Treatment of Patients with Mild to Moderate Ulcerative Colitis: A Middle East Expert Consensus.

The prevalence of ulcerative colitis (UC) in the Middle East is increasing, impacting the economic and healthcare burden. The management of patients with mild to moderate UC is still a challenge as several factors can affect optimal care, including drug choice, induction and maintenance dose, treatment optimization and de-escalation, therapy duration, monitoring, and safety profile. We conducted an expert consensus to standardize the management of patients with mild to moderate UC. Sixteen experts in inflammatory bowel diseases, through a well-established and accepted Delphi methodology, voted and approved eight statements in order to provide practical guidance to clinicians in the Middle East.

Early Renoprotective Effect of Ruxolitinib in a Rat Model of Diabetic Nephropathy.

Diabetic kidney disease (DKD) is still one of the unresolved major complications of diabetes mellitus, which leads ultimately to end-stage renal disease in both type 1 and type 2 diabetes patients. Available drugs that suppress the renin-angiotensin system have partially minimized the disease impact. Yet, there is an unmet need for new therapeutic interventions to protect the kidneys of diabetic patients. In DN, glomerular sclerosis and tubulointerstitial fibrosis are mediated through several pathways, of which JAK/STAT is a key one. The current study explored the potential renoprotective effect of the JAK1/JAK2 inhibitor ruxolitinib (at doses of 0.44, 2.2, and 4.4 mg·kg-1) compared to that of enalapril at a dose of 10 mg·kg-1, in a rat model of streptozotocin-induced diabetes mellitus over 8 weeks. The effect of ruxolitinib was assessed by determining urinary albumin/creatinine ratio, serum level of cystatin, and levels of TGF-ß1, NF-κB, and TNF-α in renal tissue homogenates by biochemical assays, the glomerular sclerosis and tubulointerstitial fibrosis scores by histological analysis, and fibronectin, TGF-ß1, and Vimentin levels by immunohistochemical staining with the respective antibodies. Our results revealed a significant early favorable effect of a two-week ruxolitinib treatment on the renal function, supported by a decline in the proinflammatory biomarkers of DKD. This pre-clinical study suggests that the renoprotective effect of ruxolitinib in the long term should be investigated in animals, as this drug may prove to be a potential option for the treatment of diabetic kidney disease.

Role of selective blocking of bradykinin receptor subtypes in attenuating allergic airway inflammation in guinea pigs.

The present study was designed to evaluate the potential role of bradykinin antagonists (R-715; bradykinin B1 receptor antagonist and icatibant; bradykinin B2 receptor antagonist) in treatment of allergic airway inflammation in comparison to dexamethasone and montelukast. R-715 as dexamethasone significantly decreased peribronchial leukocyte infiltration, bronchoalveolar lavage fluid (BALF) albumin and interleukin 1ß as well as serum OVA-specific IgE level. Also, R-715 like montelukast significantly decreased BALF cell count (total and eosinophils). Icatibant showed negative results. The current findings suggest that selective bradykinin B1 receptor antagonists may have the therapeutic potential for the treatment of allergic airway inflammation.

Can three-dimensional ultrasound measurement of fetal adrenal gland enlargement predict preterm birth?

PURPOSE: To assess the diagnostic accuracy of three-dimensional (3D) ultrasound measurements of fetal adrenal gland volume (AGV) and fetal zone enlargement (FZE) as predictors of PTB compared to measurements of cervical length (CL) and cervicovaginal fetal fibronectin (CVFF). METHOD: This prospective study included women presenting at 28-36 weeks of gestation with threatened preterm labor (TPL). Fetal AGV and FZE were measured using 3D ultrasound. Two-dimensional (2D) ultrasound was used to measure the CL. The AGV was corrected for the ultrasound-estimated fetal weight (cAGV). Qualitative CVFF detection was also performed. The diagnostic accuracy of cAGV, FZE, CL, and CVFF was compared considering preterm birth (PTB) within 7 days of recruitment as the main outcome measure. RESULTS: Seventy-five pregnant women were included in the final statistical analysis. Twenty-seven women (36 %) delivered within 7 days. cAGV and FZE had the highest sensitivities and specificities to predict PTB within 7 days when compared with CL and CVFF. Multivariate analysis, including cAGV, FZE, CL, and CVFF, revealed that cAGV and FZE were independent predictors of PTB within 7 days in the study participants. CONCLUSION: In women who presented at 28-36 weeks of gestation with TPL, cAGV and FZE can be used as independent predictors of PTB.

Bioconversion by functional P450 1A9 and P450 1C1 of Anguilla japonica.

