertoli cells prevent neuronal cell death and memory loss induced by seizures.

Epilepsy significantly reduces the patient's quality of life, and we still need to develop new therapeutic approaches to control it. Transplantation of cells such as Sertoli cells (SCs), having a potent ability to release a variety of growth and immunoprotective substances, have made them a potential candidate to deal with neurological diseases like epilepsy. Hence, this study aims to evaluate whether SCs transplant effectively protects the hippocampus astrocytes and neurons to oppose seizure damage. For this purpose, the effects of bilateral intrahippocampal transplantation of SCs were investigated on the rats with the pentylenetetrazol (PTZ) induced seizure. After one-month, post-graft analysis was performed regarding behavior, immunohistopathology, and the distribution of the hippocampal cells. Our findings showed SCs transplantation reduced astrogliosis, astrocytes process length, the number of branches, and intersections distal to the soma of the hippocampus in the seizure group. In rats with grafted SCs, there was a drop in the hippocampal caspase-3 expression. Moreover, the SCs showed another protective impact, as shown by an improvement in pyramidal neurons' number and spatial distribution. The findings suggested that SCs transplantation can potently modify astrocytes' reactivation and inflammatory responses.

Comparative RNA-Seq analysis reveals insights in Salmonella disease resistance of chicken; and database development as resource for gene expression in poultry.

Salmonella, one of the major infectious diseases in poultry, causes considerable economic losses in terms of mortality and morbidity, especially in countries that lack effective vaccination programs. Besides being resistant to diseases, indigenous chicken breeds are also a potential source of animal protein in developing countries. For understanding the disease resistance, an indigenous chicken line Kashmir faverolla, and commercial broiler were selected. RNA-seq was performed after challenging the chicken with Salmonella Typhimurium. Comparative differential expression results showed that following infection, a total of 3153 genes and 1787 genes were differentially expressed in the liver and spleen, respectively. The genes that were differentially expressed included interleukins, cytokines, NOS2, Avß-defensins, toll-like receptors, and other immune-related gene families. Most of the genes and signaling pathways involved in the innate and adaptive immune responses against bacterial infection were significantly enriched in the Kashmir faverolla. Pathway analysis revealed that most of the enriched pathways were MAPK signaling pathway, NOD-like receptor signaling pathway, TLR signaling pathway, PPAR signaling pathway, endocytosis, etc. Surprisingly some immune-related genes like TLRs were upregulated in the susceptible chicken breed. On postmortem examination, the resistant birds showed small lesions in the liver compared to large necrotic lesions in susceptible birds. The pathological manifestations and RNA sequencing results suggest a balancing link between resistance and infection tolerance in Kashmir faverolla. Here we also developed an online Poultry Infection Database (https://skuastk.org/pif/index.html), the first publicly available gene expression resource for disease resistance in chickens. The available database not only shows the data for gene expression in chicken tissues but also provides quick search, visualization and download capacity.

The Effect of Training the Nonverbal Pain Scale (NVPS) on the Ability of Nurses to Monitor the Pain of Patients in the Intensive Care Unit.

Introduction: Pain in the intensive care unit is a silent fact. Considering the positive features of the nonverbal pain scale (NVPS) in assessing the pain of non-verbal patients, this study investigates the effect of training the NVPS on the ability of nurses to monitor the pain of patients in the intensive care unit. Materials and methods: In this semi-experimental study, the effect of the NVPS training on the ability of 50 intensive care unit (ICU) nurses of Imam Khomeini Hospital affiliated to Ahvaz University of Medical Sciences was investigated. At first, the ability to diagnose the presence and intensity of pain was checked by a checklist. Then the nurses were taught how to use the scale correctly. After 2 weeks of training completion, the ability to correctly use the scale was measured again. Data analysis was performed using descriptive statistics (mean and standard deviation) and inferential statistics (McNemar, Chi-squared, paired t-test, and Fisher's exact test) in SPSS software version 16. Results: After the training on the non-verbal pain scale, there was a significant difference between the intervention and control groups in diagnosing the presence of pain related to changing the patient's position (p = 0.023). Also, nurses ability to diagnose pain intensity during airway suction increased fourfold and for physiotherapy procedures twice as much as before training. Conclusion: Nonverbal pain scale training improves ICU nurses ability in diagnosing the presence and severity of pain in nonverbal patients. How to cite this article: Ghayem H, Heidari MR, Aghaei B, Norouzadeh R. The Effect of Training the Nonverbal Pain Scale (NVPS) on the Ability of Nurses to Monitor the Pain of Patients in the Intensive Care Unit. Indian J Crit Care Med 2023;27(3):195-200.

MDV-induced differential microRNA expression in the primary lymphoid organ of thymus.

