Exploring Canine-Human Differences in Product Performance. Part II: Use of Modeling and Simulation to Explore the Impact of Formulation on Ciprofloxacin In Vivo Absorption and Dissolution in Dogs.

This study explored the in vivo performance of three oral ciprofloxacin formulations (oral solution, fast, or slow dissolving tablets) in beagle dogs. The in vivo absorption and dissolution behaviors, estimated with in silico mechanistic models, were compared to the results previously published in human volunteers. Six normal healthy male beagle dogs (five to completion) received three oral formulations and an intravenous infusion in a randomized crossover design. Plasma ciprofloxacin concentrations were estimated by tandem mass spectrometry detection. A mechanistic absorption model was used to predict the in vivo dissolution and absorption characteristics of the oral formulations. Canine ciprofloxacin absorption was constrained to the duodenum/jejunum. This absorption window was far narrower than that seen in humans. Furthermore, while substantial within-individual variability in drug absorption was seen in human subjects, a greater magnitude of variability was observed in dogs. For three sets of data, a lag time in gastric emptying was necessary to improve the accuracy of model-generated in vivo blood level profile predictions. In addition to species-associated dissimilarities in drug solubilization due to human versus canine differences in gastrointestinal fluid compositions, the far more rapid intestinal transit time and potential segmental differences in drug absorption needed to be considered during human-canine extrapolation of oral drug and drug product performance. Through the use of mechanistic models, the data generated in the human and canine studies contributed insights into some aspects of the interspecies differences to be considered when extrapolating oral bioavailability/formulation effect data between dogs and humans.

Complex formation, thermal behavior and stability competition between Cu(II) ion and Cu(0) nanoparticles with some new azo dyes. Antioxidant and in vitro cytotoxic activity.

Four triazole and thiadiazole-based azo chromophores namely [(E)-4-((1H-1,2,4-triazol-3-yl)diazenyl)benzene-1,3-diol.(HL(1)), (E)-4-((5-(methylthio)-1H-1,2,4-triazol-3-yl)diazenyl)benzene-1,3-diol.(HL(2)), (E)-4-((1,3,4-thiadiazol-2-yl)diazenyl)benzene-1,3-diol.(HL(3)) and (E)-4-((5-mercapto-1,3,4-thiadiazol-2-yl)diazenyl)benzene-1,3-diol.(HL(4))] were synthesized and characterized by elemental analyses, IR, UV-Vis as well as mass spectroscopy. Cu(II) complexes of the investigated azo dyes have been synthesized and characterized by elemental analyses, IR, electronic and ESR spectra, magnetic susceptibility and thermogravimetric analyses. The bond lengths and bond angles have been calculated to confirm the geometry of the ligands and their Cu(II) complexes. The mode of interaction of the azodyes to copper nanoparticles was described as coordination mode of charged dye molecules on the colloidal Cu(0) surface through anchoring OH(-) group. The apparent association constants of the colloidal copper nanoparticles azodye complexes in solution were evaluated using the spectral method and compared with the formation constant of the Cu(II) azo complexes. The antitumor and antioxidant activities of the synthesized azo dyes and their Cu(II) azo complexes have been evaluated.

The scientific basis for establishing solubility criteria for veterinary species.

The relationship between factors influencing the in vitro and in vivo drug solubilization is highly dependent upon the physiology of the individual animal species and the physico-chemical properties of the drug. The solubilization of a drug in an aqueous medium is controlled by the strength of the interaction between a molecule of the active pharmaceutical ingredient (API) and the molecules of the solvent. In this regard, the stronger this interaction, the greater the likelihood that the drug will go into solution. Counteracting this solubilization process is the strength of the affinity of the solute for itself, or how tightly bound the compound is to its own solid state form. The stronger affinity of a solute for itself, the more difficult it will be for that molecule to go into solubility. However, solubility is not a universal value. Rather, it should be considered from the perspective of the interactions between the API in its solid form and the solution conditions such as: pH, the nature of the primary solvent, the presence of co-solvents, the presence of solubilizing additives, the ionic strength of the medium, incubation time, and temperature. Each of these factors needs to be considered when evaluating potential interspecies differences in drug solubility.

Single incision endoscopic surgery for gynaecomastia.

