Ultramicroscopic Analysis Research Articles

Reduction of selenite to selenium nanoparticles by highly selenite-tolerant bacteria isolated from seleniferous soil

The microbial reduction of selenite to elemental selenium nanoparticles (SeNPs) is thought to be an effective detoxification process of selenite for many bacteria. In this study, Metasolibacillus sp. ES129 and Oceanobacillus sp. ES111 with high selenite reduction efficiency or tolerance were selected for systematic and comparative studies on their performance in selenite removal and valuable SeNPs recovery. The kinetic monitoring of selenite reduction showed that the highest transformation efficiency of selenite to SeNPs was achieved at a concentration of 4.24 mM for ES129 and 4.88 mM for ES111. Ultramicroscopic analysis suggested that the SeNPs produced by ES111 and ES129 had been formed in cytoplasm and subsequently released to extracellular space through cell lysis process. Furthermore, the transcriptome analysis indicated that the expression of genes involved in bacillithiol biosynthesis, selenocompound metabolism and proline metabolism were significantly up-regulated during selenite reduction, suggesting that the transformation of selenite to Se0 may involve multiple pathways. Besides, the up-regulation of genes associated with nucleotide excision repair and antioxidation-related enzymes may enhance the tolerance of bacteria to selenite. Generally, the exploration of selenite reduction and tolerance mechanisms of the highly selenite-tolerant bacteria is of great significance for the effective utilization of microorganisms for environmental remediation.

Coculture of Acinetobacter johnsonii and Shewanella putrefaciens Contributes to the ABC Transporter that Impacts Cold Adaption in the Aquatic Food Storage Environment.

Acinetobacter johnsonii and Shewanella putrefaciens were identified as specific spoilage organisms in aquatic food. The interactions among specific spoilage organisms under cold stress have a significant impact on the assembly of microbial communities, which play crucial roles in the spoilage and cold adaptation processes. The limited understanding of A. johnsonii and S. putrefaciens interactions in the cold adaptation mechanism hinders the elucidation of their roles in protein and metabolism levels. 4D quantitative proteomic analysis showed that the coculture of A. johnsonii and S. putrefaciens responds to low temperatures through ABC transporter proteins, resulting in phospholipid transport and inner membrane components. SapA and FtsX proteins were significantly upregulated, while LolC, LolD, LolE, PotD, PotA, PotB, and PotC proteins were significantly downregulated. Metabolome assays revealed that metabolites of glutathione and spermidine/putrescin were significantly upregulated, while metabolites of arginine/lysine/ornithine were significantly downregulated and involved in the ABC transporter metabolism. The results of ultramicroscopic analyses showed that the coculture of A. johnsonii and S. putrefaciens surface combined with the presence of the leakage of intracellular contents, suggesting that the bacteria were severely damaged and wrinkled to absorb metabolic nutrients and adapt to cold temperatures.

The Fate of Phosphorus in Experimental Burials: Chemical and Ultramicroscopic Characterization and Environmental Control of Its Persistency

The permanence of a buried body in soil always induces the formation of a decomposition island that can serve as a significant recording location to understand how the persistence of a clandestine grave affects soil. This study aims to analyze the elemental exchange from buried bodies to soil, with a focus on phosphorus content, and to determine the effects of environmental factors on its persistency. The experiment was carried out using eleven swine carcasses buried in an open site (northern Italy). The analyses were performed using the Olsen P method, which allowed for a recognition of the trend of the amount of phosphorus over time, due to the decomposition of phospholipids, followed by the transfer of the element from bone to soil. Additionally, microanalyses performed using a scanning electron microscope (SEM-EDS) on two different soil sample specimens (i.e., “dust” and “plug”) allowed for the identification of numerous phosphatic features (i.e., coatings, infillings, impregnations, and organo-mineral associations), which are the result of the interaction between soil and body fluids and can thus be used as indicators of the former presence of decomposing body (even in its absence). The ultramicroscopic analysis also shows increasing and decreasing amounts of P2O5 over time in the soil, which could be related to environmental conditions (i.e., soil moisture), due to the leaching of phosphorus induced by the percolation of natural rainwater. The study underlines the potential use of these methods to evaluate the possibility of a cadaver–soil linkage and of assessing the burial in the soil for a variable period. Moreover, the study may aid in analyzing the dynamics of phosphorus migration from buried bodies to soil during and after the decomposition process.