Marek's disease virus (MDV), a highly contagious cell associated virus, is the etiological agent of Marek's disease (MD), a lymphoproliferative and neuropathic disease of domestic chickens. Clinical signs of MD include transient paralysis, bursal/thymic atrophy, and T cell lymphomas. MicroRNAs (miRNAs) are short single-stranded non-coding RNAs that regulate gene expression by transcriptional suppression or mRNA degradation. Herpesviruses, including MDV, encode for miRNAs that are known to play essential roles in viral pathogenicity, oncogenesis, and evasion of immune responses. In this study, we performed miRNA sequencing in thymuses of control and MDV-infected chickens of MD-resistant (63) and susceptible (72) lines at 21 days post infection (dpi). The thymus is a lymphoid organ that undergoes severe atrophy due to MDV-induced apoptotic mediated destruction of T cells. Sequence analysis identified 658 total chicken miRNAs in the thymuses of control and MDV-infected birds of both lines. Of these, 453 were novel and 205 were known microRNAs. All novel miRNAs mapped to chicken genome with no sequence homology to existing miRNAs in the chicken miRbase. Comparative analysis between the thymuses of control and infected birds of resistant and susceptible lines identified 78 differentially expressed microRNAs that might provide insights into mechanisms of thymus atrophy.

Mutational Analysis of the VPREB1 Gene of Pre-BCR Complex in a Cohort of Sporadic Pediatric Patients With B-Cell Acute Lymphoblastic Leukemia.

During bone marrow B-cell development, the pre-B-cell receptor is formed by the association of the immunoglobulin heavy chain with a surrogate light chain, which is encoded by the VPREB1, and λ5 genes. It is known that pre-BCR signaling signifies a critical checkpoint at the pre-B-cell stage. Thus, failure pre-BCR signaling is proposed as a critical factor for the development of B-cell acute lymphoblastic leukemia (B-ALL). B­ALL is the most common pediatric cancer and is one of the leading causes of death in children. Until now, several molecular analyses were performed for genomic alterations in B-ALL, but for genomic analysis of the VPREB1 gene and its rare variations, limited studies have been conducted. In this study, using polymerase chain reaction and direct sequencing of 88 pediatric patients with B-ALL, we investigated the genomic region of the VPREB1 gene to find sequence variations of this gene. Our study presented ten homozygous and heterozygous point mutations and heterozygous nucleotide deletions, in the VPREB1 gene in 36 boys and 32 girls' patients. Our Bioinformatics assay results presented that these variations may alter the RNA folding, protein structure, and therefore probable effect on the protein function. These results propose that nucleotide changes probably contribute to B-ALL pathogenesis.

Effect of cancer treatment on sleep quality in cancer patients: A systematic review and meta-analysis of Pittsburgh Sleep Quality Index.

BACKGROUND: Sleep problems are one of the most common symptoms experienced by cancer patients. The causes of poor sleep quality might be due to treatment and its side effects. Thus, we conducted this systematic review and meta-analysis with the aims of investigating sleep quality during treatment in cancer patients. METHODS: Comprehensive search strategy was conducted in the following original databases: PubMed, Web of Science (ISI), Scopus, Embase, PsycINFO, and Ovid, from 1950 to 15th February 2021. Studies that investigated the sleep quality during treatment in cancer patients were included. Two investigators extracted all relevant data, independently. For deriving mean difference, random-effects meta-analyses were used. We assessed quality of studies by Newcastle-Ottawa Scale (NOS). RESULTS: A total of 27 studies (1884 participants) were included in the syntheses on sleep quality. The mean global Pittsburgh Sleep Quality Index (PSQI) in cancer patients before the initiation of treatment was 7.11 (95% CI: 6.48, 7.74), during 8.31 (95% CI: 6.34, 10.27), after the treatment 7.10 (95% CI: 6.54, 7.66), and finally, the mean global PSQI in the time of follow-up was estimated 7.33 (95% CI: 6.27, 8.39); all the results were meaningful (P < 0.001). The mean difference showing the quality of sleep was better before the initiation of treatment compared to after the treatment. CONCLUSION: Cancer patients who underwent cancer treatment face lots of problems and adverse effects caused by treatment. Our results revealed that cancer patients experience poor quality of sleep during the whole trajectory of cancer even after a year from the initiation of treatment. After the end of treatment, sleep quality got better compared to during the treatment and returned to before the treatment level, but it is still poor and needs more sleep-related interventions to improve.

The effect of dopaminergic neuron transplantation and melatonin co-administration on oxidative stress-induced cell death in Parkinson's disease.

