Effect of Honey and Aqueous Garlic Extracts against Short-Term Exposure of Cigarette Tobacco Smoking in Mice: Histopathological and Biochemical Investigations.

It is well known that cigarette smoking adversely affects human health and induces oxidative stress in most vital organs. This study aims to assess the biochemical, histological, and ultrastructural values of honey and garlic extracts in ameliorating the effects of short-term exposure to cigarette smoke in mice. Forty-eight mice were randomly divided into six equal groups: group I was exposed to fresh air only, group II was exposed to cigarette smoke, group III was given 0.2 ml of honey extract, group IV was exposed to cigarette smoke and was given 0.2 ml of honey extract, group V was given 0.2 ml of garlic extract, and group VI was exposed to cigarette smoke and was given 0.2 ml of aqueous garlic extract. These exposures were repeated daily for 21 consecutive days among the treated groups. By the end of the third week, the animals were euthanized by physical cervical dislocation. Blood was taken for biochemical study, and the selected organs of the liver, kidney, and jejunum were processed for histological and ultrastructural studies. The biochemical results showed that short-term exposure of experimental mice to cigarette smoking did not alter the liver function tests except for decreasing the albumin level. Moreover, cigarette smoking elevates the concentration of carbonyl protein content and cystatin C. Histologically, the use of honey and garlic showed good protection to the liver, kidney, and jejunum, which was proved by transmission electron microscopy, in addition to lowering the oxidative stress biomarkers. In conclusion, using honey and/or garlic helps protect the liver, kidney, and jejunum against the hazardous effects of cigarette smoke.

Development and In Vivo Evaluation of Sustained Release Microparticles Loaded with Levothyroxine for Hypothyroidism Treatment.

Hypothyroidism is a chronic condition combated by a daily oral supplementation of levothyroxine. In addition to the need for frequent dosing, oral administration may result in variable absorption of the drug leading to a failure in achieving normal thyroid function. Therefore, the development of a long-acting injectable system capable of delivering the drug is necessary. This work was aimed at developing sustained release microparticles loaded with levothyroxine. The microparticles were produced through the emulsification-solvent evaporation method using 2 grades of biocompatible and biodegradable polyesters: poly(ᴅ,ʟ-lactide-co-glycolide) (PLGA) and poly(ᴅ,ʟ-lactide) (PLA). Both polymers produced microparticles with very similar sizes (1.9 µm) and zeta potential values (around -22.0 mV). However, PLA microparticles had a significantly higher drug loading (6.1% vs. 4.4%, respectively) and encapsulation efficiency (36.8%, vs. 26.1%, respectively) when compared to PLGA counterparts. While both types of microparticles displayed a biphasic release pattern in vitro, a slower rate of release was observed with PLA microparticles. Moreover, a similar biphasic release pattern was found in vivo, with an initial phase of rapid release followed by a slower phase in the subsequent 10 days. These results indicate the possibility of developing levothyroxine loaded polyester microparticles as a potential long-acting thyroid hormone replacement therapy.

Optimizing formulation parameters for the development of carvedilol injectable in situ forming depots.

In situ forming depots (ISFDs) represent attractive alternatives to the conventional sustained drug delivery systems. Carvedilol, a short half-life drug used on a daily basis to manage chronic conditions, could benefit from this technology. The aim of this work was to develop, for the first time, a new injectable long-acting carvedilol-ISFD. Accordingly, 4 different grades of polyesters with varying properties as i) lactide-to glycolide ratio (polylactide-co-glycolide (PLGA) vs. polylactide (PLA)), and ii) end functionality (acid- vs. ester-capped) were utilized for the preparation of ISFD formulations. In addition, 4 different organic solvents with varying properties (i.e. N-methyl-2-pyrrolidone (NMP), dimethyl sulfoxide (DMSO), ethyl acetate, and benzyl benzoate) were also investigated. It was found that NMP and DMSO were more suitable for the formation of depots. Furthermore, all ISFD formulations demonstrated excellent encapsulation efficiency (i.e. 96-98%). Interestingly, both PLGA-based ISFDs (acid-capped and ester-capped) exhibited similar release behaviors and were able to extend carvedilol release over 30 days. On the other hand, acid-capped and ester-capped PLA-based ISFDs exhibited slower release over the 30 days with an average release of only 36% and 60%, respectively. In conclusion, the developed carvedilol-ISFDs resulted in a tunable extended-release behavior, simply by choosing the appropriate grade of polymer. These results open the door toward a novel injectable carvedilol-ISFD formulation.

