Specific Membrane Research Articles

Palmitoylation-dependent regulation of cardiomyocyte Rac1 signaling activity and minor effects on cardiac hypertrophy

S-palmitoylation is a reversible lipid modification catalyzed by 23 S-acyltransferases with a conserved zinc finger aspartate-histidine-histidine-cysteine (zDHHC) domain that facilitates targeting of proteins to specific intracellular membranes. Here we performed a gain-of-function screen in the mouse and identified the Golgi-localized enzymes zDHHC3 and zDHHC7 as regulators of cardiac hypertrophy. Cardiomyocyte-specific transgenic mice overexpressing zDHHC3 show cardiac disease, and S-acyl proteomics identified the small GTPase Rac1 as a novel substrate of zDHHC3. Notably, cardiomyopathy and congestive heart failure in zDHHC3 transgenic mice is preceded by enhanced Rac1 S-palmitoylation, membrane localization, activity, downstream hypertrophic signaling, and concomitant induction of all Rho family small GTPases whereas mice overexpressing an enzymatically dead zDHHC3 mutant show no discernible effect. However, loss of Rac1 or other identified zDHHC3 targets Gαq/11 or galectin-1 does not diminish zDHHC3-induced cardiomyopathy, suggesting multiple effectors and pathways promoting decompensation with sustained zDHHC3 activity. Genetic deletion of Zdhhc3 in combination with Zdhhc7 reduces cardiac hypertrophy during the early response to pressure overload stimulation but not over longer time periods. Indeed, cardiac hypertrophy in response to 2weeks of angiotensin-II infusion is not diminished by Zdhhc3/7 deletion, again suggesting other S-acyltransferases or signaling mechanisms compensate to promote hypertrophic signaling. Taken together, these data indicate that the activity of zDHHC3 and zDHHC7 at the cardiomyocyte Golgi promote Rac1 signaling and maladaptive cardiac remodeling, but redundant signaling effectors compensate to maintain cardiac hypertrophy with sustained pathological stimulation in the absence of zDHHC3/7.

Design, Synthesis, Molecular Modeling, Biological Activity, and Mechanism of Action of Novel Amino Acid Derivatives of Norfloxacin.

Two series of N4-substituted piperazinyl amino acid derivatives of norfloxacin (24 new compounds) were designed and synthesized to attain structural surrogates with additional binding sites and enhanced antibacterial activity. Synthesized derivatives showed increased antibacterial and antimycobacterial activity compared to their lead structure, norfloxacin. Molecular modeling studies supported the notion that the derivatives can establish additional bonds with the target enzymes gyrase and topoisomerase IV. In vitro enzyme inhibition assays confirmed that the tested compounds were significant inhibitors of these enzymes. Inhibition of gyrase and topoisomerase IV was then confirmed in living bacterial cells using bacterial cytological profiling of both Gram-negative Escherichia coli and Gram-positive Bacillus subtilis, revealing a typical topoisomerase inhibition phenotype characterized by severe nucleoid packing defects. Several derivatives exhibited additional effects on the Gram-positive cell wall synthesis machinery and/or the cytoplasmic membrane, which likely contributed to their increased antibacterial activity. While we could not identify specific cell wall or membrane targets, membrane depolarization was not observed. Our experiments further suggest that cell wall synthesis inhibition most likely occurs outside the membrane-bound lipid II cycle.

A Review of Gap Junction Protein and its Potential Role in Nervous System-Related Disease.

Gap junction (GJ) is a special cell membrane structure composed of connexin. Connexin is widely distributed and expressed in all tissues except differentiated skeletal muscle, red blood cells, and mature sperm cells, which is related to the occurrence of many genetic diseases due to its mutation. Its function of regulating immune response, cell proliferation, migration, apoptosis, and carcinogenesis makes it a therapeutic target for a variety of diseases. In this paper, the possible mechanism of its action in nervous system-related diseases and treatment are reviewed.

