Experimental Evidence on Age-related Differential Outcomes Associated With Substance Abuse.

Growing evidence indicates that adolescent substance abuse is now an alarming concern that imposes a considerable socio-economic burden on societies. On the other hand, numerous studies have shown that due to specific neurophysiological features, the brain is more vulnerable to the adverse effects of psychoactive drugs at an early age. Unfortunately, these negative effects are not limited to the period of drug use, but can persistently affect the brain's responsiveness to future exposures to the same or other types of drug. For researchers to develop pharmacological strategies for managing substance abuse disorders, they need to gain a deep understanding of the differences in behavioral outcomes associated with each type of drug across different age groups. The present study was conducted to review the experimental evidence revealing the mentioned differential effects with an emphasis on common drugs of abuse, including cocaine, nicotine, cannabis, and opioids. Although the cellular mechanisms underlying age-related effects have not been exclusively addressed for each drug, the most recent results are presented and discussed. Future studies are required to focus on these mechanisms and reveal how molecular changes during brain development can result in differential responses to drugs at the behavioral level.

Oral administration of coenzyme Q10 ameliorates memory impairment induced by nicotine-ethanol abstinence through restoration of biochemical changes in male rat hippocampal tissues.

Substance abuse among adolescents has become a growing issue throughout the world. The significance of research on this life period is based on the occurrence of neurobiological changes in adolescent brain which makes the individual more susceptible for risk-taking and impulsive behaviors. Alcohol and nicotine are among the most available drugs of abuse in adolescents. Prolonged consumption of nicotine and alcohol leads to drug dependence and withdrawal which induce various dysfunctions such as memory loss. Coenzyme Q10 (CoQ10) is known to improve learning and memory deficits induced by various pathological conditions such as Diabetes mellitus and Alzheimer's disease. In the present study we investigated whether CoQ10 treatment ameliorates memory loss following a nicotine-ethanol abstinence. Morris water maze and novel object recognition tests were done in male Wistar rats undergone nicotine-ethanol abstinence and the effect of CoQ10 was assessed on at behavioral and biochemical levels. Results indicated that nicotine-ethanol abstinence induces memory dysfunction which is associated with increased oxidative and inflammatory response, reduced cholinergic and neurotrophic function plus elevated Amyloid-B levels in hippocampi. CoQ10 treatment prevented memory deficits and biochemical alterations. Interestingly, this ameliorative effect of CoQ10 was found to be dose-dependent in most experiments and almost equipotential to that of bupropion and naloxone co-administration. CoQ10 treatment could effectively improve memory defects induced by nicotine-ethanol consumption through attenuation of oxidative damage, inflammation, amyloid-B level and enhancement of cholinergic and neurotrophic drive. Further studies are required to assess the unknown side effects and high dose tolerability of the drug in human subjects.

Omega-3 fatty acids supplementation prevents learning and memory impairment induced by chronic ethanol consumption in adolescent male rats through restoration of inflammatory and oxidative responses.

OBJECTIVE: Ethanol (Eth) intake is known to cause numerous detrimental effects on the structure and function of the brain, and it is commonly used as a psychostimulant drug by adolescents. Conversely, omega-3 (O3) can reduce the risk of cognitive decline and promote the maintenance of neurophysiological functions. In this study, we investigated the protective effects of O3 on behavioral alterations, oxidative stress, and interleukin-6 (IL-6) levels induced by chronic Eth intake during adolescence in rats. MATERIALS AND METHODS: Adolescent male rats (21 days old) were divided as follows: (1) Vehicle, (2) Eth (Eth in drinking water [20%]), (3-5) Eth + O3 (50/100/150 mg/kg), and (6) O3 (150 mg/kg). After 5 weeks, Morris water maze (MWM) and passive avoidance (PA) tests were performed, and the hippocampal and cortical levels of oxidative stress markers and inflammatory indices were measured. RESULTS: Adolescent Eth intake impairs learning and memory function in MWM and PA tests (groups × day, p < 0.05 and p < 0.001, respectively). It was shown that Eth induced oxidative stress and neuroinflammation. O3 improved learning and impairment induced by Eth by reducing the adverse effects of Eth on the oxidant/antioxidant balance in the hippocampi (for malondialdehyde [MDA]/thiol: p < 0.01, p < 0.001, respectively) and for superoxide dismutase (SOD)/catalase (CAT): p < 0.01 and p < 0.05, respectively). Furthermore, we found that O3 prevented the Eth-induced increase of hippocampal IL-6 (p < 0.001). CONCLUSION: O3 supplementation acts as an effective approach to prevent learning and memory impairments induced by chronic Eth consumption during adolescence. In this respect, the antioxidant and anti-inflammatory properties of O3 seem to be the main underlying mechanisms of neuroprotection.

