Osteomyelitis infection caused by Arcanobacterium haemolyticum in a diabetic patient: A first case report.

Arcanobacterium haemolyticum can cause deep infections, including osteomyelitis. In this study, an automated system misidentified this causal agent as Cellulomonas species but 16 s rRNA sequencing correctly identified it as A. haemolyticum. Recognizing the capability of A. haemolyticum to establish the disease is of great importance to enable accurate diagnosis and begin the suitable antibiotic therapy. Here we present the first case of successfully treated A. haemolyticum infective osteomyelitis in a 64-year-old Saudi patient with diabetes mellitus type 2 and review the characteristics of this seldom pathogenic agent.

Relative reduction of biological and phylogenetic diversity of the oral microbiota of diabetes and pre-diabetes patients.

BACKGROUND: A reciprocal relationship between oral health and systemic disease, such as type 2 diabetes, has been suggested, whereby a systemic disease is a predisposing factor for oral infection. If the infection occurs, it in turn aggravates the progression of the systemic disease. According to several studies, certain constituents of the oral microbiota are linked to diabetes, metabolic syndrome, and obesity. In the current study, we aimed to compare the microbial diversity and population structure of the oral microbiota of normoglycemic, impaired glucose tolerance (IGT), and diabetes patients. METHODOLOGY: The study followed a case-control design, with 15 type 2 diabetes patients, 10 IGT subjects, and 19 control subjects. All subjects underwent assessment of periodontitis and oral health. Saliva samples were collected, and DNA was isolated from these samples. Hypervariable regions of the 16Sr RNA gene were amplified and sequenced, and the generated sequences underwent bioinformatics analysis. Statistical analysis and diversity index calculations were made using the statistical software R, vegan R-package, and Past3.20 software. RESULTS: Overall, 551 operational taxonomic units (OTUs) were identified. Based on OTU analysis, a clear reduction of the number of species was observed in both IGT (412) and diabetes groups (372) compared with that in the normoglycemic group (502). This was associated with a similar pattern of reduction of biological diversity among the three groups. The phylogenetic diversity (PD-SBL) value in the normoglycemic group was higher than that in the diabetes group. The diabetes group exhibited the highest evenness value and the highest microbiota bacterial pathogenic content. CONCLUSION: A clear reduction of the biological and phylogenetic diversity was apparent in the diabetes and pre-diabetes oral microbiota in comparison with that in the normoglycemic oral microbiota. However, this was associated with an increase in the pathogenic content of the hyperglycemic microbiota. The results of this study may aid to better understanding of the directionality of the mysterious reciprocal relationship.

Genome sequencing and analysis of the first spontaneous Nanosilver resistant bacterium Proteus mirabilis strain SCDR1.

Background: P. mirabilis is a common uropathogenic bacterium that can cause major complications in patients with long-standing indwelling catheters or patients with urinary tract anomalies. In addition, P. mirabilis is a common cause of chronic osteomyelitis in Diabetic foot ulcer (DFU) patients. We isolated P. mirabilis SCDR1 from a Diabetic ulcer patient. We examined P. mirabilis SCDR1 levels of resistance against Nanosilver colloids, the commercial Nanosilver and silver containing bandages and commonly used antibiotics. We utilized next generation sequencing techniques (NGS), bioinformatics, phylogenetic analysis and pathogenomics in the characterization of the infectious pathogen. Results: P. mirabilis SCDR1 was the first Nanosilver resistant isolate collected from a diabetic patient polyclonal infection. P. mirabilis SCDR1 showed high levels of resistance against Nanosilver colloids, Nanosilver chitosan composite and the commercially available Nanosilver and silver bandages. The P. mirabilis -SCDR1 genome size is 3,815,621 bp. with G + C content of 38.44%. P. mirabilis-SCDR1 genome contains a total of 3533 genes, 3414 coding DNA sequence genes, 11, 10, 18 rRNAs (5S, 16S, and 23S), and 76 tRNAs. Our isolate contains all the required pathogenicity and virulence factors to establish a successful infection. P. mirabilis SCDR1 isolate is a potential virulent pathogen that despite its original isolation site, the wound, can establish kidney infection and its associated complications. P. mirabilis SCDR1 contains several mechanisms for antibiotics and metals resistance, including, biofilm formation, swarming mobility, efflux systems, and enzymatic detoxification. Conclusion: P. mirabilis SCDR1 is the first reported spontaneous Nanosilver resistant bacterial strain. P. mirabilis SCDR1 possesses several mechanisms that may lead to the observed Nanosilver resistance.

