Amplicon-Based Pyrosequencing Reveals High Diversity of Protistan Parasites in Ships' Ballast Water: Implications for Biogeography and Infectious Diseases.

Ships' ballast water (BW) commonly moves macroorganisms and microorganisms across the world's oceans and along coasts; however, the majority of these microbial transfers have gone undetected. We applied high-throughput sequencing methods to identify microbial eukaryotes, specifically emphasizing the protistan parasites, in ships' BW collected from vessels calling to the Chesapeake Bay (Virginia and Maryland, USA) from European and Eastern Canadian ports. We utilized tagged-amplicon 454 pyrosequencing with two general primer sets, amplifying either the V4 or V9 domain of the small subunit (SSU) of the ribosomal RNA (rRNA) gene complex, from total DNA extracted from water samples collected from the ballast tanks of bulk cargo vessels. We detected a diverse group of protistan taxa, with some known to contain important parasites in marine systems, including Apicomplexa (unidentified apicomplexans, unidentified gregarines, Cryptosporidium spp.), Dinophyta (Blastodinium spp., Euduboscquella sp., unidentified syndinids, Karlodinium spp., Syndinium spp.), Perkinsea (Parvilucifera sp.), Opisthokonta (Ichthyosporea sp., Pseudoperkinsidae, unidentified ichthyosporeans), and Stramenopiles (Labyrinthulomycetes). Further characterization of groups with parasitic taxa, consisting of phylogenetic analyses for four taxa (Cryptosporidium spp., Parvilucifera spp., Labyrinthulomycetes, and Ichthyosporea), revealed that sequences were obtained from both known and novel lineages. This study demonstrates that high-throughput sequencing is a viable and sensitive method for detecting parasitic protists when present and transported in the ballast water of ships. These data also underscore the potential importance of human-aided dispersal in the biogeography of these microbes and emerging diseases in the world's oceans.

The effects of prolonged darkness on temperate and tropical marine phytoplankton, and their implications for ballast water risk management.

Phytoplankton assemblages from tropical (Goa) and temperate (UK) locations were exposed to a 28 day dark period, followed by a period of re-exposure to light. During this time phytoplankton survival and changes in nutrient concentrations were mapped. The tropical plankton water samples showed high nutrient levels after the dark period which were utilised by cells during the re-exposure period. UK experiments looked at the effect of three different water types on population recovery after the 28 day dark period, and differences due to seasonal effects. The population growth observed during the re-exposure period in the tropical population was comparable to that of the temperate population. Water type affected recovery and of the three tested media fresh seawater promoted the highest levels of growth. Seasonality had a significant influence on species survival. Understanding the effects of all these factors can aid the development of effective risk assessments in ballast water management.

Sex determination in the monoecious species cucumber is confined to specific floral whorls.

In unisexual flowers, sex is determined by the selective repression of growth or the abortion of either male or female reproductive organs. The mechanism by which this process is controlled in plants is still poorly understood. Because it is known that the identity of reproductive organs in plants is controlled by homeotic genes belonging to the MADS box gene family, we analyzed floral homeotic mutants from cucumber, a species that bears both male and female flowers on the same individual. To study the characteristics of sex determination in more detail, we produced mutants similar to class A and C homeotic mutants from well-characterized hermaphrodite species such as Arabidopsis by ectopically expressing and suppressing the cucumber gene CUCUMBER MADS1 (CUM1). The cucumber mutant green petals (gp) corresponds to the previously characterized B mutants from several species and appeared to be caused by a deletion of 15 amino acid residues in the coding region of the class B MADS box gene CUM26. These homeotic mutants reveal two important concepts that govern sex determination in cucumber. First, the arrest of either male or female organ development is dependent on their positions in the flower and is not associated with their sexual identity. Second, the data presented here strongly suggest that the class C homeotic function is required for the position-dependent arrest of reproductive organs.

Separated pyelocolocutaneous diversion after bilateral ureterectomy and radical cystectomy for transitional cell carcinoma.

OBJECTIVES: To report a technique for excision of bilateral ureteral transitional cell carcinoma which preserves renal function and provides ready access for endoscopic follow-up and therapy of the renal pelvis. METHODS: The operative procedure of bilateral ureterectomy with construction of a separated pyelocolocutaneous diversion utilizing transverse colon is described and the clinical course of 4 patients is reported. RESULTS: All 4 patients maintained excellent renal function and normal electrolytes. Conduit urine cytologic studies and renal pelvis endoscopic examinations have remained normal. Parastomal herniation was the most frequently observed postoperative complication. CONCLUSIONS: This approach is well suited for the treatment of bilateral ureteral transitional cell carcinoma. Long-term complications are minimal and renal function is preserved. Periodic endoscopic visualization of each renal pelvis with resection and fulguration and/or instillation of immunotherapeutic or chemotherapeutic agents for recurrent renal pelvic lesions is readily accomplished.

Prognostic significance of tumor markers in colorectal cancer patients: DNA index, S-phase fraction, p53 expression, and Ki-67 index.

Risk of colorectal cancer recurrence has traditionally been determined by use of pathologic staging. However, it is apparent that subgroups of patients exist within tumor stages whose clinical behavior differs. This study was undertaken to identify tumor-associated factors that might be predictive of outcome in patients with intermediate stages who will benefit the most from postsurgical adjuvant therapy. Seventy patients with stage II and III colorectal cancer were assessed for DNA index, S-phase fraction, p53 expression, and Ki-67 index. Tumor recurrence was analyzed by means of nonparametric tests and Cox proportional hazard models incorporating standard clinical and pathologic criteria. Of the four prognostic markers evaluated, Ki-67 index was significantly associated with disease recurrence (P = 0.02), whereas DNA index, S-phase fraction, and p53 expression were not. After stratification by tumor stage, significant associations between Ki-67 index and disease recurrence were retained in stage II tumors (P = 0.01) but not in stage III tumors (P = 0.23). Cox proportional hazard regression analysis indicated that among stage II patients, those with a Ki-67 index >45% were associated with 6.5 times greater risk for disease recurrence than those with a Ki-67 index >/=45%. It was concluded that an elevated Ki- 67 index is associated with an increased risk of tumor recurrence in stage II colorectal cancer.

Left upper lobe mass and diffuse reticular-nodular infiltrate.

We encountered a clinical problem in a young man who presented with a left upper lobe mass and a diffuse reticular-nodular infiltrate. We thought we had appropriately applied Murphy's Law (the famed bank robber who "went where the money is"), and Ockham's Razor (the philosopher William of Ockham [1285 to 1349]-"Entities are not to be multiplied beyond necessity") as we rapidly diagnosed the lung mass with computed tomography, scintigraphy, and fine-needle aspiration. However, when his invaluable previous chest radiographs arrived, bronchoscopy with transbronchial biopsy, bronchoalveolar lavage, brushings, and postbronchoscopy sputum revealed the more ominous diagnosis in this patient. This case illustrates the complementary nature of current imaging and bronchoscopy techniques; but, even more importantly, it demonstrates the value of the history coupled with the previous radiograph. Even an unusual case can provide lessons in cost containment.