Urinary tract infections (UTIs) are among the most common bacterial infections, particularly in women, with a significant proportion developing recurrent UTIs (RUTIs). In postmenopausal women, the changes in the urogenital flora, along with physiological changes like decreased estrogen levels, contribute to an increased risk of UTIs. The repeated use of antibiotics for RUTIs leads to concerns about antibiotic resistance, making alternative treatments essential. Electrofulguration is a procedure that involves the use of high-frequency electrical currents to destroy tissue, typically abnormal or unwanted tissue. The process involves the application of an electrode to the target area, where the electrical current produces heat that causes the tissue to disintegrate or coagulate, leading to its destruction. Electrofulguration is known for its precision and ability to minimize damage to surrounding healthy tissue, making it a valuable tool for certain types of surgical procedures. In the context of RUTIs, electrofulguration involves the cauterization of the bladder lining to remove areas of inflammation, and has emerged as a potential alternative for cases where antibiotics fail.

In a new study published in Journal of Urology by Shivani Gaitonde, Alana Christie, Priya Garigipati, Feras Alhalabi, and led by Professor Philippe Zimmern from the Department of Urology at University of Texas Southwestern Medical Center, the authors evaluated the long-term outcomes of electrofulguration in the treatment of antibiotic-refractory RUTIs in a cohort predominantly composed of menopausal women. The study spanned from 2006 to 2012 and involved a follow-up period extending up to 11 years, with some participants having more than a decade of post-treatment observations. The authors included nonneurogenic women who had experienced three or more symptomatic RUTIs per year, all of whom exhibited inflammatory lesions on cystoscopy indicative of cystitis. The women chosen had a history of RUTIs that were unresponsive to conventional antibiotic treatments. Patients with less than a 5-year follow-up, significant comorbidities, or those who had undergone associated urological surgeries were excluded to maintain focus on the efficacy of electrofulguration in a specific patient population. Electrofulguration was performed as an outpatient procedure. The process they used involved the use of a fine-tip monopolar Bugbee electrode to specifically fulgurate areas of the bladder identified during cystoscopy as having chronic cystitis. The procedure aimed to disrupt and remove the nidus of infection within the bladder lining. The team collected detailed data on the number of UTI episodes, antibiotic usage, and any subsequent electrofulguration procedures post-initial treatment. They also conducted telephone interviews to capture data on UTI symptoms, recurrences, and additional treatments for patients not recently seen in clinics.

At the end of the follow-up period, the researchers observed a significant portion of the cohort (72%) experienced a complete resolution of UTI symptoms. An additional 22% of participants showed improvement, with a reduction in the frequency of UTIs, while only 6% did not benefit from the procedure. Moreover, there was a notable decrease in the reliance on continuous antibiotic therapy post-electrofulguration. Before electrofulguration, 74% of the participants were on continuous antibiotics, which reduced to 5% at the last follow-up, showcasing a substantial reduction in antibiotic dependency.  A minority of the participants (19%) underwent a repeat electrofulguration procedure, indicating that while electrofulguration was largely effective, some cases required additional interventions for optimal outcomes. In conclusion, the study by Professor Philippe Zimmern and colleagues represents an important contribution in the management of RUTIs in postmenopausal women. The very long-term outcomes of electrofulguration highlight its potential as a durable and effective alternative to antibiotics, offering hope to those suffering from antibiotic-refractory RUTIs. Future research, ideally in the form of randomized controlled trials with diverse populations, is needed to further validate these findings and explore the full potential of electrofulguration in the management of RUTIs.

Abdominal wall hernia is the result of abdominal wall muscle or connective tissue weakness that allows visceral organs to herniate through. Several prosthetic biomaterial-based devices for hernia repair are available, while more innovative and clinically efficient ones are under investigation. Among them, polypropylene meshes have been accepted for a long time as a standard element for abdominal hernia repair. These meshes are pliable, well incorporable into adjacent tissues and capable of giving robust mechanical support. However, poor flexibility, adhesion to visceral organs and inflammatory risks are problems connected with the utilization of these materials. Besides, the mesh is inert and hydrophobic, with poor cell attachment and proliferation. Ideally, the implantable mesh should trigger a minimum immune response and exhibit excellent biocompatibility to accelerate the healing process for proper integration into the body. This is why decorating the polypropylene mesh with bio-friendly materials to enhance its biocompatibility is highly desired.

