Kidney stone nano-structure — Is there an opportunity for nanomedicine development?

BackgroundKidney stone analysis techniques are well-established in the field of materials characterization and provide information for the chemical composition and structure of a sample. Nanomedicine, on the other hand, is a field with an increasing rate of scientific research, a big budget and increasingly developing market. The key scientific question is if there is a possibility for the development of a nanomedicine to treat kidney stones.Major conclusionsThe main calculi characterization techniques such as X-ray Diffraction and Fourier Transform Infrared Spectroscopy can provide information about the composition of a kidney stone but not for its nanostructure. On the other hand, Small Angle X-ray Scattering and Nitrogen Porosimetry can show the nanostructural parameters of the calculi. The combination of the previously described parameters can be used for the development of nano-drugs for the treatment of urolithiasis, while no such nano-drugs exist yet.General significanceIn this study, we focus on the most well-known techniques for kidney stone analysis, the urolithiasis management and the search for possible nanomedicine for the treatment of kidney stone disease. We combine the results from five different analysis techniques in order to represent a three dimensional model and we propose a hypothetical nano-drug with gold nanoparticles. This article is part of a Special Issue entitled "Recent Advances in Bionanomaterials" Guest Editor: Dr. Marie-Louise Saboungi and Dr. Samuel D. Bader.     

Struvite calculus in the vagina of a bottlenose dolphin (Tursiops truncatus)

On 27 January 2000, a struvite calculus was observed in the vagina during necropsy of a 138-cm-long female bottlenose dolphin (Tursiops truncatus) collected from the Stono River, South Carolina (USA). Vaginal calculi have been reported in other species of cetaceans but not in bottlenose dolphins. Urinary tract infection might have been an underlying cause of the calculus. While urinary tract inflammation was not detected by light microscopic evaluation of sections of the urinary tract, it is conceivable that sufficient time had lapsed following voiding of the calculus through the urethra for urinary tract infection to have resolved. To further define the prevalence and significance of urolithiasis, prosectors of dead stranded marine mammals are encouraged to closely observe their urinary and genital tracts for calculi and to submit them for quantitative analysis.     

A Novel Calcium Oxalate Crystal Growth Inhibitory Protein from the Seeds of Dolichos biflorus (L.)

Recurrence and persistent side effects of present day treatment for urolithiasis restrict their use, so an alternate, using phytotherapy is being sought. Dolichos biflorus seeds, which are used as dietary food in India, possess antilithiatic properties. In the present study, a novel dimeric antilithiatic protein (98 kDa) from its seeds was purified based on its ability to inhibit calcium oxalate crystallization in vitro. Amino acid analysis of Dolichos biflorus antilithiatic protein showed abundant acidic amino acids. The mascot search engine presented sequence similarity with a calcium binding protein, calnexin of Pisum sativum from the m/z data obtained by MALDI TOF mass spectrometer. Above results demonstrate the anticalcifying/antilithiatic nature of a novel protein from the seeds of Dolichos biflorus and thus open new vistas for using plant proteins as therapeutic agents to treat urolithiasis.     

Physical, X-ray diffraction and scanning electron microscopic studies of uroliths

Identification of chemical constituents of calculus is important in the diagnosis and management of urolithiasis. The compositional variability of uroliths has different etiologies and requires various modes of treatment and prophylaxis. In the present study, we report the chemical compositional analyses of calculi recovered from buck and bullock by X-ray diffraction (XRD) and energy dispersive X-ray (EDX) techniques and ultra-structure examination by scanning electron microscopy (SEM). XRD and EDX investigations conclusively established the chemical compositions of urinary calculi under investigation. The calculus from buck (sample I) had calcium oxalate monohydrate, a dominant salt phase and magnesium compound in significant amount. The calculus from bullock (sample II) had magnesium ammonium phosphate phase, with significant amount of calcium in apatite form and K+ ions. SEM study at higher magnification (X 1000) showed bipyramidal crystals in external zones of urolith (sample I). The struvite apatite calculus showed that basic unit of structure was lamination and the laminitis appeared to be made up of fine granules and high porosity. The bio-mineralization process of calculus formation was also studied, with a view to take preventive and therapeutic measures for amelioration of urinary stone diseases in animals and humans.     

