Minimally invasive quantification of lymph flow in mice and rats by imaging depot clearance of near-infrared albumin

There is a lack of available methods to noninvasively quantify lymphatic function in small experimental animals, a necessity for studies on lymphatic system pathophysiology. We present a new method to quantify lymph flow in mice and rats, based on optically monitoring the depot clearance of near-infrared fluorescently labeled albumin and subsequent calculation of removal rate constants (k). BSA was conjugated with Alexa680 NHS ester and remained stable in protein-rich solutions without free dye dissociation. To assess lymph flow, mice or rats were imaged every 30 or 60 min during a 3- to 6-h period following an intradermal injection of 0.5 or 1 μl Alexa680-albumin. Mice were awake between measurements, whereas rats were anesthetized throughout the experiment. The k, a parameter defined as equivalent to lymph flow, was calculated from the slopes of the resultant log-linear washout curves and averaged -0.40 ± 0.03 and -0.30 ± 0.02%/min for control C57BL/6 and C3H mice, respectively. Local administration of the vasoconstrictor endothelin-1 in mice led to a significant reduction in k, whereas overhydration in rats increased k, reflecting the coupling between capillary filtration and lymph flow. Furthermore, k was 50% of wild type in lymphedema Chy mice where dermal lymphatics are absent. We conclude that lymph flow can be determined as its rate constant k by optical imaging of depot clearance of submicroliter amounts of Alexa680-albumin. The method offers a minimally invasive, reproducible, and simple alternative to assess lymphatic function in mice and rats.     

Interactions among flavonoids of propolis affect antibacterial activity against the honeybee pathogen Paenibacillus larvae

Propolis is derived from plant resins, collected by honeybees (Apis mellifera) and renown for its antibacterial properties. Here we test the antibacterial effects of ethanolic extracts of propolis from different origins on Paenibacillus larvae, the bacterial pathogen that causes American Foulbrood, a larval disease that can kill the honeybee colony. All tested propolis samples inhibited significantly the growth of P. larvae tested in vitro. The extracts showed major differences in the content of total flavonoids (ranging from 2.4% to 16.4%) and the total polyphenols (ranging between 23.3% and 63.2%). We found that it is not only the content of compounds in propolis, which influences the strength of antimicrobial effects but there is also a significant interaction effect among flavonoids of the propolis extracts. We propose that interaction effects among the various chemical compounds in propolis should be taken into account when considering the antibacterial effects against honeybee pathogens.     

Endurance Training Per Se Increases Metabolic Health in Young,Moderately Overweight Men

Health benefits of physical activity may depend on a concomitant weight loss. In a randomized, controlled trial, we compared the effects of endurance training with or without weight loss to the effect of weight loss induced by an energy‐reduced diet in 48 sedentary, moderately overweight men who completed a 12‐week intervention program of training (T), energy‐reduced diet (D), training and increased diet (T‐iD), or control (C). An energy deficit of 600 kcal/day was induced by endurance training or diet in T and D and a similar training regimen plus an increased dietary intake of 600 kcal/day defined the T‐iD group. Primary end point was insulin sensitivity as evaluated by HOMA‐IR (mainly reflecting hepatic insulin sensitivity) and hyperinsulinemic, isoglycemic clamps (primarily reflecting peripheral insulin sensitivity). Body mass decreased in T and D by 5.9 ± 0.7 and 5.3 ± 0.7 kg, respectively, whereas T‐iD and C remained weight stable. Total and abdominal fat mass were reduced in an additive manner in the T‐iD, D, and T groups by 1.9 ± 0.3/0.2 ± 0.1, 4.4 ± 0.7/0.5 ± 0.1, and 7.7 ± 0.8/0.9 ± 0.1 kg, respectively. HOMA‐IR was improved in T, D, and T‐iD, whereas insulin‐stimulated glucose clearance and suppression of plasma nonesterified fatty acids (NEFAs) were increased only in the two training groups. Thus, loss of fat mass (diet or training induced) improves hepatic insulin sensitivity, whereas peripheral insulin sensitivity in skeletal muscle and adipose tissue is increased by endurance training only. This demonstrates that endurance training per se increases various metabolic health parameters and that endurance training should preferably always be included in any intervention regimen for improving metabolic health in moderately overweight men.     

Total catch of a red-listed marine species is an order of magnitude higher than official data

Accurate information on total catch and effort is essential for successful fisheries management. Officially reported landings, however, may be underestimates of total catch in many fisheries. We investigated the fishery for the nationally red-listed European lobster (Homarus gammarus) in south-eastern Norway. Probability-based strip transect surveys were used to count buoys in the study area in combination with catch per unit effort data obtained independently from volunteer catch diaries, phone interviews, and questionnaires. We estimate that recreational catch accounts for 65% of total catch in the study area. Moreover, our results indicate that only a small proportion (24%) of lobsters landed commercially were sold through the legal market and documented. Total estimated lobster catch was nearly 14 times higher than reported officially. Our study highlights the need for adequate catch monitoring and data collection efforts in coastal areas, presents a clear warning to resource managers that illegal, unreported and unregulated (IUU) fisheries in coastal areas should not be ignored, and shows the potential impact of recreational fisheries.     

