Melamine crystallization: physicochemical properties, interactions with other lithogenic salts and response to therapeutic agents.

PURPOSE: There were reports of children in the People's Republic of China being hospitalized with renal stones and/or failure by September 2008, which were caused by melamine and its co-contaminant cyanurate. We investigated the physicochemical behavior of melamine, its interaction with other endogenous urine factors and the response to therapeutic agents in the renal environment in vitro. MATERIALS AND METHODS: A mixed suspension, mixed product removal system was set up for crystallization studies of melamine in urine. Crystallization kinetic parameters, including the nucleation and growth rates, and suspension density, were determined according to crystal number and size, as measured by a Coulter particle counter. RESULTS: Melamine crystallized out from urine under normal urinary conditions (pH 5.0 to 6.5) but crystallization was strongly inhibited at pH 4.5 or lower. Melamine significantly enhanced calcium oxalate precipitation while uric acid significantly decreased melamine crystallization. Bacteria mimicking urinary tract infection promoted melamine crystallization. Clinical relevant drugs, such as citrate and bicarbonate, significantly decreased melamine crystallization. CONCLUSIONS: This implies that melamine crystallizes under normal urinary conditions and can interact with other lithogenic salts and pose a significant risk for other stones. Urinary tract infection promotes melamine crystallization. Citrate and bicarbonate therapy are effective prophylactic agents against melamine induced crystallization.

Synergistic Cytotoxic Effects of Ganoderma lucidum and Bacillus Calmette Guérin on Premalignant Urothelial HUC-PC Cells and Its Regulation on Proinflammatory Cytokine Secretion.

Bacillus Calmette-Guérin (BCG) is conventionally used as an adjuvant immunotherapy to reduce the recurrence of bladder cancer. To address the issues of efficacy and safety, an ethanol extract of Ganoderma lucidum (GLe) was evaluated for its interaction with BCG. In a model of premalignant human uroepithelial cells (HUC-PC), GLe exerted immediate cytotoxic effects while BCG showed a delayed response, given that both were immunological active in inducing the secretion of interleukin (IL)-6, IL-8, and monocyte chemotactic protein-1 (MCP-1). Synergistic cytotoxic effects were observed when cells were either coincubated with both drugs or firstly preincubated with GLe. Synergism between GLe and BCG was demonstrated to achieve a complete cytostasis in 24 hours, and such effects were progressed in the subsequent 5 days. However, the pretreatment of GLe resulted in suppression of IL-6, IL-8, and MCP-1 secretions without affecting the cytotoxicity. Given that numerous proinflammatory cytokines are associated with the high side effects toll of BCG, results herein suggested the potential implications of GL to supplement the BCG immunotherapy in bladder cancer, for better efficacy and reducing side effects.

Composition and inhibitory properties of endogenous urinary GAGS are different in subjects from two race groups with different occurrence rates of kidney stones: Pilot studies provide unique evidence in support of an inhibitory role for this group of compounds.

BACKGROUND: Calcium oxalate (CaOx) kidney stone disease is common in South African whites (W) but is rare in the black population (B). The possible role of endogenous urinary glycosaminoglycans (GAGs) has not been previously investigated in this context. AIM: To determine concentration, composition, structure and CaOx crystal-inhibiting properties of this group of compounds in ultrafiltered urine of healthy subjects from both groups. MATERIALS AND METHODS: GAGS were isolated from 24 h urine samples and were quantified and characterized by sequential precipitation, Bradford protein assay, high performance liquid chromatography, and anion exchange high performance chromatography. CaOx crystal inhibition was determined in ultrafiltered urinary fractions to which purified GAGS (PG) from each group (PGB and PGW) had been added. Nucleation, growth and aggregation were measured by Coulter particle counting, spectrophotometric assay and [14C]-oxalate deposition. RESULTS: Higher concentrations of chondroitin sulfate (CS) were found in PGB than in PGW. PGB inhibited crystallization to a greater extent than PGW. CONCLUSIONS: We attribute the stronger inhibitory effect of PGB to its higher content of CS and suggest that the superior inhibition of CaOx crystallization by PGB relative to PGW might be a contributory factor in accounting for the lower stone occurrence rate in B.

Respiratory health of road-side vendors in a large industrialized city.

