Exploring marine Lactobacillus and its protein for probiotic-based oral cancer therapy.

Cancer is a life-threatening malignancy and one of the leading global causes of human mortality. New approaches are required for cancer therapy due to the unique properties of cancer cells and the side effects of chemotherapy. Probiotics have gained significant attention in the prevention and treatment of various diseases, including cancer. Therefore, the current study aimed to investigate the anti-cancer effects of probiotics, such as marine Lactobacillus species and their proteins. Five marine Lactobacillus species were isolated and identified from the Tamil Nadu Mangrove Pichavaram (TLMP) forest and named TLMP1, TLMP2, TLMP3, TLMP4, and TLMP5. The Lactobacillus isolates, and their proteins were administered to male golden Syrian hamsters. Tumor formation was effectively controlled in hamsters treated with crude Lactobacillus, extending their lifespan. Additionally, Lactobacillus proteins demonstrated an inhibitory effect on tumor formation in the treated group compared to the control. Molecular docking analysis revealed that Lactobacillus proteins interacted significantly with the cAMP-dependent protein kinase catalytic subunit alpha. Amino acid residues LYS791, MET793, ARG841, ARG842, and LEU844 were involved in active site binding and played a crucial role in inhibiting cAMP-dependent protein kinase.

ASPAD dynamic simulation and artificial neural network for atenolol adsorption in GGSWAC packed bed column.

This study aimed to assess the dynamic simulation models provided by Aspen adsorption (ASPAD) and artificial neural network (ANN) in understanding the adsorption behavior of atenolol (ATN) on gasified Glyricidia sepium woodchips activated carbon (GGSWAC) within fixed bed columns for wastewater treatment. The findings demonstrated that increasing the bed height from 1 to 3 cm extended breakthrough and exhaustion times while enhancing adsorption capacity. Conversely, higher initial ATN concentrations resulted in shorter breakthrough and exhaustion times but increased adsorption capacity. Elevated influent flow rates reduced breakthrough and exhaustion times while maintaining constant adsorption capacity. The ASPAD software demonstrated competence in accurately modeling the crucial exhaustion points. However, there is room for enhancement in forecasting breakthrough times, as it exhibited deviations ranging from 6.52 to 239.53% when compared to the actual experimental data. ANN models in both MATLAB and Python demonstrated precise predictive abilities, with the Python model (R2 = 0.985) outperforming the MATLAB model (R2 = 0.9691). The Python ANN also exhibited superior fitting performance with lower MSE and MAE. The most influential factor was the initial ATN concentration (28.96%), followed by bed height (26.39%), influent flow rate (22.43%), and total effluent time (22.22%). The findings of this study offer an extensive comprehension of breakthrough patterns and enable accurate forecasts of column performance.

Physicochemical parameters and modes of interaction associated with the micelle formation of a mixture of tetradecyltrimethylammonium bromide and cefixime trihydrate: effects of hydrotropes and temperature.

The interaction between an antibiotic drug (cefixime trihydrate (CMT)) and a cationic surfactant (tetradecyltrimethylammonium bromide (TTAB)) was examined in the presence of both ionic and non-ionic hydrotropes (HTs) over the temperature range of 300.55 to 320.55 K. The values of the critical micelle concentration (CMC) of the TTAB + CMT mixture were experienced to have dwindled with an enhancement of the concentrations of resorcinol (ReSC), sodium benzoate (NaBz), sodium salicylate (NaS), while for the same system, a monotonically augmentation of CMC was observed in aq. 4-aminobenzoic acid (PABA) solution. A gradual increase in CMC, as a function of temperature, was also observed. The values of the degree of counterion binding (ß) for the TTAB + CMT mixture were experienced to be influenced by the concentrations of ReSC/NaBz/NaS/PABA and a change in temperature. The micellization process of TTAB + CMT was observed to be spontaneous (negative standard Gibbs free energy change (ΔG0m)) at all conditions studied. Also, the values of standard enthalpy change (ΔH0m) and entropy change (ΔS0m) were found negative and positive, respectively (with a few exceptions), for the test cases indicating an exothermic and enthalpy-entropy directed micellization process. The recommended interaction forces between the components in the micellar system are electrostatic and hydrophobic interactions. In this study, the values of ΔC0m were negative in aqueous NaBz, ReSC, and PABA media, and positive in case of NaS. An excellent compensation scenario between the enthalpy and entropy for the CMT + TTAB mixed system in the investigated HTs solutions is well defined in the current work.

