Modeling and forecasting the distribution of Vibrio vulnificus in Chesapeake Bay.

AIM: To construct statistical models to predict the presence, abundance and potential virulence of Vibrio vulnificus in surface waters of Chesapeake Bay for implementation in ecological forecasting systems. METHODS AND RESULTS: We evaluated and applied previously published qPCR assays to water samples (n = 1636) collected from Chesapeake Bay from 2007-2010 in conjunction with State water quality monitoring programmes. A variety of statistical techniques were used in concert to identify water quality parameters associated with V. vulnificus presence, abundance and virulence markers in the interest of developing strong predictive models for use in regional oceanographic modeling systems. A suite of models are provided to represent the best model fit and alternatives using environmental variables that allow them to be put to immediate use in current ecological forecasting efforts. CONCLUSIONS: Environmental parameters such as temperature, salinity and turbidity are capable of accurately predicting abundance and distribution of V. vulnificus in Chesapeake Bay. Forcing these empirical models with output from ocean modeling systems allows for spatially explicit forecasts for up to 48 h in the future. SIGNIFICANCE AND IMPACT OF THE STUDY: This study uses one of the largest data sets compiled to model Vibrio in an estuary, enhances our understanding of environmental correlates with abundance, distribution and presence of potentially virulent strains and offers a method to forecast these pathogens that may be replicated in other regions.

Insertion of Inhbb into the Inhba locus rescues the Inhba-null phenotype and reveals new activin functions.

The activins (dimers of betaA or betaB subunits, encoded by the genes Inhba and Inhbb, respectively) are TGF-beta superfamily members that have roles in reproduction and development. Whereas mice homozygous for the Inhba-null allele demonstrate disruption of whisker, palate and tooth development, leading to neonatal lethality, homozygous Inhbb-null mice are viable, fertile and have eye defects. To determine if these phenotypes were due to spatiotemporal expression differences of the ligands or disruption of specific ligand-receptor interactions, we replaced the region of Inhba encoding the mature protein with Inhbb, creating the allele Inhbatm2Zuk (hereafter designated InhbaBK). Although the craniofacial phenotypes of the Inhba-null mutation were rescued by the InhbaBK allele, somatic, testicular, genital and hair growth were grossly affected and influenced by the dosage and bioactivity of the allele. Thus, functional compensation within the TGF-beta superfamily can occur if the replacement gene is expressed appropriately. The novel phenotypes in these mice further illustrate the usefulness of insertion strategies for defining protein function.

Microduplication of Xp11.23p11.3 with effects on cognition, behavior, and craniofacial development.

We report an ~1.3 Mb tandem duplication at Xp11.23p11.3 in an 11-year-old boy with pleasant personality, hyperactivity, learning and visual-spatial difficulties, relative microcephaly, long face, stellate iris pattern, and periorbital fullness. This clinical presentation is milder and distinct from that of patients with partially overlapping Xp11.22p11.23 duplications which have been described in males and females with intellectual disability, language delay, autistic behaviors, and seizures. The duplicated region harbors three known X-linked mental retardation genes: FTSJ1, ZNF81, and SYN1. Quantitative polymerase chain reaction from whole blood total RNA showed increased expression of three genes located in the duplicated region: EBP, WDR13, and ZNF81. Thus, over-expression of genes in the interval may contribute to the observed phenotype. Many of the features seen in this patient are present in individuals with Williams-Beuren syndrome (WBS). Interestingly, the SYN1 gene within the duplicated interval, as well as the STX1A gene, within the WBS critical region, co-localize to presynaptic active zones, and play important roles in neurotransmitter release.

22q13.3 deletion syndrome: clinical and molecular analysis using array CGH.

