A Multidisciplinary Approach to the Management of Unusual Bilateral Gingival Recession Due to a Factitious Nail-Scratching Habit: A Case Report.

Self-inflicted gingival injuries have been reported in children and adolescents as a result of multiple factors such as accidental trauma, fingernail biting, sucking on objects such as pencils, or a nail-scratching habit. Management of these lesions requires a multidisciplinary approach that includes psychotherapy, behavioral counseling, and definitive treatment of oral soft tissue lesions. This paper illustrates the diagnosis and treatment of a 16-year-old male patient with a bilateral gingival recession in mandibular canines due to habitual nail scratching. A multidisciplinary approach for treatment included behavior and psychological counseling, topical application of anesthetic & antimicrobial gels, and surgical management of gingival recession defects using a modified lateral pedicle flap with and without an indigenously prepared amnion-chorion membrane. Excellent soft tissue health was found at a two-year follow-up.

The tomato EAR-motif repressor, SlERF36, accelerates growth transitions and reduces plant life cycle by regulating GA levels and responses.

Faster vegetative growth and early maturity/harvest reduce plant life cycle time and are important agricultural traits facilitating early crop rotation. GA is a key hormone governing developmental transitions that determine growth speed in plants. An EAR-motif repressor, SlERF36 that regulates various growth transitions, partly through regulation of the GA pathway and GA levels, was identified in tomato. Suppression of SlERF36 delayed germination, slowed down organ growth and delayed the onset of flowering time, fruit harvest and whole-plant senescence by 10-15 days. Its over-expression promoted faster growth by accelerating all these transitions besides increasing organ expansion and plant height substantially. The plant life cycle and fruit harvest were completed 20-30 days earlier than control without affecting yield, in glasshouse as well as net-house conditions, across seasons and generations. These changes in life cycle were associated with reciprocal changes in expression of GA pathway genes and basal GA levels between suppression and over-expression lines. SlERF36 interacted with the promoters of two GA2 oxidase genes, SlGA2ox3 and SlGA2ox4, and the DELLA gene, SlDELLA, reducing their transcription and causing a 3-5-fold increase in basal GA3/GA4 levels. Its suppression increased SlGA2ox3/4 transcript levels and reduced GA3/GA4 levels by 30%-50%. SlERF36 is conserved across families making it an important candidate in agricultural and horticultural crops for manipulation of plant growth and developmental transitions to reduce life cycles for faster harvest.

Ecological and Health Risk Assessment of Heavy Metals Bioaccumulation in Ganges Fish Near Varanasi, India.

Heavy metal contamination in river Ganga is one of the factors for deterioration in its water quality and also adds to human health risks. We designed our study to achieve a holistic approach by not only estimating the concentration of heavy metals (lead, manganese, chromium, and cadmium) in the river water at different sites based on human anthropogenic activities but also in the fishes residing in the same sites that are collected for human consumption on daily basis. We found that Ganga River in Varanasi is highly loaded with metals (PLI = 6.698). Mean concentration in water was 1.29 mg/L for Pb, 1.325 mg/L for Mn, 0.169 mg/L for Cr, and 0.161 mg/L for Cd, which were above the permissible limits stated by Environment Protection Agency (EPA) in drinking water. Fish, including exotic and invasive species, were collected from the wild and were processed for the presence of these metals in their tissues. Degree of heavy metal concentration followed liver > gills > muscles. The highest accumulation of Pb was observed in Carpio (Cyprinus carpio) liver (8.86 µg/g) and lowest in Baikari (Clupisoma garua) muscles (0.07 µg/g). Total target hazard quotient (THQ) value, i.e., hazard index (HI) showed values in following sequence: Cyprinus carpio > Oreochromis niloticus > Channa gachua > Johnius coitor > Mastacembelus armatus > Mystus tengara > Clupisoma garua. Maximum HI value was recorded in C. carpio, which is highly consumed fish by humans, hence, may be harmful to them.

