'Immune-inflammatory markers & clinical characteristics for outcomes in hospitalized SARS-CoV-2 infected patients of Pakistan: a retrospective analysis'.

OBJECTIVE: Accumulating evidence suggests the role of immune-inflammatory markers in early risk stratification and prognostication of COVID-19 patients. We aimed to evaluate their association with severity and the development of diagnostic scores with optimal thresholds in critical patients. SETTING AND PARTICIPANTS: This retrospective case study includes hospitalized COVID-19 patients from March 2019 to March, 2022, in the developing area teaching hospital in Pakistan. Polymerase chain reaction (PCR) positive patients, n = 467 were investigated for clinical outcomes, comorbidities and disease prognosis. The plasma levels of Interleukin-6 (IL-6), Lactate dehydrogenase (LDH), C-reactive protein (CRP), Procalcitonin (PCT), ferritin and Complete blood count markers were measured. RESULTS: Majority were males (58.8%) and patients with comorbidities had more severe disease. Hypertension and diabetes mellitus were the commonest comorbidities. Shortness of breath, myalgia and cough were the main symptoms. The hematological markers NLR, as well as the plasma levels of immune-inflammatory variables, IL-6, LDH, Procalcitonin, Erythrocyte sedimentation rate, Ferritin were markedly raised in severe and critical patients (p < 0.0001 for these markers). ROC analysis supports IL-6 as the most accurate marker with high prognostic relevance with proposed cut-off threshold (43 pg/ml), determining >90% of patients in terms of COVID-19 severity (AUC = 0.93, 91.7%, se; 90.3%sp). Furthermore, positive correlation with all other markers including NLR with cut-off = 2.99 (AUC = 0.87, se = 89.8%, sp = 88.4%), CRP with cut-offs at 42.9 mg/l, (AUC = 0.883, se = 89.3% and sp = 78.6%), LDH cut-off at 267µg/L, evidenced in >80% patients (AUC = 0.834 se = 84% and sp = 80%). Additionally, ESR and ferritin have the corresponding AUC 0.81 and 0.813 with cut-off at 55 mm/hr and 370, respectively. CONCLUSION: Investigating the immune-inflammatory markers can assist physicians in providing prompt treatment and ICU admission in terms of COVID-19 severity. As a result, which may reduce the overall mortality of COVID-19 patients.

Quercetin exhibits potent antioxidant activity, restores motor and non-motor deficits induced by rotenone toxicity.

The rotenone-induced animal model of Parkinson's disease (PD) has been used to investigate the pathogenesis of PD. Oxidative stress is one of the main contributors of neurodegeneration in PD. Flavonoids have the potential to modulate neuronal function and combat various neurodegenerative diseases. The pre- and post-supplementation of quercetin (50 mg/kg, p.o) was done in rats injected with rotenone (1.5 mg/kg, s.c). After the treatment, behavioral activities were monitored for motor activity, depression-like behavior, and cognitive changes. Rats were decapitated after behavioral analysis and the brain samples were dissected out for neurochemical and biochemical estimation. Results showed that supplementation of quercetin significantly (p<0.01) restored rotenone-induced motor and non-motor deficits (depression and cognitive impairments), enhanced antioxidant enzyme activities (p<0.01), and attenuated neurotransmitter alterations (p<0.01). It is suggested that quercetin supplementation improves neurotransmitter levels by mitigating oxidative stress via increasing antioxidant enzyme activity and hence improves motor activity, cognitive functions, and reduces depressive behavior. The results of the present study showed that quercetin pre-supplementation produced more significant results as compared to post-supplementation. These findings show that quercetin can be a potential therapeutic agent to reduce the risk and progression of PD.

Spirulina platensis (Blue-green algae): A miracle from sea combats the oxidative stress and improves behavioral deficits in an animal model of Schizophrenia.

Spirulina platensis (blue-green algae) is a nutritional supplement. It constitutes of high content of protein, antioxidants, various phytopigments and possesses neuroprotective activities. Schizophrenia (SZ) is recognized as a neuropsychiatric disorder in humans with a reduced lifespan followed with impairments in social as well as vocational functioning. Major psychotic symptoms of SZ cluster into three categories: positive, negative and cognitive dysfunctions. Dizocilpine recognized as one of the best drugs to mimic full spectrum of SZ can develop an animal model of the disorder. Various antipsychotics are considered as approved treatment therapy for the psychotic symptoms of SZ but they also exert adverse effects. Thus, there is an excessive need for novel treatment(s) with negligible adverse effects. Present study was designed to evaluate the neuroprotective effects of spirulina in ameliorating the psychosis- like symptoms in dizocilpine-induced rat model of SZ. Spirulina was tested at the dose of 180 mg/kg. Results showed that administration of spirulina improved behavioral deficits and combated the oxidative damage evident by a significant reduction in lipid peroxidation and increase in antioxidant level. Thus, from present findings it may be suggested that spirulina can be used as a therapy for preventive or therapeutic measures.