GOAL, SCOPE AND BACKGROUND: The literature reports a high prevalence of respiratory symptoms associated with exposure to motor vehicle exhaust emissions and people exposed to vehicle emissions are at risk of reduced lung function and cardiovascular performance. Although the effect of traffic emissions is a known risk to respiratory health, retailers are often situated along major roads in a busy urban environment to maximise customers. Shop assistants in an air-conditioned environment should be less exposed to traffic fumes and their lung function should be better preserved compared to 'unprotected' vendors exposed directly to vehicle emissions. The lung function of these two groups of workers has not previously been compared. The aims of this study are to determine if there is a difference in the concentration of respirable particles of diameter less than 10 micrometers (PM10) inside and outside air-conditioned shops along a busy major road; and to compare the lung function parameters, blood lead levels and respiratory symptoms between road-side vendors directly 'exposed' to traffic fumes and vendors working in 'protected' air-conditioned shops along the same road. METHODS: Roadside vendors (n=33) and adjacent shop assistants (n=31) were recruited for the study in a district known to have high emissions and pollution measures. All subjects were asked to complete a questionnaire providing the frequency of their respiratory symptoms. Spirometry pulmonary function tests were conducted and exhaled carbon monoxide levels (ECO), oxygen saturation and blood lead levels (BLL) were measured. PM10 level was concurrently measured in the subject's working environment. Pulmonary function data and ECO levels collected from a cohort of university staff aged > or = 30 years (n=92) at a university campus were included for comparison. RESULTS: The concentration of particulate matter (PM10) at the roadside (210 +/- 70 microg/m3) was significantly higher than inside the shops (130 +/- 40 microg/m3). There was no difference in lung function parameters or BLL between the groups, however the forced vital capacity of both groups of vendors was significantly lower than a 'control' cohort of academic institution personnel (n=92). DISCUSSION: This study illustrated that while the level of pollution improved with air conditioning, this may not be sufficient to prevent respiratory consequences. Our data suggests that a reduction in occupational exposure is not enough to protect these workers. CONCLUSIONS: Air-conditioned work environment is insufficient to protect the respiratory system against the adverse effects of exposure to vehicle emissions. RECOMMENDATIONS: The overwhelming effect of exposure during travel to and from work and at home cannot be underestimated. More stringent guidelines to control pollution appear necessary to protect the respiratory health of both shop-keepers and road-side vendors and further studies to explore the effect on respiratory health of the exposure to pollutants associated with commuting to and from work are warranted.

Electrophoretic separation and characterization of urinary glycosaminoglycans and their roles in urolithiasis.

Urinary polyanions recovered from the urine samples of kidney stone-formers and normal controls were subjected to preparative agarose gel electrophoresis, which yielded fractions 1-5 in a decreasing order of mobility. In both groups, chondroitin sulfates were identified in the fast-moving fractions and heparan sulfates in the slow-moving fractions. Furthermore, two types of heparan sulfates were identified based on their electrophoretic mobility: slow-moving and fast-moving. The fractionated urinary polyanions were then tested in an in vitro calcium oxalate crystallization assay and compared at the same uronic acid concentration, whereby, the chondroitin sulfates of stone-formers and heparan sulfates of normals enhanced crystal nucleation. Fraction 5 of the normals, containing glycoproteins (14-97 kDa) and associated glycosaminoglycans, were found to effectively inhibit crystallization. Papainization of this fraction in stone-formers revealed crystal-suppressive effects of glycoproteins, which was not seen in similar fractions of normals. It was concluded that glycoproteins could modulate the crystal-enhancing glycosaminoglycans such as chondroitin sulfates of stone-formers but not in normals. The differing crystallization activities of electrophoretic fraction 1 of normals and stone-formers revealed the presence of another class of glycosaminoglycan-hyaluronan. Hence, in the natural milieu, different macromolecules combine to have an overall outcome in the crystallization of calcium oxalate.

Folium pyrrosiae ingestion has no effect on the thermodynamic or kinetic urinary risk factors for calcium oxalate urolithiasis in healthy subjects: a poor prognosis for alternative treatment in this type of stone former.

Kidney stone disease occurs throughout the world. Conservative treatments involving herbal preparations have been used in traditional Chinese medicine. In vitro studies have suggested that Folium pyrrosiae (FP) has therapeutic potential in this context. The present study was undertaken to investigate the effects of ingested FP on urinary thermodynamic and kinetic risk factors for calcium oxalate (CaOx) stone formation in subjects from two different population groups. Healthy white (n = 9) and black (n = 9) males ingested 1.5 g FP each day for 7 days. 24 h urines (baseline and day 7) and blood samples (baseline and day 3) were collected. Urines were analyzed for lithogenic risk factors and were subjected to CaOx crystallization experiments in which the metastable limit (MSL), particle size-volume distribution and crystal deposition kinetics were determined. Urine composition values were used to calculate the relative supersaturation (RS) of CaOx and other urinary salts. Blood samples were analyzed for liver enzymes to monitor the safety of the protocol. Food diaries were recorded on days 0 and 7. Data were analyzed statistically using standard software. Nutrient intakes and the concentration of liver enzymes did not change during the study. No side effects were reported. There were no statistically significant differences in any of the thermodynamic (RS, MSL) or kinetic (particle volume-size distribution, crystal deposition rate) risk factors for CaOx stone formation in either of the groups following ingestion of FP relative to baseline values. FP does not have potential as a therapeutic agent in the management of CaOx kidney stone disease.

