Staining of Negri Bodies

The following procedure for staining Negri bodies in sections is based on methods previously described by MacNeal, by Haynes, and by Richter:

Fixation:

1. 1. Zenker's solution 4 hours at 37°C or Dominici's 3 hours.

2. 2. 70% alcohol, 12 to 18 hours at room temperature.

3. 3. 80% alcohol, about 5 to 6 hours.

4. 4. 90% alcohol, about 4 to 6 hours.

5. 5. Absolute alcohol about 16 hours.

6. 6. Ether and absolute alcohol aa, about 8 hours.

7. 7. 16 to 24 hours in the following mixture: celloidin 1 g., methyl salycilate 25 cc., abs. alcohol 25 cc., ether 25 cc.

8. 8. Chloroform and paraffin, 2 to 3 hours.

9. 10. Paraffin, 1 to 1 1/2 hours.

10. 11. Embed.

staining:

1. 1. Cut sections 4 to 5 μ.

2. 2. Bring section to water and cover with Lugol's iodine for 10 minutes.

3. 3. Decolorize with a 2% sodium thiosulfate (hypo).

4. 4. Wash thoroly with water.

5. 5. Cover with a mixture of equal parts of 0.5% phloxine and 1% eosin Y (National Aniline brand) and leave for 15 minutes.

6. 6. Wash with water and stain 2 to 5 minutes in 0.1% azure B (National Aniline).

7. 7. Wash with 96% alcohol and decolorize in a mixture of 2 parts absolute alcohol with 1 part clove oil, ordinarily for not more than 1/2 to 1 minute.

8. 8. Dehydrate rapidly, clear, and mount in Yucatan Elemi.

     

Removal of Anti-complementary Material and Australia Antigen from Immunoglobulin

Inmunoglobulin isolated from human sera, be it by the cryo-alcohol, rivanol, multi membrane electrodecantation or polyethylene glycol process, alvays contains denatured material. This may result from the influence either singly or in combination, of acme of the follwing factors:

1. inefficiency of the purification procedure;

2. surface denaturation;

3. imperfect freeze-drying of the final product; and

4. factors yet unknown vhich cause alteration in the immoglobulins or other protein components not ellminated by the purification procedures.

     

Induction of Catalase Activity by Hydrocarbons in Candida tropicalis рK 233

1. The catalase activity of Candida tropicalis pK 233 was induced by hydrocarbons but not by glucose, galactose, ethanol, acetate or lauryl alcohol.

2. The induction of the catalase activity depending upon hydrocarbons was sensitive to cycloheximide but not to chloramphenicol.

3. Glucose repressed strongly the induction of the catalase activity by hydrocarbons but galactose did not affect seriously.

4. When C. tropicalis was incubated with hydrocarbons, the appearance of microbodies was observed electronmicroscopicaliy.

     

Protective Actions of l-Aspartate from Acid or Proteolytic Inactivation

1. l-Aspartate was found to replace l-asparagine in the protective action from acid inactivation of l-asparaginase (EC 3.5.1.1) produced by Escherichia coli A–1–3 and at the same time to inhibit the proteolytic inactivation by α-chymotrypsin.

2. l-Asparaginase changed in its chromatographic properties in the presence of l-aspartate and became to be absorbed on the CM Sephadex column.

3. The sedimentation patterns of l-asparaginase at pH 3.5 were identical either in the presence or absence of l-aspartate, showing partial dissociation. But the reversibility to the active state was observed only in the enzyme dissolved in the solution containing l-aspartate.

4. l-Aspartate did not prevent the enzyme either from the dissociation into subunits or from decrease in the activity by urea.

5. High concentration of l-aspartate was shown to inhibit the l-asparagine hydrolysis reaction.

6. l-Aspartate was suggested from ORD measurements to cause changes in the higher structure as well as the ionic properties or proteolytic inactivation.

     

Ecological Risk Assessment and Quantitative Consequence Analysis

Ecological risk actually refers to two separate things. First, risk to the environment as a result of human activity. Contaminated sites are an example. Second, risk to the biota—flora, fauna, and people—as a result of environmental hazards. Geophysical risk arising from natural hazards is an example. Risk is a combination of likelihoods and consequences. This article examines methods used to quantify the consequences. At the general level, such methods are linked to the methods used to quantify the likelihoods and thus to quantify the risks. It is possible to use the existing frameworks of risk management, health risk assessment, and ecological risk analysis to develop a risk management framework that is suitable for ecological risk assessment. The framework consists of the following steps:

1. Determine concernsby using risk assessment techniques for various scenarios.

2. Identify the consequences by systematically identifying hazards.

3. Undertake calculations by using relevant models.

4. Evaluate certainties, uncertainties, and probabilities involved in the calculations of the vulnerability and of the exposure.

