UV damage to hair and the effect of antioxidants and metal chelators.

OBJECTIVE: To study the effects of addition of a redox metal, copper, antioxidants and metal chelators on the formation of free radicals in natural white Caucasian hair subsequently exposed to UV light. Three different methods, electron paramagnetic resonance (EPR), a fluorescent probe for hydroxyl radical formation (terephthalate) and free radical photoyellowing, were used. These methods utilized different UV sources and reaction conditions, and so can give insights into the different mechanisms of action occurring during UV oxidation of hair. In addition, this study demonstrates how antioxidants and chelators can be screened to determine whether they can protect hair from UV damage. RESULTS: The three methods gave somewhat different results, illustrating the importance of reaction conditions and wavelength on the photochemical mechanisms, and the efficacy of additives to influence these reactions. EPR results showed that N-acetylcysteine (NAC) pre-treatment eliminated the intensity of the signal because of sulphur and carbon free radicals in white hair both before and after exposure to UVB radiation. Doping the hair with copper ions had no effect on the intensity of the EPR signal under dry conditions. Terephthalate fluorescent probe data showed that under wet conditions, irradiation of white hair with UVA produced significant amounts of hydroxyl radicals. Pre-treatment of hair with NAC reduced the number of •OH radicals produced by natural white hair compared to an untreated control. In contrast to the EPR result, white hair doped with copper ions produced significantly higher levels of •OH radicals under wet conditions. It appears that the ability of copper ions to catalyse the photogeneration free radicals in hair is highly dependent on water content. Photoyellowing data showed a benefit for oxalic acid but no difference for NAC and an increase in yellowing for EDTA. CONCLUSION: The micro-EPR and terephthalate fluorescent probe methods are both effective techniques to study production of free radicals by hair exposed to UV light under wet and dry conditions, respectively. Both assays are simple methods for determining the effectiveness of potential protective hair treatments against UV damage, but because they assess free radical damage under dry vs wet conditions, the chemistry created on UV exposure is different. This gives insights into mechanism of action, but results may not be consistent between the two methods for actives added for reduction of UV damage. NAC pre-treatment did reduce free radical generation in UV-exposed hair under both wet and dry conditions. Photoyellowing data are more complicated as it is a less direct measure of UV damage and is highly dependent on irradiation source. Using UVB irradiation is experimentally convenient but may not be appropriate, because UVB wavelengths comprise only 0.3% of terrestrial sunlight. The photochemistry of hair exposed to sunlight involves concurrent photobleaching and photoyellowing processes and is far more complex. Under UVB irradiation conditions, oxalic acid showed a yellowing benefit.