Berberine Inhibited Growth and Migration of Human Colon Cancer Cell Lines by Increasing Phosphatase and Tensin and Inhibiting Aquaporins 1, 3 and 5 Expressions.

Introduction: Berberine is a natural isoquinoline alkaloid with anti-cancer properties. Nevertheless, the underlying mechanism of its action in human colorectal cancer (CRC) has not been thoroughly elucidated. We investigated the anti-cancer effect of berberine on HT-29, SW-480 and HCT-116 human CRC cell lines. Methods: Cell proliferation, migration and invasion were studied by MTT assay, wound healing, transwell chambers and flow cytometry. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunostaining were used to evaluate the expression of aquaporins (AQPs) 1, 3 and 5 in colon cancer cell lines before and after treatment with berberine (10, 30 and 100 µM). RT-qPCR and Western blotting were used to further explore the PI3K/AKT signaling pathway and the molecular mechanisms underlying berberine-induced inhibition of cell proliferation. Results: We demonstrated that treatment of these CRC cell lines with berberine inhibited cell proliferation, migration and invasion through induction of apoptosis and necrosis. HT-29, SW-480 and HCT-116 stained positively for AQP 1, 3 and 5, and berberine treatment down-regulated the expression of all three types of AQPs. Berberine also modulated PI3K/AKT pathway activity through up-regulating PTEN and down-regulating PI3K, AKT and p-AKT expression as well as suppressing its downstream targets, mTOR and p-mTOR at the protein level. Discussion/Conclusions: These findings indicate that berberine inhibited growth, migration and invasion of these colon cancer cell lines via down-regulation of AQP 1, 3 and 5 expressions, up-regulating PTEN which inhibited the PI3K/AKT pathway at the gene and protein levels, and that AQP 1, 3 and 5 expression level can be used as prognostic biomarkers for colon cancer metastasis.

Aqueous core microcapsules as potential long-acting release systems for hydrophilic drugs.

We have previously optimized the internal phase separation process to give rise to aqueous core microcapsules with polymeric shells composed of poly(lactide-co-glycolide) (PLGA) or poly(lactide) (PLA). In this study, the ability of these microcapsules to act as controlled release platforms of the model hydrophilic drug phenobarbital sodium was tested. Furthermore, the effect of the initial amounts of drug and water added to the system during microcapsule synthesis was investigated. Finally, the effect of varying polymer properties such as end functionalities, molecular weights, and lactide to glycolide ratios, on the characteristics of the produced microcapsules was studied. This was done by utilizing seven different grades of the polyester polymers. It was demonstrated that, within certain limits, drug loading is nearly proportional to the initial amounts of drug and water. Furthermore, drug encapsulation studies demonstrated that ester termination and increases in polymeric molecular weight result in lower drug loading and encapsulation efficiency. Moreover, drug release studies demonstrated that ester termination, increases in molecular weight, and increases in the lactide to glycolide ratio all result in slower drug release; this grants the ability to tailor the drug release duration from a few days to several weeks. In conclusion, such minor variations in polymer characteristics and formulation composition can result in dramatic changes in the properties of the produced microcapsules. These changes can be fine-tuned to obtain desirable long-acting microcapsules capable of encapsulating a variety of hydrophilic drugs which can be used in a wide range of applications.

Ameliorative effect of resveratrol against histological alterations in lung of mice induced by subchronic exposure of cigarette and waterpipe tobacco smoking / Efecto paliativo del resveratrol frente a las alteraciones histológicas en el pulmón de ratones inducidas por exposición subcrónica del tabaquismo de cigarrillos y pipas de agua

