Mitochondrial Bcl-xL promotes brain synaptogenesis by controlling non-lethal caspase activation.

Non-lethal caspase activation (NLCA) has been linked to neurodevelopmental processes. However, how neurons control NLCA remains elusive. Here, we focused on Bcl-xL, a Bcl-2 homolog regulating caspase activation through the mitochondria. We generated a mouse model, referred to as ER-xL, in which Bcl-xL is absent in the mitochondria, yet present in the endoplasmic reticulum. Unlike bclx knockout mice that died at E13.5, ER-xL mice survived embryonic development but died post-partum because of altered feeding behavior. Enhanced caspase-3 activity was observed in the brain and the spinal cord white matter, but not the gray matter. No increase in cell death was observed in ER-xL cortical neurons, suggesting that the observed caspase-3 activation was apoptosis-independent. ER-xL neurons displayed increased caspase-3 activity in the neurites, resulting in impaired axon arborescence and synaptogenesis. Together, our findings suggest that mitochondrial Bcl-xL finely tunes caspase-3 through Drp-1-dependent mitochondrial fission, which is critical to neural network design.

Pridopidine rescues BDNF/TrkB trafficking dynamics and synapse homeostasis in a Huntington disease brain-on-a-chip model.

Huntington disease (HD) is a neurodegenerative disorder caused by polyglutamine-encoding CAG repeat expansion in the huntingtin (HTT) gene. HTT is involved in the axonal transport of vesicles containing brain-derived neurotrophic factor (BDNF). In HD, diminished BDNF transport leads to reduced BDNF delivery to the striatum, contributing to striatal and cortical neuronal death. Pridopidine is a selective and potent sigma-1 receptor (S1R) agonist currently in clinical development for HD. The S1R is located at the endoplasmic reticulum (ER)-mitochondria interface, where it regulates key cellular pathways commonly impaired in neurodegenerative diseases. We used a microfluidic device that reconstitutes the corticostriatal network, allowing the investigation of presynaptic dynamics, synaptic morphology and transmission, and postsynaptic signaling. Culturing primary neurons from the HD mouse model HdhCAG140/+ provides a "disease-on-a-chip" platform ideal for investigating pathogenic mechanisms and drug activity. Pridopidine rescued the trafficking of BDNF and TrkB resulting in an increased neurotrophin signaling at the synapse. This increased the capacity of HD neurons to release glutamate and restored homeostasis at the corticostriatal synapse. These data suggest that pridopidine enhances the availability of corticostriatal BDNF via S1R activation, leading to neuroprotective effects.

Epidemiology and diagnosis of feline panleukopenia virus in Egypt: Clinical and molecular diagnosis in cats.

AIM: This work aimed to study epidemiology and diagnosis of feline panleukopenia virus (FPV) using clinical examination, direct ELISA, RNA viral isolation and identification, and knowing phylogenetic tree of our isolate. MATERIALS AND METHODS: One hundred and sixty-five cats of different ages and sex were examined. Each cat was examined clinically to detect the clinical manifestations of the disease showing symptoms suggestive of feline panleukopenia (FP) as well as ELISA, and polymerase chain reaction (PCR) amplification analyses were conducted. RESULTS: Our finding includes (a) clinical signs detected in 165 of 165 cats were in the form of lethargy, fever, anorexia, thirst, vomiting, diarrhea, dehydration, and leukopenia. (b) ELISA results revealed that 66 of all examined cats were positive for FPV. (c) The amplification products from all positive samples were confirmed as FPV (VP1) gene by nucleotide sequences analysis, in which 75 samples were positive using PCR amplification for the FPV. (d) Statistical evaluation of ELISA results in comparison to PCR findings. ELISA showed 88%, 100%, and 94.5% for sensitivity, specificity, and accuracy, respectively, while the prevalence of FP among the examined population was 45%. No effect of sex, breed, and age on ELISA results as recorded using Chi-square analysis. CONCLUSION: The results of the sequence analysis indicated that PCR products of the FPV cDNA exhibited very low variation in their nucleotide sequence of all isolates compared with the published FPV genome, which could be suggested that FPV appears to be genomically stasis compared with other Parvoviruses. The genome sequence of FPLV strain in this study has been deposited in GenBank under the accession number KY466003. Our isolate closely related 100% to isolates from Portugal, which might be the origin of infection to Egypt through importation of cats.