OBJECTIF: Étudier les effets de l'ajout d'un métal redox, d'un cuivre, d'antioxydants et de chélateurs de métaux sur la formation de radicaux libres dans les cheveux naturels blancs de Caucasiens, exposés par la suite à des rayons  à ultraviolets. Trois méthodes différentes, la résonance paramagnétique électronique (RPE), une sonde fluorescente pour la formation de radicaux hydroxyles (téréphtalate) et le photojaunissement dû aux radicaux libres ont été utilisées. Ces méthodes ont utilisé différentes sources d'UV et de conditions de réaction, et peuvent par conséquent donner un aperçu des différents mécanismes d'action qui se produisent lors de l'oxydation des cheveux aux UV. Par ailleurs, cette étude montre comment les antioxydants et les chélateurs peuvent être analysés en vue de déterminer s'ils peuvent protéger les cheveux contre les dommages causés par les UV. RÉSULTATS: Les trois méthodes ont donné des résultats quelque peu différents, ce qui illustre l'importance des conditions de réaction et de la longueur d'onde sur les mécanismes photochimiques, ainsi que l'efficacité des additifs influençant ces réactions. Les résultats de la RPE ont montré que le traitement préalable par N-acétylcystéine (NAC) éliminait l'intensité du signal en raison de la présence du soufre et des radicaux libres de carbone dans les cheveux blancs, avant et après l'exposition aux rayons UVB. Le dopage des cheveux avec des ions de cuivre n'a eu aucun effet sur l'intensité du signal RPE en condition sèche. Les données des sondes fluorescentes de téréphtalate ont montré qu'en conditions humides, l'irradiation des cheveux blancs avec des UVA produisait d'importantes quantités de radicaux hydroxyles. Le traitement préalable des cheveux par N-acétylcystéine a réduit le nombre de radicaux hydroxyles produits par les cheveux blancs naturels comparé à un témoin non traité. Contrairement au résultat de la RPE, les cheveux blancs dopés avec des ions de cuivre ont produit des concentrations beaucoup plus élevées de radicaux hydroxyles en conditions humides. Il apparaît que la capacité des ions de cuivre à catalyser les radicaux libres photogénérés dans les cheveux dépend fortement de la teneur en eau. Les données relatives au photojaunissement ont démontré un avantage pour l'acide oxalique, aucune différence pour la N-acétylcystéine et une augmentation du jaunissement pour l'EDTA. CONCLUSION: Les méthodes de sonde fluorescente micro-RPE et téréphtalate sont deux techniques efficaces d'étude de la production de radicaux libres sur les cheveux exposés aux rayons ultraviolets en conditions humides et sèches, respectivement. Les deux tests sont des méthodes simples pour déterminer l'efficacité d'éventuels traitements de protection contre les dommages causés par les UV, mais étant donné qu'ils évaluent les dommages causés par les radicaux libres dans des conditions sèches ou humides, la réaction chimique créée lors de l'exposition aux UV est différente. Cela permet de mieux comprendre le mécanisme d'action, mais les résultats peuvent ne pas être cohérents entre les deux méthodes en raison des substances actives ajoutées pour réduire les dommages causés par les UV. Le traitement préalable par N-acétylcystéine a réduit la génération de radicaux libres dans les cheveux exposés aux UV en conditions humides et sèches. Les données de photojaunissement sont plus complexes, étant donné qu'il s'agit d'une mesure moins directe des dommages causés par les UV et qu'elles dépendent fortement de la source d'irradiation. L'utilisation de l'irradiation aux UVB est convenable d'un point de vue expérimental, mais peut ne pas être appropriée, car les longueurs d'onde UVB ne représentent que 0,3 % du rayonnement solaire terrestre. La photochimie de cheveux exposés à la lumière du soleil implique des processus de photoblanchiment et de photojaunissement concomitants, et est bien plus complexe. Dans des conditions d'irradiation aux UVB, l'acide oxalique a présenté un avantage en matière de jaunissement.

Gel network shampoo formulation and hair health benefits.

OBJECTIVE: The objective of this work was to create a shampoo formula that contains a stable ordered gel network structure that delivers fatty alcohols inside hair. METHODS: X-ray diffraction (SAXS and WAXS), SEM and DSC have been used to confirm formation of the ordered Lß gel network with fatty alcohol (cetyl and stearyl alcohols) and an anionic surfactant (SLE1S). Micro-autoradiography and extraction methods using GC-MS were used to confirm penetration of fatty alcohols into hair, and cyclic fatigue testing was used to measure hair strength. RESULTS: In this work, evidence of a stable Lß ordered gel network structure created from cetyl and stearyl alcohols and anionic surfactant (SLE1S) is presented, and this is confirmed via scanning electron microscopy images showing lamella layers and differential scanning calorimetry (DSC) showing new melting peaks vs the starting fatty alcohols. Hair washed for 16 repeat cycles with this shampoo showed penetration of fatty alcohols from the gel network into hair as confirmed by a differential extraction method with GC-MS and by radiolabelling of stearyl alcohol and showing its presence inside hair cross-sections. The gel network role in delivering fatty alcohol inside hair is demonstrated by comparing with a shampoo with added fatty alcohol not in an ordered gel network structure. The hair containing fatty alcohol was measured via the Dia-stron cyclic fatigue instrument and showed a significantly higher number of cycles to break vs control. CONCLUSIONS: The formation of a stable gel network was confirmed in the formulated shampoo, and it was demonstrated that this gel network is important to deliver cetyl and stearyl alcohols into hair. The presence of fatty alcohol inside hair was shown to deliver a hair strength benefit via cyclic fatigue testing.

Mechanistic insights into the bleaching of melanin by alkaline hydrogen peroxide.