Since the use of metal nanoparticles in biological settings is limited, an alternative to decorate the implantable mesh is the use of copolymers and composites. Unfortunately, the process is cumbersome, and the materials developed are structurally complex. Another alternative is the use of bioactive materials. One example of these materials is chitosan, which exhibits excellent biocompatibility, biodegradability, nontoxicity, and antimicrobial properties.

Various crosslinkers, including formaldehyde, glyoxal and citric acid, are used to enhance the attachment of chitosan to the surface of polypropylene mesh. However, some of these crosslinkers are toxic and exhibit poor interaction with cells. Therefore, oxygen plasma treatment has been fronted as a suitable option to activate the polypropylene mesh surface to improve chitosan attachment.

Since chitosan’s antifungal and antimicrobial properties depend on various factors such as pH, molecular weight, and the microorganism cell wall properties, its sole use might be insufficient. Incorporating an active substance to improve its microbial activity is still needed. Detonation nanodiamond is an excellent example of carbon-based nanoparticles with low toxicity and multiple functional groups allowing further surface functionalization in several applications.

The application of nanodiamond and chitosan has been reported in the literature. However, the application of chitosan and hydroxylated nanodiamond composites on polypropylene mesh hasn’t been investigated for microbial and cellular activities.

Given this, Australian researchers from RMIT University: Tanushree Saha, Dr. Shadi Houshyar, Dr Satya Ranjan Sarker, Dr. Suneela Pyreddy, Dr. Chaitali Dekiwadia, Dr. Zeyad Nasa, Professor Rajiv Padhye, and Dr. Xin Wang developed a new innovative method for the designing of an antimicrobial and biocompatible polypropylene mesh via modification with bioactive chitosan and functionalized nanodiamond to accelerate the healing process after surgery and inhibit infection. Their research work has been published in the Journal of Biomedical Materials Research.

The research team used an oxygen plasma-treated polypropylene mesh and attached chitosan to its fibers. Subsequently, they loaded functionalized nanodiamonds into the chitosan-modified polypropylene fiber to provide the much-needed antibacterial properties. The authors characterized the meshes with various advanced analytical tools such as optical microscopy, XRD, water angel contact, FTIR, and SEM.

Coated-cured polypropylene mesh led to optimal chitosan attachment to the polypropylene mesh surface and a longer time in vitro. When the authors treated the chitosan-coated polypropylene mesh with a low concentration (0.3%) of functionalized nanodiamond, the cell attachment was significantly enhanced (i.e., 134%).

The authors observed that the modified polypropylene mesh with chitosan and functionalized nanodiamond exhibited superb resistance to E. coli bacteria besides having excellent fibroblast cell proliferation ability and biocompatibility. Moreover, the researchers recorded a negative effect on cell attachment in vitro when they used citric acid as a crosslinker to improve chitosan attachment to the plasma-treated polypropylene surface. From the surface characterization results of this novel polypropylene mesh modified with chitosan and functionalized nanodiamond, the authors concluded that it’s possible to adopt this new strategy to develop an efficient interactive PP hernia mesh.

The excellent performance of the newly developed by RMIT scientists of functionalized nanodiamond-treated polypropylene mesh opens new avenues for further drug loading by functionalized nanodiamonds in resolving long-standing post-operational complications and therapy.

The powerful vasodilator and neuropeptide calcitonin gene-related peptide CGRP is abundant in trigeminal ganglion neurons, and is released from the peripheral nerve and central nerve terminals as well as being secreted within the trigeminal ganglion. CGRP has a proven role in migraine and selective antagonists and antibodies are now reaching the clinic for treatment of migraine. Indeed, a new class of drugs to treat patients with frequent, episodic, and/or chronic migraine headaches acts by antagonism of the calcitonin gene-related peptide (CGRP) pathway. This is the first category of pharmaceuticals developed as targeted therapy for migraine prevention.

