Maintenance Of pH Research Articles

Impaired hippocampal plasticity associated with loss of recycling endosomal SLC9A6/NHE6 is ameliorated by the TrkB agonist 7,8-dihydroxyflavone

Proper maintenance of intracellular vesicular pH is essential for cargo trafficking during synaptic function and plasticity. Mutations in the SLC9A6 gene encoding the recycling endosomal pH regulator (Na+, K+)/H+ exchanger isoform 6 (NHE6) are causal for Christianson syndrome (CS), a severe form of X-linked intellectual disability. NHE6 expression is also downregulated in other neurodevelopmental and neurodegenerative disorders, such as autism spectrum disorder and Alzheimer's disease, suggesting its dysfunction could contribute more broadly to the pathophysiology of other neurological conditions. To understand how ablation of NHE6 function leads to severe learning impairments, we assessed synaptic structure, function, and cellular mechanisms of learning in a novel line of Nhe6 knockout (KO) mice expressing a plasma membrane-tethered green fluorescent protein within hippocampal neurons. We uncovered significant reductions in dendritic spines density, AMPA receptor (AMPAR) expression, and AMPAR-mediated neurotransmission in CA1 pyramidal neurons. The neurons also failed to undergo functional and structural enhancement during long-term potentiation (LTP). Significantly, the selective TrkB agonist 7,8-dihydroxyflavone restored spine density as well as functional and structural LTP in KO neurons. TrkB activation thus may act as a potential clinical intervention to ameliorate cognitive deficits in CS and other neurodegenerative disorders.

Protective Mechanisms of Vaginal Lactobacilli against Sexually Transmitted Viral Infections.

The healthy cervicovaginal microbiota is dominated by various Lactobacillus species, which support a condition of eubiosis. Among their many functions, vaginal lactobacilli contribute to the maintenance of an acidic pH, produce antimicrobial compounds, and modulate the host immune response to protect against vaginal bacterial and fungal infections. Increasing evidence suggests that these beneficial bacteria may also confer protection against sexually transmitted infections (STIs) caused by viruses such as human papillomavirus (HPV), human immunodeficiency virus (HIV) and herpes simplex virus (HSV). Viral STIs pose a substantial public health burden globally, causing a range of infectious diseases with potentially severe consequences. Understanding the molecular mechanisms by which lactobacilli exert their protective effects against viral STIs is paramount for the development of novel preventive and therapeutic strategies. This review aims to provide more recent insights into the intricate interactions between lactobacilli and viral STIs, exploring their impact on the vaginal microenvironment, host immune response, viral infectivity and pathogenesis, and highlighting their potential implications for public health interventions and clinical management strategies.

Optimizing Gastroesophageal Reflux Disease treatment: A call to shift from omeprazole.

Gastroesophageal reflux disease (GERD) is a disorder of the upper respiratory tract, which develops due to the regurgitation of acid from the stomach into the oesophagus. If left untreated, GERD may cause serious complications such as Barrett's oesophagus, oesophageal strictures, and oesophageal adenocarcinoma. According to a study published by Nirwan JS et al in 2020, individuals suffering from Gastroesophageal reflux disease globally is 1.03 billion.[1] Proton pump inhibitors (PPIs) are frequently used drugs for the treatment of GERD. Omeprazole belongs to PPIs class of drugs and it has been a drug of choice for GERD because it is affordable, easily available, and it can be taken along with other medicines. However, Proton pump inhibitors are not completely risk-free, chronic use of Proton Pump inhibitors has been associated with decreased calcium and magnesium absorption increasing the risk of osteoporosis and electrolyte disturbances respectively. It also increases the risk of community-acquired pneumonia [2] However, Esomeprazole the s-isomer of omeprazole yields much better results than omeprazole. It is more effective and safe in the treatment of GERD, not only does it decrease the symptoms of the disease rapidly ie. heartburn [4] but also it reduces the relapse of the symptoms twice that of omeprazole. Maintenance of intragastric pH > 4 is an effective measure for the management of gastroesophageal reflux disease, esomeprazole has demonstrated improved acid inhibition over omeprazole with a high mean percentage of 24-hour intragastric pH > 4. The healing rate for reflux esophagitis and relapse was also higher for esomeprazole than that of omeprazole.[3] According to a study, the number of individuals with GERD in the South Asia region is around 8000 per 100000 [4]. Knowing the extent of Gastroesophageal patients present in Pakistan and how rapidly this number can escalate due to poor lifestyle modifications. Omeprazole which is the most frequent drug prescribed for the treatment of GERD [5] should be discouraged and Esomeprazole being more beneficial in terms of healing time and relapse of symptoms should be promoted to prevent chronic use adverse effects of Proton pump inhibitors. Overall, such a measure will improve the quality of life of a person suffering from gastroesophageal reflux disease which otherwise may lead to more severe implications.

