High-performance Liquid Chromotography Research Articles

Isolate and Diagnose Negative Bacteria that Cause Urinary Tract Inflammation and Test their Sensitivity to Certain Antibiotics and Rheum Palmatum

The current study included the collection of 50 diarrhoea samples from UTIs, ages 25-50 years, 60% female and 40% male patients reviewed to Kirkuk city laboratories, gave 40 positive germ growth samples and 80%, while 10 other samples gave no germ growth. Isolation and diagnosis results showed that E.coli was prevalent on Proteus mirabilis and at 75%, while Proteus mirabilis was at 25% lower. The results of the study showed isolated bacterial sensitivity to a number of antibiotics and the results showed isolated bacterial sensitivity to anti-Ciprofloxacin while bacterial isolates showed the highest resistance to antibiotics. Effective compounds were identified and their concentration in the Riwas plant was determined using High Performance Liquid Chromotography HPLC (containing phenolic, turbine and alkaloid compounds, aquatic and alcoholic extracts were tested at concentrations (25), (50) and (100) and the focus (25) was on the lowest toxins.

Reproductive endocrinology, morphological traits, and sexual selection in a population of wild South African giraffes

Sex steroids are pervasive in mammals and evolutionarily conserved, but differences in the nuances of endocrine profiles characterize distinct species. Two sex steroids, testosterone and progesterone, feature prominently in the life history of mammalian taxa, but neither one has been analyzed from wild giraffes. Our study was designed to address this gap in knowledge by examining how these sex steroids are related to biological features and giraffe life history. We conducted the research at Rooipoort Nature Reserve a 44,000 ha private nature reserve in South Africa on a population of South African giraffes (Giraffa camelopardalis giraffa). Eleven adult giraffe cows and seven adult bulls were immobilized and various biological samples and morphological measurements were obtained. We analyzed both testosterone and progesterone using Ultra High Performance Liquid Chromotography – Tandem Mass Spectrometry (UHPLC-MS/MS). We found that non-pregnant females had lower progesterone concentrations than pregnant females and that those in the last trimester of pregnancy showed a slight drop in progesterone, as well as an increase in testosterone. Among males, chronological age was correlated with testicle size, testosterone concentrations, and ossicone volume. We propose that the progesterone decline functions partly to accelerate resumption of ovulation because giraffes become pregnant while lactating, and that the testosterone elevation provides an endocrine mileu for female defense of neonatal calves, given that lion predation is a major threat to calf survival. We suggest that male reproductive strategies are mediated by the age-related impacts of testosterone on growth in both body mass and ossicone volume as a consequence of sexual selection. We conclude by noting that the robust and solid ossicones of male giraffes function in mate competition as wedges that are used to topple opponents by raising their legs and placing them in an off-balanced position that can seriously injure them when falling to the ground.

Characterization of phenolic compounds and antioxidant activity in sorghum [Sorghum bicolor (L.) Moench] grains

Among cereals, sorghum, a gluten-free cereal is a rich source of bioactive polyphenols and dietary antioxidants. Sorghum polyphenolic compounds were characterized and quantified in five sorghum genotypes by high performance liquid chromatography. Grain samples of five sorghum genotypes were grounded and subjected top extraction for profile of polyphenols via High Performance Liquid Chromotography. Results showed that red and brown pericarp genotypes had higher total proanthocyanidin and total phenolic content, therefore, resulting in overall high antioxidant activities. The phenolic acids that showed the higher antioxidant activity in red (SSG 59-3) and brown (G-46) genotypes were caffeic acid, taxifolin and apigeninidin. SSG 59-3 (20.55 ± 0.11a, 45.66 ± 0.23b, 15.34 ± 0.10c) had higher antioxidant activity evaluated by DPPH, ABTS and FRAP assay. The concentrations of the sorghum-specific 3-deoxyanthocyanidins luteolinidin and apigeninidin were higher in red sorghum. Correlation analysis showed that antioxidant activity had a linear relationship with their polyphenols. Thus, our findings indicated that there is cumulative effect of both high antioxidant activity and phenolic compounds which may be exploited for human food to improve the nutritional value and health properties.