Feline panleukopenia viral infection in cats: Application of some molecular methods used for its diagnosis.

Forty diseased cats and seven healthy control cats from different sex, ages and breeds had examined clinically to confirm presence or absence of clinical symptoms of Feline panleukopenia disease (FP). Several tools including ELISA, gene expression analysis (qRT-PCR), DNA fragmentation test and apoptosis assay were conducted to determine the Feline panleukopenia disease in cat tissues. Clinical symptoms in the form of depression, fever, anorexia, vomiting, diarrhea, dehydration, anaemia and leucopenia were recorded in the diseased cats while no clinical sings were observed in control healthy cats. ELISA results showed that all of diseased (n = 40) cats were positive while control cats (n = 7) were negative for FP viral antigen. After carrying out of ELISA assay, supportive treatment trials including fluid therapy, immunostimulant, antibiotics to overcome dehydration, restoring electrolytes imbalances, combating secondary bacterial infection were conducted but all diseased cats were died and control cats exposed to soft death. Gene expression analysis detected high levels of FP viral gene in several cat tissues in which ilium exhibited high viral expression levels compared with jejunum. Also, viral expression levels in jejunum were higher than in mesenteric lymph nodes. In addition, viral expression levels were not detected in tissues of control cats. The results of the DNA fragmentation assay observed that DNA extracted from different tissues of infected cats exhibited damaged DNA bands as compared with DNA of control cats. DNA fragmentation rates in infected tissues increased significantly (P < 0.01), the highest rates were showed in ilium and jejunum tissue than in mesenteric lymph nodes. Determination of apoptosis in cat tissues showed that rate of apoptosis/necrosis increased significantly (P < 0.05) in infected cats tissues in comparison to control cats. Moreover the highest apoptotic ratios of infected cats were observed in ilium and jejunum tissues compared with mesenteric lymph nodes.

Assessment of mineral elements and heavy metals in leaves of indigenous cypress of High Atlas Mountains.

An inductively coupled plasma mass spectrometry was developed to determine 20 mineral elements and heavy metals in leaves of Cupressus atlantica Gaussen, a traditional Moroccan medicinal herb from five environmentally different sites in N'Fis valley (High Atlas Mountains). The results showed in the leaves that Ca, K, P, Mg, Na and Fe were the most abundant of the elements in all samples of the studied locality. The concentrations of trace metals from the leaves of this plant were in the order Ca>K > P > Mg>Na>Fe. The results of the mineral composition were analysed by hierarchical cluster and principal component analysis that established three statistically significant clusters.

Chemical composition and antibacterial activity of Lavandula coronopifolia essential oil against antibiotic-resistant bacteria.

The aim of this study was to analyse the composition of the essential oil (EO) of Lavandula coronopifolia from Morocco and to evaluate its in vitro antibacterial activity against antibiotic-resistant bacteria isolated from clinical infections. The antimicrobial activity was assessed by a broth micro-well dilution method using multiresistant clinical isolates of 11 pathogenic bacteria: Klebsiella pneumoniae subsp. pneumoniae, Klebsiella ornithinolytica, Escherichia coli, Enterobacter cloacae, Enterobacter aerogenes, Providencia rettgeri, Citrobacter freundii, Hafnia alvei, Salmonella spp., Acinetobacter baumannii and methicillin-resistant Staphylococcus aureus. The main compounds of the oil were carvacrol (48.9%), E-caryophyllene (10.8%) and caryophyllene oxide (7.7%). The oil showed activity against all tested strains with minimal inhibitory concentration (MIC) values ranging between 1% and 4%. For most of the strains, the MIC value was equivalent to the minimal bactericidal concentration value, indicating a clear bactericidal effect of L. coronopifolia EO.

Essential oil composition and antibacterial activity of the different parts of Thymus maroccanus Ball: an endemic species in Morocco.

Three essential oils from three samples: stems, leaves and inflorescences of Thymus maroccanus Ball, obtained by hydrodistillation, were analysed by gas chromatography and gas chromatography coupled to mass spectrometry. The oils of stems, leaves and inflorescences afforded 68, 49 and 51 constituents, respectively. Quantitative and qualitative differences have been found between the analysed parts. The most abundant components in the stems oil included carvacrol (60.8%), α-terpineol (4.7%) and p-cymene (2.4%), while carvacrol (33%), p-cymene (25.3%) and α-pinene (11.6%) were the main components in the leaves. In the inflorescences, carvacrol (84.9%) is the major component. Antibacterial activity of the T. maroccanus Ball leaves oil was tested against eight Gram-positive and Gram-negative bacteria by the disc diffusion method. The values of inhibition zones for bacterial strains, which were sensitive to the essential oil of T. maroccanus leaves, were in the range of 11-43 mm.

Chemical composition and antibacterial activities of the essential oils isolated from Juniperus thurifera L. var. Africana.

This study describes the chemical composition and antibacterial activities of essential oils of Moroccan Juniperus thurifera L. var. Africana (Cupressaceae). The essential oil of dried leaves was isolated by hydrodistillation, vapohydrodistillation and microwaves. Sixty-four compounds in J. thurifera L. var. Africana oils were identified (79.9%, 92.4% and 98.4% of the oil, respectively). The most abundant compound in J. thurifera L. var. Africana oils is sabinene (38%, 36.2% and 39.4%). Antibacterial activities of J. thurifera essential oils was tested against bacteria Gram ( - ) and Gram (+). The oil is very active against all bacteria tested except Pseudomonas, which turned out to be very resistant.

Quantitative analysis of some oligo-elements and heavy metals in some species of Thymus from Morocco.

In order to valorise natural substances, concentrations of 20 mineral elements were evaluated in five species of Moroccan thyme. These species which belong to the Lamiaceae family are Thymus leptobotrys, Thymus broussonetii, Thymus maroccanus, Thymus pallidus and Thymus satureioïdes growing in different regions of central and southern Morocco. Samples of plants were subjected to digestion and heavy metals were determined by inductively coupled plasma mass spectrometry. The highest concentrations of calcium, iron, zinc, cobalt and chromium were registered in T. broussonetii, T. pallidus, T. leptobotrys, T. maroccanus and T. satureioïdes with respective values of 1991, 423, 73, 6 and 11 mg/kg. Furthermore, silicium and boron were analysed only for the species T. broussonetii and their respective concentrations were found to be 112 and 43 mg/kg. The ultra trace elements Si, B, Ni, Ni, As, Li, V and toxic elements Cd and Pb were also evaluated. The results were treated by the method of principal components analysis.

Antibacterial, allelopathic and antioxidant activities of essential oil of Salvia officinalis L. growing wild in the Atlas Mountains of Morocco.

Salvia officinalis (Common sage, Culinary sage) is an aromatic plant that is frequently used as a spice in Mediterranean cookery and in the food industry and as a traditional medicine for the treatment of several infectious diseases. The essential oils were obtained by two different methods [hydrodistillation (HD) and microwave (Mw)] from the aerial part of S. officinalis L. growing wild in Ourika-Marrakech in Morocco. Ourika is a large zone of the Atlas Mountains which is considered as a large reserve of Flora, especially medicinal and aromatic plants. The obtained oils were analysed by gas chromatography and gas chromatography-mass spectrometry and compared with that of Tunisia. Thirty-six compounds were identified from the Mw-extracted oil which accounted for 97.32% of the total oil composition. However, 33 compounds obtained by HD representing 98.67%. The major components were trans-thujone (14.10% and 29.84%), 1,8-cineole (5.10% and 16.82%), camphor (4.99% and 9.14%), viridiflorol (16.42% and 9.92%), ß-caryophyllene (19.83% and 5.20%) and α-humulene (13.54% and 4.02%). Antibacterial, allelopathic (% germination in lettuce seeds and inhibited root growth obtained after treatment with S. officinalis oils) and antioxidant (IC50 values 22 mg/mL) activities were studied.

Antibacterial evaluation of selected Moroccan medicinal plants against Streptococcus pneumoniae

In Morocco, most people particularly in rural areas use medicinal plants to treat diseases. In order to find new antipneumococcal extracts, an ethnobotanical survey has been conducted in different regions of Marrakesh (south of Morocco). Four plants often cited by traditional practitioners (Marrubium vulgare, Thymus pallidus, Eryngium ilicifolium and Lavandula stoechas) are tested against Streptococcus pneumoniae responsible for pharyngitis, rhinitis, otitis and sinusitis infections. Aqueous and methanol extracts have been prepared and tested on S. pneumoniae collected in four regions of Marrakesh. A significant activity has been observed with methanol extracts of three plants; M. vulgare, T. pallidus and L. stoechas (MIC= 256 µg/ml)