Hydrochlorothiazide, allopurinol and magnesium oxide in the treatment of recurrent calcium urolithiasis]

The study aimed at presenting own experience in prevention of new urinary calculi in 18 patients with metabolically active calcium urolithiasis treated with hydrochlorothiazide in daily doses of 100 mg (group I) for 2 years, and in 6 patients with the same disease treated with magnesium oxide in daily doses 300 mg twice a day (group II) for average period of 10 months. In 9 patients a new calculus was formed during the treatment with hydrochlorothiazide, in 7 patients no recurrence was noted, and in 2 remaining patients the results were controversial (coral calculus). Therefore, patients were subdivided into group Ia (failure of hydrochlorothiazide therapy), and group Ib (no recurrence noted). Hydrochlorothiazide did not lead to the stable decrease in the saturation of the urine with calcium oxalate in group Ia whereas in group Ib (without recurrence of urolithiasis) the content of calcium oxalate in the urine was significantly lower than that in group Ia after a 2-year treatment with hydrochlorothiazide (p < 0.01) Recurrence of the disease was seen only in one patient of group II, i.e. treated with magnesium oxide. The treatment of the recurrent calcium urolithiasis is justified and efficient in those patients in whom therapy decreases the content of calcium oxalate in the urine.     

泌尿系结石的流行病学研究进展

尿路结石的发病率是由地理区域、种族分布、社会经济地位以及饮食习惯所决定的。在过去几十年间结石形成危险因素中的饮食习惯改变,不仅影响了结石的发病率,而且对结石的部位和化学成份的改变也产生了很大影响,但草酸钙结石的发病仍最为普遍。大量流行病学调查,阐明了结石发病机制中营养因素的重要性,并建议特定的饮食可作为结石预防和治疗最为适用的方法之一。     

Characterization and In Vitro Drug Release Studies of a Natural Polysaccharide Terminalia catappa Gum (Badam Gum)

The main objective of the present study is the physicochemical characterization of naturally available Terminalia catappa gum (Badam gum [BG]) as a novel pharmaceutical excipient and its suitability in the development of gastroretentive floating drug delivery systems (GRFDDS) to retard the drug for 12 h when the dosage form is exposed to gastrointestinal fluids in the gastric environment. As BG was being explored for the first time for its pharmaceutical application, physicochemical, microbiological, rheological, and stability studies were carried out on this gum. In the present investigation, the physicochemical properties, such as micromeritic, rheological, melting point, moisture content, pH, swelling index, water absorption, and volatile acidity, were evaluated. The gum was characterized by scanning electron microscopy, differential scanning calorimetry (DSC), powder X-ray diffraction studies (PXRD), and Fourier transform infrared spectroscopy (FTIR). Gastroretentive floating tablets of BG were prepared with the model drug propranolol HCl by direct compression methods. The prepared tablets were evaluated for all their physicochemical properties, in vitro buoyancy, in vitro drug release, and rate order kinetics. PBG 04 was selected as an optimized formulation based on its 12-h drug release and good buoyancy characteristics. The optimized formulation was characterized with FTIR, DSC, and PXRD studies, and no interaction between the drug and BG was found. Thus, the study confirmed that BG might be used in the gastroretentive drug delivery system as a release-retarding polymer.KEY WORDS: badam gum, floating, gastroretentive, propranolol HCl, Terminalia catappa     

Melanin from the Nitrogen-Fixing Bacterium Azotobacter chroococcum: A Spectroscopic Characterization

Melanins, the ubiquitous hetero-polymer pigments found widely dispersed among various life forms, are usually dark brown/black in colour. Although melanins have variety of biological functions, including protection against ultraviolet radiation of sunlight and are used in medicine, cosmetics, extraction of melanin from the animal and plant kingdoms is not an easy task. Using complementary physicochemical techniques (i.e. MALDI-TOF, FTIR absorption and cross-polarization magic angle spinning solid-state ¹³C NMR), we report here the characterization of melanins extracted from the nitrogen-fixing non-virulent bacterium Azotobacter chroococcum, a safe viable source. Moreover, considering dihydroxyindole moiety as the main constituent, an effort is made to propose the putative molecular structure of the melanin hetero-polymer extracted from the bacterium. Characterization of the melanin obtained from Azotobacter chroococcum would provide an inspiration in extending research activities on these hetero-polymers and their use as protective agent against UV radiation.     

