Molecular screening and genetic diversity of tick-borne pathogens associated with dogs and livestock ticks in Egypt.

BACKGROUND: The Middle East and North Africa (MENA) offer optimal climatic conditions for tick reproduction and dispersal. Research on tick-borne pathogens in this region is scarce. Despite recent advances in the characterization and taxonomic explanation of various tick-borne illnesses affecting animals in Egypt, no comprehensive examination of TBP (tick-borne pathogen) statuses has been performed. Therefore, the present study aims to detect the prevalence of pathogens harbored by ticks in Egypt. METHODOLOGY/PRINCIPAL FINDINGS: A four-year PCR-based study was conducted to detect a wide range of tick-borne pathogens (TBPs) harbored by three economically important tick species in Egypt. Approximately 86.7% (902/1,040) of the investigated Hyalomma dromedarii ticks from camels were found positive with Candidatus Anaplasma camelii (18.8%), Ehrlichia ruminantium (16.5%), Rickettsia africae (12.6%), Theileria annulata (11.9%), Mycoplasma arginini (9.9%), Borrelia burgdorferi (7.7%), Spiroplasma-like endosymbiont (4.0%), Hepatozoon canis (2.4%), Coxiella burnetii (1.6%) and Leishmania infantum (1.3%). Double co-infections were recorded in 3.0% (27/902) of Hy. dromedarii ticks, triple co-infections (simultaneous infection of the tick by three pathogen species) were found in 9.6% (87/902) of Hy. dromedarii ticks, whereas multiple co-infections (simultaneous infection of the tick by ≥ four pathogen species) comprised 12% (108/902). Out of 1,435 investigated Rhipicephalus rutilus ticks collected from dogs and sheep, 816 (56.9%) ticks harbored Babesia canis vogeli (17.1%), Rickettsia conorii (16.2%), Ehrlichia canis (15.4%), H. canis (13.6%), Bo. burgdorferi (9.7%), L. infantum (8.4%), C. burnetii (7.3%) and Trypanosoma evansi (6.6%) in dogs, and 242 (16.9%) ticks harbored Theileria lestoquardi (21.6%), Theileria ovis (20.0%) and Eh. ruminantium (0.3%) in sheep. Double, triple, and multiple co-infections represented 11% (90/816), 7.6% (62/816), and 10.3% (84/816), respectively in Rh. rutilus from dogs, whereas double and triple co-infections represented 30.2% (73/242) and 2.1% (5/242), respectively in Rh. rutilus from sheep. Approximately 92.5% (1,355/1,465) of Rhipicephalus annulatus ticks of cattle carried a burden of Anaplasma marginale (21.3%), Babesia bigemina (18.2%), Babesia bovis (14.0%), Borrelia theleri (12.8%), R. africae (12.4%), Th. annulata (8.7%), Bo. burgdorferi (2.7%), and Eh. ruminantium (2.5%). Double, triple, and multiple co-infections represented 1.8% (25/1,355), 11.5% (156/1,355), and 12.9% (175/1,355), respectively. The detected pathogens' sequences had 98.76-100% similarity to the available database with genetic divergence ranged between 0.0001 to 0.0009% to closest sequences from other African, Asian, and European countries. Phylogenetic analysis revealed close similarities between the detected pathogens and other isolates mostly from African and Asian countries. CONCLUSIONS/SIGNIFICANCE: Continuous PCR-detection of pathogens transmitted by ticks is necessary to overcome the consequences of these infection to the hosts. More restrictions should be applied from the Egyptian authorities on animal importations to limit the emergence and re-emergence of tick-borne pathogens in the country. This is the first in-depth investigation of TBPs in Egypt.

Oral administration of proniosomal glibenclamide formulation protects testicular tissue from hyperglycemia fluctuations and ROS via Nrf2/HO-1 pathway.