Design, Development, and In Vivo Testing of the Hydrating Effect and pH Maintenance a Cosmetic Formulation Incorporating Oils and an Extract from Peruvian Biodiversity

Design, Development, and In Vivo Testing of the Hydrating Effect and pH Maintenance a Cosmetic Formulation Incorporating Oils and an Extract from Peruvian Biodiversity

O-015 Enabling laboratory-quality male fertility analysis with 72-hour post ejaculation sample viability

Abstract Study question Can SPX72-treated semen samples sustain diagnostic viability and integrity for up to 72 hours, enabling comparable analysis of mail-in samples to fresh samples? Summary answer SPX72 preserves sperm viability, motility, and integrity up to 72 hours post-collection, equalling the diagnostic precision of fresh samples and enhancing at-home testing capabilities. What is known already Challenges with laboratory-based fertility assessments stem from logistical burdens and the necessity for timely sample delivery, often deterring male patient participation. Semen samples typically need to be assessed within 60 minutes of collection to ensure accuracy. Pre-existing solutions have failed to sufficiently prolong sample viability while matching the diagnostic accuracy of immediate laboratory analyses, an issue Sapyen's SPX72 medium addresses effectively. Study design, size, duration An exhaustive controlled study with over 100 semen samples compared SPX72-treated samples at 72 hours against fresh controls. The study incorporated actual postal logistics to emulate mail-in scenarios. Participants/materials, setting, methods A predetermined quantity of SPX72 per millilitre was added to the collected semen samples, with key indicators, including progressive and total motility, morphology, and DNA fragmentation, measured at intervals of 0, 24, 48, and 72 hours, contrasted with untreated, fresh samples. Main results and the role of chance SPX72 application preserved notable semen motility, evidenced by a mean progressive motility of 37.12% after 72 hours, a slight decrease from the initial 41.14%. This minor 4.02% reduction underscores SPX72's proficiency in maintaining diagnostic-grade motility over an extended duration. In terms of DNA integrity, SPX72 maintained DNA fragmentation at 14.78%, juxtaposed with 12.44% in control samples, a statistically insignificant variance (p-value of 1.15037E-08). This consistency in DNA fragmentation levels is pivotal for the accuracy of fertility diagnostic metrics. Additionally, SPX72 demonstrated its efficacy in stabilising the pH level of semen samples throughout the 72-hour period, ensuring an optimal environment for sperm preservation. This maintenance of pH is crucial as deviations can significantly affect sperm functionality and overall sample integrity. These results substantiate SPX72's capacity to preserve diagnostic sperm quality for up to three days, potentially redefining the logistics of male fertility testing by enabling accurate, laboratory-level assessment through a mail-in format. Limitations, reasons for caution Despite the controlled study and real-world postal validation, ongoing surveillance of sample stability in diverse global postal environments will be crucial to ensure consistent results across all geographies. Wider implications of the findings The data indicate that SPX72 sets a new standard for at-home fertility testing, matching clinical quality without the immediacy constraint. For the first time, home-based sperm diagnostics do not compromise clinical quality, revolutionising how male fertility is approached and managed. Trial registration number not applicable

Experimental Studies on the Mineral Content of Solojo Flour and Protein Isolates from Two Varieties (DAS and BS) of Nigerian Cultivated Solojo Cowpea (Vigna Unguiculata L. Walp)