This work aims to determine the roles of reactive oxygen species HO∙ and HO2- in the bleaching of melanins by alkaline hydrogen peroxide. Experiments using melanosomes isolated from human hair indicated that the HO∙ radical generated in the outside solution does not contribute significantly to bleaching. However, studies using soluble Sepia melanin demonstrated that both HO2- and HO∙ will individually bleach melanin. Additionally, when both oxidants are present, bleaching is increased dramatically in both rate and extent. Careful experimental design enabled the separation of the roles and effects of these key reactive species, HO∙ and HO2-. Rationalisation of the results presented, and review of previous literature, allowed the postulation of a simplified general scheme whereby the strong oxidant HO∙ is able to pre-oxidise melanin units to o-quinones enabling more facile ring opening by the more nucleophilic HO2-. In this manner the efficiency of the roles of both species is maximised.

Preserving fibre health: reducing oxidative stress throughout the life of the hair fibre.

Hair health is an important attribute to women globally--specifically attributes such as shine, healthy tips, frizz-free and strength. However, many women will claim to have at least moderate hair damage caused by habits and practices such as washing, combing and brushing, use of heated implements and regular use of chemical treatments. The objective of this work was to investigate two mechanisms of damage--hair colouring and UV exposure--where oxidative processes are involved. The role of copper in these oxidative processes was then investigated: its presence in hair and its consequent impact on hair damage via free radical formation. Finally, the role of chelants N,N'-ethylene diamine disuccinic acid (EDDS) and histidine in preventing free radical formation was investigated and shown to improve hair health.

Advanced hair damage model from ultra-violet radiation in the presence of copper.

OBJECTIVE: Damage to hair from UV exposure has been well reported in the literature and is known to be a highly complex process involving initiation via absorption of UV light followed by formation and propagation of reactive oxygen species (ROS). The objective of this work was to understand these mechanisms, explain the role of copper in accelerating the formation of ROS and identify strategies to reduce the hair damage caused by these reactive species. METHODS: The location of copper in hair was measured by Transmission electron microscopy-(TEM) X-ray energy dispersive spectroscopy (XEDS) and levels measured by ICP-OES. Protein changes were measured as total protein loss via the Lowry assay, and MALDI ToF was used to identify the biomarker protein fragments. TBARS assay was used to measure lipid peroxide formation. Sensory methods and dry combing friction were used to measure hair damage due to copper and UV exposure and to demonstrate the efficacy of N,N' ethylenediamine disuccinic acid (EDDS) and histidine chelants to reduce this damage. RESULTS: In this work, a biomarker protein fragment formed during UV exposure is identified using mass spectrometry. This fragment originates from the calcium-binding protein S100A3. Also shown is the accelerated formation of this peptide fragment in hair containing low levels of copper absorbed from hair during washing with tap water containing copper ions. Transmission electron microscopy (TEM) X-ray energy dispersive spectroscopy (XEDS) studies indicate copper is located in the sulphur-poor endo-cuticle region, a region where the S100A3 protein is concentrated. A mechanism for formation of this peptide fragment is proposed in addition to the possible role of lipids in UV oxidation. A shampoo and conditioner containing chelants (EDDS in shampoo and histidine in conditioner) is shown to reduce copper uptake from tap water and reduce protein loss and formation of S100A3 protein fragment. In addition, the long-term consequences of UV oxidation and additional damage induced by copper are illustrated in a four-month wear study where hair was treated with a consumer relevant protocol of hair colouring treatments, UV exposure and regular shampoo and conditioning. CONCLUSIONS: The role of copper in accelerating UV damage to hair has been demonstrated as well as the ability of chelants such as EDDS and histidine in shampoo and conditioner products to reduce this damage.

Role of copper in photochemical damage to hair.

OBJECTIVE: The objective of this work was to identify whether low levels of redox metals such as copper will accelerate damage to hair on exposure to UV irradiation and whether this damage can be prevented. METHODS: The methods used were proteomics to measure the protein damage via protein loss after different periods of exposure and mass spectroscopy methods to identify specific marker peptides that are specifically created by this type of damage. RESULTS: In this work, we have developed new insights into the mechanism of UV damage using these proteomic methods. A marker fragment in the hair protein loss extract was identified (m/z = 1279) that is unique to UV exposure and increases with time of UV exposure. We have also identified for the first time in hair the role of exogenous copper in increasing UV damage both in terms of total protein degradation and also increased formation of the marker fragment and proposed a mechanism of action. It has been demonstrated that shampoo treatment containing a chelant such as N,N'-ethylenediamine disuccinic acid (EDDS) reduced copper accumulation in hair. CONCLUSION: This work provides evidence for the role of copper in UV-induced damage to hair and strategies to reduce copper levels in hair using a chelant such as EDDS.