SUMMARY: This study aims to evaluate the ability of resveratrol (RSV) antioxidant to attenuate the oxidative stress condition induced by secondhand exposure of cigarettes and waterpipe smoking using animal model. Forty-eight mice were divided equally into six different groups, and RSV was delivered to certain groups intraperitoneally with a dose of 25 mg/kg/day. The process of smoking exposure was performed using a specialized smoking machine. The experiment duration lasts for six consecutive weeks. Five µm sections of lung were stained with hematoxylin and eosin for light microscopy, and 70 nm ultrathin sections of lung stained with uranyl acetate and lead citrate were prepared for transmission electron microscopy to observe the cellular ultrastructure. In lung, RSV reduced the deterioration and blebbing of bronchiole epithelium, reduced the inflammation, increased the surface area of alveolar sac, and reduced the thickening of alveolar walls. Also, blood vessels were less congested and less dilated with less diffusion of extravasated blood. Ultrastructural images illustrated that RSV protects the normal structure of alveolar septum, prevents DNA damage and alveolar degeneration, showed less degree of apical membrane blebbing and retained the uniform pattern of mitochondria. In conclusion, RSV has ameliorative effects against the oxidative stress condition induced by secondhand (side stream) exposure of cigarette and waterpipe tobacco smoking.

RESUMEN: Este estudio tiene como objetivo evaluar la capacidad del antioxidante resveratrol (RSV) para atenuar el estrés oxidativo inducido por la exposición de segunda mano a los cigarrillos y al tabaquismo en pipa de agua, utilizando un modelo animal. Se dividieron cuarenta y ocho ratones en seis grupos diferentes, y se administró RSV a ciertos grupos por vía intraperitoneal con una dosis de 25 mg / kg / día. El proceso de exposición al tabaquismo se realizó utilizando una máquina de fumar especializada. La duración del experimento fue de seis semanas consecutivas. Para realizar la microscopía óptica se tiñeron secciones de pulmón de cinco µm con hematoxilina y eosina, y se prepararon secciones ultrafinas de 70 nm de pulmón teñidas con acetato de uranilo y citrato de plomo para la microscopía electrónica de transmisión para observar la ultra estructura celular. En el pulmón, el RSV redujo el deterioro y la formación de ampollas del epitelio de los bronquiolos, redujo la inflamación, aumentó la superficie del saco alveolar y redujo el engrosamiento de las paredes alveolares. Además, los vasos sanguíneos se encontraron menos congestionados y menos dilatados y con menor difusión de sangre extravasada. Las imágenes ultraestructurales mostraron que el RSV protege la estructura normal del tabique alveolar, previene el daño del ADN y la degeneración alveolar, mostrando un menor grado de formación de ampollas en la membrana apical y además retuvo el patrón uniforme de las mitocondrias. En conclusión, el RSV tiene efectos de mejora contra el estrés oxidativo inducido por la exposición de segunda mano (corriente secundaria) al fumar cigarrillos y pipas de agua.

Ameliorative effects of curcumin and caffeic acid against short term exposure of waterpipe tobacco smoking on lung, heart and kidney in mice.

This study aims to evaluate the chemopreventive activity of two antioxidants (curcumin [CUM] and caffeic acid [CAF]), focusing on how these antioxidants could reduce cytotoxicity induced by short term secondhand exposure of waterpipe tobacco smoking. Forty-eight adult male BALB/c albino mice were equally divided into four groups. Antioxidants were delivered intraperitoneally, and the exposure to waterpipe smoking (WPS) was performed using a smoking machine. This experiment lasts for 14 consecutive days. Serum were collected from mice before dissection to quantify the activity of some liver enzymes, kidney function tests and proinflammatory cytokines. Lung, heart, and kidney were isolated and processed for light microscopy technique. Parallel treatment of CUM or CAF along with exposure to WPS showed less inflammation, less vacuolized, and more inflated alveoli, less deteriorations in cortex part of kidney, and less disintegration of cardiac myofibers in comparison to waterpipe only. Besides, CUM and CAF significantly reduced the activity of aspartate aminotransferase and proinflammatory cytokines. CUM and CAF were found to have anti-inflammatory and ameliorative effects against the cytotoxicity induced by exposure to waterpipe tobacco smoking, and CUM showed better chemopreventive activity than CAF.

Correction: Nanoparticle size and chemical modification play a crucial role in the interaction of nano gold with the brain: extent of accumulation and toxicity.

Correction for 'Nanoparticle size and chemical modification play a crucial role in the interaction of nano gold with the brain: extent of accumulation and toxicity' by Nouf N. Mahmoud et al., Biomater. Sci., 2020, DOI: 10.1039/c9bm02072a.

