Cold Distilled Water Research Articles

Mining the Giardia lamblia Genome for New Cyst Wall Proteins

The Giardia lamblia cyst wall (CW), which is required for survival outside the host and infection, is a primitive extracellular matrix. Because of the importance of the CW, we queried the Giardia Genome Project Database with the coding sequences of the only two known CW proteins, which are cysteine-rich and contain leucine-rich repeats (LRRs). We identified five new LRR-containing proteins, of which only one (CWP3) is up-regulated during encystation and incorporated into the cyst wall. Sequence comparison with CWP1 and -2 revealed conservation within the LRRs and the 44-amino-acid N-flanking region, although CWP3 is more divergent. Interestingly, all 14 cysteine residues of CWP3 are positionally conserved with CWP1 and -2. During encystation, C-terminal epitope-tagged CWP3 was transported to the wall of water-resistant cysts via the novel regulated secretory pathway in encystation-secretory vesicles (ESVs). Deletion analysis revealed that the four LRRs are each essential to target CWP3 to the ESVs and cyst wall. In a deletion of the most C-terminal region, fewer ESVs were stained in encysting cells, and there was no staining in cysts. In contrast, deletion of the 44 amino acids between the signal sequence and the LRRs or the region just C-terminal to the LRRs only decreased the number of cells with CWP3 targeting to ESVs and cyst wall by approximately 50%. Our studies indicate that virtually every portion of the CWP3 protein is needed for efficient targeting to the regulated secretory pathway and incorporation into the cyst wall. Further, these data demonstrate the power of genomics in combination with rigorous functional analyses to verify annotation.

PRODUÇÃO DE XILANASE POR FUNGOS FILAMENTOSOS ISOLADOS DO SOLO DA ESTAÇÃO ECOLÓGICA DE JURÉIA-ITATINS, SP, BRASIL

Os fungos vêm despertando crescentes interesses em virtude de elaborarem compostos economicamente importantes, tais como: alimentos, solventes, enzimas, vitaminas, antibióticos, ácidos graxos, dentre outros produtos, justificando sua aplicação biotecnológica. Estudos da potencialidade de espécies pertencentes as regiões ainda pouco exploradas devem ser incentivadas, como é o caso da Estação Ecológica de Juréia-Itatins (EEJI), SP, Brasil, onde há enorme biodiversidade, porém em parte ainda desconhecida. Desta forma, o presente estudo apresenta resultados preliminares da atividade enzimática da xilanase (E.C. 3.2.1.8) que foi avaliada em oitenta linhagens de fungos isolados do solo da EEJI, na região do Banhado Grande. Os fungos foram
 cultivados em farelo de trigo por quatro dias a 25oC. As enzimas liberadas no meio de cultura foram extraídas com 30 ml de água gelada e a atividade da xilanase foi avaliada
 em unidades internacionais (UI) de enzima necessária para liberar um m mol de xilose por mililitro por minuto, nas condições do ensaio. A melhor atividade da xilanase
 (1.219,8 UI g-1) foi obtida utilizando-se Trichoderma inhamatum I.

Modulation of sensory nerve function in the airways.

Several clinical studies document a greater discrimination between asthmatic and healthy subjects in bronchial responsiveness to a range of stimuli such as cold air, distilled water and sodium metabisulphite, than to conventional bronchoconstrictor agonists including histamine and methacholine. One of the mechanisms thought to account for the bronchoconstriction induced by these agents is via reflex activation of the cholinergic pathway. An increase in sensory nerve (afferent) activity in asthma might account for the increased responsiveness to these agents. If so, a number of strategies are available to inhibit the function of afferent nerves which could lead to a suppression of bronchial hyperresponsiveness, including (1) inhibition of afferent activity, (2) inhibition of neuropeptide release and (3) antagonism of tachykinin receptors. As there are numerous reviews dealing with the latter, in this review Domenico Spina, Saloni Shah and Selena Harrison focus on the first two strategies.

Fates of the Pyramidal Neurons in Neocortex After Callosotomy: Retrograde Neuronal Label with Two Fluorescent Tracers.

