Purification and characterization of two phosphoglucomutases from Lactococcus lactis subsp. lactis and their regulation in maltose- and glucose-utilizing cells.

Two distinct forms of phosphoglucomutase were found in Lactococcus lactis subsp. lactis, strains 19435 and 65.1, growing on maltose: beta-phosphoglucomutase (beta-PGM), which catalyzes the reversible conversion of beta-glucose 1-phosphate to glucose 6-phosphate in the maltose catabolism, and alpha-phosphoglucomutase (alpha-PGM). beta-PGM was purified to more than 90% homogeneity in crude cell extract from maltose-grown lactococci, and polyclonal antisera to the enzyme were prepared. The molecular mass of beta-PGM was estimated by gel filtration to be 28 kDa; its isoelectric point was 4.8. The corresponding values for alpha-PGM were 65 kDa and 4.4, respectively. The expression of both PGM enzymes was investigated under different growth conditions. The specific activity and amount of beta-PGM per milliliter of cell extract increased with time in lactococci grown on maltose, but the enzyme was absent in lactococci grown on glucose, indicating enzyme synthesis to be induced by maltose in the growth medium. When glucose was added to maltose-grown lactococci, both the specific activity and amount of beta-PGM per milliliter of cell extract decreased rapidly. This suggests that synthesis of beta-PGM is repressed by glucose in the medium. Although the specific activity of alpha-PGM did not change during growth on maltose or glucose, lactococcal strain 19435 showed a much higher specific activity of both alpha- and beta-PGM than strain 65.1 when grown on maltose.     

Transfer of the plJ101 plasmid in Streptomyces lividans requires a cis-acting function dispensable for chromosomal gene transfer

The tra gene of Streptomyces lividans plasmid plJ101 is required for both plasmid DNA transfer and plJ101-induced mobilization of chromosomal genes during mating. We show that a chromosomally inserted copy of tra mediates transfer of chromosomal DNA at high frequency but promotes efficient transfer of plasmids only when they contain a previously unknown locus, here named clt. Insertional mutation or deletion of clt from plJ101 reduced plasmid transfer mediated by either plasmid-borne or chromosomally located tra by at least three orders of magnitude, abolished the transfer-associated pocking phenomenon, and interfered with the ability of tra⁺ plasmids to promote transfer of chromosomal DNA. Our results indicate that plasmid transfer in S. lividans involves a cis-acting function dispensable for chromosomal gene transfer and imply that either the S. lividans chromosome encodes its own clt-like function or, alternatively, that transfer of plasmid and chromosomal DNA occurs by different mechanisms.     

Cholinergic innervation of the retrosplenial cortex via the fornix pathway as determined by high affinity choline uptake,choline acetyltransferase activity,and muscarinic receptor binding in the rat

The cholinergic projections from basal forebrain nuclei to the retrosplenial cortex (RSC) have previously been studied using a variety of histological approaches. Studies using acetylcholinesterase (AChE) histochemistry and choline acetyltransferase (ChAT) immunocytochemistry have demonstrated that this projection travels via the cingulum on route to the RSC. Preliminary studies from our laboratory, however, have shown that the fornix may also be involved in this projection. The present study uses the combination of pathway lesions, and the analysis of cholinergic neurochemical markers in the RSC to determine the role of the fornix in the cholinergic projection to the RSC. High affinity choline uptake (HACU) and ChAT activity were measured in the RSC of control rats, animals with cingulate lesions, and animals with fornix plus cingulate lesions. Fornix plus cingulate lesions resulted in significant deceases in HACU and ChAT activity in comparison to cingulate lesions alone. Muscarinic receptor binding was also evaluated in combination with the various lesions, and a significant increase in retrosplenial receptor binding was noted following fornix lesions. Together, these results support the concept of a fornix-mediated cholinergic pathway to the RSC.     

Is there an energy conservation “system” in brain that protects against the consequences of energy depletion?

A poorly understood marked decrease (circa 50% of control) in local cerebral glucose utilization is caused by sublethal doses of NaCN. The decrease is global, occurring in essentially all brain regions and is entirely reversible within hours, leaving no obvious pathology. This event is not unique to NaCN in so far as a strikingly similar pattern of decreased glucose utilization occus with some other toxins. Nor can it be attributed to a direct action of NaCN since local application by microdialysis to the striatum produces a global depression. These results imply that some widely distributed system or substance is involved. We speculate the existence of a system possibly related to the reticular activating system that senses a fall in energy production and acts globally to make cells quiescent and thus would give some protection from excitotoxic driven damage.Special issue dedicated to Dr. Sidney Ochs.     