We indicated that two P450s (1A9 and 1C1) from Japanese eel (Anguilla japonica) metabolized 7-ethoxycoumarin, 7-ethoxyresorufin, and flavanone. At first, we constructed expression vectors for two types of P450 (1A9 and 1C1). The reduced CO-difference spectra of Escherichia coli cells transformed with these plasmids showed Soret peaks (450 nm) that were typical of P450s. We performed bioconversion experiments in which substrates were added directly to incubation medium. The resulting metabolite(s) were extracted and analyzed by high-performance liquid chromatography and spectrofluorometer. Incubation of 50 nmol 7-ethoxyresorufin with P450 1C1 yielded 0.773 nmol of deethylated product, whereas 50 nmol 7-ethoxycoumarin resulted in 4.76 nmol. P450 1A9 metabolized 50 nmol of 7-ethoxyresorufin and 7-ethoxycoumarin to yield 6.54 and 20.9 nmol of deethylated product, respectively. Incubation of 50 nmol flavanone with P450 1C1 yielded 1.46 nmol and 0.69 nmol of products, whereas 50 nmol flavanone with P450 1A9 resulted in 1.10 nmol. In this system, 4'-hydroxy flavanones were formed by P450 1A9 and P450 1C1. P450 1A9 also metabolized 50 nmol of 17 beta-estradiol to yield 4.25 nmol of product. In this system, 2-hydroxy estradiol was formed by P450 1A9 using 17 beta-estradiol as a substrate. This study is the first to identify the substrates that P450 1C1 and 1A9 metabolize.

Complementary DNA cloning and organ expression of cytochrome P450 1C2 in carp (Cyprinus carpio).

Cytochrome P450 (CYP) genes, which make up a large gene superfamily, are known to play an important role in drug metabolism. The CYP1 family, one of the gene families of the CYP superfamily, has three subfamilies of genes whose sequences have been deposited in the GenBank/EMBL thus far: CYP1A, CYP1B, and CYP1C. Mammals as well as fish confront numerous foreign chemicals in the environment that may accumulate to toxic levels unless they are metabolized and eliminated by processes largely mediated by CYP enzymes. A new complementary DNA of the CYP1C subfamily encoding CYP1C2 was isolated from the carp liver after a single intraperitoneal injection of beta-napthoflavone (BNF). The full-length cDNA obtained contained a 5' noncoding region of 198 bp, an open reading frame of 1575 bp coding for 524 amino acids and a stop codon, and a 3' noncoding region of 531 bp. The predicted molecular weight of the protein was approximately 59.3 kDa. The amino acid sequence deduced from the carp CYP1C2 sequence showed a similarity of 76.6% with that deduced from our previously reported carp CYP1C1. It exhibited similarities of 77.3, 73.7, and 76.4% with those deduced from scup CYP1C2, scup CYP1C1, and Japanese eel CYP1C1 sequences, respectively. Carp CYP1C2 cDNA showed similarities with reported CYP1Bs of teleosts and mammals, namely, 47.6, 45.3, 45.7, 44.0, and 44.6% for carp, plaice, human, rat, and mouse CYP1B1s, respectively, while it exhibited a similarity of 49.0% with carp CYP1B2. The carp CYP1C2 sequence was aligned with the CYP1 sequences and has been deposited in the GenBank/EMBL data bank with the accession number AY437777. The phylogenetic tree constructed using fish and mammalian CYP1 sequences suggested a closer relationship of CYP1C with CYP1B than with CYP1A. The tree showed possibile existence of CYP1C subfamily genes in mammalian species. Northern blot analysis of the liver, intestines, kidneys, and gills revealed a distinct induced expression only in the kidneys, with no detectable constitutive expression in the other organs studied.

A preliminary observation on water quality and plankton of an earthen fish pond in Bangladesh: recommendations for future studies.