Oral administration of bacterial probiotics improves Helicobacter pylori-induced memory impairment in rats: Insights from behavioral and biochemical investigations.

There are numerous evidence supporting the association between Helicobacter pylori (H. pylori) infection and the occurrence of cognitive deficits in humans. In this regard, treatment of H. pylori infection has been suggested as an effective strategy to decelerate the neurodegenerative processes of memory deficits in AD patients. Numerous studies support the beneficial effects of probiotics on various pathological conditions, particularly cognitive deficits, however, this concern has not been addressed in relation to the memory impairment induced by H. pylori infection. In the present study, we aimed to reveal whether oral administration of two bacterial probiotics (including Lactobacillus rhamnosus and Lactobacillus plantarum), could ameliorate H. pylori-induced memory deficits at behavioral level in rats. Besides, cellular mechanisms were investigated by biochemical methods to find out how probiotic effects are mediated in hippocampal circuitry. Male Wistar rats were infected by H. pylori for 3 consecutive days, then probiotic treatment was done for the next 3 days and after a drug-free period (12 days), animals were assessed by Morris Water Maze and Novel Object Recognition tests. Finally, rats were euthanized by CO2 and hippocampal tissues were excised for biochemical measurements. Results indicated that H. pylori infection markedly impairs memory function in rats which is associated with alterations of oxidative, inflammatory, neurotrophic, and cholinergic markers. Interestingly, treatment with either of the probiotics alone or in combination, significantly improved the H. pylori-induced memory deficits and this was associated with restoration of balance in biochemical factors within the hippocampal neurons.

Mouse Fibroblast L929 Cell Line as a Useful Tool for Replication and Adaptation of Infectious Bursal Disease Virus.

Infectious bursal disease virus (IBDV) causes a highly contagious disease associated with immunosuppression in young chickens. Production of either egg-based or primary cell-based high-quality vaccines requires time-consuming and costly procedures. To determine a suitable cell line for IBDV replication, L929 cell line was a candidate for the growth kinetics processing of the virus. The L929 cells were proliferated in monolayer, and doubling time was calculated. Replication kinetics an IBDV isolate at the multiplicity of infection 0.1 PFU/cell were determined using virus titration. To adapt IBDV on L929 cells, seven consecutive passages were performed. Virus titer and levels of apoptosis were quantitatively analyzed at each passage. The viral VP2 gene was amplified and sequenced in three passages. An average doubling time of 21 h was estimated for monolayers of L929 cells. Although during early passages, virus growth did not produce a clear cytopathic effect (CPE), an increase in IBDV titers was observed. Serial passages led to the evidence of marked CPEs and an increase in the virus titer in the third passage. During the fourth to seventh passages, consistent CPEs characterized by the formation of granulated and round cells were evident within 24 to 48 hours post-inoculation. The titer of the virus was increased in the third passage onwards to peak in the fourth and constant at 5.9 TCID50 until the end passage. The IBDV replication in connection with DNA fragmentation and FITC, revealed the characteristic picture of apoptosis in a time-dependent manner. We found that the IBDV could easily be adapted to L929 cells, increasing virus yields by about two orders of magnitude. These results indicated that the cell line may be useful in the production of efficient virus particles.