Effect of gamma and Ultraviolet-C sterilization on BMP-7 level of indigenously prepared demineralized freeze-dried bone allograft.

The presence of bone morphogenetic proteins in demineralized freeze-dried bone allograft (DFDBA) are responsible for developing hard tissues in intraosseous defects. The most common mode of sterilization of bone allografts, i.e., Gamma rays, have dramatic effects on the structural and biological properties of DFDBA, leading to loss of BMPs. Ultraviolet-C radiation is a newer approach to sterilize biodegradable scaffolds, which is simple to use and ensures efficient sterilization. However, UV-C radiation has not yet been effectively studied to sterilize bone allografts. This study aimed to compare and evaluate the effectiveness of Gamma and Ultraviolet-C rays in sterilizing indigenously prepared DFDBA and assess their effect on the quantity of BMP-7 present in the allograft. DFDBA samples from non-irradiated, gamma irradiated, and UV-C irradiated groups were tested for BMP-7 level and samples sterilized with gamma and UV-C rays were analysed for sterility testing. The estimated mean BMP-7 level was highest in non-irradiated DFDBA samples, followed by UV-C irradiated, and the lowest in gamma irradiated samples. Our study concluded that UV-C rays effectively sterilized DFDBA as indicated by negative sterility test and comprised lesser degradation of BMP-7 than gamma irradiation.

CYP1A1 gene polymorphism and heavy metal analyses in benign prostatic hyperplasia and prostate cancer: An explorative case-control study.

INTRODUCTION: The prostatic disorder is associated with benign prostatic hyperplasia (BPH) and prostate cancer (CaP). Evidently, prevalent transcription factors and signaling pathways define their relationship. The etiology of the prostatic disorder is multifactorial including heavy metal toxicity like lead (Pb), Cadmium (Cd), and genetic factors. This study elucidates the association between heavy metal toxicity Pb, Cd, and CYP1A1 gene polymorphism with BPH and CaP. METHODS: a case-control study with (BPH, n = 104), (CaP, n = 58) and (controls, n =107) patients. Heavy metal Pb and Cd estimation by atomic absorption spectrophotometer. The polymorphism of the CYP1A1 T>C (rs4646903) gene was analyzed byPCR-RFLP. RESULT: Higher levels of Pb and Cd were found in BPH and CaP followed by the control group (P-value: < 0.05). Pb and Cd show a significant correlation among prostate volume in CaP. Additionally, PSA, IPSS score, and pre void volume were positively co-related with Pb in BPH patients. The posthoc test defines the level of Pb and Cd as significantly elevated in the mutant genotype, highest among homozygous mutant genotype of CYP1A1gene among BPH. In CaP, Pb is significantly higher among the homozygous mutant type of CYP1A1 gene. The risk is also influenced by smoking, tobacco, and alcohol. CONCLUSION: The heavy metal toxicity Pb and Cd were reported to raise the risk of BPH and CaP. However, a person with heavy metal toxicity especially in BPH has a high-risk genetic susceptibility to the CYP1A1 gene in the north Indian population.

Evaluation of heavy metal contaminants in prepared noodles: source allocation and health risk assessment.

In recent years, special attention has been given to emission research that led to the deposition of toxicants from road traffic. Thus, it is imperative to focus on heavy metal (HM) stressors in food items, their source contribution, and health risk assessment providing insight into their spatial role at the population level. In this study, heavy metal in the street vended noodles was studied while correlating the quality of noodle with different environmental origins. The samples were prepared using acid digestion and analysed by flame atomic absorption spectrophotometer, except Hg which was analysed by direct mercury analyser. The results showed that some heavy metals like Cr, Pb, Mn, Cd, and Hg exceed their permissible limits established by the international legislation for food products. In the noodle samples, the concentration of heavy metal ranged from < 0.1 to 0.904 mg/kg for Pb, < 0.09 to 0.843 mg/kg for Ni, < 0.004 to 0.201 mg/kg for Cd, < 0.0001 to 0.004 mg/kg for Hg, < 0.01 to 1.388 mg/kg for Cu, < 0.015 to 8.049 mg/kg for Mn, and < 0.02 to 16.514 mg/kg for Cr. Noodle samples vended on high traffic density streets are directly associated with increased HM content due to atmospheric deposition from the surrounding. Source apportionment study determines that HM contamination belongs to the same source of origin, except Cr. Based on the cluster analysis, these samples fall into three major groups that were further validated by the canonical discriminant function. Health risk prediction by Monte Carlo simulation revealed an elevated non-carcinogenic health hazard risk to consumers with a hazard index (HI) shift from 71 to 75%. Health hazard analysis showed that consumers of high traffic density street vended food are at higher risk of developing health-related issues. This study is important to evaluate the health risk of the population exposed to heavy metals due to ingestion of street vended food.