Studies on the Mineral Content of Solojo Flour and Protein Isolates from the two varieties (DAS and BS) of Nigeria cultivated solojo cowpeas were conducted to determine their nutritional value. These inorganic elements or minerals were classified into 3 categories, the ultra- trace minerals, which is the third category; the micro elements, also known as the trace minerals is the second category; while the first category are the macro elements, also known as major minerals. Some of the macro-elements are Ca, P, Na and Cl, the second category, micro-elements include, iron, copper, cobalt, potassium, magnesium, iodine, zinc, manganese, molybdenum, F, Cr, Se and S. Results show that the proportion of Sodium (Na) which is ingested into the body in the form of NaCl through food intake maintenance of body pH and to retain water ranged from 728.97 to 253.37 ppm (72.90 to 25.34 mg/100 g); 715.24 to 235.45 ppm; 735.28 to 270.37 ppm; 726.59 to 264.35ppm, for FFDAS, FFBS, DAS and BS respectively with all values of the germinated samples all bellow the control. While FFDAS iron content ranged from 4.25 to 13.50 mg/100 g; FFBS ranged from 3.15 to 12.56 mg/100 g; DAS ranged from 3.81 to 12.90 mg/100g; BS ranged from 3.42 to 9.40 mg/100 g. The values of the germinated flours were all greater than the ungerminated flour. Iron helps to transport oxygen round the body and also helps in red blood cells building and to convert food into needed energy by the body. While Manganese an element that is needed in micro quantity but necessary to convert food into energy, is also crucial for healthy bone and cartilage creation. Results also show that zinc quantity increased as germination proceeded and the values ranged from 38.80 ppm to 230.00 ppm (3.880 mg/100 g to 23.00 mg/100 g; 0.003880% to 0.0230%); 40.84 to 250.01 ppm; 32.85 to 93.41 ppm; 37.07 to 115.00 ppm, for FFDAS, FFBS, DAS and BS respectively. The Ca content improved significantly (p<0.05) with sprouting, the value extended from 250.56 ppm to 760.03 ppm (25.056 to 76.00 mg/100g or 0.0251 to 0.0760 %); 400.40 to 998.22 ppm; 116.87 to 195.69 ppm; 113.48 to 220.75 ppm, for FFDAS, FFBS, DAS and BS respectively. Zinc element although needed at the micro level in the body, is essential for a strong immune system to keep the body in good health. It is also crucial for the maintenance of a healthy sense of taste and odor, while Calcium is critical for strong bones and teeth, blood coagulation, and muscle tightening and relaxation. Magnesium is needed to build enzyme and antioxidants and also for healthy bones while Potassium is needed to maintain; water balance, muscle movement, and nerve impulses. It functions in conjuction with Na to regulate blood pressure.

New advances in the protective mechanisms of acidic pH after ischemia: Participation of NO

BackgroundIt has been previously demonstrated that the maintenance of ischemic acidic pH or the delay of intracellular pH recovery at the onset of reperfusion decreases ischemic-induced cardiomyocyte death. ObjectiveTo examine the role played by nitric oxide synthase (NOS)/NO-dependent pathways in the effects of acidic reperfusion in a regional ischemia model. MethodsIsolated rat hearts perfused by Langendorff technique were submitted to 40 min of left coronary artery occlusion followed by 60 min of reperfusion (IC). A group of hearts received an acid solution (pH = 6.4) during the first 2 min of reperfusion (AR) in absence or in presence of l-NAME (NOS inhibitor). Infarct size (IS) and myocardial function were determined. In cardiac homogenates, the expression of P-Akt, P-endothelial and inducible isoforms of NOS (P-eNOS and iNOS) and the level of 3-nitrotyrosine were measured. In isolated cardiomyocytes, the intracellular NO production was assessed by confocal microscopy, under control and acidic conditions. Mitochondrial swelling after Ca2+ addition and mitochondrial membrane potential (Δψ) were also determined under control and acidosis. ResultsAR decreased IS, improved postischemic myocardial function recovery, increased P-Akt and P-eNOS, and decreased iNOS and 3-nitrotyrosine. NO production increased while mitochondrial swelling and Δψ decreased in acidic conditions. l-NAME prevented the beneficial effects of AR. ConclusionsOur data strongly supports that a brief acidic reperfusion protects the myocardium against the ischemia-reperfusion injury through eNOS/NO-dependent pathways.