Encapsulation of Dicranopteris linearis extract using cellulose microparticles for antiulcer medication.

Dicranopteris linearis (DL) is a fern in the Gleicheniaceae family, locally known as resam by the Malay community. It has numerous pharmacological benefits, with antiulcer and gastroprotective properties. Peptic ulcer is a chronic and recurring disease that significantly impacts morbidity and mortality, affecting nearly 20 % of the world's population. Despite the effectiveness of peptic ulcer drugs, there is no perfect treatment for the ailment. Encapsulation is an advanced technique that can treat peptic ulcers by incorporating natural sources. This work aims to encapsulate DL extract using different types of cellulose particles by the solvent displacement technique for peptic ulcer medication. The extract was encapsulated using methyl cellulose (MC), ethyl cellulose (EC), and a blend of ethyl methyl cellulose through a dialysis cellulose membrane tube and freeze-dried to yield a suspension of the encapsulated DL extracts. The microencapsulated methyl cellulose chloroform extract (MCCH) has a considerably greater level of total phenolic (84.53 ± 6.44 mg GAE/g), total flavonoid (84.53 ± 0.54 mg GAE/g), and antioxidant activity (86.40 ± 0.63 %). MCCH has the highest percentage of antimicrobial activity against Escherichia coli (2.42 ± 107 × 0.70 CFU/mL), Bacillus subtilis (5.21 ± 107 × 0.90 CFU/mL), and Shigella flexneri (1.25 ± 107 × 0.66 CFU/mL), as well as the highest urease inhibitory activity (50.0 ± 0.21 %). The MCCH particle size was estimated to be 3.347 ± 0.078 µm in diameter. It has been proven that DL elements were successfully encapsulated in the methyl cellulose polymer in the presence of calcium (Ca). Fourier transform infrared (FTIR) analysis indicated significant results, where the peak belonging to the CO stretch of the carbonyl groups of methyl cellulose (MC) shifted from 1638.46 cm-1 in the spectrum of pure MC to 1639.10 cm-1 in the spectrum of the MCCH extract. The shift in the wavenumbers was due to the interactions between the phytochemicals in the chloroform extract and the MC matrix in the microcapsules. Dissolution studies in simulated gastric fluid (SGF) and model fitting of encapsulated chloroform extracts showed that MCCH has the highest EC50 of 6.73 ± 0.27 mg/mL with R2 = 0.971 fitted by the Korsmeyer-Peppas model, indicating diffusion as the mechanism of release.

Exploring recombinant secretory proteins from Mycobacterium tuberculosis to develop a serological platform for tuberculosis diagnosis.

The lack of a sensitive diagnostic tool for tuberculosis (TB) is the main reason for increasing cause of death in many developing countries. The routine diagnostic tests are either time-consuming or equivocal in terms of results. Hence, there is a need for quicker and accurate diagnostic tests. Certain studies have documented the usage of proteins secreted by Mycobacterium tuberculosis (MTB) in developing a sensitive tool for diagnosing TB. The study aimed to employ PPE41, MPT53, LPQH, CFP10, ESAT6 and TB18.5 proteins and analyze their usage as early diagnostic markers. The proteins were cloned, expressed, purified and applied in ELISA platforms in separate as well as combined systems to assess their early diagnostic features. The results of our study revealed that a cocktail of all six antigen combinations was identified in the maximum number of TB cases. Thus, proteins such as PPE41, MPT53, LPQH, CFP10, ESAT6, and TB18.5 incorporated detection tools could be optimized for an improvised early detection of MTB infections. Moreover, the results suggested that 95.7 % of the MTB-positive serum samples reacted with all the selected antigens of Mycobacterium tuberculosis, while the control serum samples did not react with those antigens. The hexavalent antigen system yielded a novel ELISA platform for better diagnosing MTB infections. Our study yielded a novel technology to diagnose TB, which warrants testing in clinical settings.

Antibacterial, antifungal, and antioxidant competence of Cardiospermum halicacabum based nanoemulsion and characterized their physicochemical properties.

