Production and biochemical characterization of polygalacturonases produced byAureobasidium pullulans from forest soil

The production of individual form of extracellular polygalacturonase byAureobasidium pullulans from forest soil was found to depend on the pH of cultivation medium as well as on the nitrogen source in the precultivation or cultivation medium. Polygalacturonases were purified and characterized. The pH optima of polygalacturonases produced in the first phases of cultivation (24 or 48 h) and after 10 days as well as their molecular masses, isoelectric points, action pattern and ability to cleave polymeric and oligomeric substrates were different. Generally, polygalacturonases with random action pattern (EC 3.2.1.15) were produced only in the first phases of cultivation in acidic medium. The function of these enzymes for A.pullulans in the colonization of plant material rather than in the destruction of plant was hypothesized in physiological conditions. Exopolygalacturonases (EC 3.2.1.67) with terminal action pattern were produced in later phases of growth. Oligogalacturonate hydrolase as well as strongly basic polygalacturonase with unusual action pattern on substrates were found.     

The life style of Aureobasidium pullulans and the multiple forms of its polygalacturonase

The life style of Aureobasidium pullulans on pectin medium and its production of extracellular polygalacturonases are closely related. Polygalacturonases with random action pattern (EC 3.2.1.15) were formed in the first phases of cultivation, whereas exopolygalacturonases (EC 3.2.1.67) with terminal action pattern on pectin were produced during the whole growth of this yeast-like fungus. The production and inactivation of individual enzyme forms during cultivation were strongly dependent on the pH value of the pectin medium. Various kinds of stress can support the prolongation of the phase of endo-acting enzyme production, as well as the increase of their activity.     

An ion-pair reversed-phase HPLC method for determination of fresh tissue adenine nucleotides avoiding freeze-thaw degradation of ATP

Knowledge of the energetic state of tissue is required in a wide range of experimental studies, particularly those investigating the decline and recovery of cellular metabolism after metabolic stress. Such information can be obtained from high-performance liquid chromatography (HPLC) determination of tissue levels of adenine nucleotides (ATP, ADP, and AMP) and their interrelationship in the tissue energy charge (EC). Accordingly, a large range of techniques with which to measure these molecules and their downstream metabolites have been reported. However, the accurate determination of the tissue EC also depends on the nucleotide extraction procedure given that changes in adenine nucleotide levels take place very quickly when ATPases are not inactivated immediately. In this article, we describe an ion-pair reversed-phase HPLC method by which separation of adenine nucleotides can be performed rapidly, allowing multiple analyses in 1 day, with both high sensitivity and extraction efficiency and using fresh samples, thereby avoiding freeze-thaw degradation of nucleotides. We applied this method to hippocampal brain slice extracts and show that same-day extraction and analysis results in a more accurate determination of the in situ energetic state than does the commonly used snap-freezing in liquid nitrogen.     

A comparative study of stereolithographically modelled skulls of Petralona and Broken Hill: implications for future studies of middle Pleistocene hominid evolution

Computer generated three-dimensional stereolithographic models of middle Pleistocene skulls from Petralona and Broken Hill are described and compared. The anterior cranial fossae of these models are also compared with that of another middle Pleistocene skull, Arago 21. Stereolithographic modelling reproduces not only the outer surfaces of skulls, but also features within the substance of the bones, and details of the internal braincase. The skulls of Petralona and, to a somewhat lesser degree, Broken Hill are extremely pneumatized. Previously undescribed features associated with pneumatization are detailed, along with their possible functional significance, polarity, and potential for understanding hominid cranial variation. Petralona and Broken Hill also exhibit a dramatic suite of cerebral features that is probably related to extensive pneumatization of the skull, namely frontal lobes that are tilted and located behind rather than over the orbits, laterally flared temporal lobes, marked occipital projection, and basal location of the cerebellum. Comparison of the anterior cranial fossae of Petralona, Broken Hill, and Arago 21 suggests that external resemblance of skulls may not always correlate with endocranial similarity. We believe that stereolithographic reconstructions have the potential for helping to resolve difficult questions about the origins of Neanderthal and anatomically modern people.     

Modulation of class I and class II histocompatibility antigens on human T cell lines by IFN-gamma

IFN-gamma is an immunomodulatory agent which is known to induce or enhance the expression of class II histocompatibility Ag (Ia Ag) on many lymphoid cells and cell lines of diverse origin. However, we have observed that IFN-gamma did not induce the expression of Ia Ag on Ia- human T cell lines. Neither did IFN-gamma enhance the expression of Ia Ag on Ia+ T cells. However, IFN-gamma was able to enhance the expression of class I histocompatibility Ag (HLA-A,B,C Ag) on a number of the T cell lines tested. Experiments with 125I-labeled IFN-gamma showed a relatively small degree of specific binding to these T cell lines. More extensive studies on two of the T cell lines demonstrated 1000 and 2600 IFN-gamma binding receptor sites/cell and binding affinities of 4.0 X 10(-10) M and 7.3 X 10(-10) M. Thus, although IFN-gamma can bind to human T cell lines and enhance class I histocompatibility Ag on these cells, IFN-gamma alone does not appear to regulate expression of class II histocompatibility Ag on T cell lines.     

Effect of glucose utilization inhibitor isolated from the serum and urine of patients with chronic renal failure on the free fatty acid metabolism in rat kidney cortex slices.

The authors studied interference by the inhibitor of glucose utilization with free fatty acid (FFA) metabolism in rat kidney cortex slices. They found that FFA uptake by the kidneys, their incorporation into triglycerides (TG) and oxidation to 14CO2 were lower when the inhibitor was present in the incubation medium. The findings show that the inhibitor, as well as inhibiting glucose utilization in various tissues, interferes directly with fat metabolism.     

Re-evaluation of the endocranial volume of the Guattari 1 Neandertal specimen (Monte Circeo)

The endocranial capacity of Guattari 1 originally was estimated by Sergi as approximately 1.550 cm3. Using three different approaches, a physical endocast, a stereolithographic model, and a virtual endocast, we have estimated the endocranial capacity of Guattari 1 as approximately 1.350 cm3. This paper explains our revision of the estimated endocranial volume of Guattari 1, provides a cautionary case concerning other estimates of endocranial volume, and demonstrates and encourages the use of recent advances in imaging, modeling, and analysis of endocranial volume.     

Purification of exopolygalacturonase by affinity chromatography on Concanavalin A — Bead cellulose

Summary Interaction of exopolygalacturonase isolated from carrot roots with Concanavalin A was investigated. The glycoprotein character of this enzyme was confirmed. Using affinity chromatography on Concanavalin A — triazine bead cellulose about fifty fold purification was achieved. The individual enzyme forms were not separated by this method, which might imply identical or similar pattern of n-glycosylation.