Type 2 diabetes causes high blood sugar due to insulin malfunction and is linked to male infertility. Using proniosomes can enhance the effectiveness of Glibenclamide, a medication that stimulates insulin secretion. In our study, male rats with diabetes were treated with GLB with or without proniosomal for 14 days. Proniosomal formulations maintained glucose levels prevented weight loss and showed normal testicular tissue. GLB-proniosomal reduces ROS caused by T2DM through Nrf2, HO-1 pathway and increases CAT, SOD, and GSH production in response to insulin and glucose uptake. The reference and proniosomal treatments showed CAT and SOD significant enzymatic elevation compared to the positive and negative control. CAT significantly correlated with Gpx4 expression with P = 0.0169 and r = 0.98; similarly, the enzymatic activity of SOD also showed a positive correlation between the average glucose levels (r = 0.99 and P = 0.0037). Intestinally, GSH analysis revealed that only proniosomal-GLB samples are significantly elevated from the positive control, with a P value of 0.0210. The data showed proniosomal-GLB was more effective than pure GLB, confirmed by higher Nrf2 (2.050 folds), HO-1 (2.148 folds), and GPx4 (1.9 folds) transcript levels relative to the control with less sample diversity compared to the reference samples, indicating proniosomal stabilized GLB in the blood. Administering GLB and proniosomes formulation has effectively restored testicular function and sperm production in diabetic rats by regulating ROS levels and upregulating anti-ROS in response to glucose uptake. These findings may lead to better treatments for diabetic patients who have infertility issues.

Corrigendum: The outbreak of seasonal goat's brucellosis in the Swat ecosystem of Khyber Pakhtunkhwa.

No abstract available.

Determination of dose- and time-dependent hepatotoxicity and apoptosis of Lanthanum oxide nanoparticles in female Swiss albino mice.

Nanosized lanthanum oxide particles (La2O3) are commonly utilized in various industries. The potential health risks associated with La2O3 nanoparticles, cytotoxic effects at varying doses and time intervals, and the mechanisms behind their induction of behavioral changes remain uncertain and necessitate further investigation. Therefore, this study examined in vivo hepatotoxicity, considering the quantity (60, 150, and 300 mg/kg) and time-dependent induction of reactive oxygen species (ROS) over one week or 21 days. The mice received intraperitoneal injections of three different concentrations in Milli-Q water. Throughout the experiments, no physical changes or weight loss were observed among the groups. However, after 21 days, only the highest concentration showed signs of anxiety in the activity cage (p < 0.05). Subsequently, all animals treated with La2O3 NPs exhibited a significant loss of learning and memory recall using the Active Avoidances test, after 21 days (p < 0.001). Markers for anti-reactive oxygen species (ROS) such as superoxide dismutase (SOD) were significantly upregulated in response to all concentrations of NPs after seven days compared to the control group. This was confirmed by a significant increase in glutathione peroxidase (Gpx1) and pro-apoptotic Caspase-3 expression at the lowest and highest doses. Additionally, both transcription and protein levels of the anti-apoptotic BCL-2 surpassed P53 protein in a dosage-dependent manner, indicating activation of the primary anti-apoptosis pathway. After 21 days, P53 levels exceeded BCL-2 protein levels, confirming a significant loss of BCL-2 mRNA, particularly at the 300 mg/kg concentration. Furthermore, a higher transcription level of Caspase-3, SOD, and Gpx1 was observed, with the highest values detected at the 300 mg/kg concentration, indicating the activation of cell death. Histopathological analysis of the liver illustrated apoptotic bodies resulting from La2O3 NP concentration. The investigation revealed multiple inflammatory foci, cytoplasmic degeneration, steatosis, and DNA fragmentation consistent with increased damage over time due to higher concentrations. Blood samples were also analyzed to determine liver enzymatic changes, including alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate aminotransferase (AST), and lipid profiles. The results showed significant differences among all La2O3 NP concentrations, with the most pronounced damage observed at the 300 mg/kg dose even after 21 days. Based on an animal model, this study suggests that La2O3 hepatotoxicity is likely caused by the size and shape of nanoparticles (NPs), following a dose and time-dependent mechanism that induces the production of reactive oxygen species and behavioral changes such as anxiety and memory loss.

Cytotoxicity and Radiosensitizing Potentials of Pilosulin-3, a Recombinant Ant Venom, in Breast Cancer Cells.