INTRODUCTION: Surgical excision has been an effective treatment for gynaecomastia. Recently, there has been a shift from the open approach to minimally invasive techniques. In this report we describe our technique which includes endoscopic excision and/or liposuction of gynaecomastia via a single lateral chest wall incision. METHODS: Between May 2007 and April 2010, a total of 12 gynaecomastia patients were treated with liposuction and/or endoscopic excision. Patients were divided into 3 groups: group I; liposuction only, group II; endoscopic excision plus liposuction and group III; endoscopic excision only. One 15 mm incision was made laterally at the anterior axillary line. A vacuum assisted liposuction removing the fatty tissue was performed. Then endoscopic excision of the remaining fibroglandular tissue was done under vision through the same incision. The parynchyma was then dissected into small pieces and pulled out. RESULTS: Group I had liposuction only (n = 4), group II had liposuction combined with endoscopic excision (n = 7) (58%) while group III had endoscopic excision only (n = 1). The mean operative time for liposuction and endoscopic excision was 58 min for each side. Mean hospital stay was 1.4 days. Postoperative complications included infection with abscess formation and one patient had seroma. Mean follow-up was 56 weeks. Eleven out of twelve patients (92%) were satisfied with their results. Long-term follow-up showed that results were stable over time, and no revisions were necessary. CONCLUSION: Endoscopic excision of gynaecomastia through a single lateral chest wall incision is a minimally invasive effective and safe technique for the management of gynaecomastia, with excellent aesthetic results and an acceptable complication rate.

Comparison of bovine in vivo bioavailability of two sulfamethazine oral boluses exhibiting different in vitro dissolution profiles.

The bolus (or oblet) is a dosage form that can be used for the oral administration of pharmaceutical compounds to ruminating species. Unlike traditional tablets, oral boluses may contain quantities of drug on the order of grams rather than milligrams. Due to its size, it is only recently that USP-like in vitro dissolution methods have been developed for this dosage form. However, whether or not these dissolution tests can predict product in vivo performance has yet to be determined. The importance of this issue is apparent when the U.S. Food and Drug Administration Center for Veterinary Medicine is faced with the decision of whether to require additional in vivo bioequivalence study data to support the approval of changes in product chemistry or manufacturing method. The current study was undertaken to determine whether an in vivo/in vitro correlation can be established for bovine sulfamethazine oral boluses and to acquire insight into the magnitude of changes in in vitro product performance that can occur before corresponding changes are seen in in vivo blood level profiles. Based upon the results of this investigation, it is concluded that marked changes in in vitro sulfamethazine bolus performance can be tolerated before resulting in altered in vivo blood level profiles. However, the data also suggest that rumenal absorption may occur for some compounds. Therefore the degree to which variation in product in vitro dissolution profiles can be tolerated may be compound specific.

Pancreatic stellate cells respond to inflammatory cytokines: potential role in chronic pancreatitis.

BACKGROUND: It is now generally accepted that chronic pancreatic injury and fibrosis may result from repeated episodes of acute pancreatic necroinflammation (the necrosis-fibrosis sequence). Recent studies suggest that pancreatic stellate cells (PSCs), when activated, may play an important role in the development of pancreatic fibrosis. Factors that may influence PSC activation during pancreatic necroinflammation include cytokines known to be important in the pathogenesis of acute pancreatitis, such as tumour necrosis factor alpha (TNF-alpha), and the interleukins 1, 6, and 10 (IL-1, IL-6, and IL-10). AIM: To determine the effects of these cytokines on PSC activation, as assessed by cell proliferation, alpha smooth muscle actin (alpha-SMA) expression, and collagen synthesis. METHODS: Cultured rat PSCs were incubated with cytokines for 24 hours. Cell proliferation was assessed by measuring (3)H thymidine incorporation into cellular DNA, alpha-SMA expression by western blotting, and collagen synthesis by incorporation of (14)C proline into collagenase sensitive protein. mRNA levels for procollagen alpha(1)(1) in PSCs were determined by northern and dot blotting methods. RESULTS: Expression of alpha-SMA by PSCs was increased on exposure to each of the cytokines used in the study. Stellate cell proliferation was stimulated by TNF-alpha but inhibited by IL-6, while IL-1 and IL-10 had no effect on PSC proliferation. Collagen synthesis by PSCs was stimulated by TNF-alpha and IL-10, inhibited in response to IL-6, and unaltered by IL-1. Changes in collagen protein synthesis in response to TNF-alpha, IL-10, and IL-6 were not regulated at the mRNA level in the cells. CONCLUSION: This study has demonstrated that PSCs have the capacity to respond to cytokines known to be upregulated during acute pancreatitis. Persistent activation of PSCs by cytokines during acute pancreatitis may be a factor involved in the progression from acute pancreatitis to chronic pancreatic injury and fibrosis.