The 22q13.3 deletion syndrome results from loss of terminal segments of varying sizes at 22qter. Few genotype-phenotype correlations have been found but all patients have mental retardation and severe delay, or absence of, expressive speech. We carried out clinical and molecular characterization of 13 patients. Developmental delay and speech abnormalities were common to all and comparable in frequency and severity to previously reported cases. Array-based comparative genomic hybridization showed the deletions to vary from 95 kb to 8.5 Mb. We also carried out high-resolution 244K array comparative genomic hybridization in 10 of 13 patients, that defined the proximal and distal breakpoints of each deletion and helped determine the size, extent, and gene content within the deletion. Two patients had a smaller 95 kb terminal deletion with breakpoints within the SHANK3 gene while three other patients had a similar 5.5 Mb deletion implying the recurrent nature of these deletions. The two largest deletions were found in patients with ring chromosome 22. No correlation could be made with deletion size and phenotype although complete/partial SHANK3 was deleted in all patients. There are very few reports on array comparative genomic hybridization analysis on patients with the 22q13.3 deletion syndrome, and we aim to accurately characterize these patients both clinically and at the molecular level, to pave the way for further genotype-phenotype correlations. (c) 2010 Wiley-Liss, Inc.

Drivers of germ cell maturation.

Spermatogenesis requires progression of germ line stem cells through a precisely ordered differentiation pathway to form spermatozoa. Diverse and dynamic signals from the transforming growth factor-beta (TGF-beta) superfamily influence many stages of germ cell development. For example, interactions between several TGF-beta superfamily ligands (bone morphogenetic proteins, activin, and glial-derived neurotrophic growth factor [GDNF]) appear to govern the onset of spermatogenesis, and we are exploring how germ cells interpret these competing signals. We examined the in vivo impact of activin on testis development using two mouse models, the inhba-/- mouse (which lacks the gene encoding the activin A subunit and dies at birth) and BK mice, with inhbb (encoding the activin betaB subunit) replacing inhba (which survive to adulthood and show delayed fertility onset in males). Distinct effects on Sertoli cell and germ cell populations during fetal and early postnatal development were measured. We recognize that specific proteins, including downstream targets of TGF-beta signals, such as Smads, must move into the nucleus to implement the gene transcription changes required for development. We hypothesized that changes at the level of cellular nuclear transport machinery may be required to mediate this. Examination of proteins involved in classical nuclear import, the importins, revealed that each importin has a developmentally regulated expression pattern in male germ cells. Because each importin binds a selected range of cargo proteins and mediates their nucleocytoplasmic passage, our findings suggest that each importin ferries cargo required for discrete stages of spermatogenesis.

Genital exhibitionism in women.

Historically, genital exhibitionism has usually been regarded as an exclusively male phenomenon. The authors present a case study of a female exhibitionist which is at variance with that contention. They offer a formulation of the psychodymanics of female exhibitionism, focusing on its pregenital, attention-seeking purpose. The findings from this case justify a critical reexamination of the traditional definitions of exhibitionism.

Synthesis and characterization of heteroarotinoids demonstrate structure specificity relationships.

Heteroarotinoids are synthetic retinoids derived from trans-retinoic acid and the arotinoid structures and include a heteroatom in a five- or six-membered cyclic ring. This is the first systematic study of influences of the heteroatom, ring size, number of aryl groups, and terminal side chain on retinoid receptor specificity. Two new heteroarotinoids were synthesized and characterized. Although all heteroarotinoids activated RAR receptors, two dominant associations between structure and specificity were identified across all compounds. The six-membered ring conferred increased RARbeta specificity over the five-membered ring. The sulfur atom conferred greater specificity for RARgamma than the oxygen atom. RARalpha specificity was attenuated by a combination of influences from the heteroatom and aryl groups. In summary, the heteroatom and cyclic ring size exerted dominant effects, while the number of aryl rings and terminal side chain had attenuating effects on retinoid receptor specificity of heteroarotinoids.

Synthesis, structure-activity relationships, and RARgamma-ligand interactions of nitrogen heteroarotinoids.