Venom peptides are promising agents in the development of unconventional anticancer therapeutic agents. This study explored the potential of Pilosulin-3, a recombinant peptide from the venom of the Australian jack jumper ant "Myrmecia pilosula", as a cytotoxic and radiosensitizing agent in MCF-7 and MDA-MB-231 breast cancer (BC) cell lines. Pilosulin-3's cytotoxicity was evaluated across a wide range of concentrations using a proliferation assay. Cell cycle progression and apoptosis were examined at the inhibitory concentration 25% (IC25) and IC50 of Pilosulin-3, both with and without a 4Gy X-ray irradiation dose. Radiosensitivity was assessed at IC25 using the clonogenic survival assay. The study revealed that Pilosulin-3 exerted a concentration-dependent cytotoxic effect, with IC25 and IC50 values of 0.01 and 0.5 µM, respectively. In silico screening indicated high selectivity of Pilosulin-3 peptide, which was predicted to be the most likely anticancer agent (PROB = 0.997) with low hemolytic activity (PROP = 0.176). Although Pilosulin-3 exhibited a significant (p < 0.05) G2/M cell cycle arrest in combination with radiation, there was no discernible effect on apoptosis induction or cell survival following irradiation. In conclusion, Pilosulin-3 proved to be cytotoxic to BC cells and induced a cytostatic effect (G2/M arrest) when combined with radiation. However, it did not enhance the efficacy of cell killing by irradiation. While it holds potential as a cytotoxic agent in breast cancer treatment, its application as a radiosensitizer does not find support in these results.

The outbreak of migratory goat's brucellosis in the Swat ecosystem of Khyber Pakhtunkhwa.

Brucellosis is a major threat to public health especially in developing countries including Pakistan. This study reveals the characterisation of Brucella species affecting humans and goats in the Swat region of Khyber Pakhtunkhwa, Pakistan. Blood samples were collected from shepherds and goats and analysed by Rose Bengal precipitation test (RBPT), standard plate agglutination test (SPAT), polymerase chain reaction (PCR) and Sanger sequencing of 16S rRNA gene. The findings of the study indicated 24% (36/150) and 11.3% (17/150) positivity for Brucella abortus and Brucella melitensis, respectively, in human samples. In samples of goats, 26.66% (40/150) were positive for B. abortus and 16.66% (25/150) samples were positive B. melitensis by SPAT. The species-specific PCR confirmed B. abortus in 24% (36/150) of human samples and 26.66% (17/150) of goat samples by targeting the IS711 locus. The remaining seropositive samples were confirmed as B. melitensis using IS711 M species-specific primer. The sequences of the amplified fragments of the 16S rRNA gene were blasted, and phylogenetic analysis revealed that Brucella species circulating in the Swat district were closely related to B. melitensis and B. abortus reported from India, China, Philippines, and the United States (US) showing the existence of the possible epidemiological linkage among the Brucella species. This study concluded that there was a higher prevalence of B. abortus (26.6%) in humans and goats compared to B. melitensis (16.6%). These results revealed that the Brucella species were circulating in both humans and goats in the study areas. The findings of the study concluded that B. abortus and B. melitensis were circulating in goats and shepherds with a higher prevalence of B. abortus than B. melitensis. Furthermore, the Brucella species identified in Swat were phylogenetically related to the Brucella species reported from India, China, Philippines and the US.Contribution: The proposed study covers the scope of the journal. The species of the genus Brucella affect both animals and shepherds. This study investigates the seroprevalence of brucellosis in shepherds and goats in different geographical areas in the Swat district. The phylogenetic analysis of the Brucella spp. identified in Swat showed close relationships to the Brucella species reported in India, China, Philippines and the US, which shows the possible epidemiological linkages between the Brucella spp.

Green Lead Nanoparticles Induced Apoptosis and Cytotoxicity in MDA-MB-231 Cells by Inducing Reactive Oxygen Species and Caspase 3/7 Enzymes.

Nanoparticles are widely used in the pharmaceutical, agriculture, and food processing industries. In this study, we have synthesized green lead nanoparticles (gPbNPs) by using an extract of Ziziphus spina-christi leaves and determined their cytotoxic and apoptotic effect on the human breast cancer MDA-MB-231 cell line. gPbNPs were characterized by using X-ray diffraction (XRD), energy dispersive X-ray (EDX) scanning electron microscope (SEM), and transmission electron microscope (TEM). The toxicity of gPbNPs was determined on the MDA-MB-231 cell line using MTT and NRU assays and as a result cell viability was reduced in a concentration-dependent manner. MDA-MB-231 cells were more sensitive at the highest concentration of gPbNPs exposure. In this experiment, we observed the production of intracellular ROS in cells, and induction of caspase 3/7 was higher in cells at 42 µg/ml of gPbNPs. Moreover, the Bax gene was upregulated and the Bcl-2 gene was downregulated and increased caspase 3/7 activity confirmed the apoptotic effect of gPbNPs in cells. Our observation showed that gPbNPs induced cell toxicity, increased generation of intracellular ROS, and gene expression of Bcl-2 and Bax in the MDA-MB-231 cell line. In conclusion, these findings demonstrated that gPbNPs executed toxic effects on the MDA-MB-231 cell line through activating caspase 3/7 activity.