Catalytic oligodeoxynucleotides define a key regulatory role for early growth response factor-1 in the porcine model of coronary in-stent restenosis.

Early growth response factor-1 (Egr-1) controls the expression of a growing number of genes involved in the pathogenesis of atherosclerosis and postangioplasty restenosis. Egr-1 is activated by diverse proatherogenic stimuli. As such, this transcription factor represents a key molecular target in efforts to control vascular lesion formation in humans. In this study, we have generated DNAzymes targeting specific sequences in human EGR-1 mRNA. These molecules cleave in vitro transcribed EGR-1 mRNA efficiently at preselected sites, inhibit EGR-1 protein expression in human aortic smooth muscle cells, block serum-inducible cell proliferation, and abrogate cellular regrowth after mechanical injury in vitro. These DNAzymes also selectively inhibit EGR-1 expression and proliferation of porcine arterial smooth muscle cells and reduce intimal thickening after stenting pig coronary arteries in vivo. These findings demonstrate that endoluminally delivered DNAzymes targeting EGR-1 may serve as inhibitors of in-stent restenosis.

Oral motor dysfunction and feeding difficulties in nephropathic cystinosis.

Nephropathic cystinosis is a genetic disorder in which the amino acid cystine accumulates in lysosomes, resulting in multiorgan dysfunction. Progressive neuromuscular dysfunction, with bulbar and upper extremity weakness, has been described in adults with this disorder. The purpose of the present study was to determine whether there was evidence of early bulbar involvement, suggested by feeding difficulties or oral motor dysfunction in these patients, and whether the feeding and oral motor problems were associated with other evidence of neurologic dysfunction. Twenty-two children and adolescents with nephropathic cystinosis were studied. Parents completed questionnaires on feeding history and oral motor problems. Eighteen patients were given an oral motor examination, and 14 received a complete neurologic examination. The majority of children had a history of feeding difficulties. Seven children required a gastrostomy tube. Abnormalities on oral motor examination included hypotonia, abnormal gag reflex, and throaty or congested voice. Abnormalities on neurologic examination included hypotonia, muscle weakness, gross and fine motor dysfunction, and ataxia. The results indicate that feeding difficulties and oral motor dysfunction are common in children with cystinosis and appear to correlate with the general degree of neurologic dysfunction. Long-term follow-up is necessary to determine whether the early oral motor problems predict the later development of the progressive myopathy observed in adults with cystinosis.

Does alcohol directly stimulate pancreatic fibrogenesis? Studies with rat pancreatic stellate cells.

BACKGROUND & AIMS: Activated pancreatic stellate cells have recently been implicated in pancreatic fibrogenesis. This study examined the role of pancreatic stellate cells in alcoholic pancreatic fibrosis by determining whether these cells are activated by ethanol itself and, if so, whether such activation is caused by the metabolism of ethanol to acetaldehyde and/or the generation of oxidant stress within the cells. METHODS: Cultured rat pancreatic stellate cells were incubated with ethanol or acetaldehyde. Activation was assessed by cell proliferation, alpha-smooth muscle actin expression, and collagen synthesis. Alcohol dehydrogenase (ADH) activity in stellate cells and the influence of the ADH inhibitor 4-methylpyrazole (4MP) on the response of these cells to ethanol was assessed. Malondialdehyde levels were determined as an indicator of lipid peroxidation. The effect of the antioxidant vitamin E on the response of stellate cells to ethanol or acetaldehyde was also examined. RESULTS: Exposure to ethanol or acetaldehyde led to cell activation and intracellular lipid peroxidation. These changes were prevented by the antioxidant vitamin E. Stellate cells exhibited ethanol-inducible ADH activity. Inhibition of ADH by 4MP prevented ethanol-induced cell activation. CONCLUSIONS: Pancreatic stellate cells are activated on exposure to ethanol. This effect of ethanol is most likely mediated by its metabolism (via ADH) to acetaldehyde and the generation of oxidant stress within the cells.