Copper-sulfur complexes supported by N-donor ligands: Towards models of the CuZ site in nitrous oxide reductase

The distinctive structure of the [(his)₇Cu₄(μ-S)]ⁿ⁺ cluster in the “Cu_Z” active site of nitrous oxide reductase and the intriguing mechanistic hypotheses for its catalytic reactivity provide inspiration for synthetic model studies aimed at characterizing relevant copper-sulfur compounds and obtaining fundamental insights into structure and bonding. In this brief review, we summarize such studies that have focused on the synthesis and characterization of a range of copper-sulfur complexes supported by N-donor ligands. Compounds with variable nuclearities and sulfur redox levels have been isolated, with the nature of the species obtained being dependent on the supporting ligand, sulfur source, and the reaction conditions. Spectroscopic data and theoretical calculations, often performed with a view toward drawing comparisons to oxygen analogs, have provided insight into the nature of the copper-sulfur bonding interactions in the complexes.     

Asian small-clawed otter (Aonyx cinerea) urolithiasis prevalence in North America

The prevalence of urolithiasis in the North American Asian small-clawed otter (Aonyx cinerea) population was determined through a retrospective survey. A questionnaire regarding diet and radiographic or necropsy evidence of urolithiasis was sent to the 16 North American institutions or individuals listed in the Asian Small-Clawed Otter North American Regional Studbook. Completed forms were returned by 75% of questionnaire recipients. Individual surveys or necropsy reports were received for 79.8% of the living or dead animals listed as comprising the North American population. Renal calculi were detected in 66.1%, and cystic calculi in 23.2%, of the captive adult population that had been radiographed or necropsied. All otters with cystic calculi also had renal calculi. Bilateral renal calculi occurred in 83.8% of affected otters. Both males (61.2%) and females (72%) were frequently affected. The prevalence of urolithiasis in wild-born otters was 76.7%, and in adult (≥1 year) captive-born otters it was 53.8%. The higher prevalence in wild-born otters may be a reflection of the older mean age at which they were first evaluated. In the overwhelming majority of otters with renal calculi, the calculi were multiple and diffusely distributed throughout the renal parenchyma. Renal and cystic calculi analyzed were primarily composed of calcium oxalate or urates. Glucosuria was infrequently reported. Necropsies of dependent neonates (≤ 2 months old) that died revealed cystic calculi in one animal. Renal disease was the cause of, or a contributing factor in, the deaths of all older animals whose necropsy reports were reviewed. The captive diet may be a contributing factor to urolith formation and progression.     

Structure of a New Pyrimidine from <Emphasis Type="Italic">Bacillus subtilis</Emphasis> Phage SP-15 Nucleic Acid

THE unusual modified base¹ of the nucleic acid of the large, generalized transducing phage of Bacillus subtilis SP-15 was predicted from the low denaturation temperature of the native nucleic acid. The base was detected as a fifth ultraviolet-absorbing component once the nucleic acid had been hydrolysed with formic acid; it partially replaces the thymine and represents 12 mol per cent of all the bases¹. It is responsible for the low Tm of the nucleic acid, since once all other unusual components have been removed, the remaining material melts at the same temperature as the native nucleic acid. We report here the isolation and characterization of this new pyrimidine.     

A Preformulation Study of a New Medicine for Chagas Disease Treatment: Physicochemical Characterization,Thermal Stability,and Compatibility of Benznidazole

