The Use of N-Butyl Alcohol in the Paraffin Method

Several modifications in the use of n-butyl alcohol are suggested. These modifications include a revised series of dehydration solutions for exacting work, an abbreviated schedule of limited usefulness, and a simple method for more rapid paraffin infiltration. The use of a triangular coordinate graph may be valuable in designing dehydration procedures for special purposes. Changes in the primary fixation image are significantly less severe by dehydration with butyl alcohol than with many other reagents. Such deleterious effects may be further minimized by reducing the time and temperature factors to the practical limit and by substituting acetone for ethyl alcohol in a dehydration series such as that of Zirkle.     

Risk Factors for Aspiration Pneumonia in Older Adults

BackgroundsAspiration pneumonia is a dominant form of community-acquired and healthcare-associated pneumonia, and a leading cause of death among ageing populations. However, the risk factors for developing aspiration pneumonia in older adults have not been fully evaluated. The purpose of the present study was to determine the risk factors for aspiration pneumonia among the elderly.ConclusionThe risk factors for aspiration pneumonia were sputum suctioning, deterioration of swallowing function, dehydration, and dementia. These results could help improve clinical management for preventing repetitive aspiration pneumonia.     

Protein fixation and antigen retrieval: chemical studies

Abstract

Fixation with formaldehyde is the first process to which most biopsy and necropsy specimens are exposed prior to dehydration and embedding in paraffin wax. Tissue specimens that have been fixed in formaldehyde have architectural characteristics that are familiar to virtually every pathologist and these facilitate routine diagnosis. Nevertheless, formaldehyde fixation has some deleterious effects including reduction in immunoreactivity and degradation of nucleic acids. Development of methods to counteract these deleterious effects requires an understanding of the chemical events that occur during tissue fixation and subsequent tissue processing. This short review illustrates some of the chemical consequences of formaldehyde fixation and ethanol dehydration. It also provides some insight into the molecular events accompanying heat-induced antigen retrieval.     

Rigidity and spontaneous curvature of lipidic monolayers in the presence of trehalose: a measurement in the DOPE inverted hexagonal phase

Trehalose is a sugar which plays an important protectant role in organisms against damage due to dehydration. To explore the basic molecular mechanism which governs the protective function exerted on lipid membranes, X-ray diffraction and osmotic stress experiments have been performed on l--dioleoyl-phosphatidyl-ethanolamine (DOPE) in trehalose solutions of different concentrations. In pure water, DOPE forms an inverted hexagonal (H_II) phase; in sugar solutions, a strong dehydration, which induces a large reduction of the H_II lattice parameter, has been detected, but nevertheless no phase transitions occur. Structural data, directly obtained from reconstructed electron density maps, show that the bending of the lipid monolayer induced by the sugar is coupled to changes in the DOPE molecular shape. By osmotic stress, the work required to dehydrate the monolayer has been obtained and the overall free energy described as a function of trehalose concentration. Three results should be stressed: (1) dehydration experiments performed in the presence of sugar demonstrate that the protective effect cannot be purely osmotic; (2) the pivotal surface, that location on the molecule whose area is invariant upon isothermal bending, has been analyzed by two different methods: the approach by Rand and co-workers and the approach by Templer and co-workers; in both cases its presence along the DOPE molecule has been revealed and its position estimated; (3) the spontaneous radius of curvature and the rigidity constant of the lipid monolayer, measured at the pivotal plane, changes from 3.06 nm (in pure water) to 2.82 nm (in 1.4 M trehalose), and from 0.55×10^–19 to 0.74×10^–19 J, respectively. We assume that these modifications are related to direct interactions between trehalose and DOPE that alter the interface geometry, reducing the repulsion between the polar heads. However, the increased bending rigidity also accounts for the changes of the property of the aqueous compartment, reflecting the rigidity and stiffness of the sugar matrix around and inside the lipid phase.     

Synthesis of Methylenecyclobutyl- and Cyclobutenyl Adenine,Potent Antiviral Carbocyclic Analogues of Oxetanocin A

Abstract

9-Cyclobutyladenines bearing both methylene and hydroxymethyl groups, 3 and 4, were prepared by dehydration of carbocyclic oxetanocin A (1a). Introduction of a double bond into cyclobutane ring was achieved by allylic oxidation of N ⁶-benzoyl-9-[3-methylenecyclobutyl]adenine (12), which after several steps, afforded 9-[3-(hydroxy-methyl)-2-cyclobutenyl)adenine (5).     