Revolutionizing Urolithiasis Management: A Paradigm Shift through Nutraceutical Interventions, Fluid Therapy and pH Maintenance - A Comprehensive Review

Urolithiasis is characterized by the formation of mineral stones within the urinary tract, is a significant global health concern impacting millions worldwide. This review article examines the multifaceted nature of urolithiasis, encompassing its epidemiology, etiology, diagnosis, and management strategies. Key factors influencing stone formation include biochemical changes, dietary habits, hydration levels, genetic predispositions, and environmental factors. Diagnostic imaging methods such as ultrasonography and computed tomography aids in accurate diagnosis, while treatment options range from extracorporeal shock wave lithotripsy to dietary modifications and fluid therapy. Nutraceuticals, particularly phytoconstituents rich in polyphenols and alkaloids, offer promising avenues for managing urolithiasis by inhibiting crystal formation and supporting kidney function. Moreover, optimizing fluid intake and maintaining urinary pH levels play pivotal role in preventing stone recurrence. Prospects lie in the refinement of nutraceutical formulations, the integration of wearable technology for monitoring, and comprehensive clinical investigations to establish safety and efficacy. Ultimately, this review underscores the importance of holistic approaches to mitigating the burden of urolithiasis and improving patient outcomes. Keywords: Extracorporeal Shock Wave Lithotripsy, Epidemiology, Fluid Therapy, Nutraceuticals, Urolithiasis.

Cananga odorata (Ylang-Ylang) Essential Oil Containing Nanoemulgel for the Topical Treatment of Scalp Psoriasis and Dandruff.

This research aimed to evaluate the efficacy of a nanoemulgel (NE) containing Cananga odorata (Ylang-Ylang) oil for managing scalp psoriasis and dandruff through various assessments. The study involved phytochemical screening, characterization, stability testing, in vivo performance evaluation, dermatokinetic analysis, central composite rotatable design (CCRD) optimization, in vitro release profiling, and antioxidant and antimicrobial activity assessment of the NE. The NE exhibited excellent stability and maintained physical parameters over a three-month period. In vivo studies showed no skin irritation, maintenance of skin pH (4.55 to 5.08), and improvement in skin hydration (18.09 to 41.28 AU) and sebum content (26.75 to 5.67 mg/cm2). Dermatokinetic analysis revealed higher skin retention of C. odorata in the NE (epidermis: 71.266 µg/cm2, dermis: 60.179 µg/cm2) compared to conventional formulations. CCRD optimization yielded NE formulations with the desired particle size (195.64 nm), entrapment efficiency (85.51%), and zeta potential (-20.59 mV). In vitro release studies indicated sustained release behavior, and antioxidant and antimicrobial properties were observed. This study demonstrates the stability, skin-friendliness, therapeutic benefits, and controlled release properties of the NE. The NE presents a promising option for various topical applications in treating bacterial and fungal diseases, potentially enhancing drug delivery and treatment outcomes in pharmaceuticals and cosmetics.

The electroneutral Na+-HCO3- cotransporter NBCn1 (SLC4A7) modulates colonic enterocyte pHi, proliferation, and migration.

NBCn1 (SLC4A7) is one of the two major Na+-HCO3- cotransporters in the human colonic epithelium, expressed predominantly in the highly proliferating colonocytes at the cryptal base. Increased NBCn1 expression levels are reported in tumors, including colorectal cancer. The study explores its importance for maintenance of the intracellular pH (pHi), as well as the proliferative, adhesive, and migratory behavior of the self-differentiating Caco2BBe colonic tumor cell line. In the self-differentiating Caco2BBe cells, NBCn1 mRNA was highly expressed from the proliferative stage until full differentiation. The downregulation of NBCn1 expression by RNA interference affected proliferation and differentiation and decreased intracellular pH (pHi) of the cells in correlation with the degree of knockdown. In addition, a disturbed cell adhesion and reduced migratory speed were associated with NBCn1 knockdown. Murine colonic Nbcn1-/- enteroids also displayed reduced proliferative activity. In the migrating Caco2BBe cells, NBCn1 was found at the leading edge and in colocalization with the focal adhesion markers vinculin and paxillin, which suggests that NBCn1 is involved in the establishment of cell-matrix adhesion. Our data highlight the physiological significance of NBCn1 in modulating epithelial pH homeostasis and cell-matrix interactions in the proliferative region of the colonic epithelium and unravel the molecular mechanism behind pathological overexpression of this transporter in human colorectal cancers.NEW & NOTEWORTHY The transporter NBCn1 plays a central role in maintaining homeostasis within Caco2BBe colonic epithelial cells through its regulation of intracellular pH, matrix adhesion, migration, and proliferation. These observations yield valuable insights into the molecular mechanism of the aberrant upregulation of this transporter in human colorectal cancers.