RAMPs are an example of membrane-spanning auxiliary proteins that can change the function of G protein-coupled receptors (GPCRs). RAMPs are small families of three proteins that have the potential to introduce functional variety by interacting with GPCRs. RAMPs have a single transmembrane spanning domain with an extracellular N-terminal region of 90-100 amino acids and a short intracellular C-terminal domain of 9 amino acids. Maltose-binding protein (MBP) was employed as a tagged protein in several crystal structures of the CGRP and AM receptor extracellular domain (ECD). It was predicted that N-glycosylation of the CGRP receptor ECD would inhibit MBP from docking to the altered peptide ligands.

In a new study published in the journal Peptides by Dr. Sangmin Lee from Fred Wilson School of Pharmacy at High Point University investigated the role of CGRP receptor N-glycosylation in the interaction with the MBP tag. He postulated that CLR N-glycosylation would push the MBP tag away from the CGRP receptor’s peptide binding site based on known crystal structures. According to the author N-glycosylation of CLR ECD N123 decreased MBP interaction with peptide ligands, suggesting that N-glycosylation plays a substantial role in peptide ligand binding for the MBP-tagged CGRP receptor ECD.

Dr. Lee employed FITC-labeled AM(37-52) with S45W/Q50W mutations for FP peptide binding and its affinity for the MBP tagged, or MBP-free CGRP receptor ECD (tested as RAMP1-CLR ECD fusion protein) has previously been published. Although these affinity values were obtained from two separate tests, the inclusion of the N-terminal MBP tag resulted in a 5-fold greater affinity of the peptide ligand. This matches the MBP interaction with the peptide ligands. MBP did not have any N-glycan in the crystal structures, as expected from its sequence. CLR ECD of the MBP-tagged RAMP1-CLR ECD fusion protein has three N-glycosylation sequons, which are N66, N118, and N123 of CLR ECD. The author findings show that N-glycosylation of the RAMP2-CLR ECD fusion protein increases peptide ligand binding considerably. He also demonstrated that N-glycosylation of the AM receptor ECD improved peptide ligand binding much and CLR N123 was the primary cause of this impact. He also showed that the conformational dynamics of RAMP1- or RAMP2-CLR complexes might be a significant driver of different receptor phenotypes.

According to Dr. Lee   MBP N-terminally linked to the CGRP receptor ECD offered an extra interaction site for mutant peptide ligands, increasing their affinity for the CGRP receptor ECD. However, these MBP tags can be easily removed. Moreover, CGRP can be expressed using other tags such as GST, would the affinity binding affected, something that was not studied and lacking in the study. Furthermore, the argument that MBP will affect affinity binding is not strong and it depends on expression system where glycosylation can change using different expression system. Common screening methods are are largely based on measuring CGRP-mediated changes in cAMP which will not be affected by MBP tag. Moreover, a successful advancement in the migraine treatment was the development of Erenumab which is the first FDA approved antibody therapeutic against a G-protein-coupled receptor, the canonical receptor of calcitonin gene related peptide (CGRP-R). The researchers at Amgen created a  novel, epitope-focused antigen  to reconstruct the extracellular domains of the CGRP-R in a stable conformation. The study is weak with contradictory results and we believe the published paper on the effect of MBP on binding affinity of CGRP is NOT of importance in migraine drug development.

The dopamine D2 receptor has a previously unobserved role in modulating Wnt expression and control of cell proliferation, according to a new study reported by scientists from the George Washington University and the University of Pittsburgh in Scientific Reports, which could have implications for the development of new therapeutics across multiple disciplines including nephrology, endocrinology, and psychiatry.

The Wnt/β-catenin pathway is one of the most conserved signaling pathways across species with essential roles in development, cell proliferation, and disease. Wnt signaling occurs at the protein level and via β-catenin-mediated transcription of target genes. However, little is known about the underlying mechanisms regulating the expression of the key Wnt ligand Wnt3a or the modulation of its activity.

The research team provided evidence that there is significant cross-talk between the dopamine D₂ receptor (D2R) and Wnt/β-catenin signaling pathways. Our data suggest that D2R-dependent cross-talk modulates Wnt3a expression via an evolutionarily-conserved TCF/LEF site within the WNT3A promoter. Moreover, D2R signaling also modulates cell proliferation and modifies the pathology in a renal ischemia/reperfusion-injury disease model, via its effects on Wnt/β-catenin signaling. Together, their results suggest that D2R is a transcriptional modulator of Wnt/β-catenin signal transduction with broad implications for health and development of new therapeutics.