Three heteroarotinoids containing a nitrogen atom in the first ring and a C-O linking group between the two aryl rings were synthesized and evaluated for RAR and RXR retinoid receptor transactivation, tumor cell growth inhibition, and transglutaminase (TGase) induction. Ethyl 4-(N,4,4-trimethyl-1,2,3,4-tetrahydroquinolinyl)benzoate (1) contained an N-CH(3) group and activated all retinoid receptors except for RARgamma. Inceasing the hydrophobicity around the rings with analogues ethyl 4-(N,4,4,7-tetramethyl-1,2,3, 4-tetrahydroquinolin-6-oyloxy)benzoate (2) [7-methyl group added] and ethyl 4-(4,4-dimethyl-N-isopropyl-1,2,3, 4-tetrahydroquinolin-6-oyloxy)benzoate (3) [NCH(CH(3))(2) group at C-4] increased the potency and specificity for RARalpha, RARbeta, and RXRalpha, compared to 1, but had little effect on RXRbeta and RXRgamma activation. Although 1 and 3 were unable to activate RARgamma, 2 did activate this receptor with efficacy and high potency equal to that of 9-cis-retinoic acid (9-c-RA). All three heteroarotinoids exhibited 5-8-fold greater specificities for RARbeta over RARalpha. In addition, esters 1-3 inhibited the growth of two cell lines each derived from cervix, vulvar, ovarian, and head/neck tumors with similar efficiencies to that of 9-c-RA through a mechanism independent of apoptosis. The vulvar cell lines were the most sensitive, and the ovarian lines were the least sensitive. Ester 2 was similar to 1 and 3 except that 2 was a much more potent growth inhibitor of the two vulvar cell lines, which is consistent with strong RARgamma activation by 2 (but not by 1 and 3) and the high levels of RARgamma expression in skin. All three heteroarotinoids induced production of TGase, a marker of retinoid activity in human erythroleukemic cells. Esters 2 and 3 were the more potent TGase activators than 1, in agreement with the stronger activation of the RAR receptors by 2 and 3. The biological activities of these agents, and the RARgamma potency of 2 in particular, demonstrate the promise of these compounds as pharmaceutics for cancer and skin disorders.

Heteroarotinoids inhibit head and neck cancer cell lines in vitro and in vivo through both RAR and RXR retinoic acid receptors.

A class of less toxic retinoids, called heteroarotinoids, was evaluated for their molecular mechanism of growth inhibition of two head and neck squamous cell carcinoma (HNSCC) cell lines SCC-2 and SCC-38. A series of 14 heteroarotinoids were screened for growth inhibition activity in vitro. The two most active compounds, one that contained an oxygen heteroatom (6) and the other a sulfur heteroatom (16), were evaluated in a xenograph model of tumor establishment in nude mice. Five days after subcutaneous injection of 10(7) SCC-38 cells, groups of 5 nu/nu mice were gavaged daily (5 days/week for 4 weeks) with 20 mg/kg/day of all-trans-retinoic acid (t-RA, 1), 10 mg/kg/day of 6, 10 mg/kg/day of 16, or sesame oil. After a few days, the dose of t-RA (1) was decreased to 10 mg/kg/day to alleviate the side effects of eczema and bone fracture. No significant toxic effects were observed in the heteroarotinoid groups. All three retinoids caused a statistically significant reduction in tumor size as determined by the Student t-test (P < 0. 05). Complete tumor regression was noted in 3 of 5 mice treated with t-RA (1), 4 of 5 mice treated with 16, 1 of 5 mice treated with 6, and 1 of 5 mice treated with sesame oil. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to determine that the expression levels of RARalpha, RXRalpha, and RXRbeta were similar in the two cell lines, while RARbeta expression was higher in SCC-2 over SCC-38, and RARgamma expression was higher in SCC-38 over SCC-2. Receptor cotransfection assays in CV-1 cells demonstrated that 16 was a potent activator of both RAR and RXR receptors, while 6 was selective for the RXR receptors. Transient cotransfection assays in CV-1 cells using an AP-1 responsive reporter plasmid demonstrated that t-RA (1), 6, and 16 each inhibited AP-1-driven transcription in this cell line. In conclusion, the growth inhibition activity of the RXR-selective 6 and the more potent growth inhibition activity of the RAR/RXR pan-agonist 16 implicate both RARs and RXRs in the molecular mechanism of retinoid growth inhibition. Moreover, the chemoprevention activity and the lack of toxicity of heteroarotinoids demonstrate their clinical potential in head and neck cancer chemoprevention.