Synthesis of Biomass-Derived Activated Carbons and Their Immobilization on Alginate Gels for the Simultaneous Removal of Cr(VI), Cd(II), Pb(II), As(III), and Hg(II) from Water.

Low-cost alginate gels of activated carbons were prepared, which were derived from the peels of banana and sweet lime. The synthesized carbon was activated and immobilized on alginate, producing its gel. These gels were categorized according to their methods of drying, in which air drying, freeze drying, and supercritical drying led to the formation of xerogels, cryogels, and aerogels, respectively. The gels were used for adsorption of heavy metals from their aqueous solution. The heavy metals that were targeted for removal were Pb(II), Cd(II), Cr(VI), As(III), and Hg(II). Among all the adsorbents, the alginate cryogel of sweet lime-derived activated carbon (SLACC) showed the highest removal percentage of heavy metals, and thus, it was used for batch study. The adsorption of heavy metals by SLACC was checked at different times, pH values, adsorbent doses, temperatures, and adsorbate concentrations. The study revealed that the pseudo-second-order model best described the kinetic study, while the adsorption followed the Freundlich isotherm. SLACC showed maximum adsorption capacities (q cal) of 3.71, 4.22, 20.04, 7.31, and 4.37 mg/g for Cr, Cd, Pb, As, and Hg, respectively, when 20 mg of SLACC was used for the removal of 4 ppm concentration of the targeted heavy metals from their 20 mL solution. Based on the thermodynamic study, it was found that the adsorption was spontaneous and exothermic. Furthermore, the adsorbent was also used on real water samples and showed up to 90% removal efficiency for these targeted heavy metals. SLACC was regenerated with 0.1 M ethylenediaminetetraacetic acid (EDTA) solution and reused for five cycles, in which the percentage removal of heavy metals was more than 50% till the fourth cycle. Furthermore, the leaching study showed that no toxic elements had leached from SLACC into water, making it a safe adsorbent.

Modified DLLME-GC-TQMS determination of pesticide residues in Gomti River, Lucknow, India: ecological risk assessment and multivariate statistical approach.

This research article aims to establish an easy and well-defined analytical method for detection and quantification of multiclass pesticides in Gomti river water samples because the increased agricultural activities, industrialization, and urbanization had increased the presence of pesticides in the ecosystem which causes the depletion of water quality making it a global concern. The analytical method, vortex-assisted ultrasonication-based dispersive liquid-liquid microextraction-solidification of floating organic droplets (VAUS-DLLME-SFO) was optimized using one parameter at a time approach which gave the recovery between 69.45 and 114.15%, limit of detection (LOD), and limit of quantification (LOQ) 0.0011-0.0111 µg/L and 0.0033-0.0368 µg/L, respectively, and RSD in the range of 0.75-1.29 which shows sensitivity and accuracy better than earlier reported methods. The data obtained were subjected to measurement uncertainty, risk assessment, and multivariate statistical analysis to establish the robustness of the developed analytical method. The measurement uncertainty found was concluded to be in the acceptable range for analytical results. Furthermore, the real samples were analyzed and the associated value of the risk quotient was found to be less than 1, except for aquatic invertebrates, establishing the fact that the current concentration of pesticides has no such negative threat to flora and fauna. The possible source of pesticides in the Gomti river system was established by multivariate analysis. It was thus concluded that anthropogenic activity is responsible for the variable concentration of pesticides found in the sample.

