Hypophase

Add 10 to 15 ml half-saturated NaCl solution under continued gentle shaking until the hypophase (lower phase), which contains the original organic solvent (e.g., acetone), is -50% aqueous and the chlorophylls and carotenoids are transferred to the organic epiphase. 

Transfer hypophase to a separatory funnel and extract the last traces of chlorophylls and carotenoids using 2 ml of the hydrophobic organic solvent used in step 11. 

Figure 4. Electrolyte (150 mequiv CaCl.) injected under DPL-DPPA mixed films. Kinetic curves of AV of films containing 10 wt % (upper panel) and 50 wt % (lower panel) DPPA at three different values of x (2,10, and 20 dyn/cm). Aqueous hypophase, pH 5.6, 25°C. The mixed-lipia film at the indicated pressure was spread first on distilled H 0 the electrolyte then was injected beneath at time zero. Error as in Figure 2.

In various attempts at establishing the stoichiometry of an eventual interaction of Ca++ with lecithin films, the increases in surface radioactivity observed in the presence of 45Ca++ in the aqueous phase were ascribed to adsorption of Ca+ Apart from the interpretations, the data themselves in this field are interestingly discordant (13,14,15). To appreciate the significance of the statement, we first present our own data from experiments in which the excess surface radioactivity in counts per minute (A cpm) in the presence of DPL film (cpm with DPL — cpm without DPL) are related to the hypophase pH the aqueous phase contained 150mM NaCl and 93/xM CaCl2 carrier (see Figure 6). 

The alveolar surface represents a thin liquid film formed at the interface between the alveolar gas phase and a liquid hypophase covering the epithelium. This film is stabilised by the alveolar surfactant (AS), consisting mainly of phospholipids and proteins. AS plays an important role in alveolar stabilisation in the process of breathing. It is known that AS components exist as individual molecules and as various lipid and protein/lipid micellar structures present in the so-called hypophase and, according to some researchers, form a continuous lipid monolayer at the water/air interface [e.g. 1-4]. 

Both simple and complex species (e.g., water, lipids, proteins, lipoproteins, antibodies, enzymes, viruses, and small and large electrolyte ions) may be adsorbed on or transported across the interfaces of biological membranes, membrane subunits, and particles. These translocations are associated with membrane or particle formation, fusion, exchange, and lysis. The processes may be relevant to the movement, exchange, and clearance of extracellular material at the alveolar lining layer of the lung (7). Within this framework, the role of surface and hypophase viscosity has not been investigated. 

As described for neurosporaxanthin (4) [75], exhibit carotenoid carboxylic acids acidic properties, and may therefore be converted to the methyl esters with diazomethane, Fig. 14. The polar carboxylate anion is obtained upon base treatment. Thus neurosporaxanthin (4) becomes completely hypophasic when distributed between petroleum ether and 1 % NaOH in methanol-water (9 1) [75]. 

Human a-2-macroglobulin was prepared from freshly drawn blood essentially as described by Harpel (V7) except that, rather than a KBr gradient, a sucrose density gradient (minimum 0.2 M maximum 1.0 M sucrose) was employed. After ultracentrifugation (Beckman L8-55 SW 28.1 rotor at 28,000 rpm for 16 hr), the a-2-macroglobulin activity was present largely in the hypophase. Immunodiffusion... 

In principle, then, the molar mass of the spreading material can be determined with Equation (12-1). The surface pressure and the specific surface area are measured in a Langmuir trough. Here, a fixed quantity of material spreads out over a given surface, which is separated on the one side by an easily moved float. The pressure exerted on this float by a given surface area of a given quantity of material is then the surface pressure. These measurements are not simple to carry out, since only low pressures are found with low quantities of material, and the surface of the hypophase must be meticulously clean. Therefore the method has not become a routine method for determining molar masses. 

Alkyl sulphuric acids are known to be as acidic as sulphuric acid itself and to form inorganic salts readily [13]. In the general procedure [11], the sulphur trioxide/pyridine complex, in excess, and the respective carotenol are mixed at -10°C and the reaction, monitored by TLC, is allowed to proceed at room temperature. The reaction is quenched either by the addition of 10% aqueous NaOH to ca. pH 9, or by the addition of an aqueous NaCl solution. The carotenoids are extracted with ethyl acetate (or for disulphates with chloroform-methanol) from the aqueous hypophase and separated by TLC. 