Morphologic vascular anomalies detected during migraine surgery.

Although there is increasing evidence of migraine headaches having extracranial origins, the exact mechanisms underlying the pathogenesis of surgically treated migraines continue to be poorly investigated and described. We studied the microscopic and ultrastructural characteristics of superficial temporal artery (STA) and occipital (OA) artery in the auriculotemporal and great occipital trigger points of migraine patients to determine their possible role in migraine etiopathogenesis. Fifteen biopsies, 10 of STA and 5 from OAs, were collected intraoperatively during migraine surgery and immediately processed for optical and ultramicroscopic analysis. We detected the following anomalies in all the specimens: (a) endothelial damage with internal elastic lamina fragmentation and intimal thickening; (b) marked irregularity in the shape and metachromasia of the vascular smooth muscle cells (VSMCs), separation of cells by abundant extracellular matrix and vacuoles. The electron microscopy analysis confirmed that presumed VSMCs infiltrated the intima layer revealing a consistent shift of VSMCs from contractile to synthetically active phenotypes, endosome-like organelles, multilamellar structures, abundant extracellular vacuoles filled with fine granular material and membranes, and extracellular vesicles in the matrix space surrounding synthetically active cells. Our study revealed specific alterations in the vasculature at the neurovascular bundles of the temporal and occipital trigger sites. These findings are indicative of an active involvement of the arteries in the auriculotemporal and great occipital trigger sites in evoking migraine.

Origin and Composition of Nanoparticles Induced in a Complex Plastic-Brittle Transition Process of Xiaomei Shear Zone, Hainan Island, China

Micro and nano structures of quartz schist, plagioclase granite, and granitic gneiss in the Xiaomei Shear Zone located in southeastern Hainan Island, China are observed using Scanning Electron Microscope (SEM). Ultramicroscopic analyses indicate that three types of nanoparticles were found in the samples, including spherical nanoparticles, agglomerated nanoparticles and nanofibers. These nanoparticles are mainly developed in the fracture zones. The more fractures there are, the more nanoparticles are developed. Energy dispersive spectrometer (EDS) and X-ray diffraction (XRD) are used to ascertain the composition of the nanoparticles. The results suggest that the nanoparticles are mainly composed of silicate, dolomite and calcite, rich in O, Si, Al, Ca. Based on our results, we suggest that nanoparticle formation is a complex, plastic-brittle transition process. Thermal decomposition driven by steady shearing possibly forms well-organized nanoparticles, while fast-moving dislocations by shock-like stress release possibly forms radial nanofibers.

Comparative In Vitro Studies on the Effect of Bleaching Agents on Dental Structures in Healthy and Predialysis Patients

(1) Background: What is the effect of 16% and 40% concentration bleaching agents on dental structures in healthy patients compared to predialysis patients? (2) Methods: Forty teeth were included in the study (20 from healthy patients and 20 from predialysis patients). Each group was randomly divided into another two subgroups (n-10), depending on the bleaching agent concentration (16% and 40% gels). Color parameters were registered before and after the whitening process using a spectrophotometer. To determine enamel ultramicroscopic modifications, SEM and AFM analysis were performed before and after bleaching. (3) Results: An increasing trend was identified in the average values of ∆E and ∆L within the groups of predialysis teeth between teeth whitened with 40% concentration gel and those whitened with 16% concentration gel, while for the enamel samples from healthy patients the trend was reversed. The average values for roughness in the case of the two bleaching agents in healthy and predialysis teeth presented statistically significant differences (p < 0.05). (4) Conclusions: The effects of bleaching agents are less significant on teeth from predialysis compared to healthy patients. A direct link exists in terms of the clinical effect between the concentration of the whitening gel and color modifications.