Renal-protective effect of Asparagus officinalis aqueous extract against lead-induced nephrotoxicity mouse model.

Lead is one of the cursed substances that threaten all human life. Lead poisoning can occur through food or water contaminations and it is hard to be detected. This incognito metal accumulates over time and resides in the liver, kidneys, and brain tissues leading to serious medical conditions, affecting organ functions, causing failure, kidney tubule degeneration, and destroying neuronal development. However, known metal chelators have bad negative effects. Asparagus officinalis (AO) is a promising herb; its root extract exhibited antioxidant, antiapoptotic, protective, and immunomodulatory activities. Inspired by those reasons, this study investigated to which extent Asparagus extract affected male mice's renal toxicity caused by lead acetate (LA) and antioxidant defense system. This work screened for its nephroprotective activity in four mouse groups: negative and positive control, LA group with renal injury, and diseased but pretreated mice with AO extract (AOE). Kidney index and kidney function biomarkers were evaluated. Antioxidant activities, lipid peroxidation, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), nitric oxide (NO), and reduced glutathione (GSH) were also tested. Furthermore, inflammatory cytokine (tumor necrosis factor-α (TNF-α), interleukin-1 ß (IL-1ß), and the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)), inducible nitric oxide synthase (iNOS), renal pro-apoptotic protein (Bax), antiapoptotic protein (Bcl-2), and caspase-3 levels were evaluated. The results showed that LA administration induced oxidative stress, renal inflammation, apoptosis, and renal histopathological alteration. However, due to its antioxidant activities, AOE was found to restrain oxidative stress, therefore preventing inflammation and apoptosis. Collectively, AOE perfectly clogged lead poisoning sneaking, stopped the bad deterioration, and succeeded to protect kidney tissues from toxicity, inflammation, and apoptosis.

The protective effect of apigenin against inorganic arsenic salt-induced toxicity in PC12 cells.

Poisoning by arsenic affects people worldwide, and many human illnesses and health issues, including neurotoxicity, have been linked to chronic exposure to arsenic. When exposed to arsenic, the body produces intracellular reactive oxygen species (ROS), which influence a variety of alterations in cellular activity and directly harm molecules through oxidation. Arsenic-induced lesions are improved by antioxidants with the ability to lower ROS levels. Therefore, the current research aimed to assess how well apigenin protected PC12 cells from the toxicity caused by inorganic arsenic salt (iAs). For 24 and 48 h, iAs and/or apigenin were applied to PC12 cells. Then, oxidative stress indicators like malondialdehyde (MDA), nitric oxide (NO), and ROS in addition to the enzymatic and non-enzymatic antioxidant molecules such as catalase (CAT), glutathione (GSH), and superoxide dismutase (SOD) were assessed. Moreover, after exposure to iAs, PC12 was examined for nuclear factor erythroid 2-related factor 2 (Nrf2) expression to clarify how apigenin manifests its neuroprotection. Furthermore, NF-kB p65 concentration and IL-1B, IL-6, and TNF-α mRNA expression were measured to assess neuroinflammation. Bax, caspase-3, and Bcl-2 levels were measured to investigate apigenin's potential to protect PC12 cells from iAs poisoning. The obtained results revealed that, the cell survival rate in the iAs group was significantly lower (P < 0.05), and the number of viable cells steadily increased after apigenin treatment. Furthermore, the study found that iAs decreased GSH, CAT, and SOD in the PC12 cells while increasing ROS, MDA, and NO levels. In PC12 cells, the capacity of iAs to cause oxidative stress was linked to the induction of neuroinflammation and apoptosis. Interestingly, apigenin pre-treatment of PC12 cells resulted in exceptional protection against iAs-induced neuroinflammation, oxidative stress, and apoptotic cell death. Nrf2 upregulation in PC12 cells may explain the neuroprotection effect of apigenin against iAs toxicity. In conclusion, the obtained results of the present study have clinical significance and indicate that apigenin is a promising candidate for shielding the nervous system from toxic effects caused by arsenic. These findings require further investigation using in vivo experimental models.