After Callosotomy: Retrograde Neuronal Label with Two Fluorescent Tracers Y.I. Orihara*, M. Kishikawa**, and K. Ono Departments of Pediatrics*, Physiology, and Pathology** at Scientific Data Center for the Atomic Bomb Disaster, Nagasaki University School of Medicine, Nagasaki, Japan We studied the fates of the neocortical pyramidal neurons after callosotomy by using retrograd eneuronal labeling with two fluorescent tracers, fast blue(FB) and diamidinoyel1ow(DY). All procedures were performed under sodium pentobarbital deep anaesthesia. Male sibling Wistar rats underwent the first procedure at age 2 months; injection of 7% solution of FB unilaterally throughout the frontoparietal cortex via microsyringe with a 22G needle.Two Weeks later,the rats were divided at random into two groups and underwent the second procedure; total cal1osotomy or sham operation.Four,8,and 20 weeks later, each rat underwent the third procedure; inection of 2% DY into the primary motor area opposite where the FB was injected. Five days later, they were perfused transcardiacally with physiologic saline followed by 10% neutral formalin. The brain was immediately removed from the skul1. After fixation and cryoprotection, the brain was frozen, and serial coronal sections were cut. Sections were thawed in ice cold distilled water and immediately mounted on slides. They were then cleared in xylene and coverslipped with DPX. Sections were assessed under an Olympus fluorescence microscope equipped with filter system U,providing excitation light of wavelength 360nm. Representative slices of the somatosensory area were photographed. cc-neuron

Hypertonic saline challenge in an epidemiologic survey of asthma in children.

To evaluate sensitivity and specificity of a 4.5% hypertonic saline challenge for a diagnosis of asthma, we studied 393 schoolchildren (13 to 15 yr of age) with "current wheeze," "former wheeze," and without history of wheeze in a community-based, cross-sectional survey. These children were selected from 2.836 schoolchildren in 26 schools in greater Melbourne, Australia who completed a self-administered questionnaire on respiratory symptoms, allergic rhinitis, and eczema. Three hundred eighty-two of 393 children successfully completed a 4.5% hypertonic saline challenge with increasing inhalation periods, and 365 of 393 performed a 6-min standardized, free running exercise challenge. The prevalence of bronchial hyperresponsiveness to hypertonic saline was 20.4%. Sensitivity and specificity for the hypertonic saline challenge to identify children with "current wheeze" were 47% and 92% respectively and for the exercise challenge, 46% and 88%. The agreement of response to hypertonic saline and to exercise was only moderate (kappa = 0.43). Factors associated with increased response to hypertonic saline in a multivariate logistic regression model were a history of "current wheeze," hay fever, response to the exercise challenge, female sex, and a lower baseline predicted FEV1. These results suggest that a 4.5% hypertonic saline challenge shows sensitivity and specificity similar to a standardized exercise challenge and pharmacologic challenges and a higher sensitivity than cold air hyperventilation and distilled water to identify asthma in children in a field study. Measurement of responsiveness to hypertonic saline may be of value as an objective marker of asthma to compare prevalence studies of bronchial hyperresponsiveness and of asthma over time and between countries.

Effect of Cromoglycate on Gas Changes, During Bronchial Challenge by UNCDW in Children with Asthma

Eighteen asthmatic children were challenged with ultrasonically nebulized cold distilled water (UNCDW). Blood gas composition was monitored transcutaneously (tcpO2 and tcpCO2) during and after the challenge. Assuming as basal the response to this UNCDW test, nine children (Group A) were then chosen at random to inhale cromoglycate by aerosol delivery for 8 days. Nine children (Group B), acting as a control, inhaled saline for 8 days. At the end of this therapy, each child repeated the UNCDW test. Statistical analysis with t-test for paired data was used to compare the results of each child to both tests. Mean basal tcpO2 and tcpCO2 were all within the expected normal range. In all children, both mean tcpO2 and tcpCO2 were reduced during and after UNCDW inhalation. Mean tcpCO2 values during the challenge were significantly (p < 0.001) lower than the corresponding steady state 2 rain after the UNCDW challenge, with a mean drop of −7% (2.1 S.D.). Mean tcpO2 values remained significantly decreased (p < 0.001) from the fifth mitt of the UNCDW challenge to the end of the observation period, with a mean drop of −20% (15.5 S.D.). After treatment with cromoglycate (Group A), the mean tcpCO2 values during UNCDW did not change significantly from those ofsteady state conditions: −0.8% (0.5 S.D.); whereas mean tcpO2 values decreased by −4% (4.9 S.D.). The control children treated with saline (Group B) showed mean tcpCO2 and tcpO2 values which were significantly different (p < 0.001) from those of the steady state conditions: mean drop of tcpCO2, −6% (4.2 S.D.); mean drop of tcpO2, −20% (4.7 S.D.). In conclusion, it emerges that: UNCDW induces nonspecific broncho-constriction in asthmatic children with a typical drop of tcpCO2 and tcpO2; the treatment with cromoglycate normalizes the time course of tcpCO2 (hyper-reactivity) and reduces dramatically the drop of tcpO2 time course (hyper-responsivity) during and after the UNCDW test.