Laser-assisted synthesis of nano-hydroxyapatite and functionalization with bone active molecules for bone regeneration

The main goal of bone tissue engineering research is to replace the allogenic and autologous bone graft substitutes that can promote bone repair. Owing to excellent biocompatibility and osteoconductivity, hydroxyapatite is in extensive research and high demand for both medical and non-medical applications. Although various methods have been developed for the synthesis of hydroxyapatite, in the present study we have shown the use of nanosecond laser energy in the wet precipitation method of nano-hydroxyapatite (nHAP) synthesis without using ammonium solution or any other chemicals for pH maintenance. Here, the present study aimed to fabricate the nanohydroxyapatite using a nanosecond laser. The X-ray diffraction and Fourier transform infrared spectroscopy have confirmed the hydroxyapatite formation under laser irradiation in less time without aging. A transmission electron microscopy confirmed the nano size of synthesized nHAP, which is comparable to conventional nHAP. The length and width of the laser-assisted nHAP were found to be in the range of 50–200 nm and 15–20 nm, respectively, at various laser parameters. The crystallite size obtained by Debye Scherrer formulae was found to be in the range of ∼ 16–36 nm. In addition, laser-assisted nHAP based composite cryogel (nanohydroxyapatite/gelatin/collagen I) was synthesized and impregnated with bioactive molecules (bone morphogenic protein and zoledronic acid) that demonstrated significant osteogenic potential both in vitro in cell experiment and in vivo rat muscle pouch model (abdomen and tibia muscles). Dual-energy X-ray analysis, micro-CT, and histological analysis confirmed ectopic bone regeneration. Micro-CT based histomorphometry showed a higher amount (more than 10-fold) of mineralization for animal groups implanted with composite cryogels loaded with bioactive molecules compared to only composite cryogels groups. Our findings thus demonstrate a controlled and rapid synthetic method for the synthesis of nHAP with various physical, chemical, and biological properties exhibited as comparable to conventionally synthesized nHAP.

Maintenance of an Acidic Skin Surface with a Novel Zinc Lactobionate Emollient Preparation Improves Skin Barrier Function in Patients with Atopic Dermatitis.

The skin of patients with atopic dermatitis (AD) is characterised by elevated pH. As a central homeostatic regulator, an increasedpH accelerates desquamation and suppresses lipid processing, resulting in diminished skin barrier function. The aim of this study was to determine whether a novel zinc lactobionate emollient cream can strengthen the skin barrier by lowering skin surface pH. A double-blind, forearm-controlled cohort study was undertaken in patients with AD. Participants applied the test cream to one forearm and a vehicle cream to the other (randomised allocation) twice daily for 56days. Skin surface pH and barrier function (primary outcomes) were assessed at baseline and after 28days and 56days of treatment, amongst other tests. A total of 23 adults with AD completed the study. During and after treatment, a sustained difference in skin surface pH was observed between areas treated with the test cream and vehicle (4.50 ± 0.38 versus 5.25 ± 0.54, respectively, p < 0.0001). This was associated with significantly reduced transepidermal water loss (TEWL) on the test cream treated areas compared with control (9.71 ± 2.47 versus 11.20 ± 3.62g/m2/h, p = 0.0005). Improvements in skin barrier integrity, skin sensitivity to sodium lauryl sulphate, skin hydration, and chymotrypsin-like protease activity were all observed at sites treated with the test cream compared with the control. Maintenance of an acidic skin surface pH and delivery of physiologic lipids are beneficial for skin health and may help improve AD control by reducing sensitivity to irritants and allergens.