The Use of Next-Generation Sequencing in the Identification of a Fastidious Pathogen: A Lesson From a Clinical Setup.

Clostridium haemolyticum is the causal agent of bacillary hemoglobinuria in cattle, goat, sheep, and ruminants. In this study, we report the first recorded human-infecting C. haemolyticum strain collected from an 18-year-old woman diagnosed with acute lymphoblastic leukemia. After failure of traditional techniques, only next-generation sequencing (NGS) technology in combination with bioinformatics, phylogenetic, and pathogenomics analyses revealed that our King Faisal Specialist Hospital and Research Center (KFSHRC) bacterial isolate belongs to C. haemolyticum species. KFSHRC isolate is composed of 1 chromosome and 4 plasmids. The total genome size is estimated to be 2.7 Mbp with a low GC content of 28.02%. Comparative pathogenomics analysis showed that C. haemolyticum KFSHRC isolate is a potential virulent pathogenic bacterium as it possesses the virulence factors necessary to establish an infection, acquire essential nutrients, resist antimicrobial agents, and tolerate hostile conditions both in the human host and in its surrounding environment. These factors are included in the main chromosome in addition to novel recombination of the plasmids, and they could be the reason for the incidence of that human infection. This work demonstrated the importance of using NGS in medical microbiology for pathogen identification. It also demonstrates the importance of sequencing more microbial samples and sharing this information in public databases to facilitate the identification of pathogenic microbes with better accuracy.

Genotyping of CYP2C19 polymorphisms and its clinical validation in the ethnic Arab population.

OBJECTIVES: The drug-metabolizing enzymes and transporters (DMET) Plus microarray and x-Tag assays have recently been developed for genotyping individuals in personalized medicine. Furthermore, the cytochrome 450-2C19 (CYP2C19) is a key metabolic enzyme encoded by a polymorphic gene commonly associated with diminished metabolism and variable clinical responses to several drugs in an ethnicity-dependent fashion. Therefore, validation of these clinical procedures as well as knowledge of the ethnic-specific incidences of these gene variants is prerequisite for determining their clinical relevance in any given population. METHODS: We determined the distribution of familiar CYP2C19 variants by the DMET Plus chip in 600 candidates and replicated the findings by the Affymetrix Axiom Genome-Wide Asian Structure Identification Array in 5413 individuals, all Saudis of ethic Arab origin. We then tested the robustness of employing the Luminex xMAP system clinically by comparing the results of genotyping 500 Saudi individuals visiting the Blood Bank of our institution with the findings of the two platforms. KEY FINDINGS: The DMET Plus genotyping revealed that eight of the CYP2C19 variants showed some changes. Thereby, the CYP2C19*17 exhibited the highest minor allele frequency (MAF) of 0.256, followed by the CYP2C19_801 (frequency = 0.055). Six other variants, including the CYP2C19*3, showed MAF in the range of 0.001-0.002. We replicated the frequencies of the CYP2C19*17 and CYP2C19*3, and additionally established that of the CYP2C19*2 (0.099) using the Axiom platform. The xTag genotyping also indicated that 0.834 of the 500 Saudi individuals were extensive metabolizers (*1/*1), 0.158 carried the *1/*2 genotype, 0.01% carried *2/*2 (poor metabolizers) and one each (0.2%) harboured the *1/*8, *2/*3 (intermediate metabolizers) and *8/*8 (poor metabolizers) genotypes. CONCLUSIONS: The results showed reproducible genotyping of the CYP2C19 variants in the Saudi Arab population using two Affymetrix platforms and phenotyping using the Luminex xTag assay. The prevalence of two clinically relevant genotypes (CYP2C19*2 and CYP2C19*3) were similar to other ethnic groups, while that of the CYP2C19*17 was comparably higher.

Tinospora cordifolia ameliorates urotoxic effect of cyclophosphamide by modulating GSH and cytokine levels.