This research was designed to evaluate the pharmaceutical potentials of various proportions of nanoemulsions, Cardiospermum halicacabum Nanoemulsion A and Cardiospermum halicacabum Nanoemulsion B (CHE-NE-A & CHE-NE-B) prepared from the hydroalcoholic extract of Cardiospermum halicacabum through in vitro approach, and their physicochemical properties were characterized using standard scientific analytical techniques. The physicochemical and morphological properties of CHE-NE-A and CHE-NE-B were characterized by FTIR, SEM, TEM, zeta potential, and scattering light intensity analyses. The results revealed that the size, shape, and exterior conditions of nano-droplets of the CHE-NE-A nanoemulsion were suitable as a drug carrier. The reports obtained from in vitro drug releasing potential analysis support this as well. CHE-NE-A nanoemulsion constantly removes the drug from the dialysis bag than CHE-NE-B. Moreover, the CHE-NE-A showed considerable dose-dependent antioxidant activity on DPPH, ABTS, and FRAP free radicals. CHE-NE-A and CHE-NE-B were tested for their antibacterial activity with various bacterial strains. The results demonstrated that the CHE-NE-A nanoemulsion showed remarkable antibacterial activity (zone of inhibition) against test bacterial pathogens than CHE-NE-B. The antibacterial activity of CHE-NE-A at a concentration of 200 µg mL-1was in the following order, P. aeruginosa > S. aureus > S. typhimurium > S. pneumoniae > E. coli. Furthermore, CHE-NE-A has the lowest MIC values against these test bacterial pathogens than CHE-NE-B. Moreover, the CHE-NE-A also demonstrated good antifungal activity against the test fungal pathogens such as Cryptococcus neoformans, Aspergillus niger, Candida pneumonia, and Penicillium expansum than CHE-NE-B. These results strongly suggest that the CHE-NE-A nanoemulsion possesses considerable pharmaceutical potential. Interestingly, the physicochemical properties also rope that the CHE-NE-A nanoemulsion may be considered a drug carrier and useful for drug formulation.

Association behavior and physico-chemical parameters of a cetylpyridinium bromide and levofloxacin hemihydrate mixture in aqueous and additive media.

The investigation of the micellization of a mixture of cetylpyridinium bromide (CPB) and levofloxacin hemihydrate (LFH) was carried out by a conductivity technique in aqueous and aq. additive mixtures, including NaCl, NaOAc, NaBenz, 4-ABA, and urea. The aggregation behavior of the CPB + LFH mixture was studied considering the variation in additive contents and the change in experimental temperature. The micelle formation of the CPB + LFH mixture was examined from the breakpoint observed in the specific conductivity versus surfactant concentration plots. Different micellar characteristics, such as the critical micelle concentration (CMC) and the extent of counter ion bound (ß), were evaluated for the CPB + LFH mixture. The CMC and ß were found to undergo a change with the types of solvents, composition of solvents, and working temperatures. The ΔG0m values of the CPB + LFH system in aqueous and aq. additive solutions were found to be negative, which denotes a spontaneous aggregation phenomenon of the CPB + LFH system. The changes in ΔH0m and ΔS0m for the CPB + LFH mixture were also detected with the alteration in the solvent nature and solution temperature. The ΔH0m and ΔS0m values obtained for the association of the CPB + LFH mixture reveal that the characteristic interaction forces may possibly be ion-dipole, dipole-dipole, and hydrophobic between CPB and LFH. The thermodynamics of transfer and ΔH0m-ΔS0m compensation variables were also determined. All the parameters computed in the present investigation are illustrated with proper logic.

Radiation nephropathy: Mechanisms of injury and recovery in a murine model.

BACKGROUND: Radiation nephropathy (RN) can be a severe late complication for patients treated with radiotherapy (RT) targeting abdominal and paraspinal tumors. Recent studies investigating the mechanisms of RT-mediated injury in the kidney have demonstrated that RT disrupts the cellular integrity of renal podocytes leading to cell death and loss of renal function. AIM: To determine if RT-induced renal dysfunction is associated with alterations in podocyte and glomerular function, and whether RT-induced podocyte alterations were associated with changes in the glomerular basement membrane (GBM). METHODS: C57BL/6 mice were treated with focal bilateral X-irradiation using a single dose (SD) of 4 Gy, 10 Gy, or 14 Gy or fractionated dosing (FD) of 5x6Gy or 24x2Gy. Then, 10-40 weeks after RT parameters of renal function were measured, along with glomerular filtration rate (GFR) and glomerular histology, as well as ultrastructural changes in GBM by transmission electron microscopy. RESULTS: RT treatment resulted in persistent changes in renal function beginning at 10 weeks with little recovery up to 40 weeks post RT. Dose dependent changes were seen with increasing SD but no functional sparing was evident after FD. RT-induced loss of renal function was associated with expansion of the GBM and significant increases in foot process width, and associated with significant reduction in GFR, podocyte loss, and renal fibrosis. CONCLUSION: For the first time, these data show that expansion of the GBM is one consequence of radiation injury, and disarrangement of the GBM might be associated with the death of podocytes. These data shed new light on the role podocyte injury and GBM in RT-induced renal dysfunction.