Omega-3 fatty acids prevent nicotine withdrawal-induced impairment of learning and memory via affecting oxidative status, inflammatory response, cholinergic activity, BDNF and amyloid-B in rat hippocampal tissues.

In the present study, the main objective was to reveal whether treatment by Omega-3 fatty acids could prevent the adverse effects of adolescent nicotine withdrawal on spatial and avoidance memory in male rats. For this purpose, Morris water maze and passive avoidance tests were performed on male Wistar rats and the hippocampal levels of oxidative stress markers, inflammatory indices, brain-derived neurotrophic factor, nitrite, amyloid-B and acetylcholinesterase (AChE) were measured. Moreover, density of dark neurons were assessed in CA1 and CA3 regions. Results showed that adolescent nicotine exposure followed by a period of drug cessation exacerbates the behavioral indices of learning and memory through affecting a variety of biochemical markers within the hippocampal tissues. These changes lead to elevation of oxidative and inflammatory markers, reduction of neurotrophic capacity and increased AChE activity in hippocampal tissues. In addition, it was observed that co-administration of nicotine with Omega-3 fatty acids significantly prevents nicotine withdrawal-induced adverse effects through restoration of the mentioned biochemical disturbances. Therefore, we suggest administration of Omega-3 fatty acids as a safe, inexpensive and effective therapeutic strategy for prevention of memory dysfunctions associated with nicotine abstinence during adolescence.

Preparation and Evaluation of the In situ Gel-forming Chitosan Hydrogels for Nasal Delivery of Morphine in a Single Unit dose in Rats to Enhance the Analgesic Responses.

INTRODUCTION: In this study, an in situ gel-forming chitosan hydrogel was prepared with the use of glutamate salt of chitosan (Ch-Ga), ß-glycerophosphate (Gp), and morphine (Mor). The paper is focused on in vitro physicochemical properties and in-vivo analgesic effects of the prepared chitosan hydrogel. METHOD: The thermosensitive properties of prepared chitosan hydrogel were evaluated during the different temperatures and times. The physicochemical properties of chitosan hydrogel were investigated by infrared (IR) spectroscopy and X-ray diffraction analysis (XRD). Also, its cell cytotoxicity effects were evaluated in murine NIH/3T3 normal cells. Subsequently, the distribution of chitosan hydrogel in the nasal cavity of rats and its analgesic responses were evaluated. The prepared chitosan hydrogel showed that it could be gelled at the temperature of 34 °C before leaving the nose in the shortest possible time of 30 s. RESULT: The analgesic responses of the intranasal (IN) injection of chitosan hydrogel (IN-chitosan hydrogel, 10 mg Mor/kg) in a single unit dose in rat relative to the placebo and intranasal or intraperitoneal (IP) injection of free morphine solution (IN-Free Mor or IP-Free Mor, 10 mg Mor/kg) via the hot plate test, reveal that the IN-chitosan hydrogel could induce fast analgesic effects of morphine with maximum possible effect (MPE) of 93% after 5 min compare to the IN-Free Mor and IP-Free Mor with MPE of 80% after 15 min and 66% after 30 min, respectively. Also, prolonged analgesic effects with MPE of 78 % after 6 h of injection were only seen in the IN-chitosan hydrogel injected group. The obtained fluorescent images of rat's brain injected with IN-chitosan hydrogel containing doxorubicine (Dox) as a fluorescent agent showed that the mucosal adhesive and absorption enhancer properties of IN-chitosan hydrogel resulting in longer presence of them in the nasal cavity of rats followed by more absorption of Dox from the blood vessels of olfactory bulbs with a 74% color intensity compared to the IN-Free Mor and IN-Free Dox with 15%. CONCLUSION: These data reveal that the IN-chitosan hydrogel could induce fast and prolonged analgesic effects of morphine compare to the IN/IP-Free Mor, which could be considered as an in situ gel-forming thermosensitive chitosan hydrogel for nasal delivery of wide ranges of therapeutic agents.