Protective effects of Asparagus officinalis (asparagus) against lead toxicity in mice.

One of the most prevalent harmful heavy metals is lead (Pb). It is generally recognized to be harmful to the testicles. Asparagus officinalis has many saponins, flavonoids, and other phenolics with strong antioxidant and anti-inflammatory effects. The effects of A. officinalis (asparagus) aqueous extract (AOAE) on testicular damage caused by lead acetate (PbAc) were investigated in this study. In this way, 20 mg/kg PbAc was injected intraperitoneally 2 h after mice were administered 400 mg/kg AOAE orally for 14 days. In the biochemical analysis of testicular tissue, PbAc decreased enzymatic and nonenzymatic antioxidant molecules in testicular tissue, while increasing lipid peroxidation, nitric oxide, inflammatory markers [nuclear factor kappa-B (NF-κB), tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1 ß), IL-6, and inducible nitric oxide synthase (iNOS)], and apoptotic-related proteins. Additionally, PbAc was discovered to reduce sperm motility and increase the percentage of dead sperm. However, due to its antioxidant qualities, AOAE has been found to reduce oxidative stress, therefore protecting against inflammation and apoptosis. It also allowed the AOAE sperm parameters to restore to their previous values in the control group. According to the findings, AOAE could be a natural substance that could be used to treat Pb-induced testicular toxicity; this protection may be attributed to its anti-oxidative, anti-inflammatory, and anti-apoptotic effects. However, this study warrants further works to explore in detail the underlying mechanisms of the alleviating effects of AOAE against Pb-induced toxicity and which of its active ingredients is responsible for this protection.

Role of green synthesized platinum nanoparticles in cytotoxicity, oxidative stress, and apoptosis of human colon cancer cells (HCT-116).

Progresses in the medicinal application of nanocompounds were accepted for the treatment of cancer. Nanoparticles-based therapy is of benefit for effective biodistribution and specific targeting. The current study investigated the anticancer effect of green synthesized platinum nanoparticles (PtNPs) against colon cancer cells (HCT-116). Flow cytometry and ELISA techniques were employed for detecting apoptotic and oxidative stress markers. Furthermore, PtNPs-lycopene (PtNPs-LP) on cell migration and invasion of HCT-116 cells was also examined. The PtNPs-LP was capable of diminishing cell proliferation and viability of HCT-116 cells in a dose-dependent mode. After treatment with PtNPs-LP, a significant increase in pro-apoptotic Bax and caspase-3 and a decrease in anti-apoptotic Bcl-2 was observed in treated cells that subsequently released cytochrome C into its cytoplasm, initiating cell death. Moreover, PtNPs-LP induced excessive generation of reactive oxygen species (ROS) and oxidative stress in cancer cells. In conclusion, PtNPs-LP exerts an antitumor effect against colon cancer cells via mediating important mechanisms such as cytotoxicity, apoptosis, and oxidative stress.

Evaluation of Cyto - and Genotoxic Influence of Lanthanum Dioxide Nanoparticles on Human Liver Cells.

Inorganic nanoparticles are representing an emerging paradigm in molecular imaging probe design. We have determined lanthanum oxide nanoparticles (La2O3 NPs)-induced toxicity on human livers cells for 48 hrs. Before exposure to La2O3 NPs, the size and shape of NPs were confirmed by transmission electron microscope. It was found at 32 ±1.6 nm with a sheet-like morphological structure. The viability of CHANG and HuH-7 cells was reduced as the concentration of La2O3 NPs increased. HuH-7 cells were more sensitive than CHANG cells to La2O3 NPs. We observed production of intracellular ROS in HuH-7 cells was more than CHANG cells and the LPO level was more in CHANG cells than in HuH-7 cells at 50 µg/ml of La2O3 NPs. Glutathione was decreased and catalase was increased at 50 µg/ml of La2O3 NPs. More apoptotic and necrotic cells were observed at 300 µg/ml in HuH-7 cells FACS. DNA damage was observed by the SGCE test and more DNA damage was found in CHANG cells than HuH-7 cells at 300 µg/ml La2O3 NPs over 48 hrs. Thus, study warrants the application of La2O3 NPs in daily life and provides vital information about the toxicity of La2O3 NPs.

Dose-Dependent Variation in Anticancer Activity of Hexane and Chloroform Extracts of Field Horsetail Plant on Human Hepatocarcinoma Cells.