5. Compare with criteriato assess the need for further action.

6. Determine and act on options to control, mitigate, and adapt to the risk.

7. Communicatethe results to those who need to know.

     

On the neotropical acanthocinini (coleoptera,ceramhycidae, lamiinae) some new genera and generic revisions

In Fortführung seiner Untersuchungen über neotropische Acanthocinini veröffentlicht der Verfasser hiermit Beschreibungen von folgenden neuen Gattungen:

• Nyssodrysilla nov. gen. mit N. irrorata (Melzer) aus Brasilien als Generotype, N. viliata (Melzer), comb, nov., aus Brasilien und N. lineata nov. spec, aus Peru.

• Nyssodrysola nov. gen. mit N. stictica nov. spec. aus Peru als Generotype.

• Sciadosurus nov. gen. mit S. albobrunneus nov. spec. aus Peru als Generotype.

• Acarinozineus nov. gen. mit A. striatus nov. spec. aus Peru als Generotype und A. spinicornis nov. spec, aus Mexiko.

• Alcathousites nov. gen. mit A. chaclacayoi nov. spec. aus Peru als Generotype.

• Xylergatina nov. gen. mit X. pulcher (Lane) aus Peru als Generotype.

• Xylergatoides nov. gen. mit X. asper (Bates) aus Brasilien und Argentinien als Generotype.

Ferner werden revidiert die Gattungen:

• Xylergates Bates, Generotype X. lacteus (Bates), mit Beschreibung der beiden neuen Arten X. elaineae aus Peru und X. dorotheae aus Britisch‐Guayana.

• Chaetanes Bates, Generotype C. setiger (Bates), mit Beschreibung der drei neuen Arten C. costulatus aus Peru, C. nigrobasalis aus Brasilien und C. apicalis aus Französisch‐Guayana.

• Wo es erforderlich ist, sind Bestimmungstabellen gebracht und die Arten abgebildet.

     

Participation of Mevalonate in the Biosynthetic Pathway of Ipomeamarone

1. The incorporation of mevalonate-2-¹⁴C into ipomeamarone in sweet potato root tissue infected by Ceratocystis fimbriata was demonstrated, but the rate was low when compared with acetate-2-¹⁴C. No dilution effect of mevalonate was noted during the incorporation of acetate-2-¹⁴C into ipomeamarone. This is very likely to result from the passive transfer of mevalonate into the cells.

2. No dilution effect of acetate during the incorporation of mevalonate-2-¹⁴C into ipomeamarone was noted. This indicates that mevalonate is not incorporated into ipomeamarone after its conversion to acetate.

3. Evidence for incorporation of acetate-2-¹⁴C into mevalonate was shown by the fact that the specific radioactivity of mevalonic acid benzhydrylamide was not lowered throughout repetitive crystallizations. These data also support the participation of mevalonate in ipomeamarone synthesis as an intermediate.

     

Embriologia Della “Vinca Difformis„ Pourr. (Apocinaceae)

Riassunto

L'A. ha studiato l'embriogia della Vinea difformis Pourr. ed ha potuto stabilire che:

1. l'archisporio è pluricellulare e possono svilupparis talvolta pi[ugrave] cellule madri;

2. normalmente solo una cellula madre arriva a maturità;

3. delle quattro megaspore solo una è fertile e precisamente la pi[ugrave] calazale;

4. lo sviluppo del gametofito è del tipo Normale cioè Monomegasporiale con oangio emisporiale.

Ha inoltre risontrato una anomalia di sviluppo constituita da un gametofito binucleato abnorme per ritardo delle divisioni nucleari cispetto all'acerescimento che è quello di un gametofito ottonucleato.     

Analisis de la craneometria diferencial entre la vicuña (Vicugna vicugna) y la alpaca (Lama guanicoë pacos)

Using a total of 43 craneological data obtained from any of 73 vicugna and 25 alpaca skulls, three problems were analyzed:

• Identification of skulls

• Taxonomic situation of the vicugna

• Origin of the alpaca.