Isolation of interstitial fluid from rat mammary tumors by a centrifugation method

Access to interstitial fluid is of fundamental importance to understand tumor transcapillary fluid balance, including the distribution of probes and therapeutic agents. Tumors were induced by gavage of 9,10-dimethyl-1,2-benzanthracene to rats, and fluid was isolated after anesthesia by exposing tissue to consecutive centrifugations from 27 to 6,800 g. The observed (51)Cr-EDTA (extracellular tracer) tissue fluid-to-plasma ratio obtained from whole tumor or from superficial tumor tissue by centrifugation at 27-424 g was not significantly different from 1.0 (0.92-0.99), suggesting an extracellular origin only. However, fluid collected from excised central tumor parts had a significantly lower ratio (0.66-0.77) for all imposed G forces, suggesting dilution by fluid deriving from a space unavailable for (51)Cr-EDTA. The colloid osmotic pressure in tumor fluid was generally higher than in fluid isolated from the subcutis, attributable to less selective capillaries and impaired lymphatic drainage in tumors. HPLC analysis of tumor fluid showed that low-molecular-weight macromolecules not present in arterial plasma were present in tumor fluid obtained by centrifugation and in venous blood draining the tumor, most likely representing proteins derived from tumor cells. We conclude that low-speed centrifugation may be a simple and reliable method to isolate interstitial fluid from tumors.     

Trawling damage to Northeast Atlantic ancient coral reefs

This contribution documents widespread trawling damage to cold-water coral reefs at 840-1300 m depth along the West Ireland continental shelf break and at 200 m off West Norway. These reefs are spectacular but poorly known. By-catches from commercial trawls for deep-water fish off West Ireland included large pieces (up to 1 m(2)) of coral that had been broken from reefs and a diverse array of coral-associated benthos. Five azooxanthellate scleractinarian corals were identified in these by-catches, viz. Desmophyllum cristagalli, Enallopsammia rostrata, Lophelia pertusa, Madrepora oculata and Solenosmilia variabilis. Dating of carbonate skeletons using (14)C accelerator mass spectrometry showed that the trawled coral matrix was at least 4550 years old. Surveys by remotely operated vehicles in Norway showed extensive fishing damage to L. pertusa reefs. The urgent need for deep-water coral conservation measures is discussed in a Northeast Atlantic context.     

Network organization of interstitial connective tissue cells in the human endolymphatic duct.

The human endolymphatic duct (ED) and sac of the inner ear have been suggested to control endolymph volume and pressure. However, the physiological mechanisms for these processes remain obscure. We investigated the organization of the periductal interstitial connective tissue cells and extracellular matrix (ECM) in four freshly fixed human EDs by transmission electron microscopy and by immunohistochemistry. The unique surgical material allowed a greatly improved structural and epitopic preservation of tissue. Periductal connective tissue cells formed frequent intercellular contacts and focally occurring electron-dense contacts to ECM structures, creating a complex tissue network. The connective tissue cells also formed contacts with the basal lamina of the ED epithelium and the bone matrix, connecting the ED with the surrounding bone of the vestibular aqueduct. The interstitial connective tissue cells were non-endothelial and non-smooth muscle fibroblastoid cells. We suggest that the ED tissue network forms a functional mechanical entity that takes part in the control of inner ear fluid pressure and endolymph resorption.     

Halobacterium volcanii spec. nov., a Dead Sea halobacterium with a moderate salt requirement

A halophilic bacterium was isolated from bottom sediment from the Dead Sea. The organism possessed the properties of the halobacteria, but differed from the known species in two important respects, 1) the cells were disc-shaped and often cupped when grown under optimum conditions, 2) the optimum requirements for sodium chloride was in the range 1.7–2.5 molar which is about half of that generally reported for the halobacteria. The organism was assigned to the genus Halobacterium and described as Halobacterium volcanii spec.nov. The optimum sodium chloride concentration for growth was close to that found in the Dead Sea. The tolerance for magnesium chloride was very high; the organism grew well in media containing magnesium chloride in the concentrations found in the Dead Sea. Halobacterium volcanii is therefore remarkably well fitted for life in the Dead Sea.     

Using formal logic to classify vegetation

Suggestions are made to define vegetation units by using formal logic. The characteristics that are used for the definition are logically combined using the AND, OR and NOT operators in such a way that relevés can be unequivocally allocated to vegetation units. Among all the characteristics of vegetation species groups are best suited to present allocation schemes for the Braun-Blanquet-system. It is shown that definition gaps within the system can be completely avoided by providing formal allocation rules.