Free living amoebae as opportunistic parasites in immunocompromised hosts.

The acquisition of invasiveness of mammalian tissue by free living amoebae (FLA) is an extremely noteworthy phenomenon. Many factors may be responsible for this evolutionary change in their physiology. The present work aimed to clarify the host factors in this problem. Albino mice were the laboratory animals used in this work. Non-pathogenic strains of FLA were isolated from ear, nose and pharyngeal swabs and from contact lenses of patients attending out patient clinics in Benha University Hospitals. Isolation was done by cultivation of non-nutrient agar at 37 & 43 degrees C and identification was made by flagellation test. The degree of pathogenicity of isolated strains was assessed by using animal pathogenicity test. Endoxan (0.7 mg/kg) and Prednisolone (2 mg/kg) were used for 15 days orally to suppress the immunity of clean mice before their inoculation intranasally with the isolated strains. All groups were observed for 3 weeks post-infection. Mortality rates recorded were higher in Endoxan or Prednisolone treated and Naegleria infected groups A I & B I (20 & 15% respectively) than Endoxan or Prednisolone treated and acanthamoeba infected groups A II & B II (10%) and drug control (10%). Histopathological examination of brain and meninges revealed severe changes in form of severe meningoencephalitis, severe vacuolar degeneration, and moderate haemorrhage and necrosis in brains of groups AI & BI and less severe changes in groups AII & BII (10%) form of-non granulomatous encephalitis. Drug control groups showed negligible changes. Great attention must be paid to reveal the presence of FLA before using immunosuppressive drugs in human cases.

A method for analysis of gene expression patterns.

mRNA can be copied into cDNA with the use of reverse transcriptase so that the relative abundance of individual mRNAs is reflected in the cDNA product. With further manipulation a replica of the mRNA expression pattern can be duplicated into a radioactive double-stranded DNA probe. DNA from a series of genes inserted into plasmids can be fixed to a membrane using a slot blot manifold and probed with the RNA-derived DNA probe. The intensity of the hybridization signal for a given gene is a result of its relative abundance in the RNA-derived DNA probe. Quantitation can be achieved through the use of housekeeping genes as baseline monitors. Inclusion of vector sequences can negate any spurious hybridization to vector rather than insert sequences. We have successfully used this method to obtain gene expression patterns for RNA isolated from diverse sources including rodent tissues, various cell lines, and Drosophila and Caenorhabditis elegans samples. Northern blots have verified the results obtained. The pattern of expression of many genes can be determined from as little as 10 micrograms of total RNA, making this method ideally suited for studies in which RNA is rare or in short supply.

A rare case of 'coccidioidoma' of the breast.

Coccidioidomycosis has been reported in almost every tissue of the human body. The gastrointestinal tract and breast have been uniquely spared from this fungal infection. Despite the exceedingly large number of screening mammographies that are performed every year, to our knowledge subcutaneous breast tissue involvement as the sole presentation or of secondary spread of coccidioidomycosis has not been reported. We describe a patient who showed an unusual manifestation of a coccidioidal breast nodule simulating a neoplasm. The patient had been transiently immunosuppressed by prednisone therapy for presumptive temporal arteritis.

An improved enzyme immunoassay for progesterone in human plasma.

We describe an enzyme immunoassay for progesterone in which we use a progesterone-11alpha-hemisuccinyl-horseradish peroxidase conjugate as the "label" and an antiserum raised in rabbits to a progesterone-11alpha-hemisuccinyl-bovine serum albumin conjugate. In this assay, antibody-bound and free steroid are separated by using a second antibody precipitation procedure. The assay has a lower limit of sensitivity of 10 pg/assay tube and satisfies the usual criteria of specificity, precision, and accuracy. Results obtained with a comparison radioimmunoassay and our procedure agreed well (r = 0.98). Eighty samples can be assayed per day. It is not only well suited for surveys where accurate determination of progesterone concentrations in small plasma aliquots is required, but also for monitoring ovulation induction in patients attending infertility clinics.