Horsetail fern plant is botanically known as Equisetum arvense L., and it is a good source of phenolic flavonoids, phenolic acids, and compounds. Anticancer properties of hexane and chloroform extracts of the horsetail fern plant and their mechanisms involved in the anticancer activity on human hepatocarcinoma (HuH-7) cells were examined. Cytotoxicity was evaluated by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and NRU (neutral red uptake) assays. Other parameters such as oxidative stress and apoptosis in pretreated hexane and chloroform extracts of the horsetail fern plant were examined in HuH-7 cells. The observation showed that hexane and chloroform extract of the horsetail fern plant exhibited cytotoxicity against HuH-7 cells. The value of IC50-24 h of hexane and chloroform extract of the horsetail fern plant was determined as 199.0 µg/ml and 161.90 0 µg/ml for HuH-7 cells, respectively, and on the basis of IC50 value, three acute concentrations, viz., 75% of IC50, 50% of IC50, and 25% of IC50, were determined for further study. The lower dose of extracts hexane and chloroform extract of the horsetail fern plant did not show significant toxicity. Higher concentrations of extract induced significant antioxidant effects as well as apoptosis effects. However, exposure to hexane and chloroform extract of the horsetail fern plant upregulated the expression of Bax and p53 in HuH-7 cells. These data suggest that hexane and chloroform extract of the horsetail fern plant plays a significant role in the induction of toxicity via the regulation of oxidative stress in HuH-7 cells. This work may be useful for cancer chemotherapy.

Ameliorative effect of Spirulina and Saccharomyces cerevisiae against fipronil toxicity in Oreochromis niloticus.

The ameliorative effects of Spirulina and Saccharomyces cerevisiae (S. cerevisiae) against fipronil toxicity in Nile tilapia fish were investigated. Fipronil is a kind of pesticide that is widely used in agriculture, thus this trial was conducted to evaluate the effect of fipronil on growth related parameters (final body weight, feed intake, weight gain, feed conversion ratio, specific growth rate, and protein efficiency ratio), hematology related parameters (RBCs, WBCs, hemoglobin, packed cell volume, and deferential leukocytic count), biochemistry related parameters (alanine aminotransferase, aspartate aminotransferase, total protein, albumin, urea, and creatinine), histopathology of liver, intestine, gills, and spleen, and gene expression of antioxidants, stress, inflammatory, apoptotic, and related to junction proteins genes as SOD and GPx, COX II, TNF-α, Casp-3, and Claudin-3, respectively, in Nile tilapia (Oreochromis niloticus). Four hundred and five Nile tilapia fish were distributed in a glass aquarium into nine groups according to the Spirulina and S. cerevisiae supplemented diets, with or without fipronil contaminated water. The classified groups are control, Sc: S. cerevisiae (4 g/Kg diet), Sp: Spirulina (1 g/100 g diet), Fb1: 0.0021 mg fipronil/L, ScFb1: S. cerevisiae (4 g/Kg diet) with 0.0021 mg fipronil/L, SpFb1: Spirulina (1 g/100 g diet) with 0.0021 mg fipronil/L, Fb2: 0.0042 mg fipronil/L, ScFb2: S. cerevisiae (4 g/Kg diet) with 0.0042 mg fipronil/L, and SpFb2: Spirulina (1 g/100 g diet) with 0.0042 mg fipronil/L. The results of the present investigation indicated the negative effect of fipronil on the growth performance parameters of Nile tilapia, which was confirmed by the results of hematology, biochemistry, and histopathology. In addition, the results of gene expression of antioxidants, stress, inflammatory, and apoptotic genes indicate the genotoxicity of fipronil. However, these negative effects were ameliorated by Spirulina and Saccharomyces dietary supplementation.

Proanthocyanidins alleviate pentylenetetrazole-induced epileptic seizures in mice via the antioxidant activity.