Fenomeni di Inattivazione e di Riattivazione Enzimatica in Endospermi di semi di Ricino

Abstract

Inactivation and riactivation of enzymes in endosperms of castor bean seeds. — On the basis of previous results, the possibility has been investigated of the reversible interconversion of active and inactive form of enzymes in castor bean seeds, during their development.

The results described here indicate that:

1. the activity of some glycolytic enzymes increases greatly (81% and 400% increase of, respectively, Gl-6-P-dehydrogenase and aldolase) upon incubation of dry seeds for few hours at 4 °C.

2. The decrease of enzyme activity upon dehydration of seeds and the increase during the subsequent imbibition can be shown reproducibly.

3. This same observation is made for oxygen uptake.

These results are interpreted to indicate the reversible inactivation of enzymes caused by dehydration of seeds.     

CONDENSATION OF TRIFORMYLMETHANE WITH GUANOSINE

Guanosine has been reacted with triformylmethane (TFM) in refluxing pyridine. Four different products, 4–7, were isolated by preparative RP-HPLC, and characterized by ¹H and ¹³C NMR and UV spectroscopy and mass spectrometry. One of the products, the cyclic 1:1 adduct 4, is a stable cyclic carbinolamine formed probably by cyclization of the expected aminomethylene derivative 3. Compound 4 then undergoes reversible dehydration to the fully conjugated adduct 5. The appearance of the additional adducts, 6 and 7, suggests that TFM is prone to transformations resulting in the formation of methylenemalonaldehyde (9) and 1,1,3,3-tetraformylpropane (11).     

Formation of Ni- and Zn-Sulfides in Cultures of Sulfate-Reducing Bacteria

The purpose of this study was to characterize Ni- and Zn-sulfides precipitated in sulfate-reducing bacterial cultures. Fe-free media containing 58 mM SO ₄ ^2? were amended with Ni and Zn chloride followed by inoculation. Precipitates were sampled from cultures after two weeks of incubation at 22, 45, and 60 ^° C. Abiotic controls were prepared by reacting bacteria-free liquid media with Na ₂ S solutions under otherwise identical conditions. Precipitates were collected anaerobically, freeze-dried and analyzed by x-ray diffraction (XRD), scanning electron microscopy, and for total Ni, Zn, and S. In Ni-containing media, biogenic sulfide precipitates were mostly heazelwoodite (Ni ₃ S ₂ ), whereas abiotic precipitates were mixed heazelwoodite and vaesite (NiS ₂ ). The biogenic Ni-precipitates were better crystalline than the corresponding abiotic samples. Sphalerite (ZnS) was identified by XRD in precipitates sampled from Zn-containing media. Scanning electron microscopy revealed disordered morphological features for the sulfides, which occurred mostly as aggregates of fine particles in biogenic samples, whereas abiotic precipitates contained more plate- and needle-like structures.     

Selective Synthesis and Application to the Synthesis of (E)-Fluorovinyl Nucleosides

A selective method for synthesizing (E)-fluorovinyl was developed. Novel acyclic (E)-fluorovinyl versions of neplanocin A were designed and selectively synthesized as potential antiviral agents. The condensation of the bromide 7 with the nucleosidic bases (5-FU, C, A, G) and the deprotection afforded the desired acyclic fluorovinyl nucleosides. The synthesized compounds 11, 12, 13, and 16 were evaluated for their antiviral activity. The guanine derivative 16 showed toxicity-dependent anti-HIV-1 activity in MT-4 cells.     

A benign approach to the preparation of freshwater bryozoan statoblasts for scanning electron microscopy (SEM) imaging

Abstract

Several different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanella and Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals.     

New national and regional bryophyte records, 8

Abstract

Monoclea forsteri Hook. is a thalloid liverwort species that is found in damp habitats and can, therefore, be expected to be sensitive to dehydration. It does, however, have some unique chemical constituents and anatomical features that could play a role in dealing with the adverse effects of water deficits. Corresponding to the habitat, M. forsteri lost its turgor at high relative water content (RWC≈0.90) and did not survive drying below 20% RWC. Moreover, the gametophytes showed an increase in malondialdehyde content and a depletion of the ascorbate pool during dehydration, indicating oxidative damage. Cellular constituents did not affect turgor pressure during drying and electrolyte leakage from the cells was greatly increased at RWC<0.20. Photosynthetic processes seemed not to be affected by the loss of turgor, but a decline appeared to correlate with an increase in electrolyte leakage. A speedy and fully sustained recovery from dehydration was realized from water contents above 30% and seemed only to be possible if membrane integrity could be preserved. Anatomical characteristics within M. forsteri gametophytes deserve further investigation to better understand their physiological functions.     