This work aimed the studies of physicochemical characterization, thermal stability, and compatibility of benznidazole (BNZ) drug by spectroscopy (NMR, IR), thermoanalytical (differential thermal analysis, differential scanning calorimetry, and thermogravimetry), and chromatographic (HPLC) techniques, beyond the analytical tools of Van’t Hoff equation and Ozawa model. The compatibility study was conducted by binary mixtures (1:1, w/w) of the drug with microcrystalline cellulose 102 and 250, anhydrous lactose, and sodium starch glycolate. The physicochemical characterization confirmed data reported in scientific literature, guaranteeing authenticity of the analyzed raw material. The drug melts at 191.68°C (∆H, 119.71 J g⁻¹), characteristic of a non-polymorphic raw material, and a main stage decomposition at 233.76–319.35°C (∆m, 43.32%) occurred, ending the study with almost all mass volatilized. The quantification of drug purity demonstrated a correlation of 99.63% between the data obtained by chromatographic (99.20%) and thermoanalytical technique (99.56%). The Arrhenius equation and Ozawa model showed a zero-order kinetic behavior for the drug decomposition, and a calculated provisional validity time was 2.37 years at 25°C. The compatibility study evidenced two possible chemical incompatibilities between BNZ and the tested excipients, both associated by the authors to the reaction of the BNZ’s amine and a polymer carbohydrate’s carbonile, being maillard reactions. The BNZ reaction with anhydrous lactose is more pronounced than with the sodium starch glycolate because the lactose has more free hydroxyl groups to undergo reduction by the drug. In this sense, this work guides the development of a new solid pharmaceutical product for Chagas disease treatment, with defined quality control parameters and physicochemical stability.     

Structural investigation of the capsular polysaccharide of klebsiella serotype K26

The structure of the capsular polysaccharide from Klebsiella K26 has been determined by using the techniques of methylation, periodate oxidation, partial hydrolysis, and β-elimination. N.m.r. spectroscopy (¹H and ¹³C) was used to establish the nature of the anomeric linkages and to identify oligosaccharides obtained by the different degradative techniques employed.The polysaccharide is comprised of repeating units of the heptasaccharide shown.

     

Solid-State Interactions at the Core-Coat Interface: Physicochemical Characterization of Enteric-Coated Omeprazole Pellets Without a Protective Sub-Coat

Conventionally, scanning electron or transmission microscopy, Raman and near infrared (NIR) spectroscopy, terahertz, florescence, and nuclear magnetic resonance imaging have been used to characterize functional coating structure. This study highlights the use of fluorescence microscopy to investigate the physicochemical stability and coating integrity of the commercially available enteric-coated omeprazole pellets containing a basic excipient and prepared by extrusion and spheronization or drug layering on the nonpareil seed, immediately followed by enteric coating (i.e., absence of protective sub-coat). The nature of coating interface and the likely development of an in situ interfacial layer after the application of enteric coating solution was examined using HPLC, NMR, differential scanning calorimetry (DSC), and fluorescent imaging methods. Likewise for the characterization of the solid pellet structure via fluorescence microscopy, a new approach based on fracturing technique (to avoid surface contamination) rather than microtome sectioning was used and validated. Analytical data showed that the pellets containing omeprazole remained chemically stable (>99.5% recovered). Control of the microenvironmental pH by the addition of alkalinizing excipient within a core formulation or as part of drug layering on top of nonpareil seed appears to efficiently neutralize the acidic effect of enteric coating dispersion. Fluorescence images further illustrate the absence of any discernable in situ layer formation at the coat-core interface.KEY WORDS: alkalinizing excipient, DSC, enteric coating, florescence microscopy, functional coating layer(s), HPLC, NMR, omeprazole stability     

Biofabrication of zinc oxide nanoparticles from Melia azedarach and its potential in controlling soybean seed-borne phytopathogenic fungi

Present study, report the biofabrication of zinc oxide nanoparticles from aqueous leaf extract of Melia azedarach (MaZnO-NPs) through solution combustion method and their novel application in preventing the growth of seed-borne fungal pathogens of soybean (Cladosporium cladosporioides and Fusarium oxysporum). The standard blotter method was employed to isolate fungi and was identified through molecular techniques. The characterization of MaZnO-NPs was carried out by UV–Vis spectroscopy, Fourier Transform Infrared Spectroscopy (FT-IR), X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM) equipped with Energy Dispersive Spectroscopy (EDS) and Transmission Electron Microscopy (TEM). The physicochemical characterization confirmed the particles were of high purity and nano size (30–40 nm) with a hexagonal shape. The synthesized MaZnO-NPs inhibited the growth of C. cladosporioides and F. oxysporum in a dose dependent manner. Biomass, ergosterol, lipid peroxidation, intracellular reactive oxygen species and membrane integrity determination upon MaZnO-NPs treatment offered significant activities there by confirming the mechanism of action against the test pathogens. In conclusion, due to the effectiveness of MaZnO-NPs in controlling the growth of C. cladosporioides and F. oxysporum, the synthesized MaZnO-NPs provides insight towards their potential application in agriculture and food industries.     