Massive pre-prostatic nodal metastasis from localized aggressive prostate cancer removed during robotic-assisted radical prostatectomy with extended pelvic lymph node dissection: a case report with brief literature review

BackgroundDuring robotic-assisted radical prostatectomy (RARP) for prostate cancer (PCa), few attention is given to pre-prostatic fat tissue (PPT) even during pelvic lymph node dissection (PLND). However, the rare potential involvement of PPT lymph nodes (LN) by PCa metastasis has already been reported by several authors and may influence therapeutic strategy in intermediate and high-risk patients. We present the case of a 69-year-old man who underwent RARP with extended PLND (ePLND) for aggressive PCa with massive pre-prostatic nodal metastasis, sampled during prostate biopsies. We sought to report this case for the particular preoperative images and reinforce benefits of resecting PPT during PLND for PCa.Please confirm if the author names are presented accurately and in the correct sequence (given name, middle name/initial, family name). Author 1 Given name: [Moncef] Last name [Al Barajraji].OkCase presentationA 69-year-old man consulted our department for high serum prostate specific antigen level (57 ng/mL). He had familial history of PCa only at first degree. On digital rectal evaluation, induration of left prostatic lobe was felt. Transrectal ultrasonography showed hypoechogenic lesion in left prostatic lobe with supra-centimetric nodule in PPT. Pelvic magnetic resonance revealed two lesions in the peripheral zone with a 19-mm nodule on right paramedian side of PPT (see Fig. 1). Transrectal ultrasound-guided prostate biopsies were performed, including the nodule. On left side, 2 biopsies out 6 showed Gleason 10 prostate cancer. On right side, all biopsies showed Gleason 9 prostate cancer. The PPT nodule showed Gleason 9 prostate cancer. Prostate specific membrane antigen (PSMA) positron emission tomography computed tomography scan showed hypermetabolic expression from left prostate lesions and PPT nodule. Transperitoneal RARP with ePLND was performed including PPT. Histopathological study revealed advanced prostate cancer with lymphovascular invasion and ECE (see Fig. 2). Evaluation of ePLND material showed metastasis in on pelvic LN and 23 mm nodal metastasis in PPT (see Fig. 2). Therefore, adjuvant therapy was initiated.Please check the edit made in the article title.OPkConclusionsPPT resection is not part of routine RARP with ePLND for PCa. However, this tissue might contain LN harbouring metastasis independently from pelvic LN, indicating adjuvant therapy in case of upstaging. Considering the low morbidity of resecting PPT and its facility, it should always been resected and sent for analysis in intermediate and high-risk PCa.

EFR3A: a new raft domain organizing protein?

BackgroundMembrane rafts play a crucial role in the regulation of many important biological processes. Our previous data suggest that specific interactions of flotillins with MPP1 are responsible for membrane raft domain organization and regulation in erythroid cells. Interaction of the flotillin-based protein network with specific membrane components underlies the mechanism of raft domain formation and regulation, including in cells with low expression of MPP1.MethodsWe sought to identify other flotillin partners via the immobilized recombinant flotillin-2-based affinity approach and mass spectrometry technique. The results were further confirmed via immunoblotting and via co-immunoprecipitation. In order to study the effect of the candidate protein on the physicochemical properties of the plasma membrane, the gene was knocked down via siRNA, and fluorescence lifetime imaging microscopy and spot-variation fluorescence correlation spectroscopy was employed.ResultsEFR3A was identified as a candidate protein that interacts with flotillin-2. Moreover, this newly discovered interaction was demonstrated via overlay assay using recombinant EFR3A and flotillin-2. EFR3A is a stable component of the detergent-resistant membrane fraction of HeLa cells, and its presence was sensitive to the removal of cholesterol. While silencing the EFR3A gene, we observed decreased order of the plasma membrane of living cells or giant plasma membrane vesicles derived from knocked down cells and altered mobility of the raft probe, as indicated via fluorescence lifetime imaging microscopy and spot-variation fluorescence correlation spectroscopy. Moreover, silencing of EFR3A expression was found to disturb epidermal growth factor receptor and phospholipase C gamma phosphorylation and affect epidermal growth factor-dependent cytosolic Ca2+ concentration.ConclusionsAltogether, our results suggest hitherto unreported flotillin-2-EFR3A interaction, which might be responsible for membrane raft organization and regulation. This implies participation of this interaction in the regulation of multiple cellular processes, including those connected with cell signaling which points to the possible role in human health, in particular human cancer biology.