Replacing xylene with n-heptane for paraffin embedding

Abstract

In standard histological technique, aromatic solvents such as xylene and toluene are used as clearing agents between ethanol dehydration and paraffin embedding. In addition, these solvents are used for de-waxing paraffin sections. Unfortunately, these solvents are harmful and therefore adequate substitutes would be useful. We suggest the use of n-heptane as a convenient substitute for xylene. Paraffin sections of rat tissues processed with n-heptane and stained with hematoxylin-eosin or Masson's trichrome showed proper embedment, well preserved morphology and excellent staining.     

Screening of mitochondrial mutations in Saudi women diagnosed with gestational diabetes mellitus: A non-replicative case-control study

IntroductionAmong metabolic disorders, gestational diabetes mellitus (GDM) is specified as hyperglycemia caused by glucose or carbohydrate intolerance defects. GDM is distinguished by oxidative stress, and has been connected to mitochondrial dysfunction. Previous studies have documented the relation between A12026G, A8344G and A3243G mutations in ND4, tRNA^Leu(UUR), and tRNA^Lys genes in different modes of diabetes.AimThe purpose of this study was to investigate into the relationship between GDM women and common mitochondrial mutations including A12026, A8344G, and A3243G in Saudi women.MethodsIn this case-control study, we have opted 96 GDM and 102 non-GDM pregnant women and DNA was extracted using EDTA blood and based on specific primers, Polymerase Chain Reaction was followed and then Restriction Fragment Length Polymorphism (RFLP) analysis was performed. Restriction enzymes was cross-checked with Lambda DNA and 10% of the purified PCR products were performed the Sanger sequencing analysis to reconfirm the RFLP analysis of the studied results.ResultsNone of the heterozygous and homozygous mutations were not observed in our study. All the subjects were turned to be homozygous normal genotypes.ConclusionThis study confirms that A12026, A8344G, and A3243G mutations have no role in the Saudi women with GDM.     

Structure of GlyGly peptide in the crystalline state as studied by X-ray diffraction and solid state 13C-NMR methods

A new type of crystal of glycylglycine (GlyGly) hydrate was crystallized from an aqueous solution, and the structure of the crystal has been determined by x-ray diffraction. The crystal is monoclinic, and the space group is C2/c, with the cell constants of a = 15.941(2) Å, b = 4.774(2) Å, c = 19.428(2) Å and β = 109.884(7)° at 296 K. There are eight GlyGly molecules and six water solvent in the cell. The GlyGly molecules are packed in a parallel β-sheet arrangement. The single crystal was obtained with a maximum size of 10 × 7 × 4 mm and is not stable under atmospheric conditions. The transparent crystal turned to turbid with the elapse of time. The isotropic ¹³C chemical shifts obtained from the ¹³C cross polarization magic angle spinning nmr experiments reveal that GlyGly hydrate was changed into GlyGly (form α) by dehydration. © 1998 John Wiley & Sons, Inc. Biopoly 45: 333–339, 1998     

Synthesis, structure and thermal behaviour of two magnesium complexes containing the 1,4-bis(5-tetrazolyl)benzene ligand

The reaction between the 1,4-bis(5-tetrazolyl)benzene ligand (H₂btb) and different magnesium salts allowed the recovery of two different molecular salts, namely [Mg(H₂O)₆](btb) (1) and [Mg(H₂O)₆](Hbtb)₂·2H₂O (2). These crystalline materials were fully characterized by spectroscopic, thermal and diffraction methods. In both cases the octahedrally coordinated hexaaquo Mg(II) ions are linked through several strong H-bond interactions of the OH?N type to the btb anions, completely or partially deprotonated in 1 and 2, respectively. The thermal analysis has established that, in both cases, all water molecules are lost below 200 °C. Interestingly, the process is completely reversible upon exposure to water-saturated atmosphere. Unfortunately, the anhydrous phases are amorphous, and no structural information could be derived form our PD traces, even if a number of partially crystalline intermediates, formed during the dehydration processes, were observed by thermodiffractometric methods.     