Temporally regulated overexpression of parathyroid hormone-related protein in the mammary gland reveals distinct fetal and pubertal phenotypes.

We have previously demonstrated that overexpression of parathyroid hormone-related protein (PTHrP) in the mammary glands of transgenic mice results in defects in ductal elongation and branching during puberty and in lobuloalveolar development during pregnancy. In addition, we have shown that PTHrP is necessary for the formation of the initial ductal tree during embryonic mammary development. In order to examine the effect of varying the timing of PTHrP overexpression on mammary development, we created tetracycline-regulated, K14-tTA/Tet(O)-PTHrP double transgenic mice. In this report, we document that this 'tet-off' system directs transgene expression to the mammary gland and that it is fully repressed in the presence of tetracycline. Using these mice, we demonstrate that transient overexpression of PTHrP before birth causes defects in ductal branching during puberty and that overexpression of PTHrP during puberty decreases the rate of ductal elongation. Furthermore, we demonstrate that if PTHrP overexpression is initiated after ductal morphogenesis is completed, lobuloalveolar development is unaffected. Finally, we demonstrate that the impairment in ductal elongation caused by PTHrP is associated with an increase in the basal rate of epithelial cell apoptosis in terminal end buds and a failure to increase end bud cell proliferation and decrease apoptosis in response to estrogen and progesterone.

Presenting symptoms and diagnostic lag in children with inflammatory bowel disease.

Presenting symptoms and their duration may affect the time that elapses prior to definitive diagnosis of inflammatory bowel disease (IBD). This study was undertaken to determine the mean duration of presenting symptoms and diagnostic lag in children with IBD. The medical records of all patients less than 19 years of age diagnosed with IBD at the pediatric gastroenterology clinic of Children's Hospital of Wisconsin between 1990-1995 were reviewed. The age at diagnosis, gender, presenting symptoms and duration, disease location, and diagnostic lag were analyzed. There were 91 children (49 male) diagnosed with IBD. Crohn's disease (CD) was diagnosed in 58, ulcerative colitis (UC) in 24, and indeterminate colitis in 9. The mean ages at diagnosis were 11.4 years for CD, 9.7 years for UC, and 7.8 years for indeterminate colitis. The most frequent presenting symptoms were abdominal pain, diarrhea, hematochezia, and weight loss. The average lag in diagnosis of CD was 7.1 months, which varied by disease location: small intestine 10.5 months, ileocolonic 7.5 months, and colonic 6.4 months. The average lag in diagnosis was 6.7 months for UC and 14 months for indeterminate colitis. Children presenting with growth failure had the longest diagnostic lag. (a) The elapsed time between symptom onset and the diagnosis of CD has decreased. (b) The diagnostic lag in CD decreases with distal colonic involvement. (c) Following onset of symptoms UC was diagnosed only slightly more rapidly than CD.

Central vs peripheral venous catheters in critically ill patients.

A prospective study of 2,209 intravenous catheters was performed in a multidisciplinary intensive care unit to determine when and why catheters were removed and which sites of insertion were associated with the least morbidity. Techniques of insertion were vigorously supervised. Central and peripheral catheters were cared for by identical protocols. Overt phlebitis or inflammation around the site was 14 times as common with peripheral catheters (353/1,024) than with centrally inserted central catheters (18/713), even though peripheral catheters were removed on the average at 2.9 days and centrally inserted central catheters at 6.2 days. Pneumothorax occurred in seven out of 713 patients with centrally inserted central catheterization, one with hemothorax and two with pneumothoraces requiring thoracostomy tubes. Five were treated successfully with simple catheter aspiration. Three patients out of 1,496 with peripheral or peripherally inserted central catheters required phlebectomy for suppurative thrombophlebitis. We concluded that overall morbidity in critically ill patients is lower from centrally inserted central catheters than peripheral intravenous catheters, with peripherally inserted central catheters in an intermediate position. Supervision of techniques of insertion has to be kept at a high level to keep complications of central catheterization at an acceptable level. Peripheral catheter sites would be better maintained with more frequent replacement of the catheter.