Cyclophosphamide (CP) is a commonly used anti-cancer drug which causes toxicity by its reactive metabolites. In this study we investigated the effect of Tinospora cordifolia on urotoxicity induced by acute dose of CP using Swiss albino mice model. Administration of an alcoholic extract of the plant T. cordifolia (Family: Menispermaceae) (200 mg/kg i.p.) for 5 days reduced CP (1.5 mmol/kg body wt. i.p.) induced urotoxicity as evident from the morphological analysis of bladder, decreased the relative bladder and liver weights and also decreased level of urea nitrogen and protein in blood as well as urine. Severely inflamed and dark coloured urinary bladders of the CP alone treated animals were found to be normalized by the treatment of T. cordifolia. GSH content, which was drastically reduced by CP administration in both bladder and liver was enhanced by treatment with T. cordifolia. Histopathological analysis of the bladder of CP alone-treated group showed severe necrotic damage where as the T. cordifolia-treated group showed normal bladder architecture. The lowered levels of cytokines IFN-γ, IL-2, after CP treatment were found to be increased in treated animals. At the same time the level of pro-inflammatory cytokine TNF-α, which was elevated during CP administration, was significantly reduced by extract administration. This study clearly demonstrates uroprotective role of T. cordifolia from CP induced toxicities by modulating GSH and pro-inflammatory cytokine levels.

Antiangiogenic activity of berberine is mediated through the downregulation of hypoxia-inducible factor-1, VEGF, and proinflammatory mediators.

Berberine, a naturally occurring isoquinoline alkaloid, is present in a number of important medicinal plants. Berberine has a wide range of biochemical and pharmacological effects, including anticancer effects. In this study, we elucidated the mechanism of antiangiogenic activity of berberine using in vivo and in vitro models. In vivo antiangiogenic activity was studied using B16F-10 melanoma cells and induced capillary formation in C57BL/6 mice. Berberine, at 10 mg/kg body weight, showed significant inhibition in tumor-directed capillary formation and in various proangiogenic factors, such as vascular endothelial growth factor (VEGF), and proinflammatory mediators, such as interleukin (IL)-1ß, IL-6, tumor necrosis factor alpha (TNF-α), and granulocyte macrophage colony-stimulating factor (GM-CSF), which are involved in tumor angiogenesis. At the same time, it could also increase antitumor factors, such as IL-2 and tissue-inhibitor metalloproteinase (TIMP) levels in the serum. Berberine could also inhibit endothelial motility, migration, tube formation, and vessel sprouting from rat aortic ring in vitro. Further, berberine inhibited various transcription factors involved in tumor development and angiogenesis, such as NF-ĸB, c-Fos, CREB, and ATF-2. mRNA expression levels of proangiogenic factors, such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and hypoxia-inducible factor (HIF), were also downregulated in tumor cells after treatment with berberine. Drastically elevated expressions of HIF and VEGF mRNA by tumor cells under hypoxic conditions were also decreased after treatment with berberine. This result clearly demonstrates that the antiangiogenic activity of berberine is mainly mediated through the inhibition of various proinflammatory and pro-angiogenic factors and the major ones are HIF, VEGF, COX-2, NO, NF-ĸB, and proinflammatory cytokines.

Berberine inhibits pulmonary metastasis through down-regulation of MMP in metastatic B16F-10 melanoma cells.

The present study demonstrated the potential antimetastatic and antiinvasive effect of berberine using both in vivo mouse lung metastasis and in vitro models. Administration of berberine resulted in significant suppression of B16F-10 melanoma induced tumor nodule formation and enhanced the survival of tumor-bearing mice. Berberine treatment also decreased various biochemical parameters associated with lung metastasis. These inhibitory actions may be due to the significant suppression of several signaling molecules such as ERK1/2, NF-κB, ATF-2 and CREB involved in the transcription signaling pathways for MMP gene expression. It could also inhibit the migration and invasion of highly metastatic murine melanoma cells in a dose-dependent manner in vitro. The results clearly show that berberine could significantly inhibit experimental lung metastasis produced by intravenous injection of B16F-10 melanoma cells and this effect could be linked to the down-regulation of metastasis-related signaling molecules.

Increased prevalence of rotavirus among children associated gastroenteritis in Riyadh Saudi Arabia.