Dopamine is traditionally studied in the central nervous system; however, it is increasingly implicated in regulating functions of various other organs. This new study identifies a new role for dopamine signaling via the D2 receptor outside the brain—in controlling signaling through the Wnt/β-catenin pathway, in part, through its effects on expression of Wnt3a, a key Wnt receptor ligand.

Both dopamine and the Wnt/β-catenin signaling pathways are ubiquitous across organ systems and species. Wnt signaling is essential for development and cell proliferation and is associated with a number of diseases from cancer to schizophrenia. However, little is known about the underlying mechanism regulating expression of Wnt3a, or the modulation of its activity.

The researchers found that the dopamine D2 receptor is a transcriptional regulator of Wnt signaling and this ability to modulate Wnt signaling is important for better understanding development of hypertension,” said Prasad Konkalmatt, PhD, assistant research professor of medicine at the GW School of Medicine and Health Sciences and a first author on the study.

The research team focused the study on signaling in the kidneys and in the pancreas. More broadly, the study shows that dopamine receptors can act as regulators of gene transcription and that this signaling is important in controlling cell proliferation under healthy and disease conditions.

These results were unexpected, surprising the investigators by how well conserved this dopamine regulation was across species and organs. This work also showed for the first time that lithium, one of the most commonly used psychiatric medications today, strongly increases the expression of D2 receptors, providing a new mechanism of action for this drug.

The collaborative research work by George Washington University and University of Pittsburgh opens the door to a new way of thinking about dopamine signaling and its regulation. By providing a new mechanism for the actions of lithium, we can better understand how this medication works and make better medications in the future to treat bipolar disorder and to improve the lives of the millions of people living with this illness.

The researchers also discovered that a number of common gene polymorphisms associated with hypertension and renal injury control D2 receptor expression in renal cells. This discovery provides new mechanisms and drug discovery targets for hypertension and renal injury.

The findings by Prasad Konkalmatt and colleagues have broad implications in terms of how we think about dopamine receptor signaling, especially given that the receptors are targets for diabetes and potentially for hypertension and renal injury. It also expands our understanding of this unique signaling specific to individual patients offers the promise of more effective precision medicine.

Hereditary Sensory Autonomic Neuropathy type V disorder (HSAN V) is a rare type of autosomal recessive disorder that is clinically characterized by the inability to detect the pain stimuli in adult systems. As a result, patients suffering from this condition do not have the ability to exhibit protective actions against such harmful or life-threatening conditions.

This insensitivity to pain has been attributed to a point mutation of the Nerve Growth Factor (NGF) gene in patients suffering from HSAN V. The mutation impairs the processing and secretion of NGF in adult systems. NGF is a protein that plays a critical role in both developing and mature nervous system. It is an essential component of the pain sensation machinery that exerts trophic actions on cholinergic neurons of the central nervous system. This protein also facilitates the differentiation and development of receptors and functions as an effective sensitizing and inflammatory mediator during physiological and pathological situations. Considering these aforementioned facts, researchers were surprised to discover that patients suffering from HSAN V disorder do not exhibit any form of mental retardation or cognitive impairment. Previous studies have indicated that the mutation of the NGF gene contributes to the reduced secretion of mature NGF. Thus, the clinical phenotype of patients suffering from HSAN V disorder may be due to an alteration in the signaling of the Nerve Growth Factor (NGFR^100W) protein or reduced bioavailability of mature NGF protein.

To this effect, Scuola Normale Superiore di Pisa scientists: Dr. Testa Giovanna, Calvello Mariantonietta, Cattaneo Antonino and Professor Simona Capsoni   concerted efforts to understand the effect of the reduced bioavailability of mature NGF protein on the clinical phenotype of the homozygous HSAN V condition is caused by. This effect was elucidated by generating and analyzing knock-in mice that contain human NGF^R100W alleles. The authors also demonstrated that a central mechanism linked to the NGF-dependent increase of striatal cholinergic neurons contributes to the absence of pain perception in HSAN V patients.