Characterization and Immunohistochemical Localization of the 15 kD Protein Isolated from Rat Lung Lamellar Bodies

We have characterized a protein of approximately 15 kD (lb15) derived from rat lung lamellar bodies, and then sequenced the first 42 residues. Following the normal isopycnic sucrose gradient ultracentrifugation, we diluted the band containing the crude lamellar body fraction with an equal volume of cold distilled water and further centrifuged it at 2,000 x g for 30 min to pellet a fraction of lamellar bodies. Under the electron microscope, this fraction appeared intact and highly purified. When this fraction was subjected to polyacrylamide gel electrophoresis, the major protein was one of 15 kD, regardless of whether the fraction was extracted or unextracted, reduced or unreduced; only a small amount of 35 kD protein was detected with Coomassie Blue staining. Disruption of lamellar bodies revealed that the limiting membrane was particularly enriched with lb15. Immunohistochemistry indicated that lb15 was present in lamellar bodies and tubular myelin, suggesting it was secreted along with the lipid. Amino acid analysis revealed a protein with 13.5% basic and 10.6% acidic residues. The N-terminal appeared particularly highly charged, with 32% of the charged residues in the first 14 amino acids. The lb15 protein is identical to rat lysozyme for the first 23 residues, with the important exception of residue 6, which is histidine in lb15 and cysteine in lysozyme. Residue 24 was not identified. Lb15 was also present in lavage material. We conclude that lb15 is the major protein in rat lung lamellar bodies, has a highly charged N-terminal, and shares some sequence homology with rat lysozyme.(ABSTRACT TRUNCATED AT 250 WORDS)

Production of guava seed protein isolates: Yield, composition and protein quality

AbstractThe optimum conditions were determined for the preparation of protein isolates from ground, defatted guava seed flour. The isolates were obtained by micellisation and isoelectric precipitation techniques. The isoelectric point of guava seed protein was found to be pH 4.5 with a protein solubility of more than 85% above pH 9. Using 40–80 mesh particle size and extracting at pH 10 for 30 min, an isoelectric isolate was obtained by acidification to pH 4.5 which was 85.4% of the starting protein compared to 18.3% for micellisation protein obtained by twofold dilution with cold distilled water of 5% NaCl protein extract. The isolates were easily digested by the pepsin‐pancreatin enzyme system and were comparable to casein. The amino acid composition showed that guava seed flour and isolates were deficient in sulphur‐containing amino acids and in lysine, but contained the other essential amino acids in adequate levels. The micellisation isolate had higher fat absorption and emulsion stability than the isoelectric isolate, while other functional properties were almost similar.

Histochemical Localization of Alkaline Phosphatase Activity in Leucochloridiomorpha constantiae (Trematoda) Cultivated on the Chick Chorioallantois

