Volume regulation by flounder red blood cells in anisotonic media

The nucleated high K, low Na red blood cells of the winter flounder demonstrated a volume regulatory response subsequent to osmotic swelling or shrinkage. During volume regulation the net water flow was secondary to net inorganic cation flux. Volume regulation the net water flow was secondary to net inorganic cation flux. Volume regulation after osmotic swelling is referred to as regulatory volume decrease (RVD) and was characterized by net K and water loss. Since the electrochemical gradient for K is directed out of the cell there is no need to invoke active processes to explain RVD. When osmotically shrunken, the flounder erythrocyte demonstrated a regulatory volume increase (RVI) back toward control cell volume. The water movements characteristic of RVI were a consequence of net cellular NaCl and KCl uptake with Na accounting for 75 percent of the increase in intracellular cation content. Since the Na electrochemical gradient is directed into the cell, net Na uptake was the result of Na flux via dissipative pathways. The addition of 10(-4)M ouabain to suspensions of flounder erythrocytes was without effect upon net water movements during volume regulation. The presence of ouabain did however lead to a decreased ration of intracellular K:Na. Analysis of net Na and K fluxes in the presence and absence of ouabain led to the conclusion that Na and K fluxes via both conservative and dissipative pathways are increased in response to osmotic swelling or shrinkage. In addition, the Na and K flux rate through both pump and leak pathways decreased in a parallel fashion as cell volume was regulated. Taken as a whole, the Na and K movements through the flounder erythrocyte membrane demonstrated a functional dependence during volume regulation.     

An intersexual migratory (silver) longfinned New Zealand eel and its gonadal response to treatment with salmon pituitary homogenate

An intersexual adult New Zealand longfinned eel Anguilla dieffenbachii , displaying testicular tissue within its ovaries, is described. Gonad samples were collected at 0, 21, 42 (by biopsy) and 54 days (autopsy) after the start of an experiment that aimed to induce ovarian maturation by repeated treatment with 5 mg kg⁻¹ salmon pituitary homogenate (SPH). Very small patches of testicular tissue (almost always single seminiferous tubules), not visible macroscopically, were observed on all sampling dates. Treatment with SPH resulted in advanced stages of spermatogenesis in these seminiferous tubules, synchronous with the development of oocytes, which reached the migratory nucleus stage after 53 days. This finding is discussed in relation to current knowledge on sexual development in anguillid eels.     

Biodeterioration of Mayan buildings at uxmal and tulum,Mexico

Uxmal and Tulum are two important Mayan sites in the Yucatan peninsula. The buildings are mainly composed of limestone and grey/black discoloration is seen on exposed walls and copious greenish biofilms on inner walls. The principal microorganisms detected on interior walls at both Uxmal and Tulum were cyanobacteria; heterotrophic bacteria and filamentous fungi were also present. A dark‐pigmented mitosporic fungus and Bacillus cereus, both isolated from Uxmal, were shown to be acidogenic in laboratory cultures. Cyanobacteria belonging to rock‐degrading genera Synechocystis and Gloeocapsa were identified at both sites. Surface analysis previously showed that calcium ions were present in the biofilms on buildings at Uxmal and Tulum, suggesting the deposition of biosolubilized stone. Apart from their potential to degrade the substrate, the coccoid cyanobacteria supply organic nutrients for bacteria and fungi, which can produce organic acids, further increasing stone degradation.     

Changes in steroid hormone profiles and ovarian histology during salmon pituitary-induced vitellogenesis and ovulation in female New Zealand longfinned eels, Anguilla dieffenbachii gray

To assess whether induced vitellogenesis in longfinned eels mimics that in naturally maturing conspecifics, female eels were artificially matured and steroid hormone status and oocyte cytology during oogenesis were evaluated. Successful induction of vitellogenesis was evident from the presence of yolk granules in the ooplasm of salmon pituitary homogenate (SPH)-injected, but not saline-, 17-hydroxyprogesterone-, and/or gonadotropin-releasing hormone-treated fish. In SPH-treated females, the migratory nucleus stage was reached after 33-53 days, followed by ovulation around 30 hours after induction of final maturation and ovulation. Only a portion of the germ cells matured, although resumption of vitellogenesis was seen in the majority of oocytes. In contrast, in ovaries of saline-injected controls, the most advanced oocytes were early vitellogenic. Atretic follicles were observed in ovaries of all eels, but abundance was greater in controls than in SPH-treated fish. SPH injections elevated plasma levels of estradiol-17beta and androgens, but not pregnenes, from within three days of treatment. Our results indicate that sex steroid levels in midvitellogenic hormone-treated females are similar to those in wild midvitellogenic females. In contrast, differences in yolk morphology of midvitellogenic follicles were seen between SPH-treated and wild females, especially in the second crop of midvitellogenic-sized oocytes measuring 300-400 microm in diameter. We discuss whether the observed differences affect egg quality, and perhaps explain the short life span of captive-bred eel larvae. J. Exp. Zool. 289:119-129, 2001.     

Efficient production of Arthromyces ramosus peroxidase by Aspergillus awamori

The heterologous production of Arthromyces ramosus peroxidase (ARP) was analysed in the filamentous fungus Aspergillus awamori under control of the inducible endoxylanase promoter. Secretion of active ARP was achieved up to 800 mg l(-1) in shake flask cultures. Western blot analysis showed that an rARP product of the correct molecular weight was produced. In contrast to several other studies about heterologous production of heme containing peroxidases, our results suggest that in A. awamori no heme limitation exists during overproduction of ARP.     

The production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi

In this review we will focus on the current status and views concerning the production of antibody fragments and antibody fusion proteins by yeasts and filamentous fungi. We will focus on single-chain antibody fragment production (scFv and V_HH) by these lower eukaryotes and the possible applications of these proteins. Also the coupling of fragments to relevant enzymes or other components will be discussed. As an example of the fusion protein strategy, the 'magic bullet' approach for industrial applications, will be highlighted.