The role of chelants in controlling Cu(II)-induced radical chemistry in oxidative hair colouring products.

The catalytic formation of hydroxyl radicals in oxidative hair colourant systems in the presence of added copper ions was measured and quantified using a colorimetric probe N,N'-(5-nitro-1,3-phenylene)bisglutaramide. Also monitored in the same experiments was the decomposition of hydrogen peroxide. The first set of experiments was performed using aqueous model solutions containing the key oxidant actives in a hair colourant, ammonium hydroxide and hydrogen peroxide at pH 10, with added copper and calcium ions. The second set of experiments was performed in the presence of hair containing different levels of copper in conditions very close to those found during hair colouring. Both sets of experiments demonstrate the ability of copper ions to trigger the formation of hydroxyl radicals and catalyse the decomposition of hydrogen peroxide. The ability of chelants ethylenediamine tetraacetic acid (EDTA) and N,N'-ethylenediamine disuccinic acid (EDDS) to moderate the flux of hydroxyl radicals formed in solution systems was demonstrated in the presence of copper ions alone. However, only EDDS was successful in the presence of both calcium and copper ions. This was confirmed in the hair experiments where again only EDDS was successful at preventing hydroxyl radical formation where hair is added as the source of copper and calcium ions. These results are explained using metal speciation modelling and demonstrate the importance of the chelant to be able to specifically bind and prevent the one-electron redox chemistry of copper in the presence of high levels of calcium ions as found in hair. The formation of hydroxyl radicals during the colouring process was shown to lead to hair structure damage as measured by protein loss. EDDS was demonstrated to significantly reduce cuticle damage by suppressing the formation of the hydroxyl radicals in systems with realistic concentrations of calcium and copper.

The uptake of water hardness metals by human hair.

The objective of this work was to examine the variables that influence the interaction between water hardness metals and human hair. Hair extracts various constituents from the tap water used during daily hygiene practices and chemical treatments. Calcium and magnesium metal ions are the most prevalent and give water "hardness." Inductively coupled plasma atomic emission spectroscopy (ICP-AES) was employed to quantify the metal content of hair, which was studied as a function of the following variables: hair condition (oxidative damage), level of water hardness, and water pH. We have demonstrated that these variables impact water hardness metal uptake to varying extents, and the effects are driven primarily by the binding capacity (available anionic sites) of the hair. The condition of the hair, a key representation of the binding capacity, was most influential. Interestingly, water hardness levels had only a small effect on uptake; hair became saturated with notable amounts of water hardness metals even after repeated exposure to soft water. Water pH influenced metal uptake since side chains of hair proteins deprotonate with increasing alkalinity. These insights highlight the importance to the hair care industry of understanding the interaction between water hardness metals and hair.

The structural implications of water hardness metal uptake by human hair.

Human hair can extract significant levels of calcium and magnesium, water hardness metals, from tap water immediately following chemical treatments and during hygiene practices. We have previously shown that this uptake is primarily a function of the condition of the hair. Depending on the extent of chemical damage, the hair can extract notable amounts of water hardness metals even from soft water. As water hardness metals concentrate primarily in the cuticle layers of the hair fibre, it is hypothesized that their presence will affect the structural properties that are chiefly driven by the cuticle. We examined hair mechanics and styling by technical measures of single-fibre torsional and tensile properties, combability and style retention as a function of the calcium and magnesium content of virgin and bleached hair. Our work has indicated that water hardness can affect hair properties. Fibre stiffening was induced by the presence of water hardness metals inside the fibres of both virgin and bleached hair. A reduction in combing forces was also observed, and this effect is believed to be a result of the stiffening. The style retention of virgin hair was improved by water hardness metals, whereas that of bleached hair was slightly reduced.

Liquid crystal colloidal structures for increased silicone deposition efficiency on colour-treated hair.