Selenite bioreduction and biosynthesis of selenium nanoparticles by Bacillus paramycoides SP3 isolated from coal mine overburden leachate

A native strain of Bacillus paramycoides isolated from the leachate of coal mine overburden rocks was investigated for its potential to produce selenium nanoparticles (SeNPs) by biogenic reduction of selenite, one of the most toxic forms of selenium. 16S rDNA sequencing was used to identify the bacterial strain (SP3). The SeNPs were characterized using spectroscopic (UV–Vis absorbance, dynamic light scattering, X-ray diffraction, and Raman), surface charge measurement (zeta potential), and ultramicroscopic (FESEM, EDX, FETEM) analyses. SP3 exhibited extremely high selenite tolerance (1000 mM) and reduced 10 mM selenite under 72 h to produce spherical monodisperse SeNPs with an average size of 149.1 ± 29 nm. FTIR analyses indicated exopolysaccharides coating the surface of SeNPs, which imparted a charge of −29.9 mV (zeta potential). The XRD and Raman spectra revealed the SeNPs to be amorphous. Furthermore, biochemical assays and microscopic studies suggest that selenite was reduced by membrane reductases. This study reports, for the first time, the reduction of selenite and biosynthesis of SeNPs by B. paramycoides, a recently discovered bacterium. The results suggest that B. paramycoides SP3 could be exploited for eco-friendly removal of selenite from contaminated sites with the concomitant biosynthesis of SeNPs.

Chronic pesticide exposure induced aberrant Notch signalling along the visual pathway in a murine model

Pesticides aid in crop-protection against pests and increase yield. However, the xenobiotic stress exerted by pesticides leads to the deterioration of human and animal health. There is a lacuna in our knowledge about their impact on the ocular surface The present work sheds light on this gap by analysing the deterioration of visual acuity as a consequence of pesticide induced xenobiotic stress and Notch pathway dysregulation.Alteration in the expression of vital components of the notch signalling was analyzed along the visual pathway with special focus on its two terminals-the cornea and the visual cortex, by mimicking the on-field scenario regarding chronic pesticide exposure in experimental murine model (Swiss albino mice; Mus musculus). Various aspects were taken into consideration through visual acuity tests, histological evaluations, culture analyses, wound healing assays, flowcytometric evaluation, fluorescence microscopic studies etc. Complete dysregulation of key players of the Notch signalling pathway was observed in both: cells of the ocular surface as well as those in the murine visual cortex post pesticide exposure, indicating activities relating to cell proliferation, differentiation and wound healing in the pesticide exposed samples. Ultra-microscopic analyses corroborated our findings by revealing the loss of fine neural processes in the visual cortex of the pesticide exposed murine samples, thereby hinting at delayed perception to visual stimuli. In vivo evaluations of the functional capacity of the neuroanatomical structures along the visual pathway also confirmed that pesticide exposure leads to severe damage along the various parts of the visual pathway, right from the ocular surface to the visual cortex.

Nanoparticles Study on the Indosinian Xiaomei Shear Zone in the Hainan Island, China: Implication to Developmental Stage and Formation Mechanism of Nanoparticles in a Fault Zone

Nanoparticles are widely observed in the natural shear zone and experimental slip faults, which can lubricate the fault and significantly reduce the friction coefficient during seismic slip. But it is still not clear how the nanoparticles develop during the process of sliding. Clarifying the development stage of nanoparticles in a fault zone is critical to understanding the formation mechanisms of nanoparticles and the mechanism of fault weakening from a nanoperspective. In this study, four types of nanoparticles were found in the Indosinian Xiaomei shear zone, including spherical nanoparticles, rod-like nanograins and their aggregations. Ultramicroscopic analyses indicate that polished patches are highly smooth and composed of tightly packed spherical nanoparticles and well orientated rod-like nanograins during slip at high velocities. Meanwhile, the dome nanoparticles were formed by the calcite thermal decomposition due to frictional heat during highspeed sliding. The polygonal grooves are possibly related to high temperature (>900 °C) grain boundary sliding deformation mechanisms. However, the porous and rough surfaces are accompanied by a series of holes and parallel “scratches” during a subsequent low-velocity stage. To ascertain the chemical composition of these nanoparticles, the energy dispersive spectrometer (EDS) test were conducted. The results suggest that materials rich in Fe, MgO and wollastonite are likely to form the rod-like nanograins, while materials rich in SiO2 are likely to form the spherical nanoparticles.