For rapid identification of New World cameloid skulls it is recommended to use the condilobasal length, the length to heigth ratio and the presence of the Fissura nasolacrimalis.

Some characteristics of the skulls considered essential for the evaluation of domestication processes exclude the vicugna from the alpaca's ancestors.     

Enzymatic synthesis of gallic acid from tannic acid with an inducible hydrolase of Enterobacter spp

Gallic acid acts as a precursor molecule to synthesize various tannin molecules. These are plant polyphenols and were proved to be good anti-oxidant, anti-cancerous, anti-inflammatory, anti-microbial compounds. In order to fully exploit prominent biological activities of specific tannins and to develop tannin-based new medicines, it is necessary to obtain their pure preparations with an aim of high yield and specificity. In the present study, gallic acid is synthesized by the hydrolysis of tannic acid using a microbial based transformation process. The microorganism was isolated and identified. The ability of the isolated microorganism to covert tannic acid into gallic acid was determined by HPLC and enzyme production.

• Highlights

• The present investigation signifies the role of Enterobacter spp. in various processes:

• ??To synthesize gallic acid (a precursor for food oxidant such as propyl gallate) and a bacteriostatic antibiotic (trimethoprim).

• ??To protect the environment from tannery’s discharge through the process of biodegradation.

• ??To reduce the toxicity of tannins in animal feed.

     

The Nutritional Value of the Individual Nonessential Amino Acid as the Nitrogen Source in the Chick Nutrition

The nutritional values of nonessential amino acids as the nitrogen source in the crystalline amino acid diet for the chick growth were examined. The nitrogen of the nonessential amino acids in the basal diet for chick was substituted for a nonessential amino acid to be tested on the nitrogen base. The experimental methods were the same as in the evaluation of the nutritional value of d-amino acids previously reported. Nonessential amino acids were classified into four groups.

1. Very useful nitrogen source: Glutamic acid, Aspartic acid

2. Useful nitrogen source: Alanine, Diammonium citrate

3. Insufficient nitrogen source: Glycine, Proline

4. Harmful for chick growth: Serine

At the end of experiment chicks were killed and the concentration of free amino acids in the serum were measured. The concentration of glycine and serine in the serum increased when glycine was tested, but that of serine in the serum only increased when serine was tested. This result suggested the pathway from glycine to serine was fast and the opposite one was very slow.     

Quality of Irrigation Waters of the Alkali Infested Areas in Allahabad,India

The concentration and composition of dissolved constituents in water, determine its quality for irrigation. Quality of water is important consideration in any appraisal of salinity or alkali conditions in an irrigated area. The characteristics of an irrigation water that appear to be most important in determining its quality are:

1. Total concentration of soluble salts.

2. Relative proportion of sodium to other cations.

3. Concentration of boron or other toxic elements.

4. Under certain conditions, the bicarbonate concentration as related to the concentration of calcium plus magnesium.

The total concentration of soluble salts in irrigation waters can be adequately expressed for purposes of diagnosis and classification in terms of electrical conductivity. Irrigation waters that have been used successfully for a considerable time have conductivity values less than 2,250 micro mhos/cm. Waters of higher conductivity are used occasionally, but crop production, except in unusual situations, has not been satisfactory.     

Poster Abstracts

DOCSPER—A Synthetic Lipid Fit for In Vivo Application

DOCSPER [1,3-Dioleoyloxy-2-(N⁵-carbamoyl-spermine)-propane] is a cationic amphiphile consisting of a hydrophobic 1,3 dioleylglycerol moiety and threefold positively charged spermine head group (). We optimised the 5-step-synthesis of the lipospermine and after up-scaling we have obtained sufficient amounts to initiate preclinical investigations. DOCSPER was tested for its ability to transfect eukaryotic cells in vitro. It has proven to possess high transfection efficiency in comparison to commercially available liposomal transfection agents. Furthermore, DOCSPER was extensively tested in several in vivo studies (). These studies revealed a high transfection efficiency, whereas very low toxicity levels were detected. Thus, the results clearly indicate that the cationic lipid DOCSPER is a reliable, low-risk system for broad applications in gene therapy.