Alleviation of diazepam-induced conditioned place preference and its withdrawal-associated neurobehavioral deficits following pre-exposure to enriched environment in rats.

Diazepam is one of the widely prescribed sedative drugs for the treatment of anxiety and sleep disorders. However, its continuous use can induce addiction, tolerance, and withdrawal symptoms and, therefore, the pharmacological use of diazepam is restricted. Exposure to enriched environment can reduce the addiction to stimulants including amphetamine, cocaine, and nicotine. However, the protective effect of enriched environment against preference of sedative drugs is not yet investigated. This study, therefore, determined the effects of enriched environment to prevent diazepam-preference using conditioned place preference (CPP) paradigm. Adult rats were reared in social (n = 12) or physically (n = 12) enriched environment for four weeks. Each group was then sub-divided into two groups and were administered either saline (Control; n = 6) or diazepam (1 mg/kg; n = 6) on alternate days for thirteen days. During the administration of diazepam, the CPP was conducted to monitor drug preference on 5th, 9th and 13th day of experiment. It was observed that the diazepam administration significantly (p < .01) induced preference in rats. Neurobehavioral deficits including hypolocomotor activity, depression-like behavior, impaired learning and memory functions were also observed after 24 h of drug abstinence. Exposure to enriched environment significantly reduced diazepam-preference and other neurobehavioral deficits. This study provides preliminary evidence to highlight the importance of enriched environment in the attenuation of diazepam-preference.

Administration of 5-HT-1B agonist ameliorates pseudodementia induced by depression in rats.

Major depressive disorder (MDD) is the leading cause of memory impairment in general population. The serotonin hypothesis provides a target model for the treatment of depression and depression-associated memory loss. 5-HT-1B receptor is suggested as a potential candidate in the pathophysiology of depressive illness. Dysfunction of 5-HT-1B receptors has been observed previously in depressive patients. Zolmitriptan, 5-HT-1B agonist is clinically recommended for the treatment of migraine. However, in present study this drug was tested as a potential treatment for depression and associated memory loss by altering the serotonergic function at receptor level. Rats (n=24) were equally divided into unstressed and stressed groups. Depression was induced by 19 days of restraint stress for 4 h which was followed by forced swim test and pattern separation test to assess depressive symptoms and memory impairment, respectively. The initial sign of depression-associated memory loss involves impaired pattern separation which is regarded as pseudodementia. In this study stressed rats showed depression- and pseudodementia-like symptoms. After the induction of depression, rats were treated with zolmitriptan at a dose of 0.3 mg/kg which resulted in a significant attenuation of depression and depression-associated memory impairment. Results are discussed with reference to the modulation of function of 5-HT-1B receptor following the administration of exogenous agonist.

Physical enrichment enhances memory function by regulating stress hormone and brain acetylcholinesterase activity in rats exposed to restraint stress.

To study the effects of stress on mental health activity is of great importance in neuropsychological studies as it may affect the lifelong performance related to brain and overall health and wellbeing of an individual. It is observed very often that exposure to stress during early life can alter the brain function which may reflect as cognitive disability. Impairment of memory is associated with increased oxidative stress which is due to enhanced production of free radicals that may lead to lipid peroxidation and disintegration of cell structure and functions. Exposure to enriched environment has shown to enhance spatial learning and memory, although the underlying mechanism covering the regulation of antioxidant capacity is limited. Here we investigated short and long term memory using Morris water maze before and after giving restraint stress procedure in rats exposed to social and physically enriched environment. Levels of malondialdehyde (MDA), activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and acetylcholinesterase (AChE) in brain tissue were estimated. Plasma corticosterone was also determined after decapitation. Results demonstrated that rats pre-exposed to physical along with social enrichment showed improved short and long term memory as compared to control group. However, restraint stress exerted differential effects in socially and physically enriched groups. Reduced lipid peroxidation and decreased activity of SOD, GPx and AChE were observed in physically enriched rats subjected to stress as compared to stressed rats kept in social environment. Levels of corticosterone were also found to be significantly reduced in rats kept in physically enriched environment. This study shows the beneficial effects of environmental enrichment on learning and spatial memory by reducing oxidative stress via reducing lipid peroxidation and regulation of antioxidant enzymes in rats.

Association of CACNA1C with bipolar disorder among the Pakistani population.