Prevalence of neoplasia in 10 New England populations of the soft-shell clam (Mya arenaria).

Neoplasia was a prevalent disease of the soft-shell clam and was found in widely geographically distinct sites in New England. Two types of neoplasms were recognized. Most were of hematopoietic origin, except in clams from Maine, which also had gonadal neoplasms. Both types were apparently malignant neoplasms, based on their characteristic anaplastic appearance, invasiveness, metastasis, mitotic activity, associated tissue necrosis, and mortality. Diagnosis of neoplasia in the living mollusk was achieved rapidly and accurately by cytologic examination of circulating blood. The etiology of the neoplasms was not identified. In general, nonneoplastic lesions, such as epithelial hyperplasia and accumulations of a orange-brown bodies, were more common in clams from polluted areas.

Plasma expansion in surgical patients with high central venous pressure (CVP); the relationship of blood volume to hematocrit, CVP, pulmonary wedge pressure, and cardiorespiratory changes.

There was no correlation of blood volume measurements with central venous pressure (CVP) or hematocrit determinations and only minimal suggestive trends with wedge pressure in a large series of postoperative patients; the lack of correlations emphasize the unreliability of venous pressure and hematocrit determinations to predict blood volume alterations. To evaluate the physiological problems, to define optimal therapeutic goals, and to measure therapeutic effectiveness of volume loading with an oncotically active agent, we measured the hemodynamic and oxygen transport responses to 500 ml. of 5 percent albumin given over 1 hour in 22 patients with CVP greater than 15 cm. H2O. The patients were separated into two groups according to the CVP response to volume therapy. The CVP decreased in 14 (64 percent) of these patients (Group 1), but it increased slightly but not significantly in eight (36 percent) patients (Group 2). In Group 1 patients, there was increased flow, improvement of tissue perfusion as reflected by increased oxygen consumption, and augmentation of the ventricular function. In Group 2 there were slight increases in mean flow, mean pulmonary arterial pressure, and mean transit time and slightly decreased pulmonary vascular resistance; there was appreciable improvement in left ventricular function without significant deterioration of right ventricular function. The high initial central venous pressure is not a reliable index of either hypervolemia or cardiac failure in critically ill patients. It is concluded that a trial of volume loading with an oncotically active agent with frequent auscultation of the chest and careful observation of the CVP trends will give the maximum diagnostic as well as therapeutic information.

Information needs of health care professionals in an AIDS outpatient clinic as determined by chart review.

OBJECTIVE: To examine the information needs of health care professionals in HIV-related clinical encounters, and to determine the suitability of existing information sources to address those needs. SETTING: HIV outpatient clinic. PARTICIPANTS: Seven health care professionals with diverse training and patient care involvement. METHODS: Based on patient charts describing 120 patient encounters, participants generated 266 clinical questions. Printed and on-line information sources were used to answer questions in two phases: using commonly available sources and using all available medical library sources. MEASUREMENTS: The questions were divided into 16 categories by subject. The number of questions answered, their categories, the information source(s) providing answers, and the time required to answer questions were recorded for each phase. RESULTS: Each participant generated an average of 3.8 clinical questions per chart. Five categories accounted for almost 75% of all questions; the treatment protocols/regimens category was most frequent (24%). A total of 245 questions (92%) were answered, requiring an average of 15 minutes per question. Most (87%) of the questions were answered via electronic sources, even though paper sources were consulted first. CONCLUSIONS: The participating professionals showed considerable information needs. A combination of on-line and paper sources was necessary to provide the answers. The study suggests that present-day information sources are not entirely satisfactory for answering clinical questions generated by examining charts of HIV-infected patients.