Recent studies have described the histochemical localization of hydrolytic enzymes in whole cercariae and excysted metacercariae (LeFlore and Bass, 1983, International Journal for Parasitology 13: 179-183; Fried et al., 1984, Proceedings of the Helminthological Society of Washington 51: 140-143). Less information is availble about these enzymes in whole trematode adults. In this note we report our observations on the histochemical localization of alkaline phosphatase (ALP) activity in Leucochloridiomorpha constantiae cultivated from the metacercaria to the ovigerous adult on the chick chorioallantois. Metacercariae were cultivated on the chick chorioallantois at 37.5 C for 1-4 and 6 days (Harris et al., 1972, Journal of Parasitology 58: 213216). Worms were removed from the chorioallantois, rinsed briefly in 0.85% saline, prepared whole or as 10 Mm cryostat sections fixed at 1 C for 30 min in 10% formalin buffered to pH 7.0 with sodium phosphate. The whole worms were flattened during fixation by applying gentle pressure on the coverslips. After fixation the specimens were washed with cold distilled water to remove the fixative and treated for 12 hr at 22 C by the 3-glycerophosphate calcium salt method for AlP (Gomori, 1952. Microscopic Histochemistry. University of Chicago Press). Controls consisting of specimens heat-inactivated at 90 C for 5 min prior to incubation and omission of the specific substrate from the incubation medium were negative for AlP. The testes and ovary of day-old whole worms showed an intense reaction (Fig. 1). During development the area of the gonads increased, but the intensity of the AlP reaction in these organs decreased (Figs. 2-5). Cryostat sections of 6-dayold worms showed that the AlP reaction was within the testes (Fig. 7). Halton (1967, Journal of Parasitology 53: 46-54) reported slight A1P activity in the testes of 5 of 8 digeneans examined, and noted strong phosphatase activity in the cytoplasmic membranes surrounding the gonads. The vitellaria were first visualized in whole 2-day-old worms with an intense reaction (Fig. 2). The reaction remained intense during development, although the vitelline field decreased between days 4 and 6 in whole worms (Figs. 3-5). The vitelline ducts communicating with the vitellaria were first seen on day 3 whole worms (Fig. 3), and were evident until day 6 (Figs. 36). The uterine tube was first seen on day 1 (Fig. 1) in whole worms and became more evident on days 2 and 3 (Figs. 2, 3). The uterus was filled with eggs on days 4 and 6 (Figs. 4-6). Whole worms and cryostat sections showed intense A1P reactivity in the periphery of the eggs, mainly in the eggshell material (Fig. 6). Sharma (1976, Zeitshrift fur Parasitenkunde 49: 223-231) detected A1P activity in sectioned material of vitellaria, testes, ovary, uterus and egg shell in 10 species of trematodes. A1P activity occurred in the suckers and was more intense in the acetabulum than the oral

Host-Parasite Relationships of Helminth Parasites in Leeches of the Genus Haemopis. II. Associations at the Host Species Level

Host-Parasite Relationships of Helminth Parasites in Leeches of the Genus Haemopis. II. Associations at the Host Species Level

Host-Parasite Relationships of Helminth Parasites in Leeches of the Genus Haemopis. II. Associations at the Host-Species Level

Host-Parasite Relationships of Helminth Parasites in Leeches of the Genus Haemopis. II. Associations at the Host-Species Level

78] Aldehyde dehydrogenase from bakers' yeast

Publisher Summary This chapter describes the assay method, purification, and properties of aldehyde dehydrogenase isolated from bakers' yeast. Aldehyde dehydrogenase activity is assayed at 25°C by measuring the rate of formation of nicotinamide adenine dinucleotide dehydrogenase (NADH) at 340 nm. The proteolytic products from the native enzyme are found in the process of the purification of aldehyde dehydrogenase from yeast. The preparation method in the presence of phenylmethylsulfonyl fluoride (PMSF) as a serine esterase inhibitor diminishes the amount of proteolytic degradation products. Fresh bakers' yeast is washed twice with three volumes of cold distilled water and stored at -25°C before use. The steps involved in the purification of aldehyde dehydrogenase are (1) the preparation of crude extract, (2) protamine sulfate precipitation, (3) first and second ammonium sulfate fractionation, (4) diethylaminoethyl (DEAE)-sepharose chromatography, (5) gel filtration, and (6) blue sepharose chromatography. All steps are performed at about 4°C. The purified enzyme migrates as a single component when subjected to polyacrylamide electrophoresis in the absence and in the presence of sodium dodecyl sulfate (SDS) and does not show any sign of proteolytically degraded forms.