UNLABELLED: The aim of this study is to assess the epidemiology along with the molecular structure of rotavirus causing pediatric diarrhea among Saudi patients. However, in this report we sited the epidemiological reflect coming from our project. METHODS: One thousand and seven diarrheal stool samples had been collected between Jan 1st, 2008 and OCT 31st, 2010 from hospitalized patients below the age of 5 year. Samples were then examined using Enzyme-linked immunosorbent assay (ELISA). Demographic data were collected including age, sex, date of admission and discharge. Finally, the chi-squire test, α level of significance was used to test the variables in the data. RESULTS: Of these 1007 stool samples, rotavirus was detected in 65.5% (660/1007 samples). We observed that children who are 1 year of age or less had more infection with rotavirus 81% (534/660) than those who is over 1 year of age (19%,126/660) (P = 0.000). Infections occur throughout the year with no clear significant seasonal peaks. The difference between males (57.5%, 380/660) and females (42.4%, 280/660) in terms of rotavirus positivity is statistically significant. CONCLUSIONS: The high rate of positivity, are at variance with previously published reports of rotavirus infection in Saudi Arabia since 2005 which reported a major decrease year by year in the incidence of rotavirus over; 2005, 2006 and 2008 with percentage of; 25%, 10%, 6% respectively explained by improvements in public health introduced in recent years. Our increasing rate result (65.5%) may suggest emerging of unusual serotypes, not been represent to our country earlier.

Studies on anti-metastatic and anti-invasive effects of harmine using highly metastatic murine B16F-10 melanoma cells.

Harmine is a beta-carboline alkaloid from the plant Peganum harmala. We evaluated the anti-metastatic activity of harmine using in vivo mouse lung metastasis and in vitro models. Lung metastasis was induced using B16F-10 melanoma cells in C57BL/6 mice by three different modalities of administration: simultaneous, prophylactic, and after tumor development. Harmine significantly inhibited tumor nodule formation in the lung tissue and decreased various biochemical parameters associated with lung metastasis. Higher expression levels of pro-metastatic genes such as matrix metalloproteinase-9 (MMP-9), extracellular signal[en]regulated kinase (ERK), and vascular endothelial factors (VEGFs), all of which play important roles in cancer cell migration and invasion, were observed in the metastatic group compared with normal, but were all down-regulated by treatment with harmine. Harmine was also able to inhibit tumor cell proliferation, invasion, and migration in vitro. In conclusion, harmine exerts anti-metastatic activity and this effect could be linked to the metastasis-related signaling pathway that includes ERK, VEGF, and MMPs.

Ipobscurine, an indole alkaloid from Ipomoea obscura, inhibits tumor cell invasion and experimental metastasis by inducing apoptosis.

In the present study, we demonstrate that ipobscurine, an indole alkaloid fraction isolated from Ipomoea obscura, can reduce the formation of B16F-10 melanoma-induced metastatic nodules and inhibit the proliferation, migration, and invasion of B16F-10 melanoma cells in vitro, possibly by inhibiting pro-metastatic genes such as matrix metalloproteinases (MMPs) and inflammatory mediators such as cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and pro-inflammatory cytokines. Ipobscurine may also promote apoptosis by up-regulating pro-apoptotic molecules such as caspase-3, p53, and Bax and by down-regulating anti-apoptotic Bcl-2. In addition, we have observed that ipobscurine suppresses various transcription factors such as nuclear factor-κB (NF-κB) and activator protein, which are possibly associated with the suppression of apoptosis in cancer cells. Ipobscurine has also been shown to inhibit cell growth with arrest at G1 and reduce transition to the S and G2/M phases of the cell cycle.

Augmentation of cellular immune response by Ipomoea obscura and Ipobscurine alkaloid attenuates tumor growth in mice.

The immune status of the host plays a crucial role in controling the process of carcinogenesis. General or selective activation of various immunocompetent cells and their secretory function to maintain a healthy immune status may help in cancer prophylaxis, as well as therapy. The present study focused on the effect of Ipomoea obscura and Ipobscurine on cell-mediated immune response. In this study we evaluated the effect of I. obscura and an indole alkaloid fraction from I. obscura on effector mechanisms of cell-mediated immune response by analyzing cytotoxic T lymphocyte (CTL) activity, natural killer (NK) cell activity, antibody-dependent cell-mediated cytotoxicity (ADCC), and antibody-dependent complement-mediated cytotoxicity (ACC). The effect of I. obscura and Ipobscurine on interleukin-2 (IL-2) and interferon-γ (IFN-γ) levels was also analyzed. In the in vitro and in vivo systems, I. obscura and Ipobscurine treatment augmented cell-mediated immune response by enhancing the killing activity of CTL and NK cells from splenocytes in normal as well as tumor-bearing mice. ADCC and ACC were also enhanced significantly in both normal and tumor-bearing animals after drug administration, compared with untreated control. Administration of I. obscura and Ipobscurine significantly enhanced the production of IL-2 and IFN-γ in normal as well as tumor-bearing animals. This study reveals that both I. obscura and Ipobscurine have the potential to augment immune response through the enhanced secretion of IL-2 and IFN-γ by T cells and thereby inhibit tumor growth and as an alternative medicine for cancer treatment.