Periferik Vasküler Hastalığın Tedavisinde İloprost’tun Nitrik Oksit, Asimetrik Dimetilarjinin ve Serotonin’e Etkisi

Aim: In this study, it is aimed to investigate the efficacy of iloprost use in peripheral artery disease patients on the parameters involved in endothelial functions such as circulating acetyl dimethyl arginine (ADMA), serotonin and nitric oxide (NO).
 Patients and Methods: 30 patients (19 male, 11 female, age interval 60.7 ± 13.7) who were followed-up with the diagnosis of Fontaine III-IV disease and decided to receive a medical treatment were included in the study. Iloprost infusion was initiated as a 16-hour intravenous infusion at the dose of 0.5-1.5 ng / kg / min from the forearm veins and was given for 7 days. Blood was taken for ADMA, Serotonin and NO results before and after the treatment (8th day). Total nitrite (nitrite + nitrate) concentration was measured by the modified cadmium reduction method, ADMA and serotonin levels were measured by High Performance Liquid Chromotography (HPLC) method.
 Results: ADMA value before and after the iloprost treatment was decreased after the treatment and while this was statistically significant (p=0.001), the changes in serotonin (p=0.82) and NO (P= 0.16) values were found statistically insignificant. 
 Conclisions: Peripheral arterial disease may be a parameter that can be taken into account in the evaluation of treatment for peripheral arterial disease and ADMA, which is considered to be an endothelial dysfunction indicator after iloprost treatment.

Analysis of flavonoids in Vernonia paltula by high-performance liquid chromotography

Lin-nan-yeh-chu, is the dried entire plant of Vernonia paltula (Compositae) and used as folk medicine in Taiwan. To evaluate the quality of V. paltula, a simple, rapid and accurate high-performance liquid chromatographic (HPLC) method was developed for the assay of four flavonoids apigenin (API), apigenin-7-O-glucoside (APG), luteolin (LUT) and luteolin-7-O-glucoside (LUG). The present HPLC system uses an Inertsil ODS-2 column by gradient elution with acetonitrile and 0.1% (v/v) phosphoric acid as the mobile phase. Ethyl paraben was used as an internal standard and detected at 254 nm. Regression equations revealed good linear relationships (correlation coefficients: 0.9998-0.9999) between the peak-area ratios of each constituent to ethyl paraben. The recovery of four marker constituents ranged from 89.3 to 95.6%. The contents of the four constituents in stem, flower, leaf and root parts of V. paltula have been compared. Leaf part consisted of the highest contents of flavonoids except for APG which is less than that in the flower. The root and stem only showed trace amount of APG but not the other three flavonoids. The contents of four constituents were 0.6016, 0.0042, 0.2160 and 0.0577 mg/g for apigenin-7-O-glucoside, apigenin, luteolin-7-O-glucoside and luteolin, respectively. HPLC chromatograms of another three plants of genus of Vernonia in Taiwan, V. cinerea, V. elliptica and V. gratiosa, have also been compared.

دراسة تأثير البايسنوجينول على سكر الكلوكوز والدهون في المرضى المصابين بالنوع الثاني للسكري.