Cardiorespiratory response to a new isoquinoline derivative in critically III patients.

NC 7197, a new N-substituted tetrahydroisoquinoline derivative, was given in doses of 0.2 mg/kg body weight on 26 occasions to a series of 23 critically ill postoperative and posttraumatic patients who had been in moderate or severe degrees of shock. This agent was observed to improve pressure-flow and oxygen-transport variables, including increases in cardiac index, mean arterial pressure, central venous pressure, both left and right ventricular stroke work, central blood volume, systemic vascular resistance, oxygen availability, arteriovenous oxygen content difference, and oxygen consumption, and decreases in mean transit time and pulmonary vascular resistance. Previous studies on critically ill patients have suggested that these are the most commonly desired therapeutic actions for this type of patient. The agent has pronounced inotropic effect with minimal chronotropic effects, but with higher doses, chronotropic effects as well as alpha blocking effects may occur. The optimal effects may be obtained by adjusting the dose to an appropriate therapeutic range. It is concluded that, in the dose used, this agent produced both alpha and beta adrenergic actions in critically ill patients.

Extracorporeal membrane oxygenation for massive pulmonary thromboembolism.

This study was undertaken to determine whether extracorporeal membrane oxygenation (ECMO) could modify the effects of massive lethal thromboembolism and prevent death. Twenty anesthetized dogs were prepared for venoarterial perfusion with a demand pump and membrane lung and were perfused slowly for 1 1/2 hours to lessen homologous blood shock; 1 ml per kilogram of 24-hour-old tantalum-impregnated thrombus was injected intravenously. The dogs had profound systemic hypotension with an elevated mean pulmonary artery pressure (62.9 +/-4.5 mm Hg) immediately after embolization. Control animals generally died within 15 minutes. Four of the 10 ECMO-supported animals lived for six days, at which time they were restudied and killed. Not only can ECMO maintain an animal that would otherwise die quickly of massive pulmonary thromboembolism, but such support, even though temporary, can greatly improve the chances of survival.

Treatment of metabolic alkalosis with intravenous infusion of concentrated hydrochloric acid.

A concentrated hydrochloric acid (1 N) infusion was utilized for treatment on 35 occasions of metabolic alkalosis in 24 patients. The amount of hydrochloric acid to be infused was calculated from total base excess. To avoid over-correction, two thirds of the calculated dosage of hydrochloric acid only was infused. 1 N hydrochloric acid solution was infused at a speed of 1 mEq/min through a roentgenographically confirmed central venous line. Metabolic alkalosis was successfully treated in all instances without any complication. However, increased respiratory stimulation was not demonstrated in these observations. Concentrated hydrochloric acid infusion is a safe, reliable, and effective method of rapid correction of metabolic alkalosis. Because only small volumes are needed, this method is especially useful when fluid intake must be restricted.

Automated drug dissolution monitor that uses a UV-visible diode array spectrophotometer.

A method for performing multicomponent analysis in drug dissolution testing without chromatographic separation is presented. Aliquots from dissolution vessels are automatically transferred to a UV-visible diode array spectrophotometer, spectra are measured, and the aliquots are returned to the testing vessels. A full-spectrum calibration method based on principal-component regression is used to simultaneously determine the concentrations of active ingredients and to account for interferences due to excipients in a tablet formulation. The system was evaluated with two commercial pharmaceutical formularies; the first contained pseudoephedrine hydrochloride and chlorpheniramine maleate, whereas the second was a mixture of phenylpropanolamine hydrochloride and chlorpheniramine maleate. The selections of standard mixtures for calibration and validation were based on a factorial design.

International organizations and communications.

Multidisciplinary critical care evolved from the work of Bjorn Ibsen in 1952, and from his international links, developed international organizations, and regional federations and societies. Today the international critical care community is increasingly communicating through the Internet.