Distribution and conservation status of primates in Bioko island,Equatorial Guinea

Ten species of non-human primates are indigenous to Bioko; half of these are endangered and between five and eight are endemic subspecies. Recent data on their status and distribution have been lacking. In 1986, a ten-week survey of the island was carried out to determine the distribution and status of the primates and the natural vegetation, and to evaluate the effects of man on them. This paper presents the results of that survey, gives an update of conservation achievements since 1986, and highlights current concerns. Between 1974 and 1986 it is probable that numbers of all Bioko primates rose as a result of an increase in habitat and of reduced hunting. At the time of the survey there was considerably more natural, undisturbed, vegetation remaining in Bioko tran expected. Much of this vegetation occurs within two large blocks that are of outstanding importance to the conservation of species in tropical Africa, particularly of plants and primates.     

Endophytic fungi alter foraging and dispersal by desert seed-harvesting ants

Endophytic fungi are thought to interact mutualistically with host plants by producing alkaloid metabolites that deter herbivory. Since such fungi are transmitted via seed in some grasses, the presence of endophytes may also protect plants from seed predators. We conducted seed choice experiments for two dominant seed harvesting ants, Pogonomyrmex rugosus in the Sonoran desert and Pogonomyrmex occidentalis at a higher elevation, riparian zone in Arizona, USA. Non-infected fescue (Festuca arundinacea) seeds and seeds infected with the endophytic fungus, Acremonium coenophialum, were presented to ant colonies in three different populations. Infected seeds were harvested less frequently than non-infected seed for the two populations of Pogonomyrmex rugosus but not for the population of Pogonomyrmex occidentalis. We also a conducted seed dispersal experiment for one population of Pogonomyrmex rugosus. Of the seeds that were harvested, most of the colonies discarded more infected seeds into refuse piles than expected by chance. Seeds discarded into refuse piles have greater germination success than surrounding areas. The most important interaction of endophytes and grasses may be deterrence of seed predation and enhancing the probability of germinating in favorable sites, since these processes directly increase plant fitness.     

Effects of fire on water and salinity relations of riparian woody taxa

Water and salinity relations were evaluated in recovering burned individuals of the dominant woody taxa from low-elevation riparian plant communities of the southwestern U.S. Soil elemental analyses indicated that concentrations of most nutrients increased following fire, contributing to a potential nutrient abundance but also elevated alluvium salinity. Boron, to which naturalized Tamarix ramosissima is tolerant, was also elevated in soils following fire. Lower moisture in the upper 30 cm of burned site soil profiles was attributed to shifts in evapotranspiration following fire. Higher leaf stomatal conductance occurred in all taxa on burned sites. This is apparently due to higher photosynthetic photon flux density at the midcanopy level and may be partially mitigated by reduced unit growth in resprouting burned individuals. Predawn water potentials varied little among sites, as was expected for plants exhibiting largely phreatophytic water uptake. Midday water potentials in recovering Salix gooddingii growing in the Colorado River floodplain reached levels which are considered stressful. Decreased hydraulic efficiency was also indicated for this species by examining transpiration-water potential regressions. Recovering, burned Tamarix and Tessaria sericea had enriched leaf tissue ¹³C relative to unburned controls. Higher water use efficiency following fire in these taxa may be attributed to halophytic adaptations, and to elevated foliar nitrogen in Tessaria. Consequently, mechanisms are proposed which would facilitate increased community dominance of Tamarix and Tessaria in association with fire. The theory that whole ecosystem processes are altered by invading species may thus be extended to include those processes related to disturbance.     