A gradual degeneration of the striatum and loss of nigral dopamine cells are characteristic of Parkinson's disease. Nowadays, combination therapy for neurodegenerative disease is considered. This study aimed to investigate the effects of melatonin and dopaminergic neurons derived from adipose tissue stem cells (ADSCs) in a rat model of Parkinson's disease. Parkinson's disease was induced in rats using neurotoxin 6-Hydroxydopamine. The treatment was performed using melatonin and dopaminergic neurons transplantation. Subsequently, behavioral tests, western blot analysis for Caspase-3 expression, GSH (Glutathione) content and stereology analysis for the volume and cell number of substantia nigra and striatum were performed. Treatment with melatonin and dopaminergic neuron transplantation increased the number of neurons in substantia nigra and striatum while the number of glial cell and the volume of substantia nigra and striatum did not show significant change between groups. Western blot analysis for caspase 3 indicated the significant differences between groups. The results also indicated the increased level of glutathione (GSH) content in treatment groups. this study showed that combination therapy with melatonin and dopaminergic neurons could greatly protect the neurons, reduce oxidative stress and improve the symptoms of PD.

Internal causality in agri-food Life Cycle Assessments: Solving allocation problems based on feed energy utilization in egg production.

Physical allocation in Life Cycle Assessment (LCA) should, ideally, be based on underlying causal relationships. While both cause-oriented and effect-oriented causality referred to in LCA literature are forms of external causality, internal causality addresses the actual flow of materials and inputs in a system - in other words, the real behaviour of the system under study. While a number or examples of allocation based on physical causality have been used in poultry LCAs, none of these represent the internal causality (the actual biological processes) in egg production. The current study remedies that gap by proposing such a method. Agri-food LCAs, in particular LCAs of livestock production, were used to identify existing physical allocation approaches consistent with internal causality. The most commonly used approach was found to be based on the allocation of feed energy to support the various physiological functions of the livestock species. A feed energy - Metabolizable Energy (ME) - utilization model for allocation in egg production LCAs is hence similarly proposed. Using the inventory of a previous LCA study of egg production in Canada, allocation ratios for eggs and spent hens were developed. Feed utilization models specific to each unit process were identified. The overall differences between ME utilization (∼95% eggs, 5% spent hens) and gross chemical energy content (92% eggs, 8% spent hens) for allocation were relatively small. Scenario analysis, however, showed that the allocation ratios can be considerably different if the causal relationship is interpreted differently. Differences over ∼20% was seen in a scenario which did not allocate between the co-products of each unit process in the system, but rather to the products at the end of a biological causal chain straddling multiple unit processes. The proposed approach is consistent with the interpretation of LCA as a natural sciences framework, and with the ISO 14044 multi-functionality hierarchy, because it reflects actual biological causality in egg production systems. The study results also underscore that practitioners should not only clearly justify their choice of allocation strategy, but also describe its application in detail, since small differences in methods can result in divergent outcomes.

Age Changes and Suicidal Activity in Iran Over the Past Decade: A Systematic Review and Meta-Analysis.

This review study fills an important gap by aiming to determine the age changes in attempted and completed suicide in Iran during the past decade. A systematic review of related articles in international and Iranian databases from January 2008 to January 2020 was first conducted and relevant studies were extracted based on established criteria. Results showed that the mean age of suicide in Iran is 29.8 (range 27.7 - 31.8) years old for men and 27.4 (range 25.8 - 28.9) for women (P < 0.0001). The youngest and oldest populations of completed suicide belong to eastern and central regions of the country respectively. Findings imply the critical importance of increasing awareness through educational programs and public health campaigns to increase awareness and reduce suicidal behavior in Iran.

Culture strategy as a modulator of target assessments: Functionality of suspension versus hanging drop-derived choriocarcinoma spheroids as in vitro model of embryo implantation.

The choriocarcinoma spheroid model has been amply applied to study the underlying molecular mechanism of implantation. Reproducibility and functionality of spheroid tumor models were addressed precisely. To mimic embryo-endometrium crosstalk, no functional characteristics of spheroids have been provided based on culture strategies. In this study, choriocarcinoma spheroids were provided as suspension culture (SC) or hanging drop culture (HDC). Primary assessments were performed based on morphology, cellular density, and hormonal secretion. Spheroid-endometrial cross talk was assessed as coculture procedures. Further, alkaline phosphatase (ALP) activity and expression of genes involved in attachment, invasion, and inducing migration were quantified. We found HDC spheroids provided a homogenous-shaped aggregate with a high grade of viability, cellular integration, hormonal secretion, and the dominant role of WNTs expression in their microarchitecture. SC spheroids showed a higher level of ALP activity and the expression of integrated genes in modulating attachment, invasion, and migration abilities. Spheroid confrontation assays clearly clarified the superiority of SC spheroids to crosstalk with epithelial and stromal cells of endometrium in addition to motivating an ideal endometrial response. Conclusively, culture strategies by affecting various molecular signaling pathways should be chosen precisely according to specific target assessments. Specifically, SC assumed as an ideal model in spheroid-endometrial cross talk.