Does an R2R3-MYB transcription factor affect anthurium spathe colour variation via regulation of vacuolar pH?

The anthurium is a popular cut flower worldwide having red, pink, coral, white, green, or brown spathes. There is a constant demand for new flower colours in the commercial market. Flower colour in plants is mainly determined by anthocyanins. Understanding anthocyanin variation and other factors affecting anthurium spathe colour is important for genetic engineering approaches. Therefore, our objectives were to assess the factors affecting colour variation of selected commercially available cut flower anthuriums and to determine the associated regulatory networks and transcription factors (TFs). Nineteen commercial cut flower anthurium cultivars were selected for this purpose. The colour of the spathe surface, anthocyanin location, anthocyanidin type and vacuolar pH were recorded. Anthocyanin associated Gene Network Model generation and analysis were carried out. The CIELAB colourimeter procedure indicated the colour variation among the selected 19 cultivars in terms of colour type, colour intensity, chroma, and hue angle. The location of anthocyanin was limited to mesophyll and epidermal cells. Cyanidin was detected in tested anthurium cultivars as the main anthocyanidin. The pH gradient in pigment extracts indicated a variation with a range of 4.6 to 4.94. The gene pathways of anthocyanin biosynthesis and transport were associated with that of the vacuolar pH/H+ pump according to Gene Network Model. Three pathways were regulated by an R2R3-MYB transcription factor. Although, cyanidin was the only pigment in all the tested cultivars, different pH levels by R2R3-MYB regulated V-H+ synthase was suggested to be the cause of the high colour variation in addition to the anthocyanidin type and location. Our results indicate the application of R2R3-MYB transcription factor genes for desirable vacuolar pH maintenance in genetic engineering of the blue anthurium in the future.

Ligand based pharmacophore modelling and integrated computational approaches in the quest for small molecule inhibitors against hCA IX.

Carbonic anhydrase IX is an important biomarker to fight hypoxic tumours in both initial and metastatic stages of many forms of cancer. Overexpression of hCA IX in the hypoxic environment, has an active role in pH maintenance and makes the hCA IX a better target for the inhibitors targeting specific types of cancer stages. Being a member of the carbonic anhydrase family and having sixteen isoforms, it is important to have a selective inhibition of hCA IX to limit the disruption in the biological and metabolic pathways where other isoforms of hCA are localised and to avoid the other toxicity and adverse effects we try to find selective hCA IX inhibitors from a natural derivative. In the process of finding selective hCA inhibitors we developed a pharmacophore model based on existing inhibitors with IC50 values of less than 50 nm, which is then validated with the external decoy set and used for database searching followed by virtual screening to identify the hits based on the pharmacophore fit score and RMSD. Molecular docking studies were performed to identify protein ligand interaction and molecular dynamics simulation studies to analyse the stability of the complex and DFT studies were carried out. The initial screening yielded 43 hits with the RMSD value less than 1, which when subjected to docking exhibited very good interaction with key residues ZN301, HIS94, HIS96 and HIS119. The top 4 compounds in the molecular dynamics simulation studies for 100 ns provided useful insights on the stability of the complex and the DFT studies confirmed the energy variation between HOMO and LUMO is within an acceptable range. An average binding score of -7.8 Kcal mol-1 for the lead compounds and high stability margin in the dynamics study concludes that these lead compounds demonstrated outstanding potential for hCA IX inhibitory action theoretically and that further experimental studies for selective inhibition are inevitable.

The protective effect of a new absorbent incontinence design against an alkaline pH challenge on the epidermal barrier.