تم إستخلاص البايسنوجينول (المستخلص المائي المتكون من المركبات متعددة الفينول) من لحاء الصنوبر عراقي المنشأ وتم تحليله باستخدام كروموتوغرافيا السائل ذات الكفاءة العالية المترافقة مع طيف الاشعة فوق البنفسجية عند الطول الموجي (254) نانومتر. إن تأثير البايسنوجينول على بعض المتغيرات الكيميائية الحياتية (الكلوكوز و الكوليستيرول الكلي و الكليسيريدات الثلاثية و البروتينات الدهنية عالية الكثافة و البروتينات الدهنية قليلة الكثافة و البروتينات الدهنية ضئيلة الكثافة) تم تعيينه في (30) مريضا مصابا بالسكري والذين تمت معاملتهم بعينات دوائية (500 ملغم) من البايسنوجينول ثلاث مرات يوميا لاختبار الدور الفعال للبايسنوجينول ضد شدة الاكسدة.إن المحددات الكيميائية أعلاه تم قياسها في البلازما قبل العلاج بالبايسنوجينول وبعد مرور (أسبوع) و(أسبوعين) و (ثلاثة أسابيع) من العلاج بهذا المستخلص متعدد الفينول. إن نتائجنا أظهرت أن مستوى الكلوكوز يزداد نسبيا لدى مرضى السكري بالمقارنة مع مجموعة السيطرة (p<0.001 ) ولكن بعد المعاملة بالبايسنوجينول فإن مستوى الكلوكوز ينخفض نسبيا (p<0.001) بعد أسبوع و p<0.001)) بعد أسبوعين و p<0.05)) بعد ثلاثة أسابيع من العلاج بالبايسنوجينول. إن مستوى الكوليسترول الكلي يرتفع نسبيا لدى مرضى السكري بالمقارنة مع مجموعة السيطرة (p<0.001 ) ولكن بعد العلاج بهذا المركب الفعال فإن مستوى الكوليسترول الكلي ينخفض نسبيا (p<0.001) بعد أسبوع و p<0.001)) بعد أسبوعين و p<0.01)) بعد ثلاثة أسابيع من العلاج . إن مستوى الكليسيريدات الثلاثية يزداد نسبيا لدى المصابين بالسكري مقارنة مع مجموعة السيطرة (p<0.001 ) ولكن بعد المعاملة بهذا المستخلص المضاد للاكسدة فإن مستوى الكليسيريدات الثلاثية ينخفض نسبيا (p<0.001) بعد أسبوع و p<0.001)) بعد أسبوعين و p<0.01)) بعد ثلاثة أسابيع من العلاج . إن مستوى البروتينات الدهنية عالية الكثافة ينخفض نسبيا لدى المصابين بالسكري بالمقارنة مع مجموعة السيطرة (p<0.001 )ولكن بعد العلاج بالبايسنوجينول فإن مستوى البروتينات الدهنية عالية الكثافة يزداد نسبيا (p<0.01) بعد أسبوع و p<0.01)) بعد أسبوعين و p<0.05)) بعد ثلاثة أأأاااسابيع من العلاج بالبايسنوجينول. إن مستوى البروتينات الدهنية قليلة الكثافة يزداد نسبيا لدى المصابين بالسكري بالمقارنة مع مجموعة السيطرة (p<0.001 )ولكن بعد العلاج بمستخلص البايسنوجينول فإن مستوى البروتينات الدهنية قليلة الكثافة ينخفض نسبيا (p<0.001) بعد أسبوع و p<0.001)) بعد أسبوعين و p<0.01)) بعد ثلاثة أسابيع من العلاج بالبايسنوجينول. إن مستوى البروتينات الدهنية ضئيلة الكثافة يزداد نسبيا لدى المصابين بالسكري بالمقارنة مع مجموعة السيطرة (p<0.001 )ولكن بعد العلاج بالبايسنوجينول فإن مستوى البروتينات الدهنية ضئيلة الكثافة ينخفض نسبيا (p<0.001) بعد أسبوع و p<0.001)) بعد أسبوعين و p<0001)) بعد ثلاثة أسابيع من العلاج بالبايسنوجينول.

Comparison of two different methods (turbidimetric inhibition inmunoassay and high performance liquid chromotography) for determination of HbA1c

Comparison of two different methods (turbidimetric inhibition inmunoassay and high performance liquid chromotography) for determination of HbA1c

High asymmetric dimethylarginine, symmetric dimethylarginine and L-arginine levels in migraine patients

Experimental and clinical data strongly suggests that nitric oxide (NO) plays a pivotal role in migraine. This is also supported by studies of migraine induced by substances that release NO. NO is synthesized from L-arginine by endothelial NO synthase (NOS). Asymmetric dimethylarginine (ADMA) is the major endogenous competitive inhibitor of NOS. Symmetric dimethylarginine (SDMA) is an inactive stereoisomer of ADMA. It may reduce NO production by competing with arginine for cellular uptake. The aim of this study was to measure the levels of ADMA, SDMA and L-arginine in migraine patients during the interictal period. One hundred migraine patients and 100 healthy volunteers were recruited. The patients were in the interictal period and classified into two groups as having migraine with aura and migraine without aura. Their serum ADMA, SDMA and L-arginine levels were measured by high-performance liquid chromotography (HPLC) method. ADMA, SDMA and L-arginine levels were significantly higher in migraine patients compared to the control group. But there was no difference between the patients with and without aura. These results suggest that NOS inhibitors and L-arginine/NO pathway plays an important role in migraine pathopysiology.