Effects of Ocimum basilicum L. Extract on Hippocampal Oxidative Stress, Inflammation, and BDNF Expression in Amnesic Aged Rats.

The present study was conducted to investigate the effects of Ocimum basilicum L. (OB) extract on learning and memory impairment in aged rats. Male rats were divided into the following experimental groups: Group 1 (control): including 2 months old rats, Group 2 (aged) including 2 years old rats, Groups 3-5 (aged-OB): including 2 years old rats received 50, 100, and 150 mg/kg OB for 8 weeks by oral gavage. Aging increased the delay to find the platform but, however, decreased the time spent in the target quadrant when tested by Morris water maze (MWM). Aging also reduced the latency to enter the dark chamber in the passive avoidance (PA) test compared to the control group. Moreover, interleukin-6 (IL-6) and malondialdehyde (MDA) levels were raised in the hippocampus and cortex of aged rats. In contrast, thiol levels and enzymatic activity of superoxide dismutase (SOD) and catalase (CAT) significantly reduced. In addition, aging significantly reduced BDNF expression. Finally, OB administration reversed the mentioned effects. The current research showed that OB administration improves learning/memory impairment induced by aging. It also found that this plant extract protects the brain tissues from oxidative damage and neuroinflammation.

Omega-3 fatty acids prevent nicotine withdrawal-induced exacerbation of anxiety and depression by affecting oxidative stress balance, inflammatory response, BDNF and serotonin metabolism in rats.

Adolescents are known to be more vulnerable than adults to the adverse effects of nicotine dependence. In the present study, we aimed to investigate whether adolescent nicotine exposure, followed by a period of abstinence, could affect the anxiety- and depressive-like behaviors in rats. For this purpose, behavioral assessments were carried out using open field test, elevated plus maze and forced swimming test in male rats received chronic nicotine intake during adolescence followed by a period of abstinence in adulthood, compared to their control counterparts. In addition, O3 pre-treatment was done at three different doses to reveal whether it could prevent nicotine withdrawal effects. Then, animals were euthanized and the cortical concentrations of oxidative stress markers, inflammatory indices, brain-derived neurotrophic factor, serotonin and the enzymatic activity of monoamine oxidase-A were measured. Results indicated that nicotine withdrawal exacerbates the behavioral signs of anxiety through alteration of the brain oxidative stress balance, inflammatory response and serotonin metabolism. Moreover, we found that omega 3 pre-treatment significantly prevents the nicotine withdrawal-induced complications by restoration of changes in the mentioned biochemical indices. Moreover, the improving effects of O3 fatty acids were found to be dose-dependent in all experiments. Taken together, we would like to suggest the O3 fatty acids supplementation as a safe, inexpensive and effective strategy for prevention or amelioration of detrimental effects induced by nicotine withdrawal at cellular and behavioral levels.

Acute morphine injection persistently affects the electrophysiological characteristics of rat locus coeruleus neurons.

Administration of morphine is associated with critical complications in clinic which primarily includes the development of dependence and tolerance even following a single dose (acute) exposure. Behavioral and electrophysiological studies support the significant role of locus coeruleus (LC) neurons in tolerance and dependence following chronic morphine exposure. The current study was designed to explore the electrophysiological properties of the LC neurons following acute morphine exposure. In-vitro whole-cell patch-clamp recordings were performed in LC neurons 24 h after intraperitoneal morphine injection. Acute morphine injection significantly decreased the spontaneous firing rate of LC neurons, the rising and decay slopes of action potentials, and consequently increased the action potential duration. In addition, morphine treatment did not alter the rheobase current and first spike latency while affected the inhibitory postsynaptic currents elicited in response to orexin-A. In fact, single morphine exposure could inhibit the disinhibitory effect of orexin-A on LC neurons.

A history of ethanol intake accelerates the development of morphine analgesic tolerance: A protective potential for omega-3 fatty acids.