Synthesis of an Alginate-Based Fe3O4-MnO2 Xerogel and Its Application for the Concurrent Elimination of Cr(VI) and Cd(II) from Aqueous Solution.

In this study, magnetite-manganese oxide (Fe3O4-MnO2) nanoparticles were synthesized and immobilized on alginate, producing a magnetite-manganese oxide xerogel (mMOX). This eco-friendly xerogel was used as an adsorbent of Cr(VI) and Cd(II). It was mesoporous and thermally stable, as determined by Brunauer-Emmett-Teller and thermogravimetric analysis. A scanning electron microscope coupled with an energy-dispersive X-ray system, Zetasizer, and attenuated total reflectance-Fourier transform infrared were used for characterization of adsorbents. The performance of the mMOX was investigated for the simultaneous adsorption of Cr(VI) and Cd(II) at different temperatures, pH values, contact times, initial concentrations of the adsorbate, and adsorbent doses. The developed xerogel (mMOX) showed high adsorption capacities of 3.86 mg/g for Cr(VI) and 3.95 mg/g for Cd(II) on 120 min of contact time with 5 ppm Cr(VI) and Cd(II) solution. The kinetic data fitted well with the pseudo-second order, while the Freundlich isotherm model was found to be fit for adsorption data. Thermodynamic study revealed the adsorption to be spontaneous and exothermic. The adsorbent showed useful application for real water samples by more than 75% uptake of Cr and Cd with low adsorption of Na, K, and Mg. The regeneration study indicated that the mMOX could be reused up to six cycles with more than 50% removal of Cr(VI) and Cd(II) ions from aqueous solution with minimal leaching of metal ions (Fe, Ca, Na, K, and Mn) into the solution.

Pesticide Risk and Recurrent Pregnancy Loss in Females of Subhumid Region of India.

OBJECTIVE: The objective of this study is to determine the level of pesticides and their role in cases of recurrent pregnancy loss (RPL). MATERIALS AND METHODS: This was designed as a case-control study. Gas chromatography was used to characterize the pesticide level in 70 cases and 70 controls. Case refers to women with RPL, whereas controls refer to women with full-term delivery. RESULTS: A higher level of pesticide, namely beta-hexachlorocyclohexane, malathion, chlorpyrifos, and fenvalerate was found in the case group as compared to control group (P < 0.05). CONCLUSIONS: The present study suggests that high exposure of pesticide (organochlorine and organophosphates) may increase the risk of RPL in females of the subhumid region of India.

Genetic susceptibility of CYP1A1 gene and risk of pesticide exposure in prostate cancer.

BACKGROUND: The etiology of prostate cancer (PCa) is multi-factorial including environmental and genetic factors. Present study evaluates the association between level of pesticides, stress level and CYP1A1 gene polymorphism with PCa patients. METHODS: A case control study was conducted with 102 PCa patients and age match symptomatic (n= 107) and asymptomatic benign prostatic hyperplasia (BPH, n= 70) patients. Pesticide level was characterized by Gas Chromatography. The oxidative stress and scavenging mechanisms were determined by biochemical method. Two polymorphisms of CYP1A1 gene, rs4646903 and rs1048943, were analyzed by polymerase chain reaction (PCR) followed by restriction fragment length polymorphism and allele specific PCR method. RESULTS: Higher level of pesticide namely beta-hexachlorocyclohexane (ß-HCH), Malathion, Chlorpyrifos and Fenvalerate were found in PCa group (all p value: < 0.05). Kruskal Wallis H test depicted that level of ß-HCH and Malathion significantly correlated with higher grade of PCa (all p< 0.05). The PCa Patients with simultaneously low antioxidant activity and high stress level tended to suffer worst clinical outcomes. Dominant model of rs4646903 and rs1048943 suggested that substitution is associated with a higher risk of PCa (OR: 2.2, CI: 1.6-3.8, p: 0.009 and OR: 1.95, CI: 1.1-3.4, p: 0.026; respectively) and this risk was also influenced by smoking and pesticide exposure. CONCLUSION: Environmental and genetic factors are reported to raise risk; person with high level of these pesticides especially in high risk genotype might be more susceptible to PCa.