Effect of Thuja occidentalis and its polysaccharide on cell-mediated immune responses and cytokine levels of metastatic tumor-bearing animals.

CONTEXT: Tumor microenvironment induces an active immune tolerance and escapes immune surveillance. In order to achieve an effective antitumor immune response, appropriately activated immune cells should maintain their antitumor activity to overcome the immune suppressive tumor microenvironment. OBJECTIVES: This study focuses on the effect of Thuja occidentalis L. (Cupressaceae) extract and its polysaccharide (TPS) on cell-mediated immune response (CMI) in metastasis bearing mice. MATERIALS AND METHODS: Metastasis was induced by injecting B16F-10 melanoma cells in mice through the tail vein and effector mechanisms of CMI was studied by analyzing cytotoxic T-lymphocyte (CTL) activity, natural killer (NK) cell activity, antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent complement-mediated cytotoxicity (ACC). The effect of T. occidentalis and TPS on pro-inflammatory cytokines and tissue inhibitor matrix metalloproteinases (TIMP) levels were also analyzed. RESULTS AND DISCUSSION: Administration of T. occidentalis and TPS enhanced the NK cell activity, ADCC and ACC much earlier than the control tumor-bearing animals. T. occidentalis and TPS were also found to decrease the elevated level of pro-inflammatory cytokines such as interleukin (IL)-1ß, IL-6, GM-CSF and tumor necrosis factor (TNF)-α in the serum of metastatic tumor-bearing animals. The level of antitumor factors such as IL-2 and TIMP was elevated by the treatment with T. occidentalis and TPS in the serum, which was lowered in the untreated tumor-bearing animals. CONCLUSION: This study clearly suggests that T. occidentalis and TPS effectively stimulate cell-mediated immune system and decrease pro-inflammatory cytokines, thereby inhibiting metastasis of tumor cells.

Protective role of Ipomoea obscura (L.) on cyclophosphamide-induced uro- and nephrotoxicities by modulating antioxidant status and pro-inflammatory cytokine levels.

Cyclophosphamide (CP) is widely used, alone or in combination with other chemotherapeutic agents, for the treatment of neoplastic diseases. Its urotoxicity may cause dose-limiting side-effects, for example haemorrhagic cystitis. This urotoxicity may lead to nephrotoxicity. In the present study, we investigated the protective role of Ipomoea obscura against CP-induced uro- and nephrotoxicities in animal models. Swiss albino mice were treated with an acute dose of CP (1.5 mmol/kg body wt ip) in the presence and absence of alcoholic extract of I. obscura (10 mg/kg, body wt, ip) for 5 days. The toxicities caused by CP were reversed by the extract administration as evident from the decrease in BUN, serum creatinine levels as well as an increase in body weight. A significant increase in kidney antioxidant system such as, GSH, SOD, CAT, and GPx was also observed in extract-treated animals. Histopathological analysis of urinary bladder and kidney indicated that CP-induced tissue damage was significantly reduced in animals treated with I. obscura. The lowered levels of cytokines IFN-γ and IL-2, after CP treatment were found to be increased in treated animals. At the same time the level of proinflammatory cytokine TNF-α, which was elevated during CP administration, was significantly reduced by extract administration. This study clearly demonstrates that I. obscura can ameliorate CP-induced bladder and renal toxicities by modulating antioxidant status and proinflammatory cytokine levels.

Anti-angiogenic activity of Ipomoea obscura extract and Ipobscurine-A.