An approach is described to increase the deposition efficiency of silicone conditioning actives from a shampoo on colour-treated hair via liquid crystal (LC) colloidal structures, created with a high charge density cationic polymer, poly(diallyldimethyl ammonium chloride) and negatively charged surfactants. LCs are materials existing structurally between the solid crystalline and liquid phases, and several techniques, including polarized light microscopy, small angle X-Ray analysis, and differential scanning calorimetry, were used to confirm the presence of the LC structures in the shampoo formula. Silicone deposition from the LC-containing shampoo and a control shampoo was measured on a range of hair substrates, and data from inductively coupled plasma optical emission spectroscopy analysis and ToF-SIMS imaging illustrate the enhancement in silicone deposition for the LC shampoo on all hair types tested, with the most pronounced enhancement occurring on hair that had undergone oxidative treatments, such as colouring. A model is proposed in which the LC structure deposits from the shampoo onto the hair to: (i) provide 'slip planes' along the hair surface for wet conditioning purposes and (ii) form a hydrophobic layer which changes the surface energy of the fibres. This increase in hydrophobicity of the hair surface thereby increases the deposition efficiency of silicone conditioning ingredients. Zeta potential measurements, dynamic absorbency testing analysis and ToF-SIMS imaging were used to better understand the mechanisms of action. This approach to increasing silicone deposition is an improvement relative to conventional conditioning shampoos, especially for colour-treated hair.

Copper and calcium uptake in colored hair.

During hair coloring a number of disulfide bonds in cystine are oxidized (1) to create cysteic acid, forming binding sites for metal ions such as Ca(2+ )and Cu(2+ )from tap water (2). The increased uptake of these metals can have a detrimental impact on fiber properties-for example, reducing shine and causing a poor wet and dry feel (3). In addition, the increased uptake of copper can also contribute to further fiber damage during subsequent coloring due to its ability to take part in metal-induced radical chemistry (4). It is important to know where in the fibers these metals are located in order to either effectively remove these metals or control their chemistry. Nanoscale secondary ion mass spectrometry (NanoSIMS) has been used to locate the calcium and copper within hair that has been treated with a colorant and washed multiple times in tap water containing these ions. Untreated hair is used as a baseline standard material. Images with up to 50-nm spatial resolution of the preferential locations of calcium uptake were obtained, showing a high concentration of calcium in the cuticle region of colored hair, specifically in the sulfur-rich regions (A-layer and exocuticle).

Investigations of cosmetic treatments on high-pressure differential scanning calorimetry.

High Pressure Differential Scanning Calorimetry (HPDSC) can be used to gain information on both the degree of crystallinity in the intermediate filaments (IFs) and the structural rigidity of the surrounding matrix or intermediate filament associated proteins (IFAP) of the hair cortex. We have used HPDSC to measure changes in denaturation temperature (T(D)) and enthalpy (deltaH(D)) of the crystalline components after treatment with bleach products. Literature reports suggest that a decrease in peak denaturation temperature is indicative of permanent damage to the hair. However, changing the rigidity of the matrix surrounding the IFs, by temporarily changing electrostatic interactions, should also result in a similar decrease in peak temperature. The complex nature of bleach formulations including oxidants, alkalizers and salts suggests that several of the components could have a non-permanent affect on salt bridges and hydrogen bonds and hence rigidity or viscosity of the matrix. We have compared the denaturation temperature with levels of lightening (dL) and tensile properties of the fiber after treatment both before and after removal of actives from the fiber. It is evident that the HPDSC results are strongly influenced by formulation components and that these changes are reversible with extensive washing or dialysis. Combined with tensile data, it is proposed that a decrease in T(D) and deltaH(D) following treatment with bleach products can be due to both permanent and reversible changes to either the intermediate filaments or intermediate filament associated proteins of the hair fiber.

High-pressure differential scanning calorimetry of colorant products.

High-pressure differential scanning calorimetry (HPDSC) can be used to gain information on both the degree of crystallinity in the intermediate filaments (IFs) and the structural rigidity of the surrounding matrix or intermediate filament associated proteins (IFAP) of the hair cortex (1-3). We have used HPDSC to measure changes in the denaturation temperature (T(D)) and enthalpy (DeltaH(D)) of the crystalline components after multiple treatments with permanent hair colorant products. We have observed that after three repeat treatments both the denaturation enthalpy and peak temperature are significantly decreased vs the untreated starting substrate. However, on dialysis of the fibers in deionized water this decrease is shown to be completely reversible, returning the enthalpy and temperature to that of the untreated hair. It is proposed that the decrease is due to the incorporation of formulation components such as the alkalizer and surfactants etc. and metal ions such as calcium and magnesium from the tap wash water. These components are predicted to have a non-permanent effect on the salt bridges and hydrogen bonds and hence the rigidity or viscosity of the matrix. We have compared the denaturation temperature with the tensile properties of the fiber after treatment both before and after removal of actives from the fiber.