HPTLC and GC-MS finger-printing of two potential multifunctional siddha tailams: Mathan and maha megarajanga tailam.

The Siddha system of medicine is an ancient medical lineage that is practiced primarily in the southern part of India. Siddha system of medicine has been in practice for thousands of years with documented evidence dating back to the 6th century BCE. According to siddha system of medicine's basic fundamental principle, the human body is made up of 96 thathuvam (primary components), which encompass physical, physiological, psychological, and intellectual aspects. Medicine (marunthu) is classified as a wide range of internal and external medicines. The major components of its medical formulations include plant parts, minerals and animal products. Various methods were carried out for the purification process to eliminate the toxins. Choornam, Guligai, Tailam, Parpam, Chendooram, Kattu, Pasai and Poochu are the most common medicines used in Siddha system of medicine for the treatment of various diseases. The pathophysiological classification of diseases is elaborated in detail in the classical Siddha literature. Siddha system of medicine plays an important role in protecting people from diseases such as COVID-19 by providing immune-protecting and immune-boosting medicines in today's world. Mathan tailam and maha megarajanga tailam are the two unique preparations used widely for various skin diseases including chronic wounds and burns. Scientific validation of both medicines will help in understanding their effectiveness against a typical wound condition. In the present study physio-chemical and phytochemical, HPTLC, and GC-MS analyses were carried out and discussed in detail on the multifunctional properties exhibited in the patient communities.

Picloram binds to the h1 and h4 helices of HSA domain IIIA at drug binding site 2.

Picloram (PC) is a systemic herbicide that controls herbaceous weeds and woody plants. HSA, the most abundant protein in human physiology, binds to all exogenic and endogenic ligands. PC is a stable molecule (t1/2∼157-513 days) and a potential threat to human health via the food chain. HSA and PC binding study has been done to decipher the location and thermodynamics of binding. It has been studied with prediction tools like autodocking and MD simulation and then confirmed with fluorescence spectroscopy. HSA fluorescence was quenched by PC at pH 7.4 (N state), pH 3.5 (F state), and pH 7.4 with 4.5 M urea (I state) at temperatures 283 K, 297 K, and 303 K. The location of binding was found to be interdomain between II and III which overlaps with drug binding site 2. The binding was spontaneous, and entropy-driven that show a noticeable increase in binding with the increase in temperature. No secondary structure change at the native state has been observed due to binding. The binding results are important to understand the physiological assimilation of PC. In silico predictions and the results of spectroscopic studies unambiguously indicate the locus and nature of the binding.

Empagliflozin reduces podocyte lipotoxicity in experimental Alport syndrome.

Sodium-glucose cotransporter-2 inhibitors (SGLT2i) are anti-hyperglycemic agents that prevent glucose reabsorption in proximal tubular cells. SGLT2i improves renal outcomes in both diabetic and non-diabetic patients, indicating it may have beneficial effects beyond glycemic control. Here, we demonstrate that SGLT2i affects energy metabolism and podocyte lipotoxicity in experimental Alport syndrome (AS). In vitro, we found that the SGLT2 protein was expressed in human and mouse podocytes to a similar extent in tubular cells. Newly established immortalized podocytes from Col4a3 knockout mice (AS podocytes) accumulate lipid droplets along with increased apoptosis when compared to wild-type podocytes. Treatment with SGLT2i empagliflozin reduces lipid droplet accumulation and apoptosis in AS podocytes. Empagliflozin inhibits the utilization of glucose/pyruvate as a metabolic substrate in AS podocytes but not in AS tubular cells. In vivo, we demonstrate that empagliflozin reduces albuminuria and prolongs the survival of AS mice. Empagliflozin-treated AS mice show decreased serum blood urea nitrogen and creatinine levels in association with reduced triglyceride and cholesterol ester content in kidney cortices when compared to AS mice. Lipid accumulation in kidney cortices correlates with a decline in renal function. In summary, empagliflozin reduces podocyte lipotoxicity and improves kidney function in experimental AS in association with the energy substrates switch from glucose to fatty acids in podocytes.

