Formation of Metallogenium-like structures by a manganese-oxiding fungus

Structures resembling Metallogenium spp. were observed in agar and in liquid cultures of a Mn-oxidizing basidiomycetous fungus only when Mn²⁺ was oxidized. Fungal viability was necessary for formation of the structures; Mn²⁺ concentration and the presence or absence of agar in the medium were important factors determining their morphology. Slide cultures revealed no identifiable cells in any stage of development. Fluorescent dyes that stained nucleic acids and polysaccharides in the fungal hyphae did not stain the Metallogenium-like structures. Likewise, Rhodamine 123, a fluorescent probe for membrane potential, stained fungal mitochondria, but did not stain the structures. Thin sections through the structures showed no biological membranes or other cellular features. Only the characteristic ultrastructure of biological Mn oxides were observed in serial thin sections. In agar, unfixed structures disappeared permanently during reduction of Mn oxides with hydroxylamine. Glutaraldehyde fixation stabilized these structures. Fixed structures lost most of their original phase density during reduction with hydroxylamine, but continuous microscopic observations showed that their phase density could be restored by staining with Coomassie blue. Structures that formed in liquid medium did not require stabilization with glutaraldehyde during reduction of Mn oxides. They, too, lost their original phase density during reduction with hydroxylamine; phase density could be restored by staining with cationic colloidal iron or Coomassie blue. The results suggest that the Metallogenium-like structures were formed as a result of Mn oxidation associated with exopolymers produced by the fungus.Non-standard abbreviations HEPES (N-hydroxyethylpiperazine-N-2-ethane sulfonic acid) - DAPI (4,6-diamidino-2-phenylindole) - PIPES (piperazine-N,N-bis[2-ethane sulfonic acid])     

The morphology of allograft rejection in Styela plicata (Urochordata: Ascidiacea)

Summary This study identifies three discrete processes responsible for the rejection of tunic tissue transplanted between individuals of the solitary ascidian Styela plicata. The first stage of rejection is characterized by the destruction of blood vascular components within incompatible allografts. In the second phase, dense boundaries of extracellular material are deposited between grafts and the surrounding host tunic, effectively amputating the transplanted tissues. Finally, detached transplants undergo a gradual necrosis which results in the total degeneration of extracellular graft matrices. Of these three phases, the initial cellular depletion of allografts is responsible for the immunological specificity that is characteristic of histocompatibility in S. plicata. The subsequent amputation and necrosis of extracellular graft matrices are taken to be non-specific consequences of the initial cellular reactivity.     

Intrinsic potential for high fetal hemoglobin production in a Druze family with β-thalassemia is due to an unlinked genetic determinant

Summary The mechanism for elevated production of fetal hemoglobin (Hb F) in a Druze patient with °-thalassemia intermedia was investigated. Heterozygous family members exhibited normal Hb F levels, suggesting that the increase in -gene expression in the propositus may be partly due to anemic stress. Erythroid progenitors of these family members cultured in vitro [burst forming units (erythroid); (BFUe)] showed elevated synthesis of Hb F, indicating the existence of a genetically determined intrinsic capacity for high Hb F production in this family. The propositus was found to be homozygous for a IVS2-position 1 mutation, on the background of Mediterranean haplotype I, which is not known to be linked to high Hb F production. Moreover, extensive molecular studies of the -globin gene cluster, including sequence analysis of the promoter regions of the -globin genes, did not reveal any cisacting mechanism that could account for the high Hb F production in the propositus. A young niece of the propositus with °-thalassemia major was recently discovered, who was homozygous for the same -globin allele and haplotype as the propositus. However, unlike her uncle, she does not have a high Hb F level and presents with a severe clinical course. Her inability to produce high Hb F suggests that the genetic determinant for increased -gene expression in the propositus is unlinked to the -globin gene cluster.     

New ecological information onScytalina cerdale (Pisces: Scytalinidae) from a central California rocky intertidal zone

Synopsis New information regarding the ecology ofScytalina cerdale was obtained over a four year period as a consequence of a long-term marine ecological study at the Diablo Canyon Power Plant (DCPP), San Luis Obispo County, California. Twenty intertidal fish surveys were conducted at approximately quarterly intervals, between March 1979 and June 1983, at three separate rocky shore locations (stations). During each survey, a total of 108 square meters (36 m² per station) was searched for fish during periods of low tide. A total of 280S. cerdale were collected, identified, measured, and released back into the same 4 m² area, from the same intertidal station (Diablo Cove), throughout the study period. This limited intertidal occurrence most likely reflectsS. cerdale microhabitat requirements; a combination of intertidal elevation (mean = +0.3 MLLW), substratum specificity (loose gravel, 5–10mm size range, overlying a base of sand and shell fragments), and degree of wave exposure (semi-protected). Throughout the study,S. cerdale was seasonally more abundant during summer months (June through August) and less abundant during winter months (November through February). The only exception to this abundance trend followed the 1982 winter storms, which coincided with an El Niño event, whenS. cerdale abundance uncharacteristically dropped during the subsequent 1983 spring and summer surveys. Mean total lengths did not vary greatly, reflecting the absence of early juvenile fishes, and relatively high mean fish densities (3.5 fish per m² for 20 surveys) were recorded. Qualitative comparisons betweenS. cerdale abundance and seasonal changes in water temperature indicated an inverse relationship. Gravid females, demersal egg masses, and early juvenile individuals were never observed during the four years of the study. This suggests that unlike most other intertidal fishes, particularly other blennioids,S. cerdale may not utilize the intertidal zone for reproductive and recruitment purposes; Diablo Canyon is the most southerly reported distribution for this intertidal species, and therefore, may not entirely represent the species' biology as a whole.     