Collagen, collagenase and collagenolytic activity in rat Graafian follicles during follicular growth and ovulation.

Follicles were dissected from the ovaries of immature rats at intervals after subcutaneous injection of 20 IU of pregnant mare's serum gonadotropin. A surge of luteinizing hormone was observed at 54 h and ovulation occurred at 64-66 h. The follicular volume between 36 and 48 h, then doubled again shortly before ovulation. The collagen content of the follicles increased 3-fold from 35 to 56 h, but decreased significantly (25%) from 61 to 66 h. Follicle homogenates, activated with trypsin or aminophenylmercuric acetate, digested Type I collagen at 28 degrees C to produce typical of a true collagenase. Collagenolytic activity assayed against endogenous collagen at 37 degrees C did not change significantly between 38 and 66 h.

In vitro conditions affecting the synthesis of sulfated proteoglycans by normal and rheumatoid synovial cells in culture.

In vitro conditions affecting synthesis of sulfated proteoglycans by cell suspensions derived from monolayer cell cultures of normal and rheumatoid synovial tissue were examined. The capacity of cells to synthesize proteoglycans was estimated by the incorporation of 35S--sulfate into cetylpyridinium chloride--precipitable material. Synthesis of sulfated proteoglycans was maximal during log phase, and after 2--3 hours of recovery from disaggregation. Normal synovial cells appeared to be more sensitive to changes in serum concentration than were rheumatoid synovial cells, but rheumatoid synovial cells were more sensitive to changes in cell density. The proportion of newly synthesized extracellular proteoglycans increased with the duration of incubation in 35S--sulfate.

Costs of managing patients at a Canadian rheumatic disease unit.

Financial and other costs of managing 194 inpatients and 433 outpatients referred to the rheumatology service of a Canadian general teaching hospital during 1978 were assessed. The tax financed institutional cost/outpatient visit was $102, compared to $112/inpatient day, which was 62% of the hospital's $180 cost/inpatient day. Hospital financial costs represent 78% of RDU outpatient costs and 85% of substantially lower than per diem rates. Our findings caution against relying solely on hospital data in assessing total economic costs of any diagnostic subset of patients.

Uridine diphosphate glucose dehydrogenase in rheumatoid synovial cells in culture.

Extracts containing uridine diphosphate (UDP) glucose dehydrogenase (EC 1.1.1.22) activity were prepared from rheumatoid and from normal human synovial cell lines using previously standardized techniques. Although no significant differences in the enzyme from the 2 sources were detected with respect to activity, substrate affinities, or responses to temperature and pH, these determinations have demonstrated that the enzyme is very sensitive to alterations in these parameters. The ultimate activity of the enzyme in vivo will be dependent upon the extent of the increased temperature, acidity, and altered glucose metabolism in the rheumatoid joint.

Stimulation of phospholipid labeling and steroidogenesis by luteinizing hormone in isolated bovine luteal cells.

The present study was conducted to examine the effect of LH on phospholipid metabolism in corpora luteal tissue. Collagenase-dispersed cells obtained from bovine corpora lutea of early pregnancy were incubated with 32PO4 in the presence or absence of LH and examined for their ability to incorporate this label into phospholipids. LH (1 microgram/ml) significantly increased 32P incorporation into total lipid extracts, with a time course similar to that of progesterone synthesis. This stimulation of 32P incorporation was dependent on the concentrations of LH, and this dose-response relationship correlated well with the dose response of LH-induced progesterone production. Bovine serum albumin and ACTH had no apparent effect on 32P incorporation into phospholipids or progesterone production. Separation of luteal cell phospholipid extracts by thin layer chromatography revealed that LH stimulated the incorporation 32PO4, mainly into phosphatidic acid and phosphatidylinositol, with small increases occurring in the polyphosphoinositide fraction. The LH-induced labeling of these individual phospholipids also appeared to be temporally and dose-related to the LH-induced increases in progesterone synthesis. LH had no effect on the labeling of phosphatidylcholine, phosphatidylserine, sphingomyelin, or cardiolipin. These results indicate that LH has selective effects on phospholipid metabolism in bovine luteal cells which may be a part of the mechanism of action of LH on steroidogenesis.