酵素消化法による組織内脂質同定に関する研究

Some purified esterases for lipids, e. g. mould lipase, snake venom phospholipase A2, cholesterol esterase from microorganism, were applied for the analysis of smaller fat granules in ultrastructure. The enzymes were purchased from Böhringer-Mannheim Co., and another mould lipase was donated from Drs. Tsujisaka and Iwai of Osaka Municipal Technical Research Institute. Some organs containing various amounts of lipids in rats, e. g. early and recovery stages of the Ethionine-induced fatty liver in females, the liver of fasting females, the Gentamicininjured renal cortex, the lactating mammary gland and the adrenal cortex of resting males, were perfused with a heparin-containing saline for 2 min and subsequently with 1/2 Karnovsky fixative for 3 min. Both tissue blocks, sized 0.5-1cm3, from the perfused organs and the atheromatous focus of human aorta were incubated in cold Karnovsky fixative for 1-2 hr, washed in cold cacodylate buffer with sucrose overnight, cut to 40μ-thick sections by Vibratom. The free-floating sections were treated in the following digestion media, respectively, at 37°C for 1/22 hr, while the medium was usually shaken rhythmically on the machine. The digestion medium was composed of 4m1 of 0.05 Tris buffer,1 ml of 2% calcium chloride and an enzyme. The enzyme was added as follows; 0.075 ml of lipase of Drs. Tsujisaka and Iwai at pH 7.2 for triglycerides detection,0.5 ml of snake venom phospholipase A2 of Böhringer at pH 7.5 for phospholipidsdetection, or 0.21 ml of cholesterol esterase of Böhringer at pH 7.6 for cholesterol esters detection. Duplicate control sections were treated in the reaction medium without enzyme. After treatment the sections were washed in cold distilled water, immersed in 0.1% lead nitrate for 15 min, washed again thoroughly in distilled water, post fixed in cold osmium buffer for 1.5 hr, dehydrated in cold graded ethanols according to Idelman's procedure, and finally embedded in epoxy resin. The ultrathin sections with or without metal staining were observed under an electron microscope. Derived fatty acids after the digestion with a specific enzyme were detected as lead soaps which had not been seen in the duplicate control section. These reaction products should indicate the location of the substrate lipid. Derived fatty acids could be also observed by means of Holczinger's method for fatty acids under a light microscope, when both pretreatments of removal of fatty acids and calcium in tissues should be done.Many co arse reaction products for triglycerides could be seen on newly appearing and disappearing fine lipid materials within the hepatic cells. Those for phospholipids revealed in some cytosegresomes of renal cortex, milk fat globule membranes, and biomembrane system of the examined cells, More trials are requested for ultrastructural detections of cholesterol esters, but the reaction products after cholesterol esterase digestion could be seen in adrenal cortical cells and macrophages in atheromatous foci. Both utility in histochemical analysis for the mixed phase of lipid spheres and limitation depending on the size of lipid particles were discussed on the ultracytochemical observations by this enzymic digestive method.

Dissolution and Stability of Biogenic Opal

AbstractBiogenic opal isolated from leaves of seven deciduous trees, one grass species, and surface horizons of forest and grassland soils, was examined for dissolution in hot and cold distilled water. In cold water, silica dissolution was torpid and Si concentrations remained nearly constant through a 4‐week equilibration period; Si values for all but one opal specimen were between &lt; 0.5 and 2 µg Si/ml. Dissolution over a 5‐hour period in distilled water at 90°C yielded the following Si concentrations (µg Si/ml): 4 to 5 for tree‐leaf isolates; 1 to 8 for soil isolates, 83 for the grass isolate, 0.5 for quartz, and 110 for silica gel. Opal dissolution is greater for 2–5 µm soil isolates than 5–20 µm isolates. Biogenic opal isolated from soils and deciduous tree leaves is much more stable than previously considered and exhibited Si dissolution similar to that reported for geologic opal. Further work is needed to clarify differences in opal solubilities between grass and forest origin.

Oxygen evolution from broken thylakoids fused with liposomes

Thylakoid membrane fragments obtained by hypotonic disruption of spinach chloroplasts in cold distilled water were fused with egg leicthin liposomes and tested for oxygen evolution in the presence of ferricyanide. Stability of the oxygen evolving complexes to ageing was demonstrated to be greatly enhanced by the lipsomes. Although phosphatidyl choline liposomes increased stability of oxygen evolution, phosphatidyl ethanolamine liposomes completely destroyed it. An enclosed membrane was apparently important for oxygen production. Fusion of thylakoids to liposomes may serve as a method for eventual extraction and isolation of the elusive oxygen evolving complexes.

Methyl Green-Pyronin Staining in Germinated Fruit Tree Pollen: Methods to Enhance Stainability of the Nuclei and Chromosomes

Brachet's method for the identification of tissue DNA and RNA is very specific because the nature of the stained material is confirmed by the enzymatic extraction. However, the small sized nuclei of pollen of fruit trees (Malus, Persica, Corylus) lose their affinity for methyl green after heating for 2 hours at 37°C with RNase or distilled water (as control). RNA cannot be extracted with cold perchloric acid, because of the low specificity of this extraction.In contrast, treatment with glacial acetic acid or acetic anhydride for 18 hrs, following proper alcoholic dehydration, enhances the nuclear stainability. The recommended fixative is Navashin's, with increased concentration of acetic acid up to 30% (to remove mitochondria), for 3 hrs. After thorough washing and bleaching with mixture of equal parts saturated aqueous ammonium oxalate and 3% H2O2 the slides are incubated with either enzyme or distilled water at 37°C for 2 hrs and treated with glacial acetic acid or acetic anhydride. Staining with methyl green (B. D. H.) 0.3% (w/v) and pyronin (Merck) 0.25% (w/v) in acetate buffer at pH 4.7, is performed at 37°C for 2 hrs. Differentiation is achieved by dipping in ice cold distilled water followed by a 5-40 min rinse in two changes of t-butyl alcohol.The slides are mounted in Euparal. The acetic anhydride treatment was shown to block the amino groups of basic proteins which, otherwise, would interfere with the DNA staining by methyl green.