The interplay of the inhibitory effect of nifuroxazide on NF-κB/STAT3 signaling attenuates acetic acid-induced ulcerative colitis in rats

Ulcerative colitis (UC) is a disease of increased worldwide prevalence. UC progression is associated with serious complications that leave the patient with considerable health burdens. Nifuroxazide is an oral nitrofuran antibiotic used as antidiarrheal medication. The current study places an emphasis on investigating the potential therapeutic effectiveness of nifuroxazide (10 mg/kg) and (20 mg/kg) against acetic acid (AA)-induced UC. Intra-rectal AA induced a significant colonic injury and impairment of colonic biochemical and functional incidences. Nifuroxazide in a dose-dependent manner significantly corrected UC associated injury. Macroscopic scoring of UC, serum lactate dehydrogenase (LDH) activity, C-reactive protein (CRP) titer, colon malondialdehyde (MDA) and total nitric oxide (NOx) contents significantly declined. Meanwhile, serum total antioxidant capacity (TAC) and colon catalase, superoxide dismutase (SOD) and glutathione transferase (GST) activities and reduced glutathione (GSH) concentration significantly increased in a dose-dependent way. Ultimately, histopathological, immunohistochemical and ultramicroscopic analysis of colon specimen revealed significant improvement. To pinpoint the mechanistic pathway underlying the curative effect of nifuroxazide, colon expression of NF-κB, caspase-3 was evaluated along with STAT-3 activation. Nifuroxazide induced a dose-dependent significant suppression of NF-κB and caspase-3 signaling together with STAT3 signaling. In conclusion; nifuroxazide can be proposed as a therapeutic candidate to attenuate UC and its associated symptoms. The potential underlying mechanism involves suppression of NF-κB/STAT-3/caspase- signaling.

Biofunctionalization with a TGFβ-1 Inhibitor Peptide in the Osseointegration of Synthetic Bone Grafts: An In Vivo Study in Beagle Dogs.

Objectives: The aim of this research was to determine the osseointegration of two presentations of biphasic calcium phosphate (BCP) biomaterial—one untreated and another submitted to biofunctionalization with a TGF-β1 inhibitor peptide, P144, on dental alveolus. Materials and Methods: A synthetic bone graft was used, namely, (i) Maxresorb® (Botiss Klockner) (n = 12), and (ii) Maxresorb® (Botiss Klockner) biofunctionalized with P144 peptide (n = 12). Both bone grafts were implanted in the two hemimandibles of six beagle dogs in the same surgical time, immediately after tooth extraction. Two dogs were sacrificed 2, 4, and 8 weeks post implant insertion, respectively. The samples were submitted to histomorphometrical and histological analyses. For each sample, we quantified the new bone growth and the new bone formed around the biomaterial’s granules. After optical microscopic histological evaluation, selected samples were studied using backscattered scanning electron microscopy (BS-SEM). Results: The biofunctionalization of the biomaterial’s granules maintains a stable membranous bone formation throughout the experiment timeline, benefitting from the constant presence of vascular structures in the alveolar space, in a more active manner that in the control samples. Better results in the experimental groups were proven both by quantitative and qualitative analysis. Conclusions: Synthetic bone graft biofunctionalization results in slightly better quantitative parameters of the implant’s osseointegration. The qualitative histological and ultramicroscopic analysis shows that biofunctionalization may shorten the healing period of dental biomaterials.