Adolescence is a critical life period during which significant neurodevelopmental changes occur within the central nervous system. Consistently, substance abuse in this stage has been found to induce persistent changes in brain responsiveness to future drug challenges. Nowadays, heavy episodic alcohol consumption during adolescence, also known as binge-drinking behavior, is a growing concern in modern societies. On the other hand, alcohol is well known to act as a gateway drug, that is, it promotes the individual's craving for consumption of other drugs of abuse. With this in mind, we aimed to assess whether adolescent ethanol exposure could alter the development of tolerance and dependence to morphine, as an available common opioid drug. Tail flick test was used to measure thermal nociceptive changes in adult male Wistar rats undergone ethanol/vehicle exposure during adolescence. Furthermore, morphine withdrawal syndrome was induced by naloxone injection, and behavioral signs were recorded for 20 min. It was found that adolescent ethanol intake facilitates morphine analgesic tolerance and decreases baseline latency; however, the severity of dependence is not significantly altered. Moreover, we found that 15 days of treatment with omega-3 fatty acids (O3) prevents the mentioned ethanol-induced changes suggesting a therapeutic potential for this compound. O3 supplementation, as an inexpensive and noninvasive method, may assist the clinicians to reverse the adverse effect of alcohol binge drinking on adolescents' brains and to reduce the vulnerability to drug exposure in adulthood. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Vitamin B12 administration prevents ethanol-induced learning and memory impairment through re-establishment of the brain oxidant/antioxidant balance, enhancement of BDNF and suppression of GFAP.

There are growing evidence indicating that the adolescent brain is persistently affected by the use of psychostimulant agents. In this regard, alcohol drinking has become rather common among the adolescents in many societies during the last decade. It is currently well known that long-term ethanol exposure deteriorates various cognitive functions such as learning and memory. Mechanistically, these adverse effects have been shown to be mediated by oxidative damage to central nervous system. On the other hand, Vit-B12 is known to improve cognitive performance by suppression of oxidative parameters. Thus, in the present study we aimed to test whether treatment by Vit-B12 could prevent ethanol-induced complications in mice using behavioral and biochemical methods. Different groups of male Syrian mice received ethanol, ethanol+Vit-B12, Vit-B12 alone, or saline during adolescence and then learning and memory functions were assessed by Morris water maze (MWM) and Passive Avoidance (PA) tests. Finally, mice were sacrificed for measurement of biochemical factors. Results indicated that, adolescent ethanol intake impairs learning and memory function through exacerbation of oxidative stress and Vit-B12 treatment improves these complications by re-establishment of oxidant/anti-oxidant balance in CNS. Moreover, we found that Vit-B12 prevents ethanol-induced reduction of BDNF and enhancement of GFAP and acetylcholinesterase (AChE) activity. In conclusion, it seems that Vit-B12 supplementation could be used as an effective therapeutic strategy to prevent learning and memory defects induced by chronic alcohol intake during adolescence.

Comparative investigation of analgesic tolerance to taurine, sodium salicylate and morphine: Involvement of peripheral muscarinic receptors.

Nowadays various analgesic medications are used for the management of acute and chronic pain. Among these opioid and non-steroidal anti-inflammatory drugs stand in the first line of therapy, however, prolonged administration of these substance is generally challenged by development of analgesic tolerance in patients. Therefore, it is highly valuable to find new pharmacological strategies for prolonged therapeutic procedures. In this respect, Taurine, a free amino acid, has been shown to induce significant analgesia at both spinal and peripheral levels through cholinergic mechanisms. In the present study, we used hot-plate analgesic test to investigate how taurine either as a single medication or in combination with sodium salicylate and morphine may affect both acute response to pain and development of analgesic tolerance. The effect of taurine was also tested on morphine withdrawal syndrome. Hyoscine butyl bromide was used to assess the role of muscarinic receptors in taurine-mediated effects. Finally, biochemical assay was done to reveal how the activity of brain acetylcholinesterase may change in relation with muscarinic receptor activity. Results indicated that acute administration of taurine-sodium salicylate combination causes more potent analgesia compared to the use of tau (but not SS alone) and this seems to be mediated via activity of muscarinic receptors in peripheral nervous system. Furthermore, the effect of this combination undergoes less analgesic tolerance during time. Combination of taurine and morphine is an effective strategy to attenuate both morphine analgesic tolerance and dependence and this also seems to depend on activity of muscarinic receptors, however through differential cellular mechanisms.