Nanoparticle size and chemical modification play a crucial role in the interaction of nano gold with the brain: extent of accumulation and toxicity.

The blood brain barrier (BBB) is a very selective barrier that protects the brain and the central nervous system (CNS) from the entry of harmful substances and helps regulate the exchange of different molecules and nutrients from and into the brain and the CNS. This selectivity makes delivering therapeutic and diagnostic materials across the BBB very challenging. In this study, different shapes and sizes of gold nanoparticles (GNP) were synthesized and functionalized with five different thiolated ligands to obtain GNP with various surface chemistries. The potential of GNP of different properties to be accumulated into the brain through the BBB and into other organs was investigated in a mouse model using qualitative and quantitative approaches. Gold nanorods (GNR) functionalized with 4-mercaptophenol (Mph) showed the highest penetration ability across the BBB into the brain with no significant deposition in other organs. Interestingly, increasing the size of GNR retarded their delivery into the brain, while enhancing their accumulation in other organs. On the other hand, gold nanospheres (GNS) demonstrated high deposition percentages in the brain and other organs with possible toxic effects. The properties of GNP play a crucial role in their interaction with the BBB and accumulation in the brain and other organs. Thus, GNP can be considered a promising nano-platform for drug delivery into the brain and as a photothermal-inducing agent against brain cancer.

hPL promotes osteogenic differentiation of stem cells in 3D scaffolds.

Human platelet lysate (hPL) has been considered as the preferred supplement for the xeno-free stem cell culture for many years. However, the biological effect of hPL on the proliferation and differentiation of dental stem cells combined with the use of medical grade synthetic biomaterial is still under investigation. Thus, the optimal scaffold composition, cell type and specific growth conditions, yet need to be formulated. In this study, we aimed to investigate the regenerative potential of dental stem cells seeded on synthetic scaffolds and maintained in osteogenic media supplemented with either hPL or xeno-derived fetal bovine serum (FBS). Two types of dental stem cells were isolated from human impacted third molars and intact teeth; stem cells of apical papilla (SCAP) and periodontal ligament stem cells (PDLSCs). Cells were expanded in cell culture media supplemented with either hPL or FBS. Consequently, proliferative capacity, immunophenotypic characteristics and multilineage differentiation potential of the derived cells were evaluated on monolayer culture (2D) and on synthetic scaffolds fabricated from poly 'lactic-co-glycolic' acid (PLGA) (3D). The functionality of the induced cells was examined by measuring the concentration of osteogenic markers ALP, OCN and OPN at different time points. Our results indicate that the isolated dental stem cells showed similar mesenchymal characteristics when cultured on hPL or FBS-containing culture media. Scanning electron microscopy (SEM) and H&E staining revealed the proper adherence of the derived cells on the 3D scaffold cultures. Moreover, the increase in the concentration of osteogenic markers proved that hPL was able to produce functional osteoblasts in both culture conditions (2D and 3D), in a way similar to FBS culture. These results reveal that hPL provides a suitable substitute to the animal-derived serum, for the growth and functionality of both SCAP and PDLSCs. Thus the use of hPL, in combination with PLGA scaffolds, can be useful in future clinical trials for dental regeneration.

Association between occupational lead exposure and plasma levels of selected oxidative stress related parameters in Jordanian automobile workers.

OBJECTIVES: To study any possible correlation between blood lead levels and some oxidative stress parameters of selected groups of lead-exposed automobile occupational Jordanian workers. MATERIAL AND METHODS: Blood lead levels were determined for a total of 90 male automobile workers aged within the range of 25-45 years old along with the group of 20 agematched healthy males control. To get an idea about the antioxidant status of controls and lead-exposed workers, and to estimate the oxidative stress caused by exposure to lead, we measured the hydrogen peroxide (H2O2) concentration, superoxide dismutase (SOD) activity, lipid peroxidation as malondialdehyde (MDA) and total antioxidant capacity of controls and workers. RESULTS: The study showed that lead levels in the case of workers were approximately 4-5 times as high as in controls 14.5-21 µg×dl-1 vs. 4.3 µg×dl-1, respectively. There was a significant decrease (16-25%) in the level of plasma reduced glutathione and 21-33% decrease in total antioxidant capacity (TAOC) in all worker groups, as compared to controls. The results showed that the concentration of malondialdehyde (MDA) in plasma was higher (120-333%) in the case of workers than controls being the highest in automobile electronics and the lowest - in mechanics. Furthermore, there was 149-221% increase in hydrogen peroxide (H2O2) concentration, and 26-38% increase in SOD activity in the case of workers compared to the control group. CONCLUSIONS: There is a strong evidence for the associations between occupational lead exposure and various markers of oxidative stress in Jordanian automobile occupational workers. Thus, there is an urgent need to raise awareness and to initiate suitable protection guidelines for workers. Int J Occup Med Environ Health 2018;31(4):517-525.