Ionic liquid based vortex assisted liquid-liquid microextraction combined with liquid chromatography mass spectrometry for the determination of bisphenols in thermal papers with the aid of response surface methodology.

A sensitive, rapid and efficient ionic liquid-based vortex assisted liquid-liquid microextraction (IL-VALLME) with Liquid Chromatography Mass spectrometry (LC-MS/MS) method is proposed for the determination of bisphenols in thermal paper. Extraction factors were systematically optimized by response surface methodology. Experimental factors showing significant effects on the analytical responses were evaluated using design of experiment. The limit of detection for Bisphenol-A (BPA) and Bisphenol-S (BPS) in thermal paper were 1.25 and 0.93µgkg-1 respectively. The dynamic linearity range for BPA was between 4 and 100µgkg-1 and the determination of coefficient (R2) was 0.996. The values of the same parameters were 3-100µgkg-1 and 0.998 for BPS. The extraction recoveries of BPA and BPS in thermal paper were 101% and 99%. Percent relative standard deviation (% RSD) for matrix effect and matrix match effects were not more than 10%, for both bisphenols. The proposed method uses a statistical approach for the analysis of bisphenols in environmental samples, and is easy, rapid, requires minimum organic solvents and efficient.

Determination of Cd, Cr, Pb and Ni contents among Parkinson's disease individuals: a case-control study.

BACKGROUND: Various uses of metals in industries, including the domestic sphere, agriculture, medicine and technology, have led to their wide distribution in the environment. These result in raising concerns over their potential effects on human health and the environment. Because of their high degree of toxicity, Cd, Cr and Pb are some of the priority metals that are of public health significance. The levels of Cd, Cr, Pb and Ni were measured in Parkinson's disease (PD) patients. METHODS: Blood samples were collected from 40 patients and 40 healthy controls, and stored at -80 °C until assayed. Atomic absorption spectrophotometry was used to determine the levels of metals. RESULTS: The level of Pb was significantly decreased in patients than in controls. However, the difference in the level of Ni between patients and controls failed to reach significance. Cr was not detectable in patients, but it was measurable in 12 controls (controls = 0.056-2.397 µg/ml). Similarly, Cd was not detectable in patients, but it was measurable in all the controls (controls = 0.004-1.268 µg/ml). Pb was the only metal that was found in all study participants (PD = 0.012-2.758 µg/ml and controls = 0.779-9.840 µg/ml). Ni could be measured only in six patients and in all the controls (PD = 0.154-0.754 µg/ml and controls = 0.034-1.691 µg/ml). CONCLUSION: Patients exhibited significantly decreased levels of Pb than in controls. However, Cd, Cr and Ni were too low to be measured among the patients. This indicates that these metals might play a probable role in PD.

A comprehensive toxicity study of zinc oxide nanoparticles versus their bulk in Wistar rats: Toxicity study of zinc oxide nanoparticles.