Environmental health risk factors and cutaneous leishmaniasis (CL): A case-control study in northeastern Iran.

Background & objectives: Cutaneous leishmaniasis (CL) is one of the main causes of vector-born diseases in younger population. To evaluate the association of environmental health factors on the odds of CL incidence, a case-control study was conducted in northeastern Iran. Methods: This study was conducted within 2020-2021 based on individual and household data from a tertiary referral center. Cases were patients diagnosed with CL by PCR method; controls were selected among the patients' relatives, and information was obtained from a health registry system. Demographic and socioeconomic data of 1871 subjects, included age, sex, household information and environmental health factors. Multivariable models with environmental factors in various conditions and CL were separately fit by univariate and mixed multiple unconditional logistic regression. Results: Participants included 617 cases (mean [SD] age, 13.62[13.72] years; 58.20% male) and 1264 controls (mean [SD] age, 16.45[15.44] years; 50.40% male). Results revealed that the use of well-water sources compared to surface water is significantly associated with CL (odds ratio [OR]=0.204; 95%CI, 0.13-0.33;P<0.001). Muddy houses, ruined buildings or wastelands and stagnant water, canals and rivers near the houses were also associated with CL (OR=3.85; 95%CI, 1.66-8.89; P=.002; OR=2.47; 95%CI, 1.76-3.47; P<.001). Besides, existence of pine tree was found to be a risk factor (OR=3.25; 95%CI, 2.12-4.99; P<.001) and similarly for the use of waste collection system (OR=4.43; 95%CI, 3.32-7.51; P<.001). Interpretation & conclusion: Environmental factors related to houses were significantly associated with CL and may represent the modifiable risk factors of CL disease.

Evaluation of P53 protein expression in gingival tissues of patients with chronic periodontitis by immunohistochemistry methods.

OBJECTIVE: Periodontitis is one of the most important periodontal diseases that can be affected by many factors. Although the mechanism of periodontitis development is not yet fully understood, previous studies suggest that apoptosis may be one of the pathological factors that can affect the process of the disease by destroying old and damaged cells. Low expression of P53 protein is one of the reasons for delaying cell death that allows damaged cells to survive longer and gives more time for the chance of mutations and pathogenesis. Because of the important role of P53 in gingival cells of patients with chronic periodontitis, the objective of our study is to evaluate the P53 protein expression in gingival tissues of patients with chronic periodontitis by immunohistochemistry methods. MATERIALS AND METHODS: In this cross-sectional study, 35 patients with severe to moderate chronic periodontitis (loss of attachment ≥3 mm, probing depth ≥5 mm) with no treatment and 25 people who were healthy for periodontal problems were examined. Gingival biopsies from marginal and attached gingiva were obtained, prepared, and mounted on slides. Then, the expression of P53 on each slide was evaluated by optic microscopy after using P53 antibodies and staining with hematoxylin-eosin (immunohistochemistry method). Data were analyzed using independent t-test, Mann-Whitney U-test, and Spearman correlation test using SPSS Statistics version 18.0. RESULTS: The mean ages of participants in the case and control groups were 37.58 and 32.09, respectively. Our results showed that the expression of P53 was not significant in periodontitis compared to the control group (p > .05). Also, gender could not affect the expression of P53 in both groups (p > .05), and there was no significant relationship between age and P53 gene incidence. CONCLUSION: Chronic periodontitis has no significant effect on P53 expression, so changes in apoptosis due to P53 expression in periodontitis are not significant.