Lower Selenoprotein T Expression and Immune Response in the Immune Organs of Broilers with Exudative Diathesis Due to Selenium Deficiency

The objective of the present study was to investigate whether dietary selenium (Se) deficiency would affect the expression of selenoprotein T (SelT) and immune response in the immune organs of broilers. Changes in expression of inflammatory cytokines and oxidative stress response caused by Se deficiency can lead to organism damage, which in turn leads to immune response. Sixty (1-day-old) broilers were divided into the control group and Se-deficiency group. Animal models with exudative diathesis were duplicated in the broilers by feeding them Se-deficient diet for 20days. After the Se-deficient group exhibited symptoms of exudative diathesis, all the broilers were euthanized, and their immune organs were taken for analysis. The tissues including spleen, bursa of Fabricius, and thymus were treated to determine the pathological changes (including microscopic and ultramicroscopic), the messenger RNA (mRNA) expression levels of SelT and its synthetase (SecS and SPS1), cytokine mRNA expression levels, and antioxidant status. The microscopic and ultramicroscopic analyses showed that immune tissues were obviously injured in the Se-deficient group. The mRNA expression of SelT was decreased compared with that in the control group. Meanwhile, the mRNA expression levels of SecS and SPS1 were downregulated. In the Se-deficient group, the mRNA expression levels of IL-1R and IL-1β were higher than those of three control organs. Additionally, the IL-2 and INF-γ mRNA expression levels were lower than those of the control group. The activity of CAT was decreased, and the contents of H2O2 and •OH were increased due to Se deficiency. Pearson method analysis showed that the expression of SelT had a positive correlation with IL-2, INF-γ, SecS, and SPS1 and a negative correlation with IL-1R and IL-1β. In summary, these data indicated that Se-deficient diet decreased the SelT expression and its regulation of oxidative stress, and it inhibited a pleiotropic mechanism of the immune response.

NEUROPROTECTIVE POTENTIAL OF AZADIRACHTA INDICA LEAVES IN DIABETIC RATS

Objective: Azadirachta indica is a treasure of multiple pharmacological properties and presently leaves of this plant have been explored to evaluate the neuroprotective potential in diabetic rats.Methods: Male Sprague-Dawley rats were injected with single intra peritoneal dose of streptozotocin (60mg/ Kg body weight (BW.) to develop animal model of diabetes. Post twenty one days of streptozotocin induction, animals were treated with aqueous Azadirachta indica Leaf Extract (ALE, 600mg/Kg BW.) for seven consecutive days. Followed this, all animals were evaluated for the levels of blood glucose, lipid peroxidation (LPO), C Reactive Proteins (CRP), pro oxidant biomarkers and histological changes.Results: Streptozotocin treated rats exhibited elevated levels of blood glucose, LPO, CRP and altered pro oxidant biomarkers in comparison to control rats. Additionally, histological alterations/damage was evidenced as fragmentation, vacuolization, inflammation etc. However, ALE treatment to these rats significantly decreased blood glucose levels, LPO, CRP levels and restored pro-oxidants status. Light microscopic and ultra microscopic analysis also indicated less damage, tissue architectural changes in comparison to untreated diabetic rats. Further decrease in hyperalgesia and inflammation levels; along with protective and restorative changes following ALE treatment suggested the neuroprotective potential of Azadirachta indica leaves in diabetic rats.Conclusion: The oral administration of ALE to streptozotocin induced diabetic animals resulted in neuro-protection against degenerative oxidative stress associated with metabolic and histopathological damage in the brain.Key words: Azadirachta indica, Antioxidants, Hyperalgesia, Neuroprotection

Observation on the ultrastructure morphology of HeLa cells treated with ethanol: Statistical analysis

ABSTRACTIt is estimated that 5.9% of all human deaths are attributable to alcohol consumption and that the harmful use of ethanol ranks among the top five risk factors for causing disease, disability, and death worldwide. Ethanol is known to disrupt phospholipid packing and promote membrane hemifusion at lipid bilayers. With the exception of mitochondria involved in hormone synthesis, the sterol content of mitochondrial membranes is low. As membranes that are low in cholesterol have increased membrane fluidity and are the most easily disordered by ethanol, we hypothesize that mitochondria are sensitive targets for ethanol damage. HeLa cells were exposed to 50 mM ethanol and the direct effects of ethanol on cellular ultrastructure were examined utilizing transmission electron microscopy. Our ultramicroscopic analysis revealed that cells exposed to ethanol harbor fewer incidence of apoptotic morphology; however, significant alterations to mitochondria and to nuclei occurred. We observed statistical increases in the amount of irregular cells and cells with multiple nuclei, nuclei harboring indentations, and nuclei with multiple nucleolus-like bodies. Indeed, our analysis revealed that mitochondrial damage is the most extensive type of cellular damage. Rupturing of cristae was the most prominent damage followed by mitochondrial swelling. Ethanol exposure also resulted in increased amounts of mitochondrial rupturing, organelles with linked membranes, and mitochondria localizing to indentations of nuclear membranes. We theorize that these alterations could contribute to cellular defects in oxidative phosphorylation and, by extension, the inability to generate regular levels of cellular adenosine triphosphate.