Quantitative assessment of platelet rich fibrin for the repair of extraction socket in a rat model.

The present study evaluated the quantitative effects of platelet-rich fibrin (PRF) for the repair of extraction socket in Sprague Dawley (SD) rat model by assessing several key clinical parameters. Seventy two male SD rats were subjected to surgical extraction of the maxillary right incisor. Rats were randomly divided into four groups with eighteen rats in each group based on the treatment received: extraction socket without treatment of PRF was taken as control (group I). Extraction socket implanted with 0.1, 0.2, and 0.4 mL of PRF was taken as study groups (groups II, III, and IV). The obtained results demonstrated that, low dose of PRF efficiently enhanced the natural healing cascade. Whereas, high dose interfered with native tissue contribution and altered the natural healing process. The beneficial effects of quantity-based application of PRF may raise the possibility of a new approach as complementary therapy besides conventional treatment.

SMPDL3b modulates radiation-induced DNA damage response in renal podocytes.

The kidneys are radiosensitive and dose-limiting organs for radiotherapy (RT) targeting abdominal and paraspinal tumors. Excessive radiation doses to the kidneys ultimately lead to radiation nephropathy. Our prior work unmasked a novel role for the lipid-modifying enzyme, sphingomyelin phosphodiesterase acid-like 3b (SMPDL3b), in regulating the response of renal podocytes to radiation injury. In this study, we investigated the role of SMPDL3b in DNA double-strand breaks (DSBs) repair in vitro and in vivo. We assessed the kinetics of DSBs recognition and repair along with the ATM pathway and nuclear sphingolipid metabolism in wild-type (WT) and SMPDL3b overexpressing (OE) human podocytes. We also assessed the extent of DNA damage repair in SMPDL3b knock-down (KD) human podocytes, and C57BL6 WT and podocyte-specific SMPDL3b-knock out (KO) mice after radiation injury. We found that SMPDL3b overexpression enhanced DSBs recognition and repair through modulating ATM nuclear shuttling. OE podocytes were protected against radiation-induced apoptosis by increasing the phosphorylation of p53 at serine 15 and attenuating subsequent caspase-3 cleavage. SMPDL3b overexpression prevented radiation-induced alterations in nuclear ceramide-1-phosphate (C1P) and ceramide levels. Interestingly, exogenous C1P pretreatment radiosensitized OE podocytes by delaying ATM nuclear foci formation and DSBs repair. On the other hand, SMPDL3b knock-down, in vitro and in vivo, induced a significant delay in DSBs repair. Additionally, increased activation of apoptosis was induced in podocytes of SMPDL3b-KO mice compared to WT mice at 24 h post-irradiation. Together, our results unravel a novel role for SMPDL3b in radiation-induced DNA damage response. The current work suggests that SMPDL3b modulates nuclear sphingolipid metabolism, ATM nuclear shuttling, and DSBs repair.

Physico-chemical parameters of phase separation of the mixture of BSA and triton X 100 in aqueous solutions of monohydroxy compounds.

Clouding behavior and thermodynamic properties for the TX 100 + BSA mixture were investigated in aqueous and aq. alcoholic media. In an aqueous environment, the values of cloud point (CP), at which a clear solution becomes cloudy, for TX 100 decreases with augmentation of the concentration of BSA. The reverse result was obtained in the aq. alcoholic media. In this study, we have used ethanol (EtOH), 1-propanol (1-PrOH), and 2-butanol (2-BuOH) as alcohols. The changes of CP values in alcoholic media have been obtained in the following order: CPH2O+EtOH > CPH2O+2-BuOH > CPH2O+1-PrOH. The standard free energy (∆Gco), standard enthalpy (∆Hco), and standard entropy (∆Sco) changes of clouding were derived at CP. The ΔGc0 values of TX 100 + BSA decreases in the aqueous and alcoholic media with increasing the concentration of BSA and alcohol. This process becomes endothermic in the aq. alcoholic media. Different thermodynamic properties of transfer and entropy-enthalpy compensation parameters for the phase partitioning of the TX 100 + BSA mixture have been calculated and assessed properly.

Mechanism of Multi-Organ Injury in Experimental COVID-19 and Its Inhibition by a Small Molecule Peptide.