Non-polar lipids accumulate during storage of transfusion products and do not contribute to the onset of transfusion-related acute lung injury.

The accumulation of non-polar lipids arachidonic acid, 5-hydroxyeicosatetraenoic acid (HETE), 12-HETE and 15-HETE during storage of transfusion products may play a role in the onset of transfusion-related acute lung injury (TRALI), a syndrome of respiratory distress after transfusion. We investigated non-polar lipid accumulation in red blood cells (RBCs) stored for 42 days, plasma stored for 7 days at either 4 or 20°C and platelet (PLT) transfusion products stored for 7 days. Furthermore, we investigated whether transfusion of RBCs with increased levels of non-polar lipids induces TRALI in a 'two-hit' human volunteer model. All products were produced following Dutch Blood Bank protocols and are according to European standards. Non-polar lipids were measured with high-performance liquid chromotography followed by mass spectrometry. All non-polar lipids increased in RBCs after 21 days of storage compared to baseline. The non-polar lipid concentration in plasma increased significantly, and the increase was even more pronounced in products stored at 20°C. In platelets, baseline levels of 5-HETE and 15-HETE were higher than in RBCs or plasma. However, the non-polar lipids did not change significantly during storage of PLT products. Infusion of RBCs with increased levels of non-polar lipids did not induce TRALI in LPS-primed human volunteers. We conclude that non-polar lipids accumulate in RBC and plasma transfusion products and that accumulation is temperature dependent. Accumulation of non-polar lipids does not appear to explain the onset of TRALI (Dutch Trial Register - NTR4455).

Some rape/canola seed oils: fatty acid composition and tocopherols.

Seed samples of some rape and canola cultivars were analysed for oil content, fatty acid and tocopherol profiles. Gas liquid chromotography and high performance liquid chromotography were used for fatty acid and tocopherol analysis, respectively. The oil contents of rape and canola seeds varied between 30.6% and 48.3% of the dry weight (p<0.05). The oil contents of rapeseeds were found to be high compared with canola seed oils. The main fatty acids in the oils are oleic (56.80-64.92%), linoleic (17.11-20.92%) and palmitic (4.18-5.01%) acids. A few types of tocopherols were found in rape and canola oils in various amounts: α-tocopherol, γ-tocopherol, δ-tocopherol, β-tocopherol and α-tocotrienol. The major tocopherol in the seed oils of rape and canola cultivars were α-tocopherol (13.22-40.01%) and γ-tocopherol (33.64-51.53%) accompanied by α-T3 (0.0-1.34%) and δ-tocopherol (0.25-1.86%) (p<0.05). As a result, the present study shows that oil, fatty acid and tocopherol contents differ significantly among the cultivars.

Development of Solid-Phase Extraction Using Molecularly Imprinted Polymer for the Analysis of Organophosphorus Pesticides-(Chlorpyrifos) in Aqueous Solution

A new and selective sorbent for molecularly imprinted solid-phase extraction (MISPE) was prepared to extract chlorpyrifos (CPF) residue from solutions. The extracted analyte was analyzed by high performance liquid chromotography (HPLC) coupled with photodiode array detection. To synthesize the molecularly imprinted polymers, four different pyrogens (acetonitrile, toluene, dichloromethane and chloroform) were initially studied. CPF was used as the template molecule, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the crosslinker. Thermo-polymerization method was used to produce bulk polymers. In order to determine the medium that enhances the best molecular recognition, the adsorption study of CPF to the MIPs was investigated. Both organic solvents and water were utilized as media. The acetonitrile solvent was finally selected as pyrogen for the synthesis of the polymers and water was chosen as the medium for loading the analytes into the polymers. The selectivity of the MISPE method for CPF and other pesticides in aqueous solution was also assessed.