Antigen detection and apoptosis in Mongolian gerbil’s kidney experimentally intraperitoneally infected by swine hepatitis E virus

We examined the effect of hepatitis E virus (HEV) on the renal tissue pathogenesis, morphological damages and related molecular mechanisms following swine HEV suspension intraperitoneally inoculation in Mongolian gerbils. The microscopic and ultramicroscopic analyses of kidney tissue structure were carried out at different points after inoculation of HEV. The immunohistochemistry, real-time PCR and Western blot were performed to explore the molecular mechanisms associated with HEV presence in the renal tissues. Real-time PCR revealed that the copies of HEV RNA in the kidney were detected at 7dpi, and peaked at 14dpi at a concentration was 7.18logsg−1, with detection of HEV ORF2 antigen by immunohistochemistry. Hematoxylin and eosin (HE) staining showed pathological lesions including glomerular atrophy, degeneration, edema and necrosis of renal tubular epithelial cells and Mallory and Sirius red staining indicated the presence of collagen fibers and fibrosis in kidney tissues of inoculated gerbils. Ultrastructural studies of basal membrane of renal tubules demonstrated the rough and uneven with mitochondria swelling and vacuolation in the tissues of HEV inoculated animals. Similarly, significantly higher number of (TUNEL)-positive cells were seen in renal tubule tissues compared to control group. Moreover, immuno histochemical results indicated that significant increase expression of the B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X protein (Bax), FAS and Caspase-3 in HEV inoculated Mongolian gerbils at each time points. Relative mRNA expression by real-time PCR revealed a significantly higher (P<0.05) mRNA level of BAX, Bcl-2 and caspase-3 transcription in HEV inoculated Mongolian gerbils. Our results demonstrates that activation of mitochondria and Caspase-3 protease might be induced the apoptosis which subsequently cause the necrosis and cell death of renal epithelial cells during acute phase of HEV infection in HEV inoculated Mongolian gerbils.

Plasma Blood Coagulation Without Involving the Activation of Platelets and Coagulation Factors

The objective of this study is to determine whether there is a pathway different from the natural blood coagulation process involved in the plasma‐induced activation of platelets and coagulation factors. Plasma treatment of bleeding wounds generated glomerular structures, which could be detected in high‐magnification scanning electron microscopy images, and plasma flares produced aggregation on the surface of solutions containing bovine serum albumin, ovalbumin (OVA), or similar protein material. Ultramicroscopic analysis of the plasma‐generated aggregation of OVA indicated a possible mechanism to explain the particle formation and fusion as a monotonous solid appearance. This study suggests that plasma coagulation involves aggregations of materials in blood in addition to the activation of platelets and coagulation factors by plasma treatment.

Fault mirrors along carbonate faults: Formation and destruction during shear experiments