Efficient purification of high-salt rural groundwater in the areas along the Yellow River in Henan Province, China, by hollow fiber membrane distillation process

The vast rural drinking water sources along the Yellow River basin in Henan Province mainly come from the local high-salinity groundwater, which cannot be directly drunk. Zeolitic imidazolate framework-71/polyvinylidene fluoride hollow fiber composite membrane was applied to treat rural groundwater through membrane distillation technology. The variations of water quality parameters from 12 different rural regions after membrane distillation treatment were well analyzed and compared. Membrane separation performance for water samples from different regions during the membrane distillation process was investigated, including water flux and salt rejection. During a continuous 24-h membrane distillation process for the treatment of these water samples, hollow fiber membrane exhibits high solute retention rates (>99%) for salt ions, suspended solids and total dissolved solids, which are within the limits of national standards (GB 5749-2006), meanwhile membrane permeation water flux can remain stable. After the acid-cleaning for 180 min, all hollow fiber membranes can recover water flux well with a flux recovery rate above 98%. Even if the concentration ratio continued to increase, the water production rate of membrane distillation was basically maintained between 50% and 60%, which was obviously higher than the 30% of the reverse osmosis process. The economic analysis confirmed that the water production cost varied between US$0.45 and US$0.29/m3. With the extension of membrane service life, the specific membrane cost decreased rapidly in the first 6 years and then leveled off. The proposed vacuum membrane distillation process proved to be a promising water purification method for treating rural high-salinity groundwater.

Activate the filtration capability of 3.5 nm ultra-large pores and eliminate the requirement of pore homogeneity for carbon nanotube array membranes

Vertical array carbon nanotube (VACNT) membrane is considered as one of the most promising materials for seawater desalination due to its excellent porosity, strong mechanical properties and atomic smoothness of the inner wall. However, in practical applications, the synthesis of highly oriented VACNT arrays with uniform pore sizes (< 1.1 nm) needs to be strictly controlled, and achieving high permeability-retention for large pores (> 1.1 nm) is a great challenge. Here, we use molecular dynamics simulations to introduce the oscillation paradigm into VACNT filtration membranes with a heterogeneous pore network with a wide pore size distribution (ranging from 1 nm to 3.5 nm). Notably, the oscillating heterogeneous VACNT membrane achieved ultra-high permeability of 1421.8 L/cm2/day/MPa (~5 times speed increase in comparison with homogeneous VACNT and ~ 2 times over state-of-the-art CNT membranes), while maintaining a selectivity of 97.5 %. This method activates the filtration capacity of 3.5 nm ultra-large pores, eliminating the requirement for pore homogeneity in VACNT, and achieves the effect of “killing two birds with one stone”. In addition, the filtration performance of specific heterogeneous VACNT membrane was successfully predicted by water permeability curve fitting, which provided valuable theoretical guidance for the design of high-performance VACNT reverse osmosis desalination system.

Calcifediol: Mechanisms of Action.

Due to its essential role in calcium and phosphate homeostasis, the secosteroid hormone calcitriol has received growing attention over the last few years. Calcitriol, like other steroid hormones, may function through both genomic and non-genomic mechanisms. In the traditional function, the interaction between the biologically active form of vitamin D and the vitamin D receptor (VDR) affects the transcription of thousands of genes by binding to repeated sequences present in their promoter region, named vitamin D-responsive elements (VDREs). Non-transcriptional effects, on the other hand, occur quickly and are unaffected by inhibitors of transcription and protein synthesis. Recently, calcifediol, the immediate precursor metabolite of calcitriol, has also been shown to bind to the VDR with weaker affinity than calcitriol, thus exerting gene-regulatory properties. Moreover, calcifediol may also trigger rapid non-genomic responses through its interaction with specific membrane vitamin D receptors. Membrane-associated VDR (mVDR) and protein disulfide isomerase family A member 3 (Pdia3) are the best-studied candidates for mediating these rapid responses to vitamin D metabolites. This paper provides an overview of the calcifediol-related mechanisms of action, which may help to better understand the vitamin D endocrine system and to identify new therapeutic targets that could be important for treating diseases closely associated with vitamin D deficiency.