Therapeutic Efficacy of Nigella Sativa Linn. against Antituberculosis Drug-Induced Hepatic Injury in Wistar Rats

Antituberculosis drug (ATD)-induced hepatotoxicity is a major impediment for the effective treatment of tuberculosis (TB). All first-line anti-TB medications have adverse effects that interrupt the successful completion of TB treatment. This investigation focuses on the evaluation of the protective role of Nigella sativa (NS) against liver injury caused by ATDs. Female rats were treated with ATDs for 8 weeks (3 d/wk) followed by NS for 8 weeks (3 d/wk). The antioxidant activity of NS was estimated with a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and by analyzing total phenolic contents. Qualitative characterization of active compounds of the plant was done by high-performance liquid chromotography (HPLC). ATD-induced adverse effects were associated with sharp elevation in levels of serum transaminases, albumin, cholesterol, urea, uric acid, creatinine, and blood urea nitrogen (BUN). ATDs significantly increased lipid peroxidation (LPO) and decreased enzyme activities (superoxide dismutase [SOD], catalase [CAT], adenosine triphosphatase [ATPase], and glucose-6-phosphatase [G6Pase]) in liver, indicating oxidative stress. Conjoint treatment with NS could reverse the serological biochemistry and inhibit oxidative stress by suppressing LPO and augmenting antioxidant enzyme activity toward that of the control. Histological studies support the above biochemical findings. Results indicate that NS exerts excellent hepatoprotective abilities and can be used as a supplement to improve patient adherence and reduce interruptions in treatment due to ATD-related liver injury.

Pharmacokinetics of IV Ketorolac in Horses Undergoing Orchiectomy

Ketorolac (KET) is a nonsteroidal anti-inflammatory drug for human use, with a potent analgesic activity, that is used in the relief of moderate-to-severe postoperative pain. The pharmacokinetics of KET tromethamine was evaluated after single IV injection at 0.5 mg/kg body weight, after intubation and 10 minutes before surgery, to six Arabian colts undergoing orchiectomy. Intraoperative cardiorespiratory variables were monitored. Blood samples were collected for 36 hours, and serum samples were analyzed by high performance liquid chromotography with ultraviolet-visible detection. During surgery, all monitored physiological parameters were stable. Intermittent positive pressure ventilation, and normocapnia were maintained throughout the procedure in all animals. No adverse effects were observed. The kinetics of KET was described by a two-compartment models, and also a noncompartmental analysis was performed. The distribution and elimination half-lives were t1/2λ1 0.06 ± 0.02 and t1/2λ2 0.59 ± 0.21 hours, respectively. Body clearance and mean residence time were 339.99 ± 120.19 mL/h/kg and 0.49 ± 0.22 hours, respectively. The volume of distribution at steady state and volume of distribution based on the terminal phase were 218.83 ± 134.26 mL/kg and 522.5 ± 529.3 mL/kg, respectively. The serum protein binding was 75.8 ± 2.9%. The results indicate that KET at 0.5 mg/kg IV was very rapidly eliminated and thus was likely not effective in the postoperative period. However, further studies including a control group and at higher doses are suggested to investigate the KET kinetics and the analgesic efficacy in horse and define the most appropriate dosage scheme.

Ochratoxin A levels in blood serum of Czech women in the first trimester of pregnancy and its correspondence with dietary intake of the mycotoxin contaminant

The mycotoxin ochratoxin A (OTA) can elicit a wide range of toxic properties including embryotoxicity and teratogenicity. OTA crosses the placenta at early gestation rather than in late gestation, maternal OTA exposure may represent a risk for the developing fetus. The study focuses on the assessment of OTA intake of pregnant women (aged 19–40 years) in the first trimester of pregnancy by means OTA levels in 100 blood serum samples by high-performance liquid chromotography with fluorescence detection (HPLC-FD) method and comparison with dietary OTA exposure in pregnant women. Of all, 96% tested serum samples were positive with values ranging from 0.1 to 0.35 µg/l with a mean value of 0.15 µg/l.

Herbicide Resistance Endowed by Enhanced Rates of Herbicide Metabolism in Wild Oat (Avenaspp.)

