Respiration rates of dominant hydromedusae in the North Inlet tidal estuary during winter and summer

The objective of this study was to quantify the respiratoryrequirements and oxygen consumption rates for seasonal hydromedusaepopulations in the North Inlet estuary, Georgetown, SC, USA.Respiration rates were determined for hydromedusae collectedin January 2006, 2007 and July 2006 using microrespirometry.The numerically dominant gelatinous predators were Nemopsisbachei (January 2006 and 2007) and Bougainvillia muscus (July2006). Seasonal oxygen consumption rates were low (0.80–1.27µmol O₂ g^–1 h^–1). Q₁₀ values were high duringJanuary 2006 and July 2006 (4.7), but low during January 2007(0.5), suggesting hydromedusae were sensitive to the exposuretemperatures (7–34°C) in this study and well adaptedto the temperature range encountered in this estuary. Moreover,the oxygen consumption rates during July 2006 showed significantdifferences between the two species. The allometric relationshipbetween respiration rate and hydromedusae weight differed seasonally.Our findings suggest hydromedusae can persist in hypoxic conditionsand may partially explain the frequently observed increase inthese cnidarians under conditions of low dissolved oxygen concentrations.     

Cyanobacterial-bacterial mat consortia: examining the functional unit of microbial survival and growth in extreme environments

Cyanobacterial–bacterial consortial associations are taxonomically complex, metabolically interactive, self-sustaining prokaryotic communities representing pioneer and often the only biota inhabiting extreme aquatic and terrestrial environments. Laminated mats and aggregates exemplify such communities. The fossil record indicates that these associations represent the earliest extant inhabitants and modifiers (i.e. anoxic to oxic conditions) of the Earth's biosphere. Present-day consortia flourish in physically and chemically stressed environments, including nutrient-deplete, hypersaline, calcified, desiccated and high-irradiance ecosystems ranging from the tropics to polar regions. Consortial members exhibit extensive metabolic diversification, but have remained structurally simple. Structural simplicity, while advantageous in countering environmental extremes, presents a 'packaging problem' with regard to compartmentalizing potentially cross-inhibitory aerobic versus anaerobic growth processes. To circumvent these metabolic constraints, phototrophic cyanobacteria and microheterotrophs orient along microscale chemical (i.e. O₂, pH, E _h) gradients to meet and optimize the biogeochemical processes (C, N, S cycling) essential for survival, growth and the maintenance of genetic diversity, needed to sustain life. Microscale ecophysiological, analytical, molecular (immunological and nucleic acid) techniques have helped to develop a mechanistic basis for understanding consortial growth and survival under extreme environmental conditions on Earth. Consortia are ideal model systems for developing a process-based understanding of the structural and functional requirements for life in extreme environments representative of the Earth's earliest biosphere and possibly other planets.     

Effects of low-chloride solutions on action potentials of sheep cadiac purkinje fibers

The rapid repolarization during phase 1 of the action potential of sheep cardiac purkinje fibers has been attributed to a time- and voltage-dependent chloride current. In part, this conclusion was based on experiments that showed a substantial slowing of phase 1 when larger, presumably impermeant, anions were substituted for chloride in tyrode’s solution. We have re- examined the electrical effects of low-chloride solutions. We recorded action potentials of sheep cardiac purkinje fibers in normal tyrode’s solution and in low-chloride solutions made by substituting sodium propionate, acetylglycinate, methylsulfate, or methanesulfonate for the NaCl of Tyrode’s solution. Total calcium was adjusted to keep calcium ion activity of test solutions equal to that of control solutions. Propionate gave qualitatively variable results in preliminary experiments; it was not tested further. Low-chloride solutions made with the other anions gave much more consistent results: phase 1 and the notch that often occurs between phases 1 and 2 were usually unaffected, and the action potential duration usually increased. The only apparent change in the resting potential was a transient 3-6 mV depolarization when low-chloride solution was first admitted to the chamber, and a symmetrical transient hyperpolarization when chloride was returned to normal. If a time- and voltage-dependent chloride current exists in sheep cardiac purkinje fibers, our results suggest that it plays little role in generating phase 1 of the action potential.     