Zinc oxide nanoparticles hepatotoxicity: Histological and histochemical study.

Zinc oxide nanoparticles (ZnO NPs) are widely used in industry and cosmetic products with promising investment in medical diagnosis and treatment. However, these particles may reveal a high potential risk for human health with no information about hepatotoxicity that might be associated with their exposure. The present work was carried out to investigate the histological and histochemical alterations induced in the hepatic tissues by naked 35nm ZnO NPs. Male Wistar albino rats were exposed to ZnO NPs at a daily dose of 2mg/kg for 21days. Liver biopsies from all rats under study were subjected to histopathological examinations. In comparison with the control rats, the following histological and histochemical alterations were demonstrated in the hepatic tissues of rats exposed to ZnO NPs: sinusoidal dilatation, Kupffer cells hyperplasia, lobular and portal triads inflammatory cells infiltration, necrosis, hydropic degeneration, hepatocytes apoptosis, anisokaryosis, karyolysis, nuclear membrane irregularity, glycogen content depletion and hemosidrosis. The findings of the present work might indicate that ZnO NPs have potential oxidative stress in the hepatic tissues that may affect the function of the liver. More work is needed to elucidate the toxicity and pathogenesis of zinc oxide nanoparticles on the vital organs.

Rosmarinic acid reverses the effects of metronidazole-induced infertility in male albino rats.

Rosmarinic acid (RA) is a natural antioxidant that has many biological activities. In the present study we investigated the potential of RA to reverse the negative effects of the widely used antibiotic and antiprotozoal agent metronidazole (MTZ), which is known to induce reversible male infertility. Two doses of RA (5 and 15mg kg-1) were studied in sexually mature rats with and without MTZ-induced infertility. Rats were intraperitoneally injected with 5mg kg-1 RA or 15mg kg-1 RA (in distilled water) and, 45min later, they were intraperitoneally injected with 40mg kg-1 MTZ (in distilled water). Cauda epididymidal sperm suspensions were used to assess sperm count, motility and morphology. Histological and ultrastructural studies were performed on the testes and cauda epididymidis. In rats in which infertility was not induced, neither dose of RA affected the parameters assessed. However, in sexually mature rats in which infertility was induced by 40mg kg-1 MTZ, RA at both 5 and 15mg kg-1 ameliorated the damaging effects of MTZ on final bodyweight (30 days later), sperm motility and morphology. Only 5mg kg-1 RA, and not 15mg kg-1 RA, improved the harmful effects of MTZ on the sperm count and testis ultrastructure. The findings of the present study have considerable clinical implications and suggest a possible use for RA to reverse the negative effects of MTZ on male fertility, the male reproductive system and spermatogenesis.

Effect of cigarette smoking on the structure of hepatocytes: TEM study / Efecto del tabaquismo sobre la estructura de los hepatocitos: estudio MET

The liver is one of the major organs that is indirectly affected by cigarette smoke. The aim of this project is to define the histologic and ultrastructural changes in normal liver cells after exposing animals to cigarette smoke. Thirty albino rats were exposed to cigarette smoke for 90 days, followed by morphologic examination of their livers under light microscope and electron microscope. The liver cells of cigarette smoke exposed rats showed mild swelling with increased eosinophilia. Ultrastructural examination of these cells demonstrated cytoplasm with highly proliferated and crowded mitochondria. There were many electron dense mitochondria. These mitochondria were pleomorphic in shape compared to mitochondrias of control rats. Also, loss of mitochondrial cristae and widening of the intermembranous space was noticed. It is concluded that smoking exerts cellular damage and oxidative stress on normal liver cells resulting in ultrastructural changes.