1. Groth D. et al. Int J Pharm 1998; 162:143–157.

2. Nikol S. et al. Int J Angiol 2000; 9:87–95.

3. Armeanu S. et al. Mol Ther 2000; 1(4):366–375.

     

Effects of Acetylation with Acetic Anhydricle

1. L-Asparaginase (EC 3.5.1.1) from Escherichia coli A–l–3 was acetylated using acetic anhydride as a modifying chemical. The fully acetylated L-asparaginase retained 60% of the activity of the unmodified L-asparaginase.

2. The acetylated L-asparaginase hydrolyzed D-asparagine and L-glutamine as well as L-asparagine in the same ratio as the unmodified L-asparaginase did.

3. However, the effects of pH on the activity of the acetylated L-asparaginase showed very interesting differences from that of L-asparaginase. On the other hand, both L-asparaginase and the acetylated L-asparaginase exhibited similar pH activity curves on L-glutamine hydrolysis.

4. The acetylated L-asparaginase was found to become more stable against acid or heat in the presence of L-aspartate than in its absence in the same manner as L-asparaginase was.

     

Visual shape discrimination in the fiddler crab,Uca pugilator

1. Research into the visual shape discrimination abilities of compound‐eyed animals has almost exclusively been limited to insects, the crustaceans having been virtually ignored. The two groups have many dissimilarities, having primarily adapted in different habitats to different lifestyles. Differences may exist in visual systems and visually mediated behavior.

2. Fiddler crabs (Uca pugilator), without training, differentially approached dissimilar silhouettes presented simultaneously, demonstrating visual discrimination between stationary, geometric shapes of equal‐area. The strength of response was ordered hierarchically: vertical rectangle, horizontal rectangle, triangle, square, circle.

3. Basic geometric shapes were used to facilitate replication and comparison with research findings from other species.

     

Influence d'éclairements brefs, à différentes longeurs d'onde,sur les variations circadiennes des activités enzymatiques digestives chez Palaemon serratus (Crustacea,Natantia)

Abstract

The Influence of Short Periods of Lighting with Different Wavelengths on the Orcadian Changes of the Enzymatic Digestive Activities in Palaemon serratus (Crustacea, Natantia)

The effect of light on the circadian rhythms of digestive activities in the Shrimp Palaemon serratus have been studied.

1. Continuous darkness leads in a few days to a disappearance of the variations of the circadian rhythms of digestive enzymes while these rhythms go on in continuous light.

2. Short (1 or 2 hrs) and low intensity flashes of white light are effective in bringing on the reappearance of rhythmic variations in darkness.

3. We have been able to establish an isoquantic spectrum of action of the light. Two values of wavelength appears to account for a maximum sensibility of the shrimp: one in ultraviolet light and an other one, more important, in the green (λ=544 nm).

4. In green light it is possible to obtain the same effect of light by decreasing the time of stimulation to 5 or 10 mn and in increasing the total quantity of energy. Significant responses are obtained with total energy greater than 10000 pE. cm^‐2.

     

Contributo allo studio dei fenomeni di correlazione nelle Graminacee

Summary

The Author studies the action of the presence of the ear and its parts in the growth of the stem and in the value of the water of traspiration and of assimilation in the higher leaf. The stet shows:

1. A removed ear, not yet mature, exerts a depressing effect upon the development of the stem and ucon the leaf's functions.

2. The threads and the pistils removed inhibit the enlongament of the stem and alter the functions of leaves.

3. Maturing seeds removed cause trubles on the function of the vegetative organ.

This phenomen is due to the presence of an hormonical substances elaborated in the riproduction's orgas, which are considerated the centre of functional correlation.     

Solubilization and Characterization of Ovomucin without Chemical Modification

1. The egg white, thick and thin fractions, was solubilized in 1.0% SDS solution by vigorous mixing and subjected to gel filtration on a Sepharose 4B column, eluted with 1.0% SDS. The isolated thick and thin ovomucins were found by analytical disc electrophoresis to be free from contamination with lysozyme.

2. In the velocity sedimentation the two ovomucin fractions behave similarly, both comprising at least two components with sedimentation coefficients 35 S and 30 S.

3. The chemical compositions of the two ovomucin fractions showed only notable difference in that the carbohydrate content of the thick white ovomucin was somewhat higher than that of the thin white ovomucin. The amino acid profiles of the two fractions were similar.