Effect of agitation speed on the synthesis of Mucor miehei acid protease

Different experiments using Mucor miehei CBS 370.65 were carried out to study the effect of agitation speed on the production of the mold acid protease. The experiments were conducted in shake flasks at a fixed substrate concentration of 58 g l⁻¹ of total carbohydrates and at shaker speeds from 80 to 380 rev min⁻¹. Enzyme production was found to be directly proportional to the shaker speeds, with the highest concentration of enzyme of 1,400 Soxhlet Rennet units (SU) ml⁻¹ obtained at 380 rev min⁻¹. The yield of product to substrate at 380 rev min⁻¹ was determined to be 27,081.0 SU g⁻¹ substrate and the productivity of the process was 221 SU g⁻¹ h⁻¹. Enzyme production was partially growth associated, and glucose supported both cell growth and enzyme production. Product formation and cell concentration were directly related to the rate of substrate consumption. The rate of product formation decreased when product started to accumulate, suggesting that the process was affected by feedback repression.     

Effects of 60 Hz electric and magnetic fields on maturation of the rat neopallium

This study was undertaken to determine the effects of extremely low frequency (ELF; 60 Hz) electromagnetic (EM) fields on somatic growth and cortical development, as well as biochemical and morphological maturation, of the rat neopallium. On the fifth day of pregnancy, female rats were put in pairs into plastic cages that were housed in a specially constructed apparatus for irradiation under three separate sets of combination and intensity: 1) 1 kV/m and 10 gauss; 2) 100 kV/m and 1 gauss; and 3) 100 kV/m and 10 gauss. The dams were exposed for 23 h daily, from days 5 through 19 postconception after which they were returned to cages outside the exposure apparatus until they littered. The neonates were culled to eight pups per litter. At 0 (birth), 5, 12, and 19 days postnatally, they were killed for biochemical and morphological studies. Another group of pregnant rats was sham-exposed in an identical apparatus, which was not energized, and the pups were used as controls. The irradiated rats exhibited no physical abnormalities, nor did they show brain deformities such as swelling or herniation following exposure to ELF-EM fields. There was no difference in somatic growth between control and exposed rats, but a small reduction in cortical weight was observed in rats exposed at 1 kV/m and 10 gauss, and 100 kV/m and 1 gauss, respectively. Biochemical measurements of DNA. RNA, protein, and cerebroside concentrations indicated that among the three separate exposures, only the neopallium of rats exposed at 1 kV/m and 10 gauss showed a small reduction in DNA level, as well as small reductions in RNA and protein levels. No changes were noticed in cerebroside levels in any exposed animals, and there were no differences in protein/DNA and cerebroside/DNA ratios between control and exposed rats. Morphological observations did not reveal any detectable alterations in the irradiated rats. These results indicate that exposure to ELF-EM fields caused minimal or no changes in somatic growth and cerebral development of the rat. © 1993 Wiley-Liss, Inc.     

The Biotransformation of Propylene to Propylene Oxide by Methylococcus capsulatus (Bath): 3. Reactivation of Inactivated Whole Cells to Give a High Productivity System

Methylococcus capsulatus (Bath) possesses methane monooxygenases (soluble - (sMMO) and particulate - (pMMO)) which are able to catalyse the epoxidation of propylene to propylene oxide. In a previous paper we have shown that the production of the epoxide caused a rapid inactivation of the bioconversion process (Stanley et al, 1992). This paper shows that cultures containing pMMO, inactivated by propylene oxide production, could be completely reactivated in the presence of growth substrates within 5 h after the removal of propylene oxide so long as the propylene oxide production rate was below 150 nmol min^-1 [mg dry weight cells]^-1. Reactivation under these conditions was detectable within 30 min of propylene oxide removal. On the other hand, cells inactivated by propylene oxide production rates in excess of 150 nmol min^-1 [mg dry weight]^-1 did not begin to recover activity within the 30 min period. Furthermore a lag period was observed before reactivation began which was dependent upon the initial production rate. Cultures possessing sMMO took twice as long to recover their activity compared with cells containing pMMO.

Reactivation of propylene oxide production could occur without growth, but the process required the presence of a carbon and energy source (methane or methanol), sulphur, nitrogen and oxygen, although copper (which is normally involved in pMMO activity) was not required. It was shown that de novo protein synthesis was required for reactivation of activity.

Production rates of 12 g 1^-1 d^-1 could be maintained for longer than three weeks in a single phase production process and rates up to 30 g 1^-1 d^-1 were achieved in a two stage process. Using Methylocystis parvus (OBBP) rates of up to 90 g 1^-1 d^-1 were attained over a one week period.