The present study provides a characterization of water quality and plankton samples in earthen fish pond in Rajshahi, Bangladesh. Sampling was done over a period of six months, running from October, 2004 through March, 2005. All the water quality parameters were within the optimal ranges for plankton productivity. Temperatures varied from 19.75 to 27.25 degrees C; transparency, 24.75-29.50 cm; pH, 6.62-7.85; Dissolved Oxygen (DO), 3.87-5.85 mg L(-1); free CO2 5.25-7.25 mg L(-1) and bicarbonate (HCO3) alkalinity, 81.25-147.5 mg L(-1). Analyses of plankton samples recorded a total of 5 classes phytoplankton viz.; Bacillariophyceae, Chlorophyceae, Cyanophyceae, Dinophyceae, Euglenophyceae and 2 classes of zooplankton; Crustacea and Rotifera. The phytoplankton population was comprised of 17 genera belonging to Cyanophyceae (5 classes, 34.47%), Bacillariophyceae (3, 13.87%), Cyanophyceae (3, 34.48%), Euglenophyceae (3, 10.68%) and 1 to Dinophyceae (6.50%). The zooplankton population consisted of 10 genera belonging to Rotifera (4, 40.13%) and Crustacea (6, 59.87%). Phytoplankton and zooplankton abundance varied from 60800 to 239400 units/l and 7620 to 12160 units/l, respectively. It is concluded that the phytoplankton groups provide the main support for earthen pond aquaculture in the pond compared to zooplankton classes. The information provides for more research to compare water quality and pond plankton characteristics in earthen aquaculture systems with and without fish stocking. Further studies on the seasonal changes of water quality parameters and its effects on plankton production in the fish ponds and all year extended monitoring is recommended in future studies.

A preliminary observation on the pond culture of European eel, Anguilla anguilla (Linnaeus, 1758) in Egypt: recommendations for future studies.

An experiment was conducted to assess the potential of the European eel, Anguilla anguilla for earthen pond aquaculture without supplementary feeding at Lake Manzala, Egypt. Juvenile A. anguilla of mean length 11.7 cm and 2.4 g weight were stocked in earthen ponds measuring 3 feddans (about 12,600 m2) and 1 m deep. Stocking was done in May 2003 at a rate of 5000 fish feddan(-1) in a polyculture system including tilapia and mullets and fed mainly on natural occurring prey (natural spawned tilapia) and small shrimp. The eels were culture for a period of 2 years, May 2003 to April 2005. Sampling for growth and survival were evaluated yearly. At the end of the culture period, the gross weight of the harvested eels was measured and the net pond production calculated by the difference between weight stocked and weight harvested. Temperature varied from 11.5 to 28.2 degrees C and 12.2 to 29.3 degrees C; P(H), 7.3 to 8.9 and 7.5 to 8.8; Dissolved Oxygen (DO), 5.2 to 9.8 mg L(-1) and 4.1 to 8.3 mg L(-1); and Salinity, 2.5 to 5.5 psu and 3.0 to 6.8 psu for first year and second year, respectively. At the end of the culture period, A. anguilla attained average weight of 121.4 g fish(-1) at the end of the first year and a weight range of 152.5 to 430 g fish(-1) with an average of 280.36 g fish(-1) at the end of the second year. Survival rate ranged from 91% during the first year to 100% during the second year. Net eel production was 540.18 kg feddan(-1) at the end of the first year and 723.36 kg feddan(-1) at the end of the second year. Daily increments in weight per fish were 0.33 and 0.44 for first and second year, respectively. This experiment demonstrated the possibility of cultivation of eels as well as the higher growth rate in earthen ponds. The aquaculture strategy of eel with high stocking densities through low cost artificial feeds are recommended in future studies.

Estrogen-responsive element (ERE)-like motifs affect the 3-methylcholanthrene induction of eel CYP1A gene.

Some forms of cytochrome P450 (CYP) genes are known to be induced by xenobiotics such as dioxins. Induction of the CYP1A gene is mediated by an aryl hydrocarbon receptor (AHR) which binds to a specific nucleotide sequence called a dioxin-responsive element (DRE) located in the 5' enhancer region of the gene. Functional analysis of the regulatory region of the eel CYP1A gene had shown that a 654-bp region near the basal promoter, containing no DREs but three motifs that resemble estrogen-responsive element (ERE) halfsites, contributes substantially to the induced expression. Considering the importance of non-DRE elements in CYP1A gene induction, we investigated the role of ERE-like motifs using a point mutation technique. The regulatory region of the eel CYP1A gene was identified by creating mutations in all three ERE half-sites simultaneously or individually. The regulatory region constructs were cloned upstream of the luciferase gene in an expression vector which was microinjected into medaka ova. Expression of luciferase as a foreign gene in medaka fry exposed to 3-methylcholanthrene (3-MC)-treated feed was measured by competitive PCR analysis. Mutation in all the three ERE half-sites reduced expression to 10%. Mutation in ERE(-2) or ERE(-3) reduced the expression to 50%, whereas mutation in ERE(-1) did not affect the induction. The two functional half-sites of the eel CYP1A gene are palindromic to each other and appear to constitute the full-ERE.

Complementary DNA cloning and constitutive expression of cytochrome P450 1C1 in the gills of carp (Cyprinus carpio).