The biochemical basis of resistance to the acetyl-coenzyme A carboxylase (ACCase)-inhibiting herbicide diclofop-methyl was investigated in a resistant wild oat population (R1), which does not exhibit a resistant ACCase. Rates of foliar uptake and translocation of [14C]-diclofop were the same in the R1 vs. susceptible (S) populations. However, the level of phytotoxic diclofop acid was always found to be lower in the R1 vs. S plants, with a concomitant higher level (up to 1.7-fold) of nontoxic polar diclofop metabolites in R1 relative to the S plants. These results indicate that a non–target-site-based mechanism of enhanced rate of diclofop acid metabolism confers resistance in population R1. Moreover, the high-performance liquid chromotography elution profile of the major diclofop metabolites in R1 is similar to that of wheat, suggesting resistance in individuals of population R1 involves a wheat-like detoxification system mediated by cytochrome P450 monooxygenases. In addition, lower level of tissue diclofop acid was also observed using nonradioactive ultra-performance liquid chromatography–mass spectrometry analysis in resistant individuals of three other resistant wild oat populations (R2, R3, and R4) known to posses ACCase gene resistance mutations. These results establish that either one or at least two independent resistance mechanisms (target-site ACCase resistance mutations and non–target-site enhanced rates of herbicide metabolism) can be present in individual wild oat plants.

Effects of highly hygroscopic excipients on the hydrolysis of simvastatin in tablet at high relative humidity

Effects of highly hygroscopic sorbitol, citric acid, sodium carboxymethyl cellulose or polyvinylpolypyrrolidone, on the hydrolysis of simvastatin in tablets at 25°/90% RH were studied. The simvastatin tablets were prepared by direct powder compression. Simvastatin and its hydrolyte, simvastatin acid, were quantitatively analysed by high performance liquid chromotography. The hygroscopicity, water swelling ratio, water solubility and pH of the four hygroscopic excipients were investigated. During the investigation period, the weight gain of sorbitol or citric acid increased faster than that of polyvinylpolypyrrolidone or sodium carboxymethyl cellulose at 25°/90% RH, accordingly, the moisture sorption of the tablets containing citric acid or sorbitol (T-3 or T-6) were more than that of the tablets containing sodium carboxymethyl cellulose or polyvinylpolypyrrolidone (T-4 or T-5). The increase of simvastatin acid content with time at 25°/90% RH for the tablets was in the following order: T-6 < T-4 < T-3 < T-5. The effects of the four excipients on the hydrolysis of simvastatin in tablet were related to not only their hygroscopicity but also their other properties, such as moisture retention capacity and pH. Sorbitol as hygroscopic excipient in tablet can most effectively prevent the hydrolysis of simvastatin in tablet.

Hydrolysis of Whey Protein Isolate Using Subcritical Water

Hydrolyzed whey protein isolate (WPI) is used in the food industry for protein enrichment and modification of functional properties. The purpose of the study was to determine the feasibility of subcritical water hydrolysis (SWH) on WPI and to determine the temperature and reaction time effects on the degree of hydrolysis (DH) and the production of peptides and free amino acids (AAs). Effects of temperature (150 to 320 °C) and time (0 to 20 min) were initially studied with a central composite rotatable design followed by a completely randomized factorial design with temperature (250 and 300 °C) and time (0 to 50 min) as factors. SWH was conducted in an electrically heated, 100-mL batch, high pressure vessel. The DH was determined by a spectrophotometric method after derivatization. The peptide molecular weights (MWs) were analyzed by gel electrophoresis and mass spectrometry, and AAs were quantified by high-performance liquid chromotography. An interaction of temperature and time significantly affected the DH and AA concentration. As the DH increased, the accumulation of lower MW peptides also increased following SWH (and above 10% DH, the majority of peptides were <1000 Da). Hydrolysis at 300 °C for 40 min generated the highest total AA concentration, especially of lysine (8.894 mg/g WPI). Therefore, WPI was successfully hydrolyzed by subcritical water, and with adjustment of treatment parameters there is reasonable control of the end-products.

Simultaneous determination of bufadienolides in the traditional Chinese medicine preparation, liu-shen-wan, by liquid chromatography.