The purpose of this study was to characterize the zinc oxide nanoparticles (ZnO-NPs) and their bulk counterpart in suspensions and to access the impact of their acute oral toxicity at doses of 300 and 2000 mg/kg in healthy female Wistar rats. The hematological, biochemical, and urine parameters were accessed at 24 and 48 h and 14 days posttreatment. The histopathological evaluations of tissues were also performed. The distribution of zinc content in liver, kidney, spleen, plasma, and excretory materials (feces and urine) at 24 and 48 h and 14 days posttreatment were accessed after a single exposure at dose of 2000 mg/kg body weight. The elevated level of alanine amino transferase, alkaline phosphatase, lactate dehydrogenase, and creatinine were observed in ZnO-NPs at a dose of 2000 mg/kg at all time points. There was a decrease in iron levels in all the treated groups at 24 h posttreatment as compared to control groups but returned to their normal level at 14 days posttreatment. The hematological parameters red blood cells, hemoglobin, hematocrit, platelets, and haptoglobin were reduced at 48 h posttreatment at a dose of 2000 mg/kg ZnO-NPs and showed hemolytic condition. All the treated groups were comparable to control group at the end of 14 days posttreatment. The zinc concentration in the kidney, liver, plasma, feces, and urine showed a significant increase in both groups as compared to control. This study explained that ZnO-NPs produced more toxicological effect as compared to their bulk particles as evidenced through alteration in some hemato-biochemical parameters and with few histopathological lesions in liver and kidney tissues.

Phyto-extraction of heavy metals and biochemical changes with Brassica nigra L. grown in rayon grade paper mill effluent irrigated soil.

In this study, distribution of metal accumulation and their biological changes of Indian mustard plants (Brassica nigra L.) grown in soil irrigated with different concentration of rayon grade paper effluent (RGPE, 25%, 50%, 75%, 100%, v/v) were studied. A pronounced effect was recorded at 50% (v/v) RGPE on germination of seeds, amylase activity and other growth parameters in Indian mustard plants. An increase in the chlorophyll and protein contents was also recorded at <50% (v/v) RGPE followed by a decrease at higher concentrations of RGPE (>75%). A significant increase lipid peroxidation was recorded, which was evidenced by the increased malondialdehyde (MDA) content in shoot, leaves and seeds in tested plant at all the concentrations of RGPE. This Indian mustard plants (Brassica nigra L.) are well adapted for tolerance of significant amount of heavy metals due to increased level of antioxidants (cysteine and ascorbic acid) in root shoot and leaves of treated plants at all concentration of RGPE. Moreover, it is also important that RGPE should be treated to bring down the metal concentration well within the prescribed limit prior to use in agricultural soil for ferti-irrigation.

Fast agitated directly suspended droplet microextraction technique for the rapid analysis of eighteen organophosphorus pesticides in human blood.

A new sample preparation technique named as fast agitated directly suspended droplet microextraction (FA-DSDME) was proposed as an improved version of directly suspended droplet microextraction (DSDME) for the extraction and pre-concentration of wide-range organophosphorus pesticides (OPPs) from human blood prior to liquid chromatography tandem mass spectrometric (LC-MS/MS) analysis. In this method, instead of protecting the unwanted rupturing of extraction droplet (organic solvent), it was deliberately splintered into fine droplets by providing automated high-speed agitation to the biphasic extraction system (extraction solvent and sample solution). Fine organic droplets were then recollected into one, not by using a centrifuge machine but just by giving a very slow stirring to the bottom of the extraction system. The present method has surmounted the problem of prolonged extraction time associated with old DSDME. Under optimum extraction conditions, the method showed good sensitivity with low detection limits ranging from 0.0009 to 0.122µgL(-1). Mean recoveries were achieved in the range of 86-109% at three levels of spiking concentration (low, middle and high) from linearity range of individual analyte. Intra-day and inter-day precisions were ≤4.68 and ≤9.57 (%RSD) respectively. Enrichment factor (EF) for each analyte varied from 30 to 132 which prove the ability of this technique to pre-concentrate the extracted analytes up to a good extent. The sample matrices have shown an insignificant influence on method's sensitivity. The proposed method may find immense use in epidemiological, toxicological, regulatory and forensic laboratories.

In vitro toxicity evaluation of low doses of pesticides in individual and mixed condition on human keratinocyte cell line.