Herbal preparations affect the kinetic factors of calcium oxalate crystallization in synthetic urine: implications for kidney stone therapy.

Herbal remedies are increasingly being considered as suitable long-term treatments for renal dysfunction. The objective of the present study was to investigate the effect of some herbal extracts, all previously identified in published studies as influencing kidney stone formation, on the crystallization characteristics of calcium oxalate (CaOx) in synthetic urine (SU). Five herbal extracts were selected for the study: Folium pyrrosiae, Desmodium styracifolium, Phyllanthus niruri, Orthosiphon stamineus and Cystone(®). Concentrated stock solutions of each herbal extract were prepared and were tested at their recommended dosages in in vitro crystallization studies in SU. CaOx crystallization experiments were performed in which the metastable limit (MSL), average particle size, and nucleation and growth rates were determined. The CaOx MSL of SU was unaltered by the five herbal extracts. Three of the herbs (Desmodium styracifolium, Orthosiphon stamineus and Cystone(®)) significantly reduced the average particle size of precipitated crystals relative to undosed SU. All of the extracts increased the rate of nucleation and decreased the rate of growth significantly in SU. Cystone(®) showed the greatest effect on the measured risk factors. It is concluded that all of the herbs have the potential to serve as inhibitors of calcium oxalate stone formation and warrant investigation in clinical trials.

Does guasha offer hepatoprotective effect to chronic inactive hepatitis B carriers? A built-in design to control subject expectation.

BACKGROUND: A previous case study showed that Guasha, an ancient manual therapeutic technique, could exert hepatoprotective effect in a human chronic active hepatitis B carrier (active-CHB) by modulating the liver enzymes, cytokines, and heme oxygenase-1 (HO-1). The present study serves as a control to the aforementioned case report. The controls were chronic inactive carriers (inactive-CHB) and noncarriers of hepatitis B (NCs). Besides showing a difference in biochemical markers between controls and the previously reported active-CHB case, the asymptomatic condition in both inactive- and active-CHB offers an excellent control for the patient's expectation about Guasha's efficacy. The purpose of this case study was to investigate whether hepatoprotective biochemical markers previously measured in active-CHB in response to Guasha were also present in controls. PARTICIPANTS AND METHODS: Four inactive-CHB and nine NC participants were included. Each participant received a 15-minute Guasha treatment. Blood samples were obtained immediately before Guasha (day 0) and after Guasha (days 2, 5, and 7). Biochemistry values for liver function, HO-1, and T-helper (Th) cytokines were determined from blood tests. Neither the participants nor the investigator who administered Guasha were aware of the blood test results until after all data were collected for all participants. RESULTS: In both inactive-CHB and NC participants, liver function, serum HO-1, and Th1/Th2 cytokines did not significantly differ before and after Guasha. CONCLUSIONS: In contrast to results in active-CHB patients, Guasha did not induce any significant modulation of liver enzymes, HO-1, or cytokines in inactive-CHB and NC participants. The current results suggest that a Guasha-induced hepatoprotective effect depends on the inflammatory event or clinical stage of chronic hepatitis B. Because both active and inactive carriers were completely unaware of their liver status at the time of receiving Guasha, the research protocol is effective in discounting the model that attributes the Guasha therapeutic efficacy to a placebo effect due to participants' expectations.

Urinary glycosaminoglycans and glycoproteins in a calcium oxalate crystallization system.

This study measures the effects of total urinary glycosaminoglycans (GAGs), glycoproteins (GPs) and individual GAGs on the nucleation rates (Bo), growth rates (G) and suspension densities (Mт) of calcium oxalate (CaOx) crystallization by the mixed suspension mixed product removal (MSMPR) system. Total urinary GAGs, glycoproteins and individual GAGs including heparan sulfate (HS), chondroitin sulfate (CS) and Hyaluronic acid (HA) were added into the artificial urine (AU) and then introduced into the MSMPR test chamber and the crystal sizes and numbers were analyzed by a particle counter. The effects of added GAGs and GPs on CaOx crystallization were reflected by the changes on the crystallization indexes including the Bo, G and Mт of CaOx that were calculated based on the crystal size and numbers. Total urinary GAGs showed no statistical significance on CaOx crystallization. However, individual GAGs such as HA, CS and HS enhanced Bo and suppressed the G when measured individually. CS and HS enhanced the Mт while HA shown no significant change in the Mт of CaOx. Total urinary GPs showed an increase in the G and Mт of crystals. Although total urinary GAGs showed no statistically significant effect on CaOx crystallization, individual GAGs (CS, HS) promoted the CaOx crystallization by increasing the suspension density of smaller crystals, indicative of reduced risk of stones while HA showed no significance in the M(T) of CaOx formed. Urinary GPs indicated increased sizes and M(T) suggesting larger crystals and/or aggregates.