Fluorometric estimation of surface associated microbial abundance

Surface associated microbes have historically been difficult to accurately and effectively enumerate. In the current study, we propose a rapid and simple method for estimating abundance of surface associated microbial cells by fluorescence of SYBRGreen stained bacteria and in vivo chlorophyll a fluorescence of benthic diatoms in 24 and 48-well microtiter plates. The effectiveness of this high-throughput technique is demonstrated by assessing sensitivity of a clinical strain of Vibrio cholerae, a benthic bacterial isolate and the benthic microalgae Cylindrotheca closterium to three antibiotics — tylosin, lincomycin and ciproflaxacin. We report on the significant linear relationships between spectral chl a fluorescence and cell abundance and between microalgal growth rates derived from cell counts and fluorescence. Additionally, we provide a simplified and improved method for preparation of a silica gel matrix (SGM), which is an ideal plating media for fluorescence applications. These findings indicate that spectrofluorometry is an inexpensive tool for rapidly estimating abundance of surface associated microbiota and can be employed for assessing antibiotic sensitivity.     

COMBINING NEW TECHNOLOGIES FOR DETERMINATION OF PHYTOPLANKTON COMMUNITY STRUCTURE IN THE NORTHERN GULF OF MEXICO 1

In situ analysis of phytoplankton community structure was determined at five stations along the Texas Gulf coast using two instruments, the Fluoroprobe and FlowCAM. Results were compared with traditional methods to determine community structure (pigment analysis and microscopy). Diatoms and small nanoplankton (most likely haptophytes) dominated the phytoplankton community at all stations. Estimated chl concentrations for diatoms+dinoflagellates obtained via the Fluoroprobe were not significantly different for three of the five stations sampled when compared with HPLC‐chemical taxonomy analysis, whereas the concentrations of green algal and cryptophyte chl were overestimated. The FlowCAM estimates of overall nanoplankton and microplankton cell abundance were not significantly different when compared with epifluorescence microscopy, and recorded images of phytoplankton cells provided a representative population of the phytoplankton community at each station. The Fluoroprobe and FlowCAM, when used in tandem, are potentially capable of determining the general characteristics of phytoplankton community structure in situ and could be an important addition to biological observing systems in the coastal ocean.     

Quantitative patterns of Hsps in tubular adenoma compared with normal and tumor tissues reveal the value of Hsp10 and Hsp60 in early diagnosis of large bowel cancer

Large bowel carcinogenesis involves accumulation of genetic alterations leading to transformation of normal mucosa into dysplasia and, lastly, adenocarcinoma. It is pertinent to elucidate the molecular changes occurring in the pre-neoplastic lesions to facilitate early diagnosis and treatment. Heat shock proteins (Hsps), many of which are molecular chaperones, are implicated in carcinogenesis, and their variations with tumor progression encourage their study as biomarkers. There are many reports on Hsps and cancer but none to our knowledge on their systematic quantification in pre-neoplastic lesions of the large bowel. We performed immunohistochemical determinations of Hsp10, Hsp60, Hsp70, and Hsp90 in biopsies of large bowel tubular adenomas with moderate grade of dysplasia and compared to normal mucosa and adenocarcinoma with a moderate grade of differentiation (G2). A significant elevation of Hsp10 and Hsp60 only, i.e., in the absence of elevation of Hsp70 or Hsp90, in both epithelium and lamina propria was found in tubular adenoma by comparison with normal mucosa. In contrast, adenocarcinoma was characterized by the highest levels of Hsp10 and Hsp60 in epithelium and lamina propria, accompanied by the highest levels of Hsp70 only in epithelium and of Hsp90 only in lamina propria, by comparison with normal and tubular adenoma counterparts. Hsp10 and Hsp60 are promising biomarkers for early diagnosis of tubular adenoma and for its differentiation from more advanced malignant lesions. Hsp10 and Hsp60 may be implicated in carcinogenesis from its very early steps and, thus, are potentially convenient targets for therapy.