A water drop-shaped slingshot in plants: geometry and mechanics in the explosive seed dispersal of Orixa japonica (Rutaceae)

Background and AimsIn angiosperms, many species disperse their seeds autonomously by rapid movement of the pericarp. The fruits of these species often have long rod- or long plate-shaped pericarps, which are suitable for ejecting seeds during fruit dehiscence by bending or coiling. However, here we show that fruit with a completely different shape can also rely on pericarp movement to disperse seeds explosively, as in Orixa japonica.MethodsFruit morphology was observed by hard tissue sectioning, scanning electron microscopy and micro-computed tomography, and the seed dispersal process was analysed using a high-speed camera. Comparisons were made of the geometric characteristics of pericarps before and after fruit dehiscence, and the mechanical process of pericarp movement was simulated with the aid of the finite element model.Key ResultsDuring fruit dehydration, the water drop-shaped endocarp of O. japonica with sandwich structure produced two-way bending deformation and cracking, and its width increased more than three-fold before opening. Meanwhile the same shaped exocarp with uniform structure could only produce small passive deformation under relatively large external forces. The endocarp forced the exocarp to open by hygroscopic movement before seed launching, and the exocarp provided the acceleration for seed launching through a reaction force.ConclusionsTwo layers of water drop-shaped pericarp in O. japonica form a structure similar to a slingshot, which launches the seed at high speed during fruit dehiscence. The results suggest that plants with explosive seed dispersal appear to have a wide variety of fruit morphology, and through a combination of different external shapes and internal structures, they are able to move rapidly using many sophisticated mechanisms.     

Influence of an additional amino group on the potency of aminoadamantanes against influenza virus A. II - Synthesis of spiropiperazines and in vitro activity against influenza A H3N2 virus

Spiro[piperidine-2,2′-adamantane] 4 is one of the most potent synthetic anti-influenza A aminoadamantanes or other cage structure amines tested so far. Based on previous results Tataridis et al. (2007) [5h] which demonstrate the boost of in vitro potency by the presence of an additional amino group, we examined whether the incorporation of a second amino group into this heterocycle would increase the anti-influenza A virus activity. The new synthetic molecules 5–7 are capable of forming two hydrogen bonds within the receptor. We identified the diamino derivatives 5 and 6, which are active against influenza A H3N2 virus although less potent than amantadine and its equipotent spiropiperidine 4.     

SYNTHESIS AND ANTI-HIV ACTIVITY OF THYMIDINE ANALOGUES BEARING A 4′-CYANOVINYL GROUP AND SOME DERIVATIVES THEREOF

Treatment of 3′-O-tert-butyldimethylsilyl-2′,5′-dideoxy-5′-oxothymidine (4) with potassium or magnesium nitromethanide afforded in good yield the resolvable epimeric mixture of the expected blocked nitronucleosides 5 which upon dehydration led to the corresponding E-nitroenofuranosylthymidine 6. Nucleophilic attack of cyanide onto the nitrovinyl group led to a nucleoside analogue bearing a terminal 1-cyanovinyl group (7), a soft electrophilic group which, upon reaction with benzeneselenol, underwent a conjugate addition to the phenylselenonucleoside derivative 9. All these compounds, eventually de-O-silylated, were subject of in vitro biological testing, some exhibiting interesting cytotoxic or antiviral properties.     

Managing Diabetes in the Heat: Potential Issues and Concerns

ObjectiveTo review issues surrounding manage- ment of diabetes mellitus during times of extreme high temperatures.MethodsMaterials used for this article were identi- fied through a search of MEDLINE publications from 1966 to 2009. We chose English-language articles by using terms that cross-referenced diabetes mellitus, hot tempera- ture, heat, desert, and insulin.ResultsPersons with diabetes may have greater sus- ceptibility to adverse effects from heat (ie, increased num- ber of emergency department visits and hospitalizations, increased occurrence of dehydration and electrolyte abnor- malities, and higher death rate) than persons without dia- betes. Alterations in glucose homeostasis may occur, and changes in insulin kinetics and stability are possible. The impact of heat exposure on equipment performance (eg, glucometers) must be considered.ConclusionsHaving diabetes places a person at risk for heat-related health problems. Physicians must be aware of possible complications that diabetic patients may encounter in summer heat to prevent problems. Patient educational materials should be developed relating to self- management skills in the heat, and the topic should be in- cluded in standard diabetes education programs when ap- plicable. (Endocr Pract. 2010;16:506-511)