Experimental data on lithium salts: From neuroprotection to multi-organ complications.

Lithium-salts stand on the first line of therapy for the management of specific psychiatric conditions, mainly bipolar mood disorder. It is also known to protect the brain against neurodegenerative processes such as Alzheimer's disease. Despite the mentioned merits, recent studies have revealed that high dose or prolonged lithium intake deteriorate the function of multiple key organs including heart, ovaries, thyroid gland and kidneys. Mechanistically, both positive and negative effects of lithium are mediated through methylation of ß-catenin nuclear-binding proteins which is potentiated by lithium-induced inhibition of GSK-3 or inositol monophosphatase. The current study briefly reviews the recent experimental data on lithium therapy considering both positive (i.e., neuroprotective) and negative aspects. In this regard, the question is that whether doses of lithium administered in experimental research are comparable with the therapeutic doses, as currently prescribed in clinical practice. It should be noted that the experimental data on animal studies, as widely reviewed here, could not be directly generalized to clinic. This is mainly because lithium doses applied in animal models are usually higher than therapeutic doses, however, there are evidence indicating that even animal to human translated doses of lithium, cause serious complications and this has been reported by meta-analyses on human studies. Therefore, we suggest the clinicians to use lithium-salts with precaution particularly in pregnancy and precisely adjust lithium concentration considering the patient's general health status to avoid lithium toxicity. Indeed, alternative approaches are recommended when the subject is pregnant, prolonged therapy is required or specific organ dysfunction is diagnosed.

Evaluation of BHK-21 Cell Line for Specific Viruses by Two Different PCR Methods.

The selection of suitable cell cultures for use in the biological industry as well as research and diagnostic studies is critical. One of the factors affecting cell culture that can affect the results of studies is the contamination with viral agents. Therefore, efforts to preserve the health of cell cultures from contamination sound logical, and the use of virus-free cells is vital in research and diagnostic studies as well as in the manufacturing industries. For this purpose, it is crucial to use fast and correct diagnostic methods to detect the presence of critical viral contaminants in cells. Moreover, the use of susceptible diagnostic methods is also doubly important, especially in the case of contaminants that may remain hidden. Therefore, in this study, the BHK-21C5 cell line, one of the most widely used cells in the production and quality control of biological products and virological studies, was examined in terms of contamination with the most important viruses such as BVD and BLV. Detection of possible contaminants by using two-step RT-PCR to detect the 5' UTR portion of the BVD virus. Moreover, Nested PCR was carried out to detect the BLV virus using the gp51 env gene region. Also, an Flk cell line and Hela cell line that were consistently contaminated with the BLV virus were used as positive controls in Nested PCR. Due to the absence of bands in the BHK-21C5 cell column and the bandwidth observed in the positive control column (BVDV) in the range of 283 bp, non-contaminating of the cell clone with the BVD virus was proved. Also, no band was observed in well related to BHK-21C5 cell, and no cell clone contamination with BLV was confirmed, and in wells related to the positive controls (Flk and Hela cells), the bands have seen in the range 444bp. So, the results showed that no obvious or covert viral contamination effect was observed in the cell clone studied. Hence, the use of this cell line seems unobstructed in the quality control and production of biological products and research and diagnostic studies.

A Review of the Establishment of the Seed Lot System in the Production of Biological Products and Its Importance.

Today, due to the importance of diseases controlled by vaccination, the production of biological products is of great importance in ensuring public health, so producing high-quality biological products plays an important role in maintaining public health. One of the most important principles of manufacturing high-quality and efficient biological products is using suitable seeds based on standard principles. To produce a suitable seed for continuous use for mass production of biological products, the seed must be defined according to certain principles and foundations. These principles are formed in the form of a seed lot system. In this review article, all the requirements for the establishment of a seed to produce a biological product include general seed information, including basic and seed information and microorganisms, seed-specific information including seed passage levels, propagation method, seed storage conditions, coding and labeling, identification information and design of suitable laboratories for passage, seed propagation, and storage, determination of seed characteristics including all necessary tests to determine seed identity, purity, potency, efficacy, stability, safety and also all the necessary information for documenting and storing seeds has been studied. Also, this study discusses the method of preparing all the necessary information for establishing a seed lot system, especially determining the characteristics of seeds. Based on the study's results, a complete and comprehensive seed lot system has been formed that can be used to prepare, propagate, passage, and store seeds used in the production of biological products.