Severe disease from SARS-CoV-2 infection often progresses to multi-organ failure and results in an increased mortality rate amongst these patients. However, underlying mechanisms of SARS- CoV-2-induced multi-organ failure and subsequent death are still largely unknown. Cytokine storm, increased levels of inflammatory mediators, endothelial dysfunction, coagulation abnormalities, and infiltration of inflammatory cells into the organs contribute to the pathogenesis of COVID-19. One potential consequence of immune/inflammatory events is the acute progression of generalized edema, which may lead to death. We, therefore, examined the involvement of water channels in the development of edema in multiple organs and their contribution to organ dysfunction in a Murine Hepatitis Virus-1 (MHV-1) mouse model of COVID-19. Using this model, we recently reported multi-organ pathological abnormalities and animal death similar to that reported in humans with SARS-CoV-2 infection. We now identified an alteration in protein levels of AQPs 1, 4, 5, and 8 and associated oxidative stress, along with various degrees of tissue edema in multiple organs, which correlate well with animal survival post-MHV-1 infection. Furthermore, our newly created drug (a 15 amino acid synthetic peptide, known as SPIKENET) that was designed to prevent the binding of spike glycoproteins with their receptor(s), angiotensin- converting enzyme 2 (ACE2), and carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) (SARS-CoV-2 and MHV-1, respectively), ameliorated animal death and reversed altered levels of AQPs and oxidative stress post-MHV-1 infection. Collectively, our findings suggest the possible involvement of altered aquaporins and the subsequent edema, likely mediated by the virus-induced inflammatory and oxidative stress response, in the pathogenesis of COVID- 19 and the potential of SPIKENET as a therapeutic option.

Compounds targeting OSBPL7 increase ABCA1-dependent cholesterol efflux preserving kidney function in two models of kidney disease.

Impaired cellular cholesterol efflux is a key factor in the progression of renal, cardiovascular, and autoimmune diseases. Here we describe a class of 5-arylnicotinamide compounds, identified through phenotypic drug discovery, that upregulate ABCA1-dependent cholesterol efflux by targeting Oxysterol Binding Protein Like 7 (OSBPL7). OSBPL7 was identified as the molecular target of these compounds through a chemical biology approach, employing a photoactivatable 5-arylnicotinamide derivative in a cellular cross-linking/immunoprecipitation assay. Further evaluation of two compounds (Cpd A and Cpd G) showed that they induced ABCA1 and cholesterol efflux from podocytes in vitro and normalized proteinuria and prevented renal function decline in mouse models of proteinuric kidney disease: Adriamycin-induced nephropathy and Alport Syndrome. In conclusion, we show that small molecule drugs targeting OSBPL7 reveal an alternative mechanism to upregulate ABCA1, and may represent a promising new therapeutic strategy for the treatment of renal diseases and other disorders of cellular cholesterol homeostasis.

Noninvasive assessment of radiation-induced renal injury in mice.

PURPOSE: The kidney is a radiosensitive late-responding normal tissue. Injury is characterized by radiation nephropathy and decline of glomerular filtration rate (GFR). The current study aimed to compare two rapid and cost-effective methodologies of assessing GFR against more conventional biomarker measurements. METHODS: C57BL/6 mice were treated with bilateral focal X-irradiation (1x14Gy or 5x6Gy). Functional measurements of kidney injury were assessed 20 weeks post-treatment. GFR was estimated using a transcutaneous measurement of fluorescein-isothiocyanate conjugated (FITC)-sinistrin renal excretion and also dynamic contrast-enhanced CT imaging with a contrast agent (ISOVUE-300 Iopamidol). RESULTS: Hematoxylin and eosin (H&E) and Periodic acid-Schiff staining identified comparable radiation-induced glomerular atrophy and mesangial matrix accumulation after both radiation schedules, respectively, although the fractionated regimen resulted in less diffuse tubulointerstitial fibrosis. Albumin-to-creatinine ratios (ACR) increased after irradiation (1x14Gy: 100.4 ± 12.2 µg/mg; 6x5Gy: 80.4 ± 3.02 µg/mg) and were double that of nontreated controls (44.9 ± 3.64 µg/mg). GFR defined by both techniques was negatively correlated with BUN, mesangial expansion score, and serum creatinine. The FITC-sinistrin transcutaneous method was more rapid and can be used to assess GFR in conscious animals, dynamic contrast-enhanced CT imaging technique was equally safe and effective. CONCLUSION: This study demonstrated that GFR measured by dynamic contrast-enhanced CT imaging is safe and effective compared to transcutaneous methodology to estimate kidney function.