The bufadienolide compounds (bufalin, cinobufagin and resibufogenin), major constituents of Chansu in Liu-Shen-Wan (LSW), were determined by reverse phase high performance liquid chromatography. The procedure involves a preliminary extraction of the bufadienolides from LSW with chloroform using ultrasonication and subsequent evaporation to dryness of the chloroform extract. The residue of the chloroform extract was dissolved in methanol and separated on a Merck LiChrosorb RP-18 column. Methanol: water (74:26) was used as mobile phase. The compounds were satisfactorily separated with good chromatographic peaks. Good coefficients of correlation (r > 0.999) were obtained from the calibration of peak areas with concentrations for the 3 bufadienolides. Results of analysis showed that there were differences between the contents of bufadienolides in 11 LSW samples of different origin available to the public in Hong Kong where at present there is no legal control over the sale of traditional Chinese medicines. The variability of quantities of bufadienolides in Chansu may be a hazard to the public.

Histatins are the major wound‐closure stimulating factors in human saliva as identified in a cell culture assay

Wounds in the oral cavity heal much faster than skin lesions. Among other factors, saliva is generally assumed to be of relevance to this feature. Rodent saliva contains large amounts of growth factors such as epidermal growth factor (EGF) and nerve growth factor (NGF). In humans, however, the identity of the involved compounds has remained elusive, especially since EGF and NGF concentrations are approximately 100,000 times lower than those in rodent saliva. Using an in vitro model for wound closure, we examined the properties of human saliva and the fractions that were obtained from saliva by high-performance liquid chromotography (HPLC) separation. We identified histatin 1 (Hst1) and histatin 2 (Hst2) as major wound-closing factors in human saliva. In contrast, the d-enantiomer of Hst2 did not induce wound closure, indicating stereospecific activation. Furthermore, histatins were actively internalized by epithelial cells and specifically used the extracellular signal-regulated kinases 1/2 (ERK1/2) pathway, thereby enhancing epithelial migration. This study demonstrates that members of the histatin family, which up to now were implicated in the antifungal weaponry of saliva, exert a novel function that likely is relevant for oral wound healing.

A comparison of three methods of measuring plasma glycosylated hemoglobin

Background: The current clinical dilemma of diabetes mellitus is prevention of micro- and macrovascular complications. These complications are directly linked to the degree of hyperglycaemia in diabetics. For long-term control of the glycaemic state, measurement of glycohaemoglobin in blood is essential, and glycohaemoglobin harmonization is encouraged for significant inter-laboratory variability. Objectives: To evaluate the performance of plasma glycosylated hemoglobin (HbA1c) assay on Primus PDQ and Bio-Rad D10 analyzers with respect to imprecision against the HbA1c goal, recovery and correlation to the HbA1c test on Cobas Integra 800 automated analyzer. Methods: PDQ employs high performance liquid chromotography (HPLC) boronate affinity chromatography. D10 uses HPLC cation-exchange chromatography. Cobas Integra 800 uses an immunoturbidimetric method. Imprecision studies were performed using patient samples and commercial control sera. Each sample was assayed 20 times and run every morning and afternoon for 5 consecutive days to evaluate between-run data. Recovery was assessed using a patient’s sample with low HbA1c level, admixed with another sample containing high HbA1c level. Comparison was made using 60 patient samples on PDQ and D10 using Cobas Integra 800 as the reference system. Ten venous samples from each group of patients with thalassaemia and uraemia were assayed for HbA1c on Cobas Integra 800, PDQ and D10. Results: The within-run coefficients of variation (CVs) on patient samples were 0.5 % and 0.6 % for PDQ and 1.1% and 1.3 % for D10. The between-run CVs were 1.0 % and 0.8 % for PDQ and 0.3 % and 1.2 % for D10. PDQ yielded CVs of 1.1 % and 0.7 % for level 1 (normal) and level 2(abnormal) controls respectively while D10 demonstrated CVs of 0.5 % for each level of controls. The between-run CVs for normal and abnormal controls were 1.7 % and 0.8 % respectively for PDQ, and 1.0 % and 0.7 % respectively for D10. Mean recovery for PDQ and D10 were 99.7 % and 101.7 % respectively. Both PDQ and D10 correlated well with Cobas Integra 800 [Cobas800]. HbA1c method by PDQ correlated well with that by D10. Conclusion: Both the HPLC systems demonstrated acceptable data, good recovery and are comparable with our current operating system (Cobas Integra 800). Both these systems appear to be satisfactory analytical alternatives to the Cobas Integra 800.