The induced toxicity of three pesticides (alpha-Hexachlorocyclohexane: α-HCH; Parathion methyl:PM; Carbofuran: CN) in single and four possible combination on human keratinocyte cell line have been investigated. There was no significant change in toxicity (cyto and genotoxicity) on cell line exposed by individual pesticides except α-HCH. But, a synergistic effect was observed when we tested mixture of pesticides. The intracellular ROS and cytotoxicity assay revealed maximum reduction in cell viability (60%) was found in tri mixture of pesticides. All the possible combination of these pesticides demonstrated genotoxic activity in terms of olive tail moment and % tail DNA on cell line at low concentration. The order of toxicity was ranked as α-HCH+PM+CN>α- HCH+CN>PM+CN>α-HCH+PM. Our results call for more research to be undertaken in order to understand the mechanisms behind the synergy observed and quantify the extent of its environmental impacts.

Developmental exposure to As, Cd, and Pb mixture diminishes skeletal growth and causes osteopenia at maturity via osteoblast and chondrocyte malfunctioning in female rats.

We studied the effect of metal mixture (MM), comprising As, Cd, and Pb, in developing female rat skeleton from gestation day 5 until postnatal day 60 (P-60). MM resulted in synergistic inhibition in viability and differentiation of osteoblasts in vitro, likely induced by reactive oxygen species. MM, administered at their most frequently occurring concentrations present in the groundwater of India, i.e., As: 0.38 ppm, Pb: 0.22 ppm, and Cd: 0.098 ppm or 10× of the ratio to developing rats, exhibited a synergistic decrease in ex vivo mineralization of bone marrow stromal (osteoprogenitor) cells. MM group showed a dose-dependent attenuation in weight and axial lengths and shortening of tibias at P-60. Furthermore, the growth plate was shortened, which was associated with shorter proliferative and hypertrophic zones, decreased parathyroid hormone-related protein and Indian hedgehog expression in the chondrocytes, reduced primary and secondary spongiosa, and hypomineralized osteoids-a major characteristic of osteomalacia. In addition, compared with the control, MM-treated rats were clearly osteopenic based on bone mineral density, microarchitecture, biomechanical strength, and particularly the biochemical profile, that suggested high turnover bone loss. Finally, in comparison to the control, the fracture-healing ability of MM group was delayed and accompanied by inferior quality of the healed bone. Together, these data demonstrated that the mixture of As, Cd, and Pb induced synergistic toxicity to developing skeleton, thereby diminishing modeling-directed bone accrual, inducing osteopenia and dampening fracture healing.

Optimization and validation of an extraction method for the analysis of polycyclic aromatic hydrocarbons in chocolate candies.

UNLABELLED: Chocolate is a key ingredient in many foods such as milk shakes, candies, bars, cookies, and cereals. Chocolate candies are often consumed by mankind of all age groups. The presence of polycyclic aromatic hydrocarbons (PAHs) in chocolate candies may result in health risk to people. A rapid, precise, and economic extraction method was optimized and validated for the simultaneous determination of polycyclic aromatic hydrocarbons in chocolate candy by high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GS-MS) as a confirmatory technique. The method was optimized by using different solvents for liquid-liquid extraction, varying volume of de-emulsifying agent, and quantity of silica gel used for purification. The HPLC separation of 16 PAHs was carried out by C-18 column with mobile phase composed of acetonitrile : water (70 : 30) in isocratic mode with runtime of 20 min. Limit of detection, limit of quantification (LOQ), and correlation coefficients were found in the range of 0.3 to 4 ng g⁻¹, 0.9 to 12 ng g⁻¹, and 0.9109 to 0.9952, respectively. The exploration of 25 local chocolate candy samples for the presence of PAHs showed the mean content of benzo[a]pyrene as 1.62 ng g⁻¹, which representing the need to evaluate effective measures to prevent more severe PAHs contamination in chocolate candies in future. PRACTICAL APPLICATION: Chocolate is one of the most favorite food items among people, especially children. Chocolate candies are often consumed by mankind of all age groups. Chocolate candies are often consumed by children in large quantities. The presence PAHs in chocolate candies may result in health risk to people. In the present study, a precise and cost effective rapid method was employed for the determination of PAHs, which can be employed for daily routine analysis of PAHs in chocolate products.