Rapid, Sustainable, and Versatile Strategy Towards Fabricating Superhydrophobic Cotton Textile Membranes for Separation of Emulsified and Stratified Oil/Water Mixtures

Special wettability membranes represent one of the smart, advanced, and energy-efficient solutions for the separation of contaminated oily wastewater as well as the treatment of crude oil spills. To increase sustainability and facilitate the large-scale applications of the superwetting membranes, novel renewable materials combined with rapid fabrication approaches should be explored. This study describes a simple and rapid strategy for fabricating superhydrophobic cotton textile (SCT) membranes via a solution immersion strategy from a few renewable and low-cost materials. The versatile fabrication approach was extended to fabricate other types of porous 2D/3D superhydrophobic substrates including metal mesh and melamine sponge. The fabricated SCT was employed for the gravity-driven continuous separation of immiscible oil/water mixtures with a high flux of 62500 ± 650 L. m−2. h−1 and high separation efficiency of > 99%. The SCT was employed for the demulsification of oil-in-water and water-in-oil emulsions as well as for the cleanup of crude oil spills. The fabricated SCT membranes exhibited high chemical and mechanical durability as well as excellent recyclability. Thus, the presented fabrication strategy offers a versatile, practical, and sustainable pathway toward the rapid construction of superhydrophobic porous membranes for efficient and diverse oil/water separations for the treatment of stratified and emulsified oily wastewater effluents and crude oil spills.

Rational design of a super anti-fouling PVDF-PC-TBT@X membrane for emulsion/dye wastewater separation

The exploration of membrane separation materials with special wettability has been widely focused in the field of oily wastewater treatment. However, serious membrane fouling and functional singularity are still bottlenecks in existing methods. In this paper, a proanthocyanidin (PC)-Ti composite network was constructed on the polyvinylidene fluoride (PVDF) membrane through a layer-by-layer self-assembly process. The modified membrane exhibits superhydrophilic-underwater superoleophobicity with good resistance to oil fouling. The separation flux and efficiency of the oil-in-water emulsion could reach 969.6 L·m−2h−1 and 99.2 %, respectively. After severe chemical stability, mechanical stability, and long-term exposure tests, the emulsion separation flux and efficiency still reached 841.7 L·m−2h−1 and 99 %, showing excellent stability and weatherability. In addition, the membrane can filter both oils and dyes in the water, realizing the simultaneous removal of multi-component pollutants in oily wastewater (dye removal rate exceeds 99.9 %). Through a simple desorption process, the membrane can be recycled. The preparation strategy of the special wettable membrane provided in this study has potential application in practical oily wastewater treatment.

Prostate volume analysis in image registration for prostate cancer care: a verification study

Combined magnetic resonance imaging (MRI) and positron emission tomography/computed tomography (PET/CT) may enhance diagnosis, aid surgical planning and intra-operative orientation for prostate biopsy and radical prostatectomy. Although PET-MRI may provide these benefits, PET-MRI machines are not widely available. Image fusion of Prostate specific membrane antigen PET/CT and MRI acquired separately may be a suitable clinical alternative. This study compares CT-MR registration algorithms for urological prostate cancer care. Paired whole-pelvis MR and CT scan data were used (n = 20). A manual prostate CTV contour was performed independently on each patients MR and CT image. A semi-automated rigid-, automated rigid- and automated non-rigid registration technique was applied to align the MR and CT data. Dice Similarity Index (DSI), 95% Hausdorff distance (95%HD) and average surface distance (ASD) measures were used to assess the closeness of the manual and registered contours. The automated non-rigid approach had a significantly improved performance compared to the automated rigid- and semi-automated rigid-registration, having better average scores and decreased spread for the DSI, 95%HD and ASD (all p < 0.001). Additionally, the automated rigid approach had similar significantly improved performance compared to the semi-automated rigid registration across all accuracy metrics observed (all p < 0.001). Overall, all registration techniques studied here demonstrated sufficient accuracy for exploring their clinical use. While the fully automated non-rigid registration algorithm in the present study provided the most accurate registration, the semi-automated rigid registration is a quick, feasible, and accessible method to perform image registration for prostate cancer care by urologists and radiation oncologists now.

Total-body PET/CT with half-dose [68Ga]Ga-PSMA-11 for biochemical recurrent prostate cancer: comparable diagnostic value to short axial field-of-view PET/CT with full-dose [68Ga]Ga-PSMA-11.