Many single nucleotide polymorphisms (SNPs) have been identified for Bipolar disorder (BD), but association between SNPs and BD can vary depending on the population tested. SNPs rs10994336 and rs9804190 in ANK3 and rs1006737 in CACNA1C have emerged as the most highly replicated SNPs significantly associated with BD. The aim of the present study was to assess the association of these SNPs with BD in the Pakistani population, which has never before been examined. A total of 120 BD and 120 control individuals from Pakistan were examined in this analysis. Genotyping results indicated that rs1006737 in CACNA1C was significantly associated with BD, while rs10994336 or rs9804190 in ANK3 was not significant when examined individually. However, risk score assessment found that the presence of two or more risk alleles was significantly associated with disease, indicating that risk alleles from ANK3 and CACNA1C may additively contribute to BD. A protein-protein interaction network was generated using STRING to probe the relationship between ANK3 and CACNA1C interactors and their associations with BD. While none of the interactors are directly linked to BD, they play a role in pathways linked to BD, including oxytocin and dopamine signaling pathways. Collectively, these results reveal a significant association of CACNA1C with BD among the Pakistani population, extending results from other ethnic groups to the Pakistani population for the first time.

Enriched environment palliates nicotine-induced addiction and associated neurobehavioral deficits in rats.

This study was designed to investigate the role of enriched environment in preventing and/or reducing the neurobehavioral deficits produced after nicotine administration in albino Wistar rats. Equal numbers of rat in two groups were either placed in social environment (control group) or social along with physically enriched environment for four weeks before the administration of nicotine. Exposure to different environmental conditions was followed by the intraperitoneal injection of nicotine at the dose of 0.6 mg/kg for seven consecutive days during which addictive behavior was monitored using conditioned placed preference paradigm. Behavioral responses to locomotor activity, anxiety and retention of short term memory were investigated in control and nicotine injected groups exposed to different environments. Results of this study showed that the rats pre-exposed to physical along with social enrichment exhibited a decrease in drug seeking behavior, hyper locomotion, anxiogenic effects along with improvement of working memory as compared to control and nicotine injected groups that were kept in social environment alone. This behavioral study suggests that the exposure to physical enrichment along with socialization in young age can later reduce the chances of compulsive dependence on nicotine and related neurobehavioral deficits.

Nootropic and anti-stress effects of rice bran oil in male rats.

Rice bran oil (RBO) is an important product of rice bran. It is considered to be one of the most important nutritious oil due to its favorable fatty acid composition and unique composition of naturally occurring biologically active antioxidant compounds. This study was designed to monitor the effects of oral intake of RBO on stress response in rats. RBO was extracted using hexane. Rats were divided into Control and test (RBO-treated). RBO-treated rats were given 0.2 ml/day RBO for 6 weeks. Food intake and body weight changes were monitored weekly. After 6 weeks open field activity and Morris Water Maze (MWM) test were performed. Results showed that weekly cumulative food intake but not body weight were lower in RBO-treated rats during 1st to 5th week of treatment, which were normalized at the end of treatment. Exploratory activity of RBO-treated rats in an open field was increased. Spatial memory in Morris water maze was enhanced in RBO-treated than control rats. An episode of 2 h restraint stress decreased the 24 h food intake of both control and RBO-treated animals. Behavioral deficits were lower in RBO-treated rats. Exposure of 2 h restraint stress increased brain serotonin (5-hydroxytryptamine: 5-HT) metabolism. These increases were lower in RBO-treated restrained than their respective control animals. Serotonergic neurotransmitter mechanism is implicated in stress. The findings of the study show beneficial effects of RBO in learning and memory functions. Moreover, the study also highlights the attenuating effect of RBO on stress induced behavioral and neurochemical effects in rats.

Preventive effects of curcumin against drug- and starvation-induced gastric erosions in rats.

The present study was designed to investigate the gastroprotective, analgesic, and antipyretic effects of curcumin (Cur), the major constituent of turmeric. Acetylsalicylic acid (ASA) was used in this study as a standard drug for comparison. The analgesic activity was measured using the Hot-Plate Test. The antipyretic and antiulcer effects were assessed using yeast-induced pyrexia and gastric ulceration, respectively. Curcumin (100 mg/kg) injected intra-peritoneally 1 hr prior to the Hot-Plate Test showed significant analgesic activity expressed by both parameters: an increase in latency time and a reduction in paw licking as compared to the controls. In the animal model of pyrexia, curcumin (100 mg/kg injected intra-peritoneally) exhibited a significant reduction in the rectal temperature after 1 hr, 2 hrs, 4 hrs, and 5 hrs of treatment, indicating the antipyretic effect of curcumin. Rats with orally administered curcumin (200 mg/kg) did not show any lesions on the inner lining of the stomach after a 16 hr fast, indicating the gastroprotective effects of curcumin as compared to saline- and acetylsalicylic acid-administered rats. The significantly low ulcer index in curcumin-treated rats following starvation highlights the gastroprotective characteristics of curcumin.