The authors observed that most of the NGF^R100W/R100W mice that were born alive with normal size experienced severe weight loss and failed to survive beyond the first month of age. Although the treatment of these mice with wild type NGF restored the bodyweight of the mice, completely reversed their lethal phenotype, and enabled them to survive till the end of the treatment, the increase in body weight of NGF^R100W/R100W mice was less than that of the wild type mice.

Furthermore, the researchers discovered that the density of cholinergic neurons increased in the basal forebrain and striatum of NGF^R100W/R100W mice while the density of cholinergic neurons in homozygous mice was unaffected in the nucleus basalis and medial septum of Meynert.

Professor Simona Capsoni and her colleagues’ findings show that the postnatal lethal phenotype of homozygous NGF^R100W/R100W mice is caused by the systemic NGF haploinsufficiency, which is triggered by the reduced secretion of NGF^R100W protein. These findings will serve as an important point of reference for further studies on the interplay between NGF signaling and pain modulation in adult systems. The research work is now published in Biochemical and Biophysical Research Communications.

Glioblastoma multiforme (GBM) is the most common type of malignant tumor that affects the central nervous system. It is considered to be the most aggressive type of tumor due to its high proliferation rate and high mortality rates of patients with GBM. Patients suffering from this type of brain tumor often exhibit resistance to various therapeutic solutions. Despite the fact that the first-line treatment for GBM combines radiotherapy, chemotherapy and surgery, patients die within a year of initial diagnosis.

A commonly used drug for the treatment of GBM is temozolomide. Temozolomide causes cell death by undergoing methylation and inducing DNA degradation in the cell. Although temozolomide undergoes rapid absorption and irreversible conversion to its active metabolite (i.e. 5 (3-methyltriazen-1-yl) imidazole-4-carboxamide metabolite (MTIC)) after oral administration, the pharmacological efficacy of the drug is affected by its low bioavailability. The low bioavailability of temozolomide has been attributed to its low permeability through tumor cell membranes, the blood-brain barrier, and the multidrug resistance mechanism of the p-glycoprotein pump that is found in the cell membrane of GBM. Although many researchers have indicated that the ability of MTIC to interact with blood-barriers and biological membranes will contribute significantly to its pharmacological activity, studies are yet to be carried out to investigate the interaction between temozolomide and/or its active metabolite with biological membranes. Hence, this study aimed to investigate the molecular interactions between temozolomide and its active metabolite with a mimetic biomembrane model.

Recently, Maria João Ramalho, Stéphanie Andrade, Manuel Álvaro Neto Coelho, Joana Angélica Loureiro, led by Dr. Maria do Carmo Pereira from the Laboratory for Process Engineering, Environment, Biotechnology and Energy (LEPABE) in Portugal, demonstrated the molecular interactions between temozolomide and its active metabolite MTIC with the lipidic components of a mimetic biomembrane model composed of DMPC and cholesterol. The authors used various biophysical parameters to evaluate the drugs’ partition coefficient, their preferential location within the membrane and their effects on membrane properties.

Professor Maria do Carmo Pereira and her research team observed that the interactions between MTIC and the biomembrane model are mainly due to the electrostatic and ion-dipole forces between the polar head of the phospholipids and the negatively charged MTIC species, while temozolomide molecules interact through hydrophobic interactions with the acyl chains of the phospholipid in the membrane. They also observed that the drug’s partition is dependent on the composition of lipid in the biomembrane model.

Although both temozolomide and its active metabolite MTIC produced a perturbation in the fluidity of the membrane, the temozolomide exhibited a higher affinity for the membrane and penetrated the bilayer in a greater extent compared to its active metabolite MTIC. The researchers found that temozolomide significantly decreased the cooperativity of the biomembrane model.

Dr. Maria do Carmo Pereira and her research team provided compelling evidence that the composition of membranes affects drugs partition, and the bio-distribution of drugs is dependent on its physicochemical properties as well as the characteristics of the membrane. These findings will advance further studies on the design of new therapeutic molecules and development of strategies to enhance the bioavailability and pharmacological efficacy of drugs. The research work is now published in European Journal of Pharmaceutics and Biopharmaceutics.