The objective of this study was to evaluate the diagnostic performance and image quality of total-body positron emission tomography/computed tomography (PET/CT) imaging using a half-dose of [68Ga]Ga-prostate specific membrane antigen ([68Ga]Ga-PSMA) radiotracer, compared to conventional short axial field-of-view PET/CT imaging using a full dose of [68Ga]Ga-PSMA. This retrospective study enrolled 52 patients with biochemical recurrent (BCR) prostate cancer after radical prostatectomy who underwent total-body PET/CT with a half-dose (0.9-1.1MBq/kg) of [68Ga]Ga-PSMA. These patients were matched by baseline characteristics to another 52 BCR patients after prostatectomy who underwent conventional PET/CT with a full dose (1.8-2.2MBq/kg) of [68Ga]Ga-PSMA. The half-dose group was further divided into 5-min (G5) and 2-min (G2) acquisition subgroups. Image quality was assessed through subjective analysis using a 5-point scale and objective measurements of standard uptake value maximum (SUVmax), standard uptake value mean (SUVmean), background variation (BV) of the liver, blood pool, and parotid glands. Additionally, SUVmax and tumor-to-background ratio (TBR) were calculated for lesions. No significant difference in subjective image quality was found between the G2 and full-dose groups (p > 0.05). PET/CT image quality was significantly higher for the G5 versus G2 (p < 0.001) and full-dose groups (p < 0.001). TBR did not differ between the G2 and full-dose groups (4.23 ± 5.21 vs 4.22 ± 3.97, p = 0.99). Liver BV was significantly lower for G2 versus full-dose groups (0.16 ± 0.03 vs 0.20 ± 0.05, p < 0.001). Total-body PET/CT with a half-dose [68Ga]Ga-PSMA yields image quality superior or comparable to that of conventional PET/CT. The utilization of total-body [68Ga]Ga-PSMA PET/CT meets the diagnostic demands of BCR patients, particularly those who exhibit reduced tolerance to prolonged horizontal positioning and scan durations, while simultaneously reducing radiation exposure for the subjects.

Locations of membrane protein production in a cyanobacterium.

Cyanobacteria show an unusually complex prokaryotic cell structure including a distinct intracytoplasmic membrane system, the thylakoid membranes that are the site of the photosynthetic light reactions. The thylakoid and plasma membranes have sharply distinct proteomes, but the mechanisms that target proteins to a specific membrane remain poorly understood. Here, we investigate the locations of translation of thylakoid and plasma membrane proteins in the model unicellular cyanobacterium Synechococcus elongatus PCC 7942. We use fluorescent in situ hybridization to probe the locations of mRNAs encoding membrane-integral proteins, plus Green Fluorescent Protein tagging of the RplL subunit to reveal the location of ribosomes under different conditions. We show that membrane-integral thylakoid and plasma membrane proteins are translated in different locations. Thylakoid membrane proteins are translated in patches at the innermost thylakoid membrane surface facing the nucleoid. However, different proteins are translated in different patches, even when they are subunits of the same multiprotein complex. This implies that translation is distributed over the proximal thylakoid surface, with newly inserted proteins migrating within the membrane prior to incorporation into complexes. mRNAs encoding plasma membrane proteins form patches at the plasma membrane. Ribosomes can be observed at similar locations near the thylakoid and plasma membranes, with more ribosomes near the plasma membrane when conditions force rapid production of plasma membrane proteins. There must be routes for ribosomes and mRNAs past the thylakoids to the plasma membrane. We infer a system to chaperone plasma membrane mRNAs to prevent their translation prior to arrival at the correct membrane. IMPORTANCE Cyanobacteria have a complex and distinct membrane system within the cytoplasm, the thylakoid membranes that house the photosynthetic light reactions. The thylakoid and plasma membranes contain distinct sets of proteins, but the steps that target proteins to the two membranes remain unclear. Knowledge of the protein sorting rules will be crucial for the biotechnological re-engineering of cyanobacterial cells, and for understanding the evolutionary development of the thylakoids. Here, we probe the subcellular locations of the mRNAs that encode cyanobacterial membrane proteins and the ribosomes that translate them. We show that thylakoid and plasma membrane proteins are produced at different locations, providing the first direct evidence for a sorting mechanism that operates prior to protein translation.