OBJECTIVE: Ipomoea obscura (L.) Ker-Gawl, is a medicinal herb with indole alkaloids as an active constituent. In this study, we investigated the anti-angiogenic activity of I. obscura extract and one of its major compounds Ipobscurine-A (IPO-A). METHODS: In vivo angiogenesis was induced by injecting B16F10 melanoma cells intradermally on the shaven ventral skin of C57BL/6 mice. In vitro experiments were conducted using human umbilical vein endothelial cells. RESULTS: I. obscura and IPO-A significantly inhibited endothelial cell proliferation, migration, invasion, and tube formation in vitro. Vascular endothelial growth factor (VEGF)-induced sprouting of endothelial cells from rat aorta ex vivo was also inhibited. A marked decrease in the production of matrix metalloproteinases (MMPs) and the expressions of VEGF, cyclooxygenase-2, and nitric oxide synthase by B16F10 cells were observed after the treatment with the extract or IPO-A. Intraperitoneal administration of the extract significantly inhibited B16F10 melanoma cell line-induced neo-vessel formation in C57BL/6 mice in vivo. Analysis of serum cytokine profile clearly showed that extract significantly reduced the elevated levels of pro-inflammatory cytokines such as interleukins (IL)-1ß, IL-6, tumor necrosis factor-α, and granulocyte-monocyte colony stimulating factor and the most potent angiogenic factor VEGF in animals. Serum NO level was also found to be significantly lowered by the administration of the extract. Anti-angiogenic factors such as TIMP-1 and IL-2 level were elevated in the extract-treated animals. CONCLUSION: These data clearly demonstrate that I. obscura extract and IPO-A inhibit the tumor-specific angiogenesis by downregulating pro-angiogenic factors such as MMP, VEGF, and pro-inflammatory mediators and upregulating anti-angiogenic factors such as IL-2 and TIMP-1.

Evaluation of the anti-inflammatory and anti-tumor effect of Ipomoea obscura (L) and its mode of action through the inhibition of proinflammatory cytokines, nitric oxide and COX-2.

Ipomoea obscura (L) is a widely used medicinal plant. In this study, we investigated its anti-inflammatory and anti-tumor effect using in vitro and in vivo models. Methanolic extract of I. obsucra (10 mg/kg b.wt) was given interaperitoneally before inducing inflammation (both acute and chronic) and tumor to mice. I. obscura produced significant inhibition of 55.6%, 42%, and 65% in the paw edema of animals induced by carrageenan, dextran, and formalin respectively. The extract was also a potent inhibitor of lipopolysaccharide (LPS)-induced NO, CRP, and proinflammatory cytokine production via gene expression in peritoneal macrophages. TNF-α production by macrophage culture treated with LPS was found to be significantly inhibited by I. obscura. The extract was 100% toxic at a concentration of 500 µg/mL for both Dalton's lymphoma ascites (DLA) and Ehrlich ascites carcinoma (EAC) cells. The extract was also found to inhibit tumor cell proliferation in a dose and time-dependent manner. It could also inhibit solid tumor development in mice induced with DLA cells and increased life span of mice bearing EAC tumor to 83% and 53.8%, respectively. This anti-inflammatory effect of the extract is assumed to result mainly from the inhibition of some key enzymes and mediators involved in the inflammation and/or cell signaling pathways such as iNOS, COX-2, and proinflammatory cytokines. This anti-inflammatory property might be the reason for its anti-tumor effects.

Ipomoea obscura ameliorates cyclophosphamide-induced toxicity by modulating the immune system and levels of proinflammatory cytokine and GSH.

Ipomoea obscura L. is a widely used medicinal plant. The objective of this study was to investigate its protective activity against cyclophosphamide (CTX)-induced toxicity in mouse models. Swiss albino mice were treated intraperitoneally with CTX (25 mg/kg body weight) along with I. obscura extract (10 mg/kg body weight) for 10 days. Extract significantly reduced myelosuppression caused by CTX and improved the relative organ weight, total white blood cell count, and bone marrow cellularity. The elevated levels of parameters related to pathophysiology of the liver, namely glutamate pyruvate transaminase, alkaline phosphatase, and lipid peroxidation, were significantly reduced by extract treatment. Reduction of liver and intestinal glutathione levels of CTX-treated animals was reversed by I. obscura. The lowered levels of cytokines, namely IFN-γ, IL-2, and granulocyte-monocyte colony-stimulating factor after CTX treatment were found to be increased in I. obscura treated animals. Treatment with I. obscura could also decrease the level of proinflammatory cytokine TNF-α. The data suggested that I. obscura can act as a potent chemoprotective agent and can be used as an adjuvant in chemotherapeutic applications.

Molecular epidemiology of human astrovirus infections in Saudi Arabia pediatric patients.