Discovery and characterization of a novel extremely acidic bacterial N-glycanase with combined advantages of PNGase F and A

Peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidases [PNGases (peptide N-glycosidases), N-glycanases, EC 3.5.1.52] are essential tools in the release of N-glycans from glycoproteins. We hereby report the discovery and characterization of a novel bacterial N-glycanase from Terriglobus roseus with an extremely low pH optimum of 2.6, and annotated it therefore as PNGase H⁺. The gene of PNGase H⁺ was cloned and the recombinant protein was successfully expressed in Escherichia coli. The recombinant PNGase H⁺ could liberate high mannose-, hybrid- and complex-type N-glycans including core α1,3-fucosylated oligosaccharides from both glycoproteins and glycopeptides. In addition, PNGase H⁺ exhibited better release efficiency over N-glycans without core α1,3-fucose compared with PNGase A. The facile expression, non-glycosylated nature, unusual pH optimum and broad substrate specificity of this novel type of N-glycanase makes recombinant PNGase H⁺ a versatile tool in N-glycan analysis.     

Hydrologic Drivers and Seasonality of Dissolved Organic Carbon Concentration, Nitrogen Content, Bioavailability, and Export in a Forested New England Stream

We present the results of a full year of high-resolution monitoring of hydrologic event-driven export of stream dissolved organic matter (DOM) from the forested Bigelow Brook watershed in Harvard Forest, Massachusetts, USA. A combination of in situ fluorescent dissolved organic matter (FDOM) measurement, grab samples, and bioassays was utilized. FDOM was identified as a strong indicator of concentration for dissolved organic carbon (DOC, r ² = 0.96), dissolved organic nitrogen (DON, r ² = 0.81), and bioavailable DOC (BDOC, r ² = 0.81). Relationships between FDOM and concentration were utilized to improve characterization of patterns of hydrological event-driven export and the quantification of annual export. This characterization was possible because DOM composition remained relatively consistent seasonally; however, a subtle shift to increased fluorescence per unit absorbance was observed for summer and fall seasons and percent BDOC did increase slightly with increasing concentrations. The majority of export occurred during pulsed hydrological events, so the greatest impact of bioavailable exports may be on downstream aquatic ecosystems. Export from individual events was highly seasonal in nature with the highest flow weighted mean concentrations (DOC_FW) being observed in late summer and fall months, but the highest total export being observed for larger winter storms. Seasonal trends in DOC export coincide with weather driven changes in surface and subsurface flow paths, potential for depletion and rebuilding of a flushable soil organic matter pool, and the availability of terrestrial carbon sources such as leaf litter. Our approach and findings demonstrate the utility of high frequency FDOM measurement to improve estimates of intra-annual temporal trends of DOM export.     

<Emphasis Type="Italic">In Vitro</Emphasis> and <Emphasis Type="Italic">Ex Vivo</Emphasis> Evaluations of Lipid Anti-Cancer Nanoformulations: Insights and Assessment of Bioavailability Enhancement

Lipid-based nanoformulations have been extensively investigated for improving oral efficacy of plethora of drugs. Chemotherapeutic agents remain a preferred option for effective management of cancer; however, most chemotherapeutic agents suffer from limitation of poor oral bioavailability that is associated with their physicochemical properties. Drug delivery via lipid-based nanosystems possesses strong rational and potential for improving oral bioavailability of such anti-cancer molecules through various mechanisms, viz. improving their gut solubilisation owing to micellization, improving mucosal permeation, improving lymphatic uptake, inhibiting intestinal metabolism and/or inhibiting P-glycoprotein efflux of molecules in the gastrointestinal tract. Various in vitro characterization techniques have been reported in literature that aid in getting insights into mechanisms of lipid-based nanodevices in improving oral efficacy of anti-cancer drugs. The review focuses on different characterization techniques that can be employed for evaluation of lipid-based nanosystems and their role in effective anti-cancer drug delivery.     