Bile Acid Network and Vascular Calcification-Associated Diseases: Unraveling the Intricate Connections and Therapeutic Potential.

Bile acids play a crucial role in promoting intestinal nutrient absorption and biliary cholesterol excretion, thereby protecting the liver from cholesterol accumulation and bile acid toxicity. Additionally, bile acids can bind to specific nuclear and membrane receptors to regulate energy expenditure and specific functions of particular tissues. Vascular calcification refers to the pathological process of calcium-phosphate deposition in blood vessel walls, which serves as an independent predictor for cardiovascular adverse events. In addition to aging, this pathological change is associated with aging-related diseases such as atherosclerosis, hypertension, chronic kidney disease, diabetes mellitus, and osteoporosis. Emerging evidence suggests a close association between the bile acid network and these aforementioned vascular calcification-associated conditions. Several bile acids have been proven to participate in calcium-phosphate metabolism, affecting the transdifferentiation of vascular smooth muscle cells and thus influencing vascular calcification. Targeting the bile acid network shows potential for ameliorating these diseases and their concomitant vascular calcification by regulating pathways such as energy metabolism, inflammatory response, oxidative stress, and cell differentiation. Here, we present a summary of the metabolism and functions of the bile acid network and aim to provide insights into the current research on the profound connections between the bile acid network and these vascular calcification-associated diseases, as well as the therapeutic potential.

Microalgae cell adhesions on hydrophobic membrane substrates using quartz crystal microbalance with dissipation.

Microalgal cell adhesion and biofilm formation are affected by interactions between microalgae strains and membrane materials. Variations of surface properties of microalgae and membrane materials are expected to affect cell-membranes and cell-cell interactions and thus initial microalgal cell adhesion and biofilm formation rates. Hence, it should be possible to identify the dominant mechanisms controlling microalgal cell adhesion and biofilm formation. The effects of surface properties of three different microalgal strains and three different types of membrane materials on microalgal cell adhesion and biofilm formation were systematically investigated in real time by monitoring changes in the oscillation frequency and dissipation of the quartz crystal resonator (QCM-D). The results revealed that in general a higher surface free energy, more negative zeta potential, and higher surface roughness of membrane materials positively correlated with a larger quantity of microalgae cell deposition, while a more hydrophilic microalgae with a larger negative zeta potential preferred to attach to a more hydrophobic membrane material. The adhered microalgal layers exhibited viscoelastic properties. The relative importance of these mechanisms in controlling microalgae cell attachment and biofilm formation might vary, depending on the properties of specific microalgae species and hydrophobic membrane materials used.

1787P Prostate specific membrane antigen positron emission tomography (PSMA PET)-directed clinical outcomes in metastatic hormone-sensitive prostate cancer (mHSPC): Implications for the STAMPEDE2 trial design

1787P Prostate specific membrane antigen positron emission tomography (PSMA PET)-directed clinical outcomes in metastatic hormone-sensitive prostate cancer (mHSPC): Implications for the STAMPEDE2 trial design

Analysing the tumor transcriptome of prostate cancer to predict efficacy of Lu-PSMA therapy

Rationale177Lu-PSMA ([177Lu]Lutetium-PSMA-617) therapy is an effective treatment option for patients with prostate specific membrane antigen (PSMA)-positive metastatic castration-resistant prostate cancer, but still shows a non-responder rate of approximately 30%. Combination...

Tumoral Ki67 and PSMA Expression in Fresh Pre-PSMA-RLT Biopsies and Its Relation With PSMA-PET Imaging and Outcomes of PSMA-RLT in Patients With mCRPC.

