Bacillus anthracis edema toxin inhibits hypoxic pulmonary vasoconstriction via edema factor and cAMP-mediated mechanisms in isolated perfused rat lungs

The most important findings here are edema toxin’s potent adenyl cyclase activity can interfere with hypoxic pulmonary vasoconstriction, an action that could worsen hypoxemia during invasive anthrax infection with lung involvement. These findings, coupled with other studies showing that lethal toxin can disrupt pulmonary vascular integrity, indicate that both toxins can contribute to pulmonary pathophysiology during infection. In combination, these investigations provide a further basis for the use of antitoxin therapies in patients with worsening invasive anthrax disease.

Neuroprotective potential of adenyl cyclase/cAMP/CREB and mitochondrial CoQ10 activator in amyotrophic lateral sclerosis rats

Aim: To investigate neuroprotective potential of of forskolin (FSK) in combination with solanesol (SNL) along with clinically proven drugs (riluzole, baclofen, citalopram) on behavioral, molecular and neurochemical alterations in methyl mercury-induced amyotrophic lateral sclerosis (ALS) rats. Background: ALS is a motor neuron disease in which oxidative stress is the principle mechanism ofneuronal death which can be mimicked by the dominant mutations in an antioxidant enzyme SOD-1. Due to MeHg neurotoxicity, behavioral and neurochemical alterations occur in rats. During ALS mitochondrial CoQ10 dysfunctioning and downregulation of adenyl cyclase/CREB are major pathological hallmark for neurodegeneration in ALS. Clinically proven drug therapy comes with limited therapeutic involvement, and is used as approachable therapy in ALS patients. Objective: Therefore, current research explores the up-regulation of adenyl cyclase/cAMP/CREB by FSK 30, 60 mg/kg in combination with mitochondrial ETC-coenzyme-Q10 precursor SNL 15, 30 mg/kg can be a preventive therapeutic approach to overcome the ALS like symptoms. Method: MeHg (5 mg/kg) is a neurotoxic compound that leads to ALS like behavioral & neurochemical alterations. Results: Chronic treatment with the combination of FSK 30,60 mg/kg and SNL 15,30 mg/kg alone and along with standard drugs citalopram (5 mg/kg), riluzole (5 mg/kg) and baclofen (3 mg/kg) increased the adenyl cyclase and mitochondrial CoQ10 and ETC-complexes enzyme levels and shows the neuroprotective potential by significantly improving the cognitive deficitslocomotion, , grip strength, and restoration of neurochemicals alterations along with reducing the level of inflammatory mediators and oxidative stress in ALS rats. Conclusion: Thus, we concluded that FSK in combination with SNL along with standard drugs can be a possible therapeutic approach for the treatment of ALS.

Exploring molecular approaches in Amyotrophic lateral sclerosis: Drug targets from clinical and pre-clinical findings

: Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disease (MND) characterised by the death of upper and lower motor neurons (corticospinal tract) in the motor cortex, basal ganglia, brain stem, and spinal cord. The patient experiences the sign and symptoms between 55 to 75 years of age included impaired motor movement, difficulty in speaking and swallowing, grip loss, muscle atrophy, spasticity and sometimes associated with memory and cognitive impairments. Median survival is 3 to 5 years after diagnosis and 5 to 10% beyond 10 years of age. The limited intervention of pharmacologically active compounds that are used clinically is majorly associated with the narrow therapeutic index. Pre-clinically established experimental models where neurotoxin methyl mercury mimics the ALS like behavioural and neurochemical alterations in rodents associated with neuronal mitochondrial dysfunctions and downregulation of adenyl cyclase mediated cAMP/CREB is the main pathological hallmark for the progression of ALS in central as well in the peripheral nervous system. Despite the considerable investigation into neuroprotection, it still constrains treatment choices to strong care and organization of ALS complications. Therefore, current review specially targeted in the investigation of clinical and pre-clinical features available for ALS to understand the pathogenic mechanisms and to explore the pharmacological interventions associated with up-regulation of intracellular adenyl cyclase/cAMP/CREB and mitochondrial-ETC coenzyme-Q10 activation as a future drug target in the amelioration of ALS mediated motor neuronal dysfunctions.

Kadar Kortisol Plasma pada Dispepsia Fungsional dengan Gangguan Psikosomatik

Pendahuluan. Peningkatan kortisol akan merangsang produksi asam lambung dan dapat menghambat prostaglandin E yang merupakan inhibitor enzim adenyl cyclase dalam sel parietal yang melindungi mukosa lambung. Nilai kortisol plasma yang tinggi dikaitkan dengan stres psikologis yang dapat memicu keluhan dispepsia. Penelitian ini dilakukan untuk mengetahui kadar kortisol plasma pada penderita dispepsia fungsional dengan gangguan psikosomatik.Metode. Studi deskriptif analitik dengan desain cross sectional dilakukan pada pasien dispepsia yang dipilih menggunakan teknik non random consecutive sampling. Setelah diagnosis dispepsia, depresi, kecemasan, dan keduanya (depresi – kecemasan), semua subjek diperiksa gastroskopi dan kortisol plasma pada pagi dan sore hari.Hasil. Selama periode penelitian, didapatkan sampel sebanyak 40 pasien dengan dispepsia dan gangguan psikosomatik dan 10 pasien dengan dispepsia tanpa gejala psikosomatis sebagai kelompok kontrol dengan rentang usia 20 - 40 tahun. Rerata kadar kortisol di pagi hari pada pasien dengan gangguan psikosomatik adalah 322,33 nmol/L (simpang baku [SB] 166,92 nmol/L) sedangkan pada kelompok normal 188,82 nmol/L (SB 103,14 nmol/L). Sementara itu, median kortisol sore hari kelompok dengan psikosomatisk adalah 136,25 nmol/L (rentang 17,8-494,1 nmol/L) dan pada kelompok normal 91,40 nmol/L (rentang 10,6-291,6 nmol/L). Berdasarkan hasil uji statistik didapatkan bahwa terdapat peningkatan kortisol pagi hari yang signifikan antara kelompok depresi (rerata 338,82 nmol/L (SB 166,82 nmol/L) dengan kelompok normal, dengan nilai p=0,013. Sedangkan, nilai kortisol sore hari antara kelompok dengan gangguan psikosomatik tidak memiliki perbedaan yang signifikan secara statistik (p>0,05).Simpulan. Nilai kortisol plasma pagi hari pada dispepsia fungsional dengan gangguan psikosomatik (depresi) secara signifikan lebih tinggi daripada kelompok tanpa gangguan psikosomatik. Kata Kunci: Dispepsia fungsional, gangguan psikosomatik, kortisol plasmaLevel of Plasma Cortisol in Functional Dyspepsia with Psychosomatic DisorderIntroduction. Increasing of cortisol will stimulate the production of gastric acid and can inhibit prostaglandin E which is an inhibitor of the enzyme adenyl cyclase in parietal cells that is protective of gastric mucosa. High value of plasma cortisol is associated with psychological stress which can trigger the complaints of dyspepsia. This study aims to find out the level of plasma cortisol in dyspepsia functionalMethods. An analytic descriptive - cross sectional study with nonrandom consecutive sampling was conducted among dyspepsia patients. After diagnosis of functional dyspepsia, depression, anxiety and both of depression – anxiety, all subjects were examined for gastroscopy and plasma cortisol in the morning and evening.Results. There were 40 patients with dyspepsia and psychosomatic disorders and 10 patients with dyspepsia without psychosomatic symptom as control group with an age range of 20 – 40 years old. The mean value of cortisol level in the morning among subjects with psychosomatic disorder was 322.33 nmol/L (SD 166.92 nmol/L) whereas in the normal group was 188.82 nmol/L (SD 103.14 nmol/L). The median value of cortisol level in the evening among subjects with psychosomatic disorder was 136.25 nmol/L (range 17.8-494.1 nmol/L) whereas in the normal group was 91.40 nmol/L (range 10.6-291.6 nmol / L). Statistical analysis showed that there was a significant increased in the morning cortisol between sujects with depression (mean value = 338.82 nmol/L (SD 166.82 nmol/L) compared to normal group (p value=0.013). Meanwhile, there difference in afternoon cortisol between subjects with psychosomatic disorder and the normal group was not statistically significant (p>0.05).Conclusion. The level of morning plasma cortisol in functional dyspepsia with psychosomatic patients (depression) was significantly higher than patients without psychosomatic disorder.