Characterizing the BHK-21 C5 cell line and determining cellular sensitivity to rubella virus compared with the routine cell (RK13).

The World Health Organization has strict rules and recommendations on the selection and use of cell substrates in laboratories. Given the widespread use of safe and secure cell substrates in the production and quality control of viral vaccines and also the high demand for vaccines against viral diseases, obligating the selection of a suitable cell substrate for cultivation and production of biological products. Animal cell lines play a valuable role in the preparation and propagation of viral seeds; thus, the current study used the BHK-21 cell line among others for viral checking with the aim of replacing the BHK-21 C5 cell line with the RK13 cell line to investigate the cytopathic effects of the rubella virus. To this end, attempts were made to determine the characteristics of the BHK-21 C5 cell line including cell growth characteristics and sterility tests to validate its safety and security. Then, by culturing the cells in a 96-well microplate, titration of the rubella virus was subsequently performed by preparing serial dilutions of the virus from 10-1 to 10-5 and inoculated to cell lines in order to compare the sensitivity of BHK-21 C5 and RK13 cell lines to rubella virus. Data analysis according to the results of the tests by ahead default, p-value < 0/05 was equal to p-value = 0.01 based on SPSS analysis with the paired-sample t-test. In addition, the box-plot diagram indicated a significant difference between these cell lines. Based on the results, the BHK-21 C5 cell line seems to be more sensitive to the rubella virus than others. Therefore, it can be used for production and quality control of the vaccine and in research and diagnosis of rubella.

COVID-19, body mass index and cholesterol: an ecological study using global data.

BACKGROUND: Coronavirus disease 2019 (COVID-19) is now globally considered a serious economic, social and health threat. A wide range of health related factors including Body Mass Index (BMI) is reported to be associated with the disease. In the present study, we analyzed global databases to assess the correlation of BMI and cholesterol with the risk of COVID-19. METHODS: In this ecological study, we used age-standardized BMI and cholesterol levels as well as the incidence and mortality ratio of COVID-19 at the national-levels obtained from the publicly available databases such as the World Health Organization (WHO) and NCD Risk Factor Collaboration (NCD-RisC). Bivariate correlation analysis was applied to assess the correlations between the study variables. Mean differences (standard deviation: SD) of BMI and cholesterol levels of different groups were tested using independent sample t-test or Mann-Whitney rank test as appropriate. Multivariable linear regression analysis was performed to identify variables affecting the incidence and mortality ratio of COVID-19. RESULTS: Incidence and mortality ratio of COVID-19 were significantly higher in developed (29,639.85 ± 20,210.79 for cases and 503.24 ± 414.65 for deaths) rather than developing (8153.76 ± 11,626.36 for cases and 169.95 ± 265.78 for deaths) countries (P < 0.01). Results indicated that the correlations of BMI and cholesterol level with COVID-19 are stronger in countries with younger population. In general, the BMI and cholesterol level were positively correlated with COVID-19 incidence ratio (ß = 2396.81 and ß = 30,932.80, p < 0.01,| respectively) and mortality ratio (ß = 38.18 and ß = 417.52, p < 0.05,| respectively) after adjusting for socioeconomic and demographic factors. CONCLUSION: Countries with higher BMI or cholesterol at aggregate levels had a higher ratios of COVID-19 incidence and mortality. The aggregated level of cholesterol and BMI are important risk factors for COVID-19 major outcomes, especially in developing countries with younger populations. We recommend monitoring and promotion of health indicices to better prevent morbidity and mortality of COVID-19.