Cytochrome P450 (CYP) enzymes constitute a multigene family of many endogenous and xenobiotic substances. The CYP1 family is of particular interest in environmental toxicology because its members are dominant in the metabolism of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and aryl amines. A new complementary DNA of the CYP1C subfamily encoding CYP1C1 was isolated from carp liver after intraperitoneal injection of beta-napthoflavone (BNF). The full-length cDNA obtained contained a 5' noncoding region of 244 bp, an open reading frame of 1572 bp coding for 524 amino acids, a stop codon, and a 3' noncoding region of 965 bp. The predicted molecular weight of the protein was approximately 59.3 kDa. The deduced amino acid sequence of this cDNA was 82.1% and 80.2% similar to Japanese eel and scup CYP1C1 sequences, respectively, while it exhibited a similarity of 74.9% with the scup CYP1C2 sequence. The deduced amino acid sequence of carp CYP1C1 showed similarities with those of the reported CYP1B1s of teleosts and mammals of 48.4, 48.8, 48.2, 48.6, 45.3, and 45.5% for carp CYP1B1, carp CYP1B2, plaice CYP1B1, and human, rat, and mouse CYP1B1, respectively. The phylogenetic tree constructed using fish and mammalian CYP1 sequences suggested a closer relationship of the CYP1C subfamily to CYP1B than to CYP1A. The tree showed the possibility of the existence of CYP1C subfamily genes in mammalian species. Northern blot analysis for the liver, intestine, gills, and kidney showed no detectable induced expression but constitutive expression in the gill organs.

cDNA cloning, sequence analysis and expression of 3-methylcholanthrene-inducible cytochrome P450 1B1 in carp (Cyprinus carpio).

Cytochrome P450 (CYP) genes, which make up a large gene superfamily, are known to play an important role in drug metabolism. The levels of expression of CYP genes in the tissue of fish inhabiting polluted areas have been used extensively in biomonitoring studies as indicators of dioxin pollution. Complementary DNA of cytochrome CYP1B1 was isolated from carp (Cyprinus carpio) liver 24 h after the injection of 3-methylcholanthrene (3-MC). The full length cDNA obtained contained a 5' noncoding region of 178 bp, an open reading frame of 1593 bp coding for 530 amino acids and a stop codon, and a 3' noncoding region of 1599 bp. The predicted molecular weight of the encoded protein was approximately 60.2 kDa. The deduced amino acid sequence exhibited an identity of 60.6% with reported CYP1B sequences of plaice CYP1B, and of 52.4, 51.4, and 50.3% with human, rat, and mouse CYP1B1s, respectively. It exhibited similarities of 48.4 and 47.3% with scup CYP1C2 and -1C1 sequences. The percent identities with CYP1A sequences showed lower values in the range from 35.3 to 39.5% with mammals and teleosts. The phylogenetic tree of the CYP1 family members constructed by the protein maximum likelihood method indicates that the carp CYP1B1 and plaice CYP1B share a common ancestry with the mammalian CYP1B1s. Carp treated with 3-MC showed expression of CYP1B1 in liver, intestine and gills with distinct induction except for the gills that showed marked constitutive expression. The presence of two successive signals in treated gills at low stringency hybridization may suggest the existence of another CYP1B member that is expressed in the gills of carp.

Isolation of cDNA of novel cytochrome P450 1B gene, CYP1B2, from Carp (Cyprinus carpio) and its induced expression in gills.

Cytochrome P450 (CYP) enzymes constitute a multigene family of many endogenous and xenobiotic substances. The CYP1 family is of particular interest in environmental toxicology because its members are dominant in the metabolism of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and aryl amines. A new complementary DNA of the CYP1B subfamily encoding CYP1B2 was isolated from carp liver after intraperitoneal injection with 3-methylcholanthrene (3-MC). The obtained full-length cDNA contained a 5'noncoding region of 161 bp, an open reading frame of 1593 bp coding for 530 amino acids and a stop codon, and a 3'noncoding region of 1457 bp. The predicted molecular weight of the protein was approximately 60.2 kDa. The deduced amino acid sequence of this cDNA was 91% similar to that of our previously reported carp CYP1B1; its similarities with those of the reported CYP1B1s of teleosts and mammals were 60.8, 54.4, 50.8, and 51.4% for plaice, human, rat, and mouse, respectively. The phylogenetic tree of fish and mammalian CYP1 sequences constructed by the protein maximum-likelihood method suggested a relatively recent divergence of CYP1B2 from CYP1B1 in the ancestor of carp and closely related species. Despite the structural similarity of CYP1B2 with CYP1B1, which showed induced expression in 3-MC-treated liver, intestine, and gills with marked constitutive expression in gills, CYP1B2 revealed induced expression in gills but not in liver or intestine, and no detectable constitutive expression in the tissues studied.