Earlier work Tayeb et al. [Tayeb et al. (2008): J Med Virol 80: 1919-1929] set out to study the epidemiology of diarrhea viruses in pediatric populations. The study addressed initially rotavirus, enteric adenovirus, and astrovirus but was later expanded to include norovirus (NoV). Viruses were sought in fecal specimens and characterized for genotype using molecular methods (PCR, RT-PCR, and RFLP) for the first time in KSA. The survey focused on three locations; Jeddah, Makkah, and Riyadh. During the Hajj, the chief population fluxes are via Jeddah to Makkah. One thousand samples were obtained from children (aged 6 years or less) presenting with diarrhea and thus representing community acquired rather than nosocomial infections. Rotavirus was identified in 6% of the samples followed by NoV accounted for 3.5%, astrovirus 1.9%, and adenovirus 1.4%. Rotavirus G9 was characterized for the first time in Saudi Arabia. Adenoviruses were confirmed and further typed using hexon-specific PCR and RFLP. These data were published by Tayeb et al. [Tayeb et al. (2008): J Med Virol 80: 1919-1929]. However, the nature of astrovirus identified was not investigated further. Therefore, more analysis details are appropriate for astrovirus in the Kingdom. As an extension of earlier work carried out on astrovirus serotype distribution in the Kingdom [Tayeb et al. (2008): J Med Virol 80: 1919-1929], a major objective of the project is to use molecular methods to determine the distribution of astrovirus genotype. Such data will help to provide valuable insights into genetic identities and possible sources of virus strains involved not only in pediatric gastroenteritis but also possible outbreaks in the community.

Harmine inhibits tumour specific neo-vessel formation by regulating VEGF, MMP, TIMP and pro-inflammatory mediators both in vivo and in vitro.

Harmine is a beta-carboline alkaloid present in medicinal plants such as Peganum harmala that have been used as folk medicine in anticancer therapy. In this study, we demonstrated the anti-angiogenic activity of harmine using in vivo and in vitro assay systems. In vivo anti-angiogenic activity was studied using B16F-10 melanoma cells which induced capillary formation in C57BL/6 mice. Intraperitoneal administration of harmine at 10 mg/kg body weight significantly decreased tumour directed capillary formation. A drastic elevation in serum pro-angiogenic factors such as vascular endothelial growth factor (VEGF), nitric oxide (NO) and pro-inflammatory cytokines in angiogenesis induced animals was significantly decreased by harmine treatment. At the same time harmine increased anti-tumour factors like interleukin-2 (IL-2) and tissue inhibitor metalloprotease (TIMP). Moreover nuclear factor (NF)-κB and other transcription factors like CREB, ATF-2 involved in tumour development and angiogenesis were also inhibited by harmine. Various in vitro assays also supported the anti-angiogenic activity of harmine. It reduced proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVEC). Direct treatment of the harmine also inhibited microvessel outgrowth from the rat aortic ring. Production of other factors by tumour cells which are involved in angiogenesis like cyclooxygenase (COX-2), inducible nitric oxide synthase (iNOS) and matrix metalloproteases (MMPs) were also decrease by the treatment with harmine. Our data suggest that harmine may be a strong angiogenic inhibitor with the ability to decrease the proliferation of vascular endothelial cells and to reduce expression of various pro-angiogenic factors.

Ipomoea obscura (L.) enhances the functions of immunological effector cells, inhibits proinflammatory cytokines and nitric oxide production by LPS induced macrophages.

Most of the synthetic chemotherapeutic agents available today are immunosuppressant, cytotoxic and exerts variety of side effects. Botanical based immunomodulators are often employed as supportive or adjuvant therapy to overcome the undesired effects of cytotoxic chemotherapeutic agents and to restore normal health. The methanolic extract of traditionally important medicinal plant Ipomoea obscura exhibited immunomodulatory activity in BALB/c mice. Intraperitoneal administration of five doses of the extract (10 mg/kg body wt) was found to enhance the total WBC count (13912 cells/mm(3)) on the 12(th) day, bone marrow cellularity (28.9 x 10(6)cells/femur) and number of alpha-esterase positive cells (1246 cells/4000 cells). Treatment with the extract along with the antigen, sheep red blood cells (SRBC), produced an enhancement in the circulating antibody titer and the number of plaque forming cells (PFC) in the spleen. Maximum number of PFC (267.6 PFC/10(6) spleen cells) was obtained on the 6(th) day. At the same time administration of Ipomoea obscura extract significantly reduced the elevated levels of proinflammatory cytokines and nitric oxide production by lipopolysaccharide stimulated macrophages. These results indicate the immunomodulatory activity of the alcoholic extract of Ipomoea obscura.