Incontinence-associated dermatitis (IAD) is one of the most common complications of incontinence. Improved diaper designs can minimize the occurrence of IAD. To develop a novel diaper design to minimize the damaging effects of incontinence on the epidermal barrier. An optimized diaper design was tested for surface dryness (ie, rewet), maintenance of a skin-adapted surface pH of 5.5, and ability to protect epidermal barrier function from an alkaline pH 10.7 challenge. The diapers released a mean (standard deviation [SD]) of 1.2 (0.2) mg/cm2 of solution under pressure after the first loading and a mean of 2.9 (1.7) mg/cm2 after the second loading. The surface pH remained between 4.5 and 5.5 over 5 hours. In healthy skin, transepidermal water loss (TEWL) increased by a mean of 3.43 (4.67) g/m2/h after the alkaline urine solution challenge with the new diaper design versus a mean of 8.38 (5.67) g/m2/h with a cellulose patch (P < .001) as a control. The mean erythema readings were 1.18 (1.30) g/m2/h for the new design and 2.56 (1.25) g/m2/h for the cellulose patches (P < .001). The new diaper design minimizes rewetting, maintains an acidic surface, and protects the epidermal barrier against an alkaline pH challenge. This design may help prevent IAD.

Can hypoxia marker carbonic anhydrase IX serve as a potential new diagnostic marker and therapeutic target of non-small cell lung cancer?

Lung cancer represents the leading cause of cancer-related deaths. Non-small cell lung cancer (NSCLC), the most common form of lung cancer, is a molecularly heterogeneous disease with intratumoral heterogeneity and a significant mutational burden associated with clinical outcome. Tumor microenvironment (TME) plays a fundamental role in the initiation and progression of primary de novo lung cancer and significantly influences the response of tumor cells to therapy. Hypoxia, an integral part of the tumor microenvironment and a serious clinical phenomenon, is associated with increased genetic instability and a more aggressive phenotype of NSCLC, which correlates with the risk of metastasis. Low oxygen concentration influences all components of TME including the immune microenvironment. Hypoxia-inducible pathway activated in response to low oxygen supply mediates the expression of genes important for the adaptation of tumor cells to microenvironmental changes. A highly active transmembrane hypoxia-induced metalloenzyme - carbonic anhydrase IX (CAIX), as a part of transport metabolon, contributes to the maintenance of intracellular pH within physiological values and to the acidification of the extracellular space. CAIX supports cell migration and invasion and plays an important role in NSCLC tumor tissue and pleural effusion. Due to its high expression, it also represents a potential diagnostic differential biomarker and therapeutic target in NSCLC. To test new potential targeted therapeutic compounds, suitable models are required that more faithfully simulate tumor tissue, TME components, and spatial architecture.

Acid adaptation alters Streptococcus mutans drug susceptibility profile

The ability of Streptococcus mutans, the primary causative agent of dental caries, to generate and tolerate an acidic environment is a leading factor in its capacity to outcompete other organisms within the oral cavity. The acid tolerance response (ATR) involves coordination of multiple pathways to upregulate macromolecule repair, maintenance of cytosolic pH, and restructuring the membrane. In this study, we asked if the changes associated with adaptation to a low pH environment would result in detectable changes to drug susceptibility. Accordingly, we screened using the Selleck library of 853 FDA approved drugs for agents that exhibit antimicrobial activity toward planktonic cultures of S. mutans or disrupt biofilm formation in growth medium titrated to pH 5 or in the presence of hydrogen peroxide. Results revealed that a total of 182 compounds displayed antimicrobial properties toward the test strain in at least one screening condition. Our findings provide evidence that the S. mutans ATR is characterized by a concerted shift in drug susceptibility. The presence of an intact ATR, associated with growth in continuous culture at pH 5, leads to increased sensitivity to 22 compounds and decreased sensitivity to a separate set of 20 compounds. Disruption of the ATR in a mutant lacking the fabM gene resulted in a susceptibility profile that was distinct from the result of our results. This suggests that loss of unsaturated fatty acids produces a physiological change that is not only measurable, but perhaps more significant than the process of ATR. Additional tests revealed that S. mutans exhibits a distinct drug susceptibility profile compared to a closely related, non-cariogenic bacterium, S. gordonii. Overall, shifts in S. mutans susceptibility patterns based on pH of growth medium or type of culture (batch vs. continuous culture), indicate that the ATR can be tracked using changes in drug susceptibility profile. Furthermore, our results provide clues that compounds could be useful in treatment of dental caries as well as providing molecular tools to study shifts in physiology of disease-associated bacteria.