Nutritional and immunological characteristics of fresh and refrigerated stored human milk in Hong Kong: a pilot study.

BACKGROUND: The nutritional and immunological qualities of human milk from Hong Kong mothers were profiled. METHODS: A total of 25 colostrum (≤ 3 days postpartum) and 11 mature (30-45 days postpartum) milk specimens were collected from healthy Chinese women in Hong Kong. Parameters including total protein, whey, casein, triglycerides, lactose, lysozyme, secretory immunoglobulin A, lactoferrin and antibacterial activity of human milk were quantified. RESULT: Breast milk of Hong Kong Chinese mothers is nutritionally comparable to that of western mothers for colostrum and mature milk, with protein (25.0±11.3 g/l vs 10.1±1.4 g/l) and lactose (44.2±7.5 g/l vs 66.6±5.5 g/l) for growth and energy supply in infants, respectively. The milk of Hong Kong mothers is however characterized by its exceptionally high levels of sIgA (806.3±792.6 mg/dl and 1545.9±334.6 mg/dl) and low levels of triglycerides (10.8±9.3 mmol/l and 11.1±7.7 mmol/l). The human milk in Hong Kong was also shown to be superior to bovine formula milk, particularly in terms of lower total protein level, the high whey to casein protein ratio, rich immunological active contents, and significant antibacterial activity against 2 common Escherichia coli strains. CONCLUSIONS: Preliminary data also indicated that mature milk could be refrigerated for at least 3 days in order to maintain the major nutritional constituents and antibacterial activity.

Guasha-induced hepatoprotection in chronic active hepatitis B: a case study.

BACKGROUND: Heme oxygenase-1 (HO-1) has demonstrated hepatoprotective effect in animal hepatitis models. HO-1 was also reported to be upregulated with Guasha, an ancient therapeutic technique which applies instrument assisted press-stroking to treat many disorders. METHODS: We report a case on the changes of liver function, plasma HO-1 and T-helper (Th) cytokine balance in a chronic active hepatitis B carrier before and after Guasha. The patient presented with increased activities of liver enzymes (ALT and AST), indicating inflammatory damage in liver before Guasha. RESULTS: Forty-eight hours after receiving Guasha, the patient showed changes in a number of serum markers: a decline of liver enzymes (ALT and AST) indicating reduced chronic inflammation, an elevated plasma HO-1, and a modulation of T-helper (Th)1/Th2 balance. CONCLUSIONS: Guasha was shown to transiently reduce the inflammatory markers of liver injury in human, together with an enhancement of HO-1 which might be responsible for the hepatoprotective action.

The initial and subsequent inflammatory events during calcium oxalate lithiasis.

BACKGROUND: Crystallization is believed to be the initiation step of urolithiasis, even though it is unknown where inside the nephron the first crystal nucleation occurs. METHODS: Direct nucleation of calcium oxalate and subsequent events including crystal retention, cellular damage, endocytosis, and hyaluronan (HA) expression, were tested in a two-compartment culture system with intact human proximal tubular HK-2 cell monolayer. RESULTS: Calcium oxalate dihydrate (COD) was nucleated and bound onto the apical surface of the HK-2 cells under hypercalciuric and hyperoxaluric conditions. These cells displayed mild cellular damage and internalized some of the adhered crystals within 18h post-COD-exposure, as revealed by electron microscopy. Prolonged incubation in complete medium caused significant damage to disrupt the monolayer integrity. Furthermore, hyaluronan disaccharides were detected in the harvested media, and were associated with HAS-3 mRNA expression. CONCLUSION: Human proximal cells were able to internalize COD crystals which nucleated directly onto the apical surface, subsequently triggering cellular damage and HAS-3 specific hyaluronan synthesis as an inflammatory response. The proximal tubule cells here demonstrate that it plays an important role in facilitating urolithiasis via endocytosis and creating an inflammatory environment whereby free hyaluronan in tubular fluid can act as crystal-binding molecule at the later segments of distal and collecting tubules.