The role of oxidative stress in the initiation and progress of epilepsy is well established. Proanthocyanidins (PACs), a naturally occurring polyphenolic compound, have been reported to possess a broad spectrum of pharmacological and therapeutic properties against oxidative stress. However, the protective effects of proanthocyanidins against epilepsy have not been clarified. In the present study, we used the pentylenetetrazole (PTZ)-induced epilepsy mouse model to explore whether proanthocyanidins could help to reduce oxidative stress and protect against epilepsy. Mice were allocated into four groups (n = 14 per each group): control, PTZ (60 mg/kg, intraperitoneally), PACs + PTZ (200 mg/kg, p.o.) and sodium valproate (VPA) + PTZ (200 mg/kg, p.o.). PTZ injection caused oxidative stress in the hippocampal tissue as represented by the elevated lipid peroxidation and NO synthesis and increased expression of iNOS. Furthermore, depleted levels of anti-oxidants, GSH, GR, GPx, SOD, and CAT also indicate that oxidative stress was induced in mice exposed to PTZ. Additionally, a state of neuroinflammation was recorded following the developed seizures. Moreover, neuronal apoptosis was recorded following the development of epileptic convulsions as confirmed by the elevated Bax and caspase-3 and the decreased Bcl2 protein. Moreover, AChE activity, DA, NE, 5-HT, brain-derived neurotrophic factor levels, and gene expression of Nrf2 have decreased in the hippocampal tissue of PTZ exposed mice. However, pre-treatment of mice with PACs protected against the generation of oxidative stress, apoptosis, and neuroinflammation in the PTZ exposed mice brain as the biomarkers for all these conditions was bought to control levels. In addition, the gene expression of Nrf2 was significantly upregulated following PACs treatment. These results suggest that PACs can ameliorate oxidative stress, neuroinflammation, and neuronal apoptosis by activating the Nrf2 signaling pathway in PTZ induced seizures in mice.

Application of Green Technologies in Design-Based Extraction of Celastrus paniculatus (Jyotishmati) Seeds, SEM, GC-MS Analysis, and Evaluation for Memory Enhancing Potential.

Background: The Celastrus paniculatus (CP), commonly known as Jyotishmati, is considered as "elixir of life" by Indian people for the prevention or management of many ailments. The seed powder and its extract have widely used commercially for the preparation of various Ayurvedic formulations for the improvement of memory. CP seeds were generally extracted by conventional extraction methods (CEMs) which are assumed to impact environment burden and also produce low extract yield. Green extraction with possible improvement in extract yield has always been the need of hour for selected medicinal plant. Objective: In the present research, we aimed to optimize the different extraction factors in microwave and ultrasound-based extraction. The various extracts obtained in conventional and green methods are also evaluated for the possible improvement in memory enhancing potential. Materials and Methods: The selected medicinal herb was extracted by CEM (maceration and percolation). In green methods such microwave-assisted extraction (MAE) and ultrasound assisted-extraction (UAE), various parameters were optimized using Box-Behnken design coupled with response surface methodology. The scanning electron microscopy (SEM) and gas chromatography-mass spectroscopy (GC-MS) analyses were also done to confirm the possible improvement in concentration of plant actives. The Swiss albino mice were used to evaluate memory enhancing potential of different extracts. Results: At the optimized conditions MAE and UAE the extraction yield, total phenolic content (TPC) and Total flavonoid content (TFC) are significantly improved. The GC-MS analysis further confirms the improvement in concentration of certain fatty acid esters, pilocarpine, and steroidal compounds in optimized extracts. The optimized extracts also exhibited the significant improvement in behavioral parameters, oxidative stress-induced parameters, and acetylcholinesterase inhibitory potential. Discussion and Conclusion: From the results, we can say that the application of green technologies in design-based extraction of selected herb not only significantly reduces the extraction time but also improves the extract yield and concentration of plant actives. In nutshell, it can be concluded that the green approaches for extraction of seeds of Celastrus paniculatus could be scale up at a commercial level to meet the rising demand for herbal extract.

Insecticidal Effect of Wild-Grown Mentha pulegium and Rosmarinus officinalis Essential Oils and Their Main Monoterpenes against Culex pipiens (Diptera: Culicidae).

The present study investigates the insecticidal effect of plant extract such as Mentha pulegium and Rosmarinus officinalis essential oils and some of their major compounds; these plants are well known for their many biological activities. The fumigant toxicity was evaluated, using glass jars, against female adults of Culex pipiens that constitute a mosquito vector of important diseases such as the West Nile virus. The adulticidal test showed that both essential oils and monoterpenes presented an insecticidal effect better than the chemical insecticide (Deltamethrin). The highest mortality percentages for the two essential oils have occurred at 312.5 µL/L air (between 56.14 ± 1.7% and 97.71 ± 3.03% after 24 h and 48 h of treatment). Moreover, all tested monoterpenes (carvone, R(+)-pulegone, 1,8-cineole, camphor and α-pinene) have produced high mortalities that varied depending on the time of the treatment and the concentrations used. Lethal concentrations (LC50) obtained for the essential oils and the main compounds have also varied according to the exposure time. M. pulegium and R. officinalis essential oil exhibited the lowest LC50 values after 24 h (72.94 and 222.82 µL/L air, respectively) and after 48 h (25.43 and 55.79 µL/L air, respectively) while the pure molecules revealed the lowest LC50 values after 48 h (between 84.96 and 578.84 µL/L air). This finding proves that the two essential oils and their main compounds have an insecticidal potential, which could help to develop natural toxic fumigants that may be used as an eco-friendly alternative in integrated and sustainable vector management.