Struvite Urolithiasis in Long–Evans Rats

Struvite urinary calculi, which are composed of magnesium, ammonium, and phosphate, can cause complications including sepsis and renal failure. Struvite calculi were identified within the urinary bladder and renal pelvis of 2 Long-Evans rats that died within days after arrival from a commercial vendor. The remaining rats in the shipment were screened by physical examination, radiography, and ultrasonography, revealing an additional 2 animals that were clinically affected. These rats were euthanized, necropsied, and yielded similar findings to those from the first 2 rats. In addition, urine samples had an alkaline pH and contained numerous bacteria (predominantly Proteus mirabilis), leukocytes, and crystals. All calculi were composed completely of struvite. Another 7 rats in the shipment had alkaline urine with the presence of blood cells; 6 of these rats also had abundant struvite crystals, and P. mirabilis was cultured from the urine of 3 rats. Further investigation by the vendor identified 2 of 100 rats with struvite calculi from the same colony. Although no specific cause could be implicated, the fact that all the affected rats came from the same breeding area suggests a genetic or environmental triggering event; a contribution due to diet cannot be ruled out. Our findings suggest that the affected rats had metabolic disturbances coupled with bacterial infection that predisposed them to develop struvite calculi. During sudden increases of struvite urinary calculi cases in rats, urine cultures followed by appropriate surgical intervention and antibiotic therapy is warranted. Additional factors, including diet, merit attention as well.Struvite, also known as triple phosphate, is a crystalline substance composed of magnesium ammonium phosphate (MgNH₄PO₄• 6 H₂0)¹⁰ that was first identified in the 18th century. This mineral is what consolidates into urinary calculi, or stones, both in humans and animals. Other frequently encountered types of urinary stones include calcium oxalate, calcium phosphate, uric acid, and cysteine.¹⁷ Although only 2% to 3% of stones from humans are composed of struvite,⁴³ struvite calculi are important clinically because they can lead to sepsis and renal failure.⁴³In both humans and animals, stones in the urinary tract can obstruct the urine outflow, with subsequent extreme pain, hydronephrosis, and (possibly) rupture of the urinary bladder. In addition, disruption of kidney function can lead to metabolic imbalances, such as uremia, seizures, depression, anorexia, dehydration, even coma and death.^1,8 Here we present a case study describing the spontaneous presentation of struvite urolithiasis in a recently imported cohort of rats and related significant clinical findings.     

Physicochemical and Mineralogical Characterization of Uranium-Contaminated Soils

Physicochemical and mineralogical properties of the contaminants should be taken into account to decide a remediation strategy for a given radionuclide because development and optimization of soil remedial technologies are based on physicochemical and mineralogical separation techniques. The objectives of this study are to (1) demonstrate how a priori physicochemical and mineralogical characterization of soil contaminants can direct the development of remediation strategies and their performance evaluation for soil treatments and (2) understand the nature of uranium contamination and its association with the soil matrix by chemical extractions. This study examined two U-contaminated sites (K311 and K1300) at the DOE K-25 site, presently located at East Tennessee Technology Park, Oak Ridge, Tennessee. Uranium concentrations of the soils ranged from 1499 to 216,413 Bq kg^?1 at both sites. Scanning electron microscopy with backscattered electron spectroscopy and X-ray diffraction analysis showed that the dominant U phases are U oxides (schoepite), U-Ca-silicate (uranophane) and U silicate (coffinite) from the K311 site soils, whereas U-Ca-oxide and U-Ca-phosphate dominate in the K1300 site soils. Sodium carbonate/bicarbonate leaching was effective on the K1300 site soils, whereas citric acid leaching is effective on the K311 site soils. Sequential leaching showed that the majority of the uranium in the contaminated soils was contained in carbonate minerals (45%) and iron oxides (40%). Conventional leaching showed that citric acid treatment was most effective on the K311 site soils, whereas the sodium carbonate/ bicarbonate treatment was most effective on the K1300 site soils.