Discoidin domain receptor 1 activation links extracellular matrix to podocyte lipotoxicity in Alport syndrome.

BACKGROUND: Discoidin domain receptor 1 (DDR1) is a receptor tyrosine kinase that is activated by collagens that is involved in the pathogenesis of fibrotic disorders. Interestingly, de novo production of the collagen type I (Col I) has been observed in Col4a3 knockout mice, a mouse model of Alport Syndrome (AS mice). Deletion of the DDR1 in AS mice was shown to improve survival and renal function. However, the mechanisms driving DDR1-dependent fibrosis remain largely unknown. METHODS: Podocyte pDDR1 levels, Collagen and cluster of differentiation 36 (CD36) expression was analyzed by Real-time PCR and Western blot. Lipid droplet accumulation and content was determined using Bodipy staining and enzymatic analysis. CD36 and DDR1 interaction was determined by co-immunoprecipitation. Creatinine, BUN, albuminuria, lipid content, and histological and morphological assessment of kidneys harvested from AS mice treated with Ezetimibe and/or Ramipril or vehicle was performed. FINDINGS: We demonstrate that Col I-mediated DDR1 activation induces CD36-mediated podocyte lipotoxic injury. We show that Ezetimibe interferes with the CD36/DDR1 interaction in vitro and prevents lipotoxicity in AS mice thus preserving renal function similarly to ramipril. INTERPRETATION: Our study suggests that Col I/DDR1-mediated lipotoxicity contributes to renal failure in AS and that targeting this pathway may represent a new therapeutic strategy for patients with AS and with chronic kidney diseases (CKD) associated with Col4 mutations. FUNDING: This study is supported by the NIH grants R01DK117599, R01DK104753, R01CA227493, U54DK083912, UM1DK100846, U01DK116101, UL1TR000460 (Miami Clinical Translational Science Institute, National Center for Advancing Translational Sciences and the National Institute on Minority Health and Health Disparities), F32DK115109, Hoffmann-La Roche and Alport Syndrome Foundation.

A Multimodality Image Guided Precision Radiation Research Platform: Integrating X-ray, Bioluminescence, and Fluorescence Tomography With Radiation Therapy.

PURPOSE: Small animal irradiation is crucial to the investigation of radiobiological mechanisms. The paradigm of clinical radiation therapy is trending toward high-precision, stereotactic treatment. However, translating this scheme to small animal irradiation is challenging owing to the lack of high-quality image guidance. To overcome this obstacle, we developed a multimodality image guided precision radiation platform. METHODS AND MATERIALS: The platform consists of 4 modules: x-ray computed tomography (CT), bioluminescence tomography (BLT), fluorescence molecular tomography (FMT), and radiation therapy. CT provides animal anatomy and material density for radiation dose calculation, as well as body contour for BLT and FMT reconstruction. BLT and FMT provide tumor localization to guide radiation beams and molecular activity to evaluate treatment outcome. Furthermore, we developed a Monte Carlo-based treatment planning system (TPS) for 3-dimensional dose calculation, calibrated it using radiochromic films sandwiched in a water-equivalent phantom, and validated it using in vivo dosimeters surgically implanted into euthanized mice (n = 4). Finally, we performed image guided irradiation on mice bearing orthotopic breast and prostate tumors and confirmed radiation delivery using γH2AX histology. RESULTS: The Monte Carlo-based TPS was successfully calibrated by benchmarking simulation dose against film measurement. For in vivo dosimetry measured in the euthanized mice, the average difference between the TPS calculated dose and measured dose was 3.86% ± 1.12%. Following the TPS-generated treatment plan, we successfully delivered 20 Gy dose to an animal bearing an orthotopic prostate tumor using 4 BLT-guided radiation beams and 5 Gy dose to an animal bearing an orthotopic breast tumor using a single FMT-guided radiation beam. γH2AX histology presented significantly more DNA damage in irradiated tumors and thus validated the dose delivery accuracy. CONCLUSIONS: Combined with Monte Carlo TPS, this multimodality CT/BLT/FMT image guided small animal radiation platform can specifically localize tumors, accurately calculate dose distribution, precisely guide radiation delivery, and molecularly evaluate treatment response. It provides an advanced toolset for radiobiology and translational cancer research.