Behavior of captive-raised rattlesnakes (Crotalus enyo) as a function of rearing conditions

Two litters of rattlesnakes (Crotalus enyo; n = 6 per litter) were raised in large cages and small plastic boxes, respectively. No significant differences in exploration of a novel environment were observed between the litters, or between either of them and a group of wild-caught C. enyo, suggesting that the common practice of rearing baby snakes in small cages has no debilitating consequences on these measures. In the absence of differences between the litters, the two samples were pooled and additional analyses revealed the existence of reliable individual variation. Experiment 2 compared the two litters and the wild-caught snakes on measures of prey-directed behavior. The two captive-reared litters did not differ, but both exhibited lower levels of strike-induced chemosensory searching than did wild-caught snakes.     

Tired,weak, or in need of rest: fatigue among general practice attenders.

OBJECTIVES--To determine the prevalence and associations of symptoms of fatigue. DESIGN--Questionnaire survey. SETTING--London general practice. PARTICIPANTS--611 General practice attenders. MAIN OUTCOME MEASURES--Scores on a fatigue questionnaire and reasons given for fatigue. RESULTS--10.2% Of men (17/167) and 10.6% of women (47/444) had substantial fatigue for one month or more. Age, occupation, and marital status exerted minor effects. Subjects attributed fatigue equally to physical and non-physical causes. Physical ill health, including viral infection, was associated with more severe fatigue. Women rather than men blamed family responsibilities for their fatigue. The profile of persistent fatigue did not differ from that of short duration. Only one person met criteria for the chronic fatigue syndrome. CONCLUSIONS--Fatigue is a common complaint among general practice attenders and can be severe. Patients may attribute this to physical, psychological, and social stress.     

Interaction of alpha-N-Acetyl-beta-endorphin and calmodulin

Acetylation at the -amino terminal is a common post-translational modification of many peptides and proteins. In the case of the potent opiate peptide -endorphin, -N-acetylation is a known physiological modification that abolishes opiate activity. Since there are no known receptors for -N-acetyl--endorphin, we have studied the association of this peptide with calmodulin, a calcium-dependent protein that binds a variety of peptides, phenothiazines, and enzymes, as a model system for studying acetylated endorphin-protein interactions. Association of the acetylated peptide with calmodulin was demonstrated by cross-linking with bis(sulfosuccinimidyl)suberate; like -endorphin, adducts containing 1 mol and 2 mol of acetylated peptide per mole calmodulin were formed. Some of the bound peptides are evidently in relatively close proximity to each other since, in the presence of amidated (i.e., lysine-blocked) calmodulin, cross-linking yielded peptide dimers. The acetylated peptide exhibited no appreciable helicity in aqueous solution, but in trifluoroethanol (TFE) considerable helicity was formed. Also, a mixture of acetylated peptide and calmodulin was characterized by a circular dichroic spectrum indicative of induced helicity. Empirical prediction rules, applied earlier to -endorphin, suggest that residues 14–24 exhibit -helix potential. This segment has the potential of forming an amphipathic helix; this structural unit is believed to be important in calmodulin binding. The acetylated peptide was capable of inhibiting the calmodulin-mediated stimulation of cyclic nucleotide phosphodiesterase (EC 3.1.4.17) activity with an effective dose for 50% inhibition of about 3 µM; this inhibitory effect was demonstrated using both an enzyme-enriched preparation as well as highly purified enzyme. Thus, acetylation at the -amino terminal of -endorphin, although abolishing opiate activity, does not interfere with the binding to calmodulin. Indeed, -endorphin and the -N-acetylated peptide behave very similarly with respect to calmodulin association.Portions of this work are in partial fulfillment of the requirements for the Ph.D. degree from Vanderbilt University.     

Atrial natriuretic peptide in the central nervous system of the rat

1. Studies of the presence of atrial natriuretic peptide immunoreactivity and receptor binding sites in the central nervous system have revealed unusual sites of interest. 2. As a result, numerous studies have appeared that indicate that brain atrial natriuretic peptide is implicated in the regulation of blood pressure, fluid and sodium balance, cerebral blood flow, brain microcirculation, blood-brain barrier function, and cerebrospinal fluid production. 3. Alteration of the atrial natriuretic peptide system in the brain could have important implications in hypertensive disease and disorders of water balance in the central nervous system.     

The causes and consequences of deforestation among the prehistoric Maya

The collapse of the Classic Maya state is investigated from an ecological perspective. Settlement and palynological data from the Maya center of Copan, Honduras, are presented which indicate that substantial clearing of the upland pine forest had occurred prior to and during the abandonment of that urban center. A comparative use- rate analysis suggests that the increased clearing of pine was primarily caused by demands for domestic fuel wood by an expanding urban population. This forest mismanagement is directly linked to accelerated erosion rates which are considered primary elements in the collapse of the Maya state.