Telomerase-associated apoptotic events by mushroom ganoderma lucidum on premalignant human urothelial cells.

The chemopreventive effects of Ganoderma lucidum was tested, using a tumorigenic transformable human urothelial cell (HUC-PC) model. These in vitro data show that G. lucidum can inhibit the viability and growth of HUC-PC. This could be explained by a concomitant induction of apoptosis and inhibition of telomerase activity. Significant exteriorization of phosphatidylserine was detected by Annexin-V on cell surface, and the cells subsequently lost membrane integrity for uptake of 7-amino-actinomycin D dye. Additionally, the levels of hydrogen peroxide and 8-hydroxy-2'-deoxyguanosine (8-OHdG) production of the apoptotic cells were significantly increased. The induction of apoptosis and suppression of telomerase activity help to explain the anti-HUC-PC growth properties; however, the induction of oxidative stress requires further study. This study strongly suggests that G. lucidum is a potential source of chemopreventive agents for bladder cancer based on its effectiveness in controlling the premalignant urothelial cell growth and carcinogen-induced transformation.

Mechanism of pajama material on stratum corneum water content under mild cold conditions: explored by hierarchical linear regression.

OBJECTIVE: The aim of this study was to explore the effect of fabric material worn during nocturnal sleep on stratum corneum water content (SCWC) under mild cold conditions. METHOD: A controlled cross-over study has been designed to investigate systematically the effects of two kinds of pajama materials, cotton and polyester, on SCWC. Factors such as pajama fabric material, subjective perception and sleep quality were examined. Hierarchical linear regression was applied to assess the association between the factors and SCWC. RESULTS: Fabric material and subjective perception of coldness significantly predicted SCWC in the wear trial. Polyester fabric had a negative effect on SCWC compared with cotton, and subjective perception of coldness also had a negative effect on SCWC. CONCLUSION: The mechanisms by which pajama fabrics affect the SCWC could be due to the different thermal/hygroscopicity properties of the polyester and cotton fabric and the integral involvement of physiological and neurophysiological processes.

Chinese herbal medicines and their efficacy in treating renal stones.

In herbal treatment of kidney stones, antilithics are used to "dissolve" the stones or aid their passing to guard against further retention. Diuretic action is also needed to increase the amount of fluid going through the kidneys and flush out the deposits. Previous clinical studies have shown that herbal medicines and their concoctions could be used to inhibit calcium oxalate crystallization. However, the pharmacodynamics and in-vitro effects of such medicines have not been established. Five Chinese herbal medicines were selected based on their usefulness in treating stone disease. A 96-well plate oxalate-induced turbidity in artificial urine was used to evaluate the efficacies of the different herbal medicines on calcium oxalate crystallization. The metastable limit was determined and the nucleation rate was derived from 12-min time-course measurement of turbidity at 405 nm. Phase-contrast microscopy was used to visualize the crystals. The results showed that with increasing concentrations of herbal extracts, smaller calcium oxalate crystal sizes were observed. Overall, the five herbal medicinal extracts tested were able to promote nucleation of calcium oxalate crystals while at the same time decreasing the size. This in-vitro crystallization confirms that prophylaxis of renal stones could be achieved by reducing overall supersaturation through promotion of small crystal nucleates and concomitant pharmacological diuretic action of herbal medicines. Clinical studies will provide more definitive conclusions.

Anticancer effects of Ganoderma lucidum: a review of scientific evidence.

"Lingzhi" (Ganoderma lucidum), a popular medicinal mushroom, has been used in China for longevity and health promotion since ancient times. Investigations into the anticancer activity of lingzhi have been performed in both in vitro and in vivo studies, supporting its application for cancer treatment and prevention. The proposed anticancer activity of lingzhi has prompted its usage by cancer patients. It remains debatable as to whether lingzhi is a food supplement for health maintenance or actually a therapeutic "drug" for medical proposes. Thus far there has been no report of human trials using lingzhi as a direct anticancer agent, despite some evidence showing the usage of lingzhi as a potential supplement to cancer patients. Cellular immune responses and mitogenic reactivity of cancer patients have been enhanced by lingzhi, as reported in two randomized and one nonrandomized trials, and the quality of life of 65% of lung cancer patients improved in one study. The direct cytotoxic and anti-angiogenesis mechanisms of lingzhi have been established by in vitro studies; however, clinical studies should not be neglected to define the applicable dosage in vivo. At present, lingzhi is a health food supplement to support cancer patients, yet the evidence supporting the potential of direct in vivo anticancer effects should not be underestimated. Lingzhi or its products can be classified as an anticancer agent when current and more direct scientific evidence becomes available.