Glossy, light reflective surfaces are commonly exposed in carbonate fault-zones. It was suggested that such surfaces, recently termed Fault Mirrors (FMs), form during seismic slip. Ultramicroscopic analyses indicate that FMs are highly smooth and composed of a cohesive thin layer of nano-size grains. We explore here mechanisms of formation and destruction of FMs by shear experiments that were conducted on three types of limestone which were sheared at wide range of slip-velocities of v=0.001–0.63 m/s, and normal stress up to 1.57 MPa. The experiments showed that FMs started to develop as local patches when the slip velocity exceeded a critical value of 0.07 m/s. The area coverage by FM patches increases systematically with increasing velocity, reaching in a few cases ∼100% coverage. The measured quasi-steady-state friction coefficient, μss, was inversely correlated with the FM coverage: μss∼0.8 for no-FM, at v<0.07 m/s, and μss∼0.4 for 50% FM coverage at v∼0.6 m/s. Further, in a series of slip-velocity alternation between low and high values, the FMs which formed at a high-velocity stage were destroyed during a subsequent low-velocity stage. Our analyses of the experimental thermal conditions and ultramicroscopy imaging of the FMs suggest that the FMs form by sintering of gouge nanograins during shear. We propose that formation/destruction of FMs in high/low slip-velocity reflects a competition between brittle and ductile processes: FMs form in a ductile mode, and are destroyed by brittle wear. Shear heating during high velocity leads to ductile deformation and sintering so that FM construction rate exceeds brittle FM destruction rate. Based on our results, we suggest that, at least for shallow faults, the presence of extensive FM coverage along natural carbonate faults indicates that the fault segment slipped at seismic velocities and experienced dynamic weakening.

Hypoelectrolytic isoosmotic solution for infusion prevents saline‐induced ultrastuructural artifacts of renal biopsy specimens

Artifacts in the process of specimen preparation are frequent in ultrastructural evaluation of renal biopsy. We hypothesized that the common practice of wrapping kidney biopsy specimens in saline-soaked gauze to prevent the drying of the specimens could be the major factor of artifacts. In this study, whole kidneys from two male Sprague-Dawley rats were used. Before fixation, fresh small cubes of kidney tissue were macerated in saline (Saline group) or hypoelectrolytic isoosmotic solution for infusion (HISI group) (Sorita T3 or SOLDEM 3A) for 10 or 30 min. Then, the specimens were processed by 1% OsO(4) in 0.1 M phosphate buffer (pH 7.4) and embedded by EPON 812 for ultramicroscopic analysis. In the Saline group, ultrastructural examination revealed swollen podocyte, swollen capillary protuberance of the mesangium into the glomerular capillary loop, tubular cells with swollen mitochondria and microvilli, and the smooth muscle cells in the arteriolar wall with marked vacuolar degeneration were detected after 10 min maceration in saline and these findings become more pronounced after 30 min maceration. However, in the HISI group, these artifacts were not identified or limited within 30 min. It is postulated that HISI solution could prevent the artifacts, and be used for soaking and wrapping instead of physiologic saline solution.

Self-assembling triazolophanes: from croissants through donuts to spherical vesicles.

Macrocyclic compounds M1-M3 with different ring sizes containing amide and triazole units were synthesized. These triazolophanes displayed a variety of self-assembled structures such as hemi-toroids, toroids, and vesicles in a concentration dependent manner. Detailed ultramicroscopic and crystallographic investigations delineated a hierarchical mechanism of self-assembly.

Ameliorative effects of phyllanthin on carbon tetrachloride-induced hepatic oxidative damage in mice

ObjectiveTo evaluate the liver protecting efficacy of phyllanthin, a lignin, isolated from the leaves of Phyllanthus amarus using mice model. MethodsPhyllanthin was orally administered with or without CCl4 for 30 d. Serum levels of hepatic marker enzymes namely alanine transaminase and aspartate transaminase were evaluated. Oxidative stress was ascertained by measuring hepatic lipid peroxidation levels and by estimating non-enzymatic antioxidants such as glutathione, total ascorbic acid, enzymatic antioxidants namely catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione transferase. Histopathological and ultramicroscopic analyses were also carried out. ResultsOral administration of CCl4 caused significant increase in lipid peroxidation. The hepatic levels of both non-enzymatic antioxidants and enzymatic antioxidants were significantly lowered in CCl4-treated mice as compared to control. Treatment with phyllanthin significantly mitigated these changes in the CCl4-treated mice. Histopathological and ultramicroscopic studies correlated well with the biochemical findings, as phyllanthin treatment reversed the alterations induced by the toxin and the subcellular features of phyllanthin treated mice were similar to those present in the normal mouse liver. ConclusionsThis study reports the in vivo anti-hepatotoxic potential of this isolated molecule phyllanthin, which may be responsible for the liver protecting property of Phyllanthus amarus.