El hígado es uno de los principales órganos indirectamente afectado por el humo del cigarrillo. El objetivo de este trabajo fue definir los cambios histológicos y ultraestructurales de las células normales del hígado después de exponer a los animales al humo del cigarrillo. Treinta ratas albinas fueron expuestas al humo de cigarrillo durante 90 días, seguido de un examen morfológico de los hígados bajo microscopio de luz y microscopio electrónico. Las células hepáticas de las ratas expuestas al humo de cigarrillo mostraron una leve inflamación con un aumento de la eosinofilia. En el examen ultraestructural de estas células se observó el citoplasma mitocondrial altamente proliferado y saturado. Se observó gran cantidad de mitocondrias electrón-densas y éstas presentaban forma pleomórfica en comparación con las mitocondrias del grupo control. Además, se observó pérdida de las crestas mitocondriales y ensanchamiento del espacio intermembranoso. Se concluye que el tabaquismo ejerce daño celular y estrés oxidativo en las células hepáticas normales, lo que resulta en la aparición de cambios ultraestructurales.

Structural characterization of rat ventricular tissue exposed to the smoke of two types of waterpipe.

OBJECTIVES: this study focused on the effect of waterpipe smoke exposure toxicity on the structure of albino rat's ventricular tissue and their recovery. MATERIALS AND METHODS: Albino rats were divided into three groups: control, flavored, and unflavored. The control group was exposed to normal air while the flavored and unflavored groups were exposed to waterpipe smoke for a period of 90 days. Each group was followed by a period of 90 days of fresh air exposure. Following each period, the ventricular tissue was removed for biochemical and histopathological studies. RESULTS: The ventricular tissues of waterpipe exposed rats showed some degree of separation between cardiac muscle fibers, infiltration of lymphocytes, and congestion of blood vessel. Also, thin cross sections of ventricular cells revealed pleomorphic mitochondria with partially disrupted cristae, partial disruption of the myofibrils, and deposited toxic materials. The unflavored waterpipe has more deleterious effects on heart ventricular tissues than the flavored one. Waterpipe smoke didn't induce apoptosis in the ventricular tissue. We also found very high levels of plasma thiocyanate after exposure to smoke in the flavored and unflavored groups, while the control group showed no increase. After the recovery period, those tissues showed partial recovery. CONCLUSION: Waterpipe smoke induces structural changes in the heart ventricle tissues, causing a negative impact on the capacity of the cardiac muscle for pumping blood and may lead to heart attack due to accumulation of free radicals and tissue inflammation. Cessation of smoking is important in returning most of these changes to their normal structure.

Lactic dehydrogenase as a biochemical marker of adverse pregnancy outcome in severe pre-eclampsia.

BACKGROUND: Pre-eclampsia is a specific disease of pregnancy with multisystem complications. We conducted this study to examine the relationship between lactic dehydrogenase concentration and the severity of the disease and the occurrence of its complications. MATERIAL/METHODS: One hundred eleven pre-eclamptic women (49 with mild and 62 with severe pre-eclampsia) and 60 healthy normotensive controls were studied prospectively at the King Hussein Medical Center between January and December 2002. Demographic, hemodynamic, and laboratory data were compared among the three groups. The symptoms and complications of severe pre-eclampsia along with fetal outcome were analyzed according to the levels of LDH (<600, 600-800, and >800 IU/l). One-way analysis of variance (ANOVA) and the chi-square test were used to compare the results. Differences were considered significant when p<0.05. RESULTS: The incidence of severe pre-eclampsia was 1.3%. Severely pre-eclamptic patients were significantly younger, with low gravidity and parity. On the other hand, they had significantly increased systolic and diastolic pressure and liver enzymes, uric acid, urine albumin, and LDH levels. The symptoms and complications of pre-eclampsia along with perinatal mortality were increased significantly in patients with LDH >800 IU/l compared with those who had lower levels. CONCLUSIONS: Lactic dehydrogenase is a useful biochemical marker that reflects the severity of and the occurrence of complications of pre-eclampsia. Identification of high-risk patients with elevated levels of lactic dehydrogenase, their close monitoring, and prompt, correct management may prevent these complications, with a subsequent decrease in maternal and fetal morbidity and mortality.