The profound effects of glucocorticoids in immunosuppressive and anti-inflammatory therapy are due to their potent modulation of diverse physiological processes. These effects are mediated by their ability to inhibit the synthesis of proinflammatory mediators by monocytes/macrophages and induce changes in the process of phagocytosis, cell survival, and proliferation. In spite of the benefits associated with the use of exogenous glucocorticoids, their long-term use causes negative side-effects in patients. This has necessitated the investigation of the mode of action of glucocorticoids on immune cells, particularly the regulation of survival/death signaling pathways in monocytes. Thus, this study proposed a novel mechanism of action, in which glucocorticoids participate in the selective regulation of inflammatory processes.

Dr. Adrian Achuthan and colleagues at the University of Melbourne in Australia demonstrated that the addition of glucocorticoids resulted in the inhibition of ERK/12 phosphorylation, which triggers the apoptosis of monocytes treated with Granulocyte Macrophage Colony Stimulating Factor (GM-CSF). The work is published in the peer-reviewed journal, Cell Death and Disease.

The research team observed that the monocytes co-treated with GM-CSF and dexamethasone (Dex) showed increased numbers of cell death compared to monocytes treated with only GM-CSF. The pretreatment of GM-CSF-treated monocytes and GM-CSF and Dex-treated monocytes with mifepristone (a glucocorticoid receptor antagonist) resulted in the abrogation of glucocorticoid-induced apoptosis.

Their study also revealed that the presence of GM-CSF or Macrophage Colony Stimulating Factor, induced the activation of ERK1 and basal ERK2 phosphorylation while the co-treatment of dexamethasone caused a decrease in the activity of GM-CSF induced ERK1, basal ERK2 phosphorylation and reduced levels of RSK activity. The pre-treatment of co-treated monocytes with mifepristone restored the induction of ERK1 and ERK2 basal phosphorylation and resulted in an increase in RSK activities.

Furthermore, the authors observed that the induction of ERK1 activity and ERK2 basal phosphorylation did not result in the abrogation of pro-apoptotic Bad protein phosphorylation in monocytes treated with GM-CSF alone, M-CSF alone and GM-CSF and dexamethasone. However, the inhibition of ERK1 activity and ERK2 basal phosphorylation resulted in the abrogation of Bad phosphorylation in monocytes co-treated with GM-CSF and Dex. There was a strong correlation between the increase in bad phosphorylation and a decrease in apoptotic monocytes.

Adrian Achuthan and colleagues study successfully identified a novel molecular mechanism of glucocorticoids-induced apoptosis of proinflammatory monocytes via the inhibition of ERK activity. The detailed findings explicate the clinical significance of selective glucocorticoids-mediated apoptosis of proinflammatory monocytes/macrophages in the treatment of autoimmune and inflammatory diseases.

Cardiovascular disease is the leading cause of death in patients with chronic kidney disease. More than half of the patients with kidney diseases develop vascular complications at some stage of their disease progression. Hyperphosphatemia, high serum inorganic phosphate (Pi) levels, is a common biochemical abnormality in kidney patients and can exert damaging effects on endothelial cells lining the blood vessels.

Microparticles, small 100-1000nm in diameter cell membrane fragments, are released from cells under stress. While hyperphosphatemia can exert its damaging effect on cells of the vasculature by inducing soft-tissue calcification, we have proposed that Pi can exert functional effects on the cells of the blood vessels (especially endothelial cells), resulting in generation of MPs by a direct inhibition of phosphoprotein phosphatases resulting in cytoskeletal protein dysregulation and subsequent MP formation.

The mechanism of this Pi-induced cell stress and microparticle formation is elusive. In this study we have shown a novel mechanism by which Pi induces cell-stress, resulting in the generation of pro-coagulant forms of endothelial MPs which may contribute to acute occlusive events.

The major finding of our study is that inorganic phosphate concentrations similar to those observed in hyperphosphatemia in kidney patients trigger an acute release of pro-coagulant microparticles from human endothelial cells. This is of direct interest to nephrologists because it provides a molecular basis for the observed link between hyperphosphatemia and cardiovascular disease in kidney patients. It is also of interest to a wider audience because the proposed mechanism provides a widely applicable explanation for pathological effects of phosphate excess in mammalian cells. Our study provides a novel pathologic link between hyperphosphatemia, generation of MPs, and thrombotic risk.