Prostate specific membrane antigen (PSMA) directed radioligand therapy (RLT) is a novel therapy for metastatic castration-resistant prostate cancer (mCRPC) patients. However, it is still poorly understood why approximately 40% of the patients does not respond to PSMA-RLT. The aims of this study were to evaluate the pretreatment PSMA expression on immunohistochemistry (IHC) and PSMA uptake on PET/CT imaging in mCRPC patients who underwent PSMA-RLT. We correlated these parameters and a cell proliferation marker (Ki67) to the therapeutic efficacy of PSMA-RLT. In this retrospective study, mCRPC patients who underwent PSMA-RLT were analyzed. Patients biopsies were scored for immunohistochemical Ki67 expression, PSMA staining intensity and percentage of cells with PSMA expression. Moreover, the PSMA tracer uptake of the tumor lesion(s) and healthy organs on PET/CT imaging was assessed. The primary outcome was to evaluate the association between histological PSMA protein expression of tumor in pre-PSMA-RLT biopsies and the PSMA uptake on PSMA PET/CT imaging of the biopsied lesion. Secondary outcomes were to assess the relationship between PSMA expression and Ki67 on IHC and the progression free survival (PFS) and overall survival (OS) following PSMA-RLT. In total, 22 mCRPC patients were included in this study. Nineteen (86%) patients showed a high and homogenous PSMA expression of >80% on IHC. Three (14%) patients had low PSMA expression on IHC. Although there was limited PSMA uptake on PET/CT imaging, these 3 patients had lower PSMA uptake on PET/CT imaging compared to the patients with high PSMA expression on IHC. Yet, no correlation was found between PSMA uptake on PET/CT imaging and PSMA expression on IHC (SUVmax: R2=0.046 and SUVavg: R2=0.036). The 3 patients had a shorter PFS compared to the patients with high PSMA expression on IHC (HR: 4.76, 95% CI: 1.14-19.99; P=.033). Patients with low Ki67 expression had a longer PFS and OS compared to patients with a high Ki67 expression (HR: 0.40, 95% CI: 0.15-1.06; P=.013) CONCLUSION: The PSMA uptake on PSMA-PET/CT generally followed the PSMA expression on IHC. However, heterogeneity may be missed on PSMA-PET/CT. Immunohistochemical PSMA and Ki67 expression in fresh tumor biopsies, may contribute to predict treatment efficacy of PSMA-RLT in mCRPC patients. This needs to be further explored in prospective cohorts.

Identification of the immunogenic membrane proteins, Catalase, PgbA, and PgbB, as potential antigens against Helicobacter pylori.

This study aims to investigate the specific membrane antigens that are targeted by antibodies raised against H. pylori. Bovine milk antibodies were prepared using whole H. pylori, purified membrane proteins, or both. Enzyme-linked immunosorbent assay (ELISA) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiments revealed that these immunogens triggered anti-H. pylori antibody production in milk. The highest antibody titer was induced by the mixture of whole bacteria and purified membrane proteins. The antibodies induced by mixed immunogens significantly inhibited H. pylori growth in vitro and were used to identify catalase, plasminogen-binding protein A (PgbA), and PgbB via western blotting, immunoprecipitation and two-dimensional western blotting (2D-WB) followed by LC-MS/MS. The immunogenicity of PgbA and PgbB was verified in mice vaccinated with their B-cell epitope vaccines. Following prophylactic vaccination of C57BL/6 mice, each of the three antigens alone and their combination reduced the weight loss in mice, increased antibody titers, and relieved the inflammatory status of the gastric mucosa following H. pylori infection. Catalase, PgbA, and PgbB could serve as valuable membrane antigens for the development of anti-H. pylori immunotherapies.

Enhancing ultrafiltration performance for dairy wastewater treatment using a 3D printed turbulence promoter

Dairy factories annually generate an increasing amount of wastewater, which can cause eutrophication due to high concentrations of amino acids and lipids. To address this issue, membrane technology has emerged as a promising solution, but membrane fouling remains a significant challenge, since it can cause decreased flux, decrease membrane rejection performance, and increased energy demand. This study aimed to reduce membrane fouling by integrated a three-dimensional printed (3DP) turbulence promoter into an ultrafiltration dead-end cell and varying stirring speeds. Two mathematical models, Hermia and resistance-in-series, were used to analyze the fouling process. According to both models, the cake layer formation model indicated the most prevalent fouling mechanism. Specific energy demand, permeate flux, membrane rejection, and membrane reversible and irreversible resistances were measured, calculated, and compared. The results suggest that the combination of an integrated 3DP turbulence promoter and high stirring speeds can effectively reduce membrane fouling in a dairy wastewater treatment module.