Plant-Derived Smoke Solution Alleviates Cellular Oxidative Stress Caused by Arsenic and Mercury by Modulating the Cellular Antioxidative Defense System in Wheat.

Heavy metal stress is a significant factor in diminishing crop yield. Plant-derived smoke (PDS) has been used as a growth promoter and abiotic stress alleviator for the last two decades. Although the roles of PDS have been determined in various plants, its role in ameliorating heavy metal stress in wheat has not been reported so far. Therefore, the present work was conducted to investigate the effect of smoke solution extracted from a wild lemongrass Cymbopogon jwarancusa (C. jwarncusa) on physiological and biochemical features of wheat under arsenic (As) and mercury (Hg) stress. The results showed that higher concentrations of As and Hg pose inhibitory effects on wheat seed germination and seedling growth, including shoot/root length and shoot/root fresh weight. Photosynthetic pigments, such as chlorophyll a and b and carotenoids, were significantly decreased under As and Hg stress. Importantly, the levels of H2O2, lipid peroxidation, and TBARS were increased in wheat seedlings. The activity of antioxidant enzymes, such as CAT, was decreased by As and Hg stress, while the levels of SOD, POD, and APX antioxidant enzymes were increased in root and shoot. Interestingly, the application of PDS (2000 ppm), individually or in combination with either As or Hg stress, enhanced wheat seed germination rate, shoot/root length, and shoot/root fresh weight. However, the levels of H2O2, lipid peroxidation, and TBARS were decreased. Similarly, the levels of SOD, POD, and APX were decreased by PDS under As and Hg stress, while the level of CAT was enhanced by PDS under As and Hg stress. Interestingly, the levels of chlorophyll a and b, and total carotenoids were increased with the application of PDS under As and Hg stress. It is concluded that PDS has the capability to alleviate the phytotoxic effects of As and Hg stress in wheat by modulating the antioxidative defense system and could be an economical solution to reduce the heavy metal stress in crops.

Chemical Composition and Antimicrobial Activity of Essential Oil of Fruits from Vitex agnus-castus L., Growing in Two Regions in Bulgaria.

The chemical composition of Vitex agnus-castus L. (Verbenaceae family) fruits, collected from two regions in Bulgaria (south-central and north-east Bulgaria), was investigated. The content of proteins (5.3-7.4%), carbohydrates (73.9-78.8%), fiber (47.2-49.9%), ash (2.5-3.0%), essential oils (0.5%), and vegetable oil (3.8-5.0%) were identified in the fruits. The composition of the essential oils (EOs) of Vitex fruits from both regions was determined; the main compounds were 1,8-cineole (16.9-18.8%), α-pinene (7.2-16.6%), sabinene (6.7-14.5%), and bicyclogermacrene (7.3-9.0%), but significant differences in the quantitative and qualitative composition of EOs between the regions were found. The EOs of plants from north-east Bulgaria demonstrated antimicrobial activity against the pathogenic species Salmonella abony, Staphylococcus aureus, and Bacillus subtilis, but the Gram-negative bacteria Esсherichia coli and Pseudomonas aeruginosa exhibited resistance to the oil. Linoleic acid predominated in vegetable oil from both regions, followed by oleic acid. ß-sitosterol and γ-tocopherol were the main components in the sterol and tocopherol fraction of the lipids. Phosphatidic acids were the main components in the vegetable oil from north-east Bulgaria, while in the vegetable oil from south-central Bulgaria, all phospholipids were found in almost the same quantity. Overall, significant differences were observed in the chemical composition (proteins, carbohydrates, ash and moisture) of the fruits from the two regions of Bulgaria, as well as in the content of the main components of their essential and vegetable oils.