Effect of Calcium Bicarbonate on Disinfection by Halogens

The inorganic entities in water comprise a significant portion of chemicals in water and their effects upon the disinfection process should be fully evaluated. This investigation was undertaken to determine possible effects of various inorganic chemical additives in water on the efficacy of disinfection by halogens. The chemical used in this experimentation was calcium bicarbonate. Halogen concentrations used were on an equal equivalent weight basis. The test microorganism was \IEscherichia coli\N. The exposure substrate was cold sterile distilled water to which appropriate quantities of calcium bicarbonate were added. All bacterial exposures to disinfectant were performed at pH values 5.0 and 7.0 and a temperature of 2-3°C. Bacterial death kinetics are presented in comparisons of varying additive conditions for each halogen and also for constant additive conditions for all halogens used. A hampering effect of calcium on the disinfection efficacy of chlorine, bromine and iodine was found at the conditions of the experimentation.

A new method of labeling human chorionic gonadotropin for physiologic studies.

A new technique for labeling human chorionic gmadotropin (HCG) is described. Tritium was introduced into the molecule by reduction with tritiated borohydride after oxidation with periodate. Immunologic and biologic activity was preserved. MATERIALS AND METHODS Labeling. Oxidation of 19.6 mg of HCG in 4 ml of 0.1 M Na acetate, pH 5.6, was carried out with 50 μmoles potassium periodate at room temperature. After 30 minutes, 100 umoles glucose was added to reduce excess periodate. After dialyzing overnight in 0.1 M K2HPO4, pH 7.5, at 3 C, one-half of the preparation was reduced with 9.25 umoles of tritiated potassium borohydride (324 μCi/μmole) dissolved in 0.2 ml of 0.01 M KOH. To assure reduction of all oxidized groups, an additional 37 umoles of unlabeled borohydride was added. After adjusting the pH (5.0 to 5.5) with acetic acid, the preparation was dialyzed in cold distilled water overnight and lyophilized. All preparations were assayed by either the mouse uterine weight (MUW) or ventral prostate weight...

EFFECT OF MODE OF REHYDRATION ON TEXTURAL PARAMETERS OF PRECOOKED FREEZE-DRIED SLICED BEEF.

Rehydration procedure had a significant effect on microscopic appearance, manner of liquid release on mechanical chewing, numerical values for the mechanical parameters of texture, correlations between objective measurements (Warner-Bratzler Tenderometer, Kramer Shear Press and General Foods Texturometer) and the ability of instrumental measurements to show grade and cooking level effects. To a lesser extent, it also affected residual errors and coefficients of variation. As compared to cold distilled water, rehydration in warm 1% NaCl appeared to toughen the meat fibers while allowing more water to be absorbed intrafibrally. This water did not release easily on mechanical chewing. Overnight equilibration of drained samples increased the diameter of meat fibers, roughness of those rehydrated in the hot salt solution and smoothness of those rehydrated in cold distilled water. Best approximation to the microscopic appearance of fresh cooked beef was obtained by 20 min rehydration in room temperature distilled water followed by overnight equilibration in the refrigerator. This procedure also offered good differentiation of meat grade and cooking time effects by instrumental texture measurements.

Ganglion cells in the human ciliary body.

Ganglion cells have been identified in the human ciliary body by histochemical demonstration of the cholinesterase in their cytoplasm. Routine histological preparations as well as silver stains reveal occasional bipolar or multicentric ganglion cells in the choroid of the normal human eye.^1-4No ganglion cells have been found in the normal iris. Occasional ganglion cells have been recognized in the normal human ciliary body with routine stains as well as with silver techniques.^1,5Technical difficulties, however, have prevented a complete histological identification of these cells in the ciliary body. <h3>Method</h3> The following modification of the histochemical staining technique for cholinesterase by Gerebtzoff⁶was used in this study: human eyes were fixed immediately after removal in neutral Formalin at 10 C for 5 to 24 hours. Frozen sections were cut at a thickness of 20μ and placed into cold distilled water. The water was changed three times over