Accordingly, a hypothesis was formulated, which seeks the theoretical rationale of appearance of the specific flow structures inside the liquid layer. It is known, that under certain circumstances, the surface tension variations may lead to the flow instability and to the induction of convection cells [e.g., 5, 6]. Our preliminary theoretical analysis of the hydrodynamic stability of the system [7] indicated that it is possible, that for the Reynolds numbers exceeding the critical value, convection cells inside the hypophase can be formed. This should lead to the significant increase of the mass transfer rate. 

First, a high content of capsanthin (70) (Figure 20) was identified in their hypophasic carotenoids from red spice paprika extract (PMl). 

Second, their hypophasic fraction carotenoids from orange extract (PM3) and apple extract (PM4) contained mainly violaxanthin (8), zeaxanthin (9) and lutein (6). 

Among total 5 extract carotenoids of hypophasic paprika extract (PMl), epiphasic paprika extract (PM2), hypophasic orange extract (PM3), hypophasic apple extract (PM4) and epiphasic apple extract (PM5), and 1 carotene of P-carotene (P,P-carotene, 2), carotenoid of hypophasic apple extract (PM4) showed the potent anti-human immunodeficiency vims (anti-HIV) activity when compared to metronidazole (71) (Figure 20) of an antiprotozoal dmg, antitrichomonal agent or antibacterial agent, however, other 4 extract carotenoids were inaetive for human immunodeficiency vims (HIV). 

Fourth, on the antitumor activity) for the tumor cells, the hypophasic orange extract (PM3) and hypophasic apple extract (PM4) showed slightly their higher cytotoxicity against four the human tumor cells such as squamous cell carcinoma HSC-2, HSC-3, submandibular gland carcinoma HSG and human promyelocytic leukemic HL-60 cells when compared to their cytotoxic activity of three normal human oral cells such as gingival fibroblast HGF, pulp cell HPC and periodontal ligament fibroblast HPLF. 

Fifth, from their electron spin resonance (ESR) spectroscopy, hypophasic paprika carotenoid (PMl) and epiphasic paprika extract carotenoid (PM2) scavenged efficiently l,l-diphenyl-2-picrylhydrazyl (DPPH) radical, whereas singlet oxygen was also quenched efficiently by epiphasic apple extract carotenoid (PM5) and epiphasic paprika extract carotenoid (PM2). 

The filtrates are combined in a 500-mL decanting funnel and treated with 70 mL of hexane to extract the fatty matter. The mixture is shaken for 1 min and allowed to stand until complete separation of phases. The upper layer, slightly yellow, retains lipids, carotenes, and di-esterified xanthophyUs. TheN,N-DMF hypophase, containing the rest of the pigments in solution, is treated twice more with hexane to remove completely any lipid remains. The three phases of hexane are combined and washed with50mLofMA -DMF. 

The extracellular fluid lining the respiratory tract and airspaces exists as a continuum from larynx to alveolus. It has a complex composition and a structure that varies from site to site that reflects its many functions. In the airways it consists of sol and gel layers surmounted by a surfactant film of unknown composition at the air-hquid interface. In the alveolus, the extracellular fluid consists of a thin hypophase covered by a dipalmitoyl phosphatidylcholine (DPPC)-rich surfactant film. These surfactant films are the first point of interaction between inhaled particles and the host and influence their deposition, clearance, and toxicity (1). 

The lamellar body contents are secreted from the apices of the type 11 cells into the alveoli. Secretion appears to involve membrane fusion and exocytosis into the hypophase between the cell and the air surface. After their release, the lamellar bodies become hydrated in the aqueous subphase and unravel into a matrix of tubular myelin (Fig. 2). Tubular myelin is hypothesized to release phospholipid spontaneously to the air-liquid interface where the phospholipids reside in thermodynamically favorable orientation with the acyl chains extending into the air phase, whereas the head groups remain submerged in the aqueous phase. After one or several respiratory cycles, the surfactant material is then forced back into the aqueous phase, where the surfactant forms bilayer vesicles. The vesicles may then enter into either a recycling or degradative pathway (31). 

Figure 3 Scanning electron micrograph of a rat lung following fixation with a nonaqueous osmium-fluorocarbon mixture. Surface details of the alveoli, for example microvilli on type II cells or pores of Kohn, are not seen beeause of the preservation of the surfaetant and assoeiated hypophase as a eontinuous film (magnification X 1000).

An aqueous hypophase lies beneath the surface film and above the epithelium. Its thickness may vary depending on its location within the alveolus (74). The hypophase has an amorphous structure and contains proteins, proteoglycans, lipoproteins, lipid micelles, and tubular myelin (78). 

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