A Comparative Kidney Transcriptome Analysis of Bicarbonate-Loaded insrr-Null Mice.

The maintenance of plasma pH is critical for life in all organisms. The kidney plays a critical role in acid-base regulation in vertebrates by controlling the plasma concentration of bicarbonate. The receptor tyrosine kinase IRR (insulin receptor-related receptor) is expressed in renal β-intercalated cells and is involved in alkali sensing due to its ability to autophosphorylate under alkalization of extracellular medium (pH > 7.9). In mice with a knockout of the insrr gene, which encodes for IRR, urinary bicarbonate secretion in response to alkali loading is impaired. The specific regulatory mechanisms in the kidney that are under the control of IRR remain unknown. To address this issue, we analyzed and compared the kidney transcriptomes of wild-type and insrr knockout mice under basal or bicarbonate-loaded conditions. Transcriptomic analyses revealed a differential regulation of a number of genes in the kidney. Using TaqMan real-time PCR, we confirmed different expressions of the slc26a4, rps7, slc5a2, aqp6, plcd1, gapdh, rny3, kcnk5, slc6a6 and atp6v1g3 genes in IRR knockout mice. Also, we found that the expression of the kcnk5 gene is increased in wild-type mice after bicarbonate loading but not in knockout mice. Gene set enrichment analysis between the IRR knockout and wild-type samples identified that insrr knockout causes alterations in expression of genes related mostly to the ATP metabolic and electron transport chain processes.

Advances in the Development of Carbonic Anhydrase Inhibitors as New Antiprotozoal Agents.

Parasitic diseases are a public health problem despite the existence of drugs for their treatment. These treatments have variable efficacy and, in some cases, serious adverse effects. There has been interest in the enzyme carbonic anhydrase (CA) in the last two decades since it is essential in the life cycle of various parasites due to its important participation in processes such as pyrimidine synthesis, HCO3 - transport across cell membranes, and the maintenance of intracellular pH and ion transport (Na+, K+, and H+), among others. In this review, CA was analyzed as a pharmacological target in etiological agents of malaria, American trypanosomiasis, leishmaniasis, amoebiasis, and trichomoniasis. The CA inhibitors´ design, binding mode, and structure-activity relationship are also discussed. According to this review, advances in discovering compounds with potent inhibitory activity suggest that CA is a candidate for developing new antiprotozoal agents.

Vacuolar H+-ATPase and BZR1 form a feedback loop to regulate the homeostasis of BR signaling in Arabidopsis

Brassinosteroid (BR) is a vital plant hormone that regulates plant growth and development. BRASSINAZOLE RESISTANT 1 (BZR1) is a key transcription factor in BR signaling, and its nucleocytoplasmic localization is crucial for BR signaling. However, the mechanisms that regulate BZR1 nucleocytoplasmic distribution and thus the homeostasis of BR signaling remain largely unclear. The vacuole is the largest organelle in mature plant cells and plays a key role in maintenance of cellular pH, storage of intracellular substances, and transport of ions. In this study, we uncovered a novel mechanism of BR signaling homeostasis regulated by the vacuolar H+-ATPase (V-ATPase) and BZR1 feedback loop. Our results revealed that the vha-a2 vha-a3 mutant (vha2, lacking V-ATPase activity) exhibits enhanced BR signaling with increased total amount of BZR1, nuclear-localized BZR1, and the ratio of BZR1/phosphorylated BZR1 in the nucleus. Further biochemical assays revealed that VHA-a2 and VHA-a3 of V-ATPase interact with the BZR1 protein through a domain that is conserved across multiple species. VHA-a2 and VHA-a3 negatively regulate BR signaling by interacting with BZR1 and promoting its retention in the tonoplast. Interestingly, a series of molecular analyses demonstrated that nuclear-localized BZR1 could bind directly to specific motifs in the promoters of VHA-a2 and VHA-a3 to promote their expression. Taken together, these results suggest that V-ATPase and BZR1 may form a feedback regulatory loop to maintain the homeostasis of BR signaling in Arabidopsis, providing new insights into vacuole-mediated regulation of hormone signaling.