Neuroprotective methodologies in treatment of Multiple Sclerosis: Current status of Clinical and Pre-clinical findings

: Multiple sclerosis an idiopathic and autoimmune associated motor neuron disorder that attacks myelinated neurons in specific brain regions of young people, especially females; characterized by oligodendrocytes destruction causes demyelination, neuroinflammation, mitochondrial abnormalities, oxidative stress and neurotransmitter deficits associated with motor and cognitive dysfunctions, vertigo and muscle weakness. The limited intervention of pharmacologically active compounds like interferon-β, mitoxantrone, fingolimod and monoclonal antibodies used clinically are majorly associated with adverse drug reactions. Pre-clinically, gliotoxin ethidium bromide mimics the behavioral and neurochemical alterations in multiple sclerosis like experimental animals associated with the down regulation of adenyl cyclase/cAMP/CREB further responsible for varieties of neuropathogenic factors. Despite the considerable investigation of neuroprotection in curing multiple sclerosis, some complications still constrained. As of now approachable drugs give only symptomatic alleviation but don’t end the development of illness. In this way, the advancement of unused helpful techniques remains neglected restorative requires. The limitations of current steady treatment may be because of their activity at one of many neurotransmitters included or their failure to up direct signaling flag bearers detailed to have a vital part in neuronal sensitivity, biosynthesis of neurotransmitters and its discharge, development and separation of neuron, synaptic versatility and cognitive working. Therefore, the current review strictly focused on the exploration of various clinical and pre-clinical features available for multiple sclerosis to understand the pathogenic mechanisms and to introduced pharmacological interventions associated with the upregulation of intracellular adenyl cyclase/cAMP/CREB activation to ameliorate multiple sclerosis like features.

The activation of the chymotrypsin-like activity of the proteasome is regulated by soluble adenyl cyclase/cAMP/protein kinase A pathway and required for human sperm capacitation

One of the first events of mammalian sperm capacitation is the activation of the soluble adenyl cyclase/cAMP/protein kinase A (SACY/cAMP/PKA) pathway. Here, we evaluated whether the increase in PKA activity at the onset of human sperm capacitation is responsible for the activation of the sperm proteasome and whether this activation is required for capacitation progress. Viable human sperm were incubated with inhibitors of the SACY/cAMP/PKA pathway. The chymotrypsin-like activity of the sperm proteasome was evaluated using a fluorogenic substrate. Sperm capacitation status was evaluated using the chlortetracycline assay and tyrosine phosphorylation. To determine whether proteasomal subunits were phosphorylated by PKA, the proteasome was immunoprecipitated and tested on a western blot using an antibody against phosphorylated PKA substrates. Immunofluorescence microscopy analysis and co-immunoprecipitation (IPP) were used to investigate an association between the catalytic subunit alpha of PKA (PKA-Cα) and the proteasome. The chymotrypsin-like activity of the sperm proteasome significantly increased after 5 min of capacitation (P < 0.001) and remained high for the remaining incubation time. Treatment with H89, KT5720 or KH7 significantly decreased the chymotrypsin-like activity of the proteasome (P < 0.001). IPP experiments indicated that PKA inhibition significantly modified phosphorylation of proteasome subunits. In addition, PKA-Cα colocalized with the proteasome in the equatorial segment and in the connecting piece, and co-immunoprecipitated with the proteasome. This is the first demonstration of sperm proteasome activity being directly regulated by SACY/PKA-Cα. This novel discovery extends our current knowledge of sperm physiology and may be used to manage sperm capacitation during assisted reproductive technology procedures.

The Adenylate Cyclase (CyaA) Toxin from Bordetella pertussis Has No Detectable Phospholipase A (PLA) Activity In Vitro

The adenylate cyclase (CyaA) toxin produced in Bordetella pertussis is the causative agent of whooping cough. CyaA exhibits the remarkable capacity to translocate its N-terminal adenyl cyclase domain (ACD) directly across the plasma membrane into the cytosol of eukaryotic cells. Once translocated, calmodulin binds and activates ACD, leading to a burst of cAMP that intoxicates the target cell. Previously, Gonzalez-Bullon et al. reported that CyaA exhibits a phospholipase A activity that could destabilize the membrane to facilitate ACD membrane translocation. However, Bumba and collaborators lately reported that they could not replicate these results. To clarify this controversy, we assayed the putative PLA activity of two CyaA samples purified in two different laboratories by using two distinct fluorescent probes reporting either PLA2 or both PLA1 and PLA2 activities, as well as in various experimental conditions (i.e., neutral or negatively charged membranes in different buffers.) However, we could not detect any PLA activity in these CyaA